ZNF718, HOXA4, and ZFP57 are differentially methylated in periodontitis in comparison with periodontal health: Epigenome-wide DNA methylation pilot study.

OBJECTIVE: To investigate the differences in the epigenomic patterns of DNA methylation in peripheral leukocytes between patients with periodontitis and gingivally healthy controls evaluating its functional meaning by functional enrichment analysis. BACKGROUND: The DNA methylation profiling of peripheral leukocytes as immune-related tissue potentially relevant as a source of biomarkers between periodontitis patients and gingivally healthy subjects has not been investigated. METHODS: A DNA methylation epigenome-wide study of peripheral leukocytes was conducted using the Illumina MethylationEPIC platform in sixteen subjects, eight diagnosed with periodontitis patients and eight age-matched and sex-matched periodontally healthy controls. A trained periodontist performed the clinical evaluation. Global DNA methylation was estimated using methylation-sensitive high-resolution melting in LINE1. Routine cell count cytometry and metabolic laboratory tests were also performed. The analysis of differentially methylated positions (DMPs) and differentially methylated regions (DMRs) was made using R/Bioconductor environment considering leukocyte populations assessed in both routine cell counts and using the FlowSorted.Blood.EPIC package. Finally, a DMP and DMR intersection analysis was performed. Functional enrichment analysis was carried out with the differentially methylated genes found in DMP. RESULTS: DMP analysis identified 81 differentially hypermethylated genes and 21 differentially hypomethylated genes. Importantly, the intersection analysis showed that zinc finger protein 718 (ZNF718) and homeobox A4 (HOXA4) were differentially hypermethylated and zinc finger protein 57 (ZFP57) was differentially hypomethylated in periodontitis. The functional enrichment analysis found clearly immune-related ontologies such as "detection of bacterium" and "antigen processing and presentation." CONCLUSION: The results of this study propose three new periodontitis-related genes: ZNF718, HOXA4, and ZFP57 but also evidence the suitability and relevance of studying leukocytes' DNA methylome for biological interpretation of systemic immune-related epigenetic patterns in periodontitis.

Effect of bovine lactoferrin on prevention of late-onset sepsis in infants <1500 g: a pooled analysis of individual patient data from two randomized controlled trials.

We previously conducted two randomized controlled trials with bovine lactoferrin (bLF) for the prevention of late-onset sepsis (LOS) in infants with a birth weight <2500 g (Study 1) and <2000 g (Study 2). The aim of this study was to determine the preventative effects of bLF on culture-proven or probable LOS in infants with a birth weight <1500 g from both studies, and to determine the effect of bLF in relation to intake of human milk. Both trial designs had similar inclusion and exclusion criteria, the same dose of bLF [200 mg·(kg body mass)-1·day-1], and used the same control (maltodextrin). We fitted multivariate Cox regression models to estimate the effect of bLF on the risk of development of the composite outcome, adjusting for covariates. We included 335 neonates with a mean birth weight of 1162 ± 244 g; 27.5% were <1000 g. There were 33 first episodes of LOS in the bLF treatment group and 48 in the control group (19.5% vs. 28.9%). bLF had a protective effect on the risk of development of LOS [hazard ratio (HR) = 0.64; %95 CI = 0.41-0.99; p = 0.048]; particularly among infants weighing <1000 g [HR = 0.46; %95 CI = 0.22-0.96; p = 0.039] and infants with a low intake of human milk [HR = 0.40; %95 CI = 0.19-0.84; p = 0.015]. Therefore, bLF supplementation protects infants <1500 g from LOS, particularly those infants not receiving human milk.

Randomized Controlled Trial of Bovine Lactoferrin for Prevention of Sepsis and Neurodevelopment Impairment in Infants Weighing Less Than 2000 Grams.

OBJECTIVES: To determine the effect of bovine lactoferrin on prevention of late-onset sepsis (LOS) and neurodevelopment delay. STUDY DESIGN: Randomized, double-blind, controlled trial in neonates with a birth weight of 500-2000 g in 3 neonatal units in Lima, Peru, comparing bovine lactoferrin 200 mg/kg/day with placebo administered for 8 weeks. The primary outcome was the first episode of culture-proven LOS or sepsis-associated death. Neurodevelopment delay was assessed by the Mullen Scales at 24 months corrected age. RESULTS: Of the 414 infants enrolled, 209 received bovine lactoferrin and 205 received placebo. LOS or sepsis-associated death occurred in 22 infants (10.5%) in the bovine lactoferrin group vs 30 (14.6%) in the placebo group; there was no difference after adjusting for hospital and birth weight; hazard ratio 0.73 (95% CI, 0.42-1.26). For infants with birth weights of <1500 g the hazard ratio was 0.69 (95% CI, 0.39-1.25). The mean age-adjusted normalized Mullen composite score at 24 months was 83.3 ± 13.6 in the bovine lactoferrin group vs 82.6 ± 13.1 in the placebo group. Growth outcomes and rehospitalization rates during the 2-year follow-up were similar in both groups, except for significantly less bronchiolitis in the bovine lactoferrin group (rate ratio, 0.34; 95% CI, 0.14-0.86). CONCLUSIONS: Supplementation with bovine lactoferrin did not decrease the incidence of sepsis in infants with birth weights of <2000 g. Growth and neurodevelopment outcomes at 24 months of age were similar. Neonatal bovine lactoferrin supplementation had no adverse effects. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01525316.

[Intraventricular hemorrhage and periventricular leukomalacia in low birth-weight neonates in three hospitals in Lima, Peru]. / Hemorragia intraventricular y leucomalacia periventricular en neonatos de bajo peso al nacer en tres hospitales de Lima, Perú.

To describe the frequency and severity of intraventricular hemorrhage and periventricular leukomalacia in low birth-weight neonates in three hospitals in Lima, Peru, 385 newborn babies weighing under 2,000 g at birth were evaluated between May 2012 and July 2014. Brain ultrasounds were obtained at 40 weeks' gestation, 3-5 days of life, and 3-4 weeks of life. Intraventricular hemorrhage occurred in 19.2% of neonates weighing under 1,500 g and was severe (grade III or with periventricular hemorrhagic infarction) in 9.6% of neonates under 1,500 g. Mortality in infants with intraventricular hemorrhage was 47.1%, while periventricular leukomalacia was found in 5.4% of neonates 1,500 g and under; both diagnoses were more frequent in lower-weight babies. The frequency of intraventricular hemorrhage is similar to that reported in other countries; however, severity and mortality are greater.

Con el objetivo de describir la frecuencia y severidad de la hemorragia intraventricular y leucomalacia periventricular en neonatos de bajo peso en tres hospitales de Lima, Perú se evaluaron 385 neonatos menores de 2000 g de peso al nacer durante mayo del 2012 a julio del 2014. Se obtuvo ultrasonidos cerebrales a las 40 semanas de gestación, 3-5 días de vida y 3-4 semanas de vida. Hemorragia intraventricular se presentó en 19,2% neonatos con menos de 1500 g y fue severa (grado III o con infarto hemorrágico periventricular) en 9,6% neonatos menores de 1500 g. La mortalidad en neonatos con hemorragia intraventricular fue de 47,1%, se encontró leucomalacia periventricular en 5,4% de los neonatos menores de 1500 g. Ambos diagnósticos fueron más frecuentes en neonatos con menor peso. La frecuencia de hemorragia intraventricular es similar a lo reportado en otros países; sin embargo, la severidad y mortalidad es mayor.

Hemorragia intraventricular y leucomalacia periventricular en neonatos de bajo peso al nacer en tres hospitales de Lima, Perú / Intraventricular hemorrhage and periventricular leukomalacia in low birth-weight neonates in three hospitals in Lima, Peru

RESUMEN Con el objetivo de describir la frecuencia y severidad de la hemorragia intraventricular y leucomalacia periventricular en neonatos de bajo peso en tres hospitales de Lima, Perú se evaluaron 385 neonatos menores de 2000 g de peso al nacer durante mayo del 2012 a julio del 2014. Se obtuvo ultrasonidos cerebrales a las 40 semanas de gestación, 3-5 días de vida y 3-4 semanas de vida. Hemorragia intraventricular se presentó en 19,2% neonatos con menos de 1500 g y fue severa (grado III o con infarto hemorrágico periventricular) en 9,6% neonatos menores de 1500 g. La mortalidad en neonatos con hemorragia intraventricular fue de 47,1%, se encontró leucomalacia periventricular en 5,4% de los neonatos menores de 1500 g. Ambos diagnósticos fueron más frecuentes en neonatos con menor peso. La frecuencia de hemorragia intraventricular es similar a lo reportado en otros países; sin embargo, la severidad y mortalidad es mayor.

ABSTRACT To describe the frequency and severity of intraventricular hemorrhage and periventricular leukomalacia in low birth-weight neonates in three hospitals in Lima, Peru, 385 newborn babies weighing under 2,000 g at birth were evaluated between May 2012 and July 2014. Brain ultrasounds were obtained at 40 weeks' gestation, 3-5 days of life, and 3-4 weeks of life. Intraventricular hemorrhage occurred in 19.2% of neonates weighing under 1,500 g and was severe (grade III or with periventricular hemorrhagic infarction) in 9.6% of neonates under 1,500 g. Mortality in infants with intraventricular hemorrhage was 47.1%, while periventricular leukomalacia was found in 5.4% of neonates 1,500 g and under; both diagnoses were more frequent in lower-weight babies. The frequency of intraventricular hemorrhage is similar to that reported in other countries; however, severity and mortality are greater.

Efectos de la terapia periodontal con antibióticos tópicos en el control glicémico de pacientes diabéticos: revisión sistemática / Effects of periodontal therapy with topical antibiotics in the glycemic control of diabe tic patients: systematic review

Background: Periodontal disease is considered as a diabetes complication and has been suggested that periodontal treatment plus antibiotics should reduce glycated hemoglobin A, by reducing local production of pro inflammatory substances. Objective: To evaluate diabetic patients with periodontal disease under periodontal treatment plus topical antibiotics and reduction of HbA1c, compared to diabetic patients under periodontal treatment without antibiotics. Materials and Methods: Using PUBMED, SCOPUS, WEB OF SCIENCE, EMBASE and Google Scholar data bases, were screened documents from 2008 to 2018. The documents included were the clinical studies, which included non-surgical periodontal treatment plus topical antibiotics, whose outcomes included the HbA1c report. Two independent researchers evaluate title; abstract and bias risk with Downs Black scale and Cochrane tool. Documents with a score higher than 15 on average by the two evaluators were included. Results: Five articles, which find inclusion criteria, were identified. Two documents failed to demonstrate statistically significant effect when compared to non-surgical periodontal therapy alone. Conclusion: In general a modest reduction of HbA1c was identified when using antibiotic therapy.

Antecedentes: La enfermedad periodontal es considerada una complicación del paciente diabético y se ha sugerido que su tratamiento con o sin antibióticos podría aportar a la reducción de la hemoglobina glucosilada, al reducir la producción local de sustancias pro-inflamatorias. Objetivo: Evaluar pacientes diabéticos con enfermedad periodontal que recibieron terapia periodontal no quirúrgica (TPNQ) más antibióticos tópicos y la reducción de la hemoglobina glucosilada, comparados con pacientes que recibieron TPNQ sin antibióticos tópicos. Materiales y métodos: Se utilizaron bases de datos MEDLINE vía PubMed, WEB OF SCIENCE, SCOPUS, EMBASE y Google Scholar utilizando vocabulario controlado y limitando la búsqueda a los años 2008-2018. Los documentos incluidos fueron los estudios clínicos, que incluían tratamiento periodontal no quirúrgico mas antibióticos tópicos, cuyos desenlaces incluyeran el reporte de HbA1c. Estos fueron analizados por dos evaluadores independientes, para identificación de riesgo de sesgo con la escala Downs Black y herramienta Cochrane. Se incluyeron los documentos con puntuación mayor a 15 en promedio de los dos evaluadores. Resultados: Se identificaron 5 documentos que cumplen con los criterios de inclusión. Dos documentos no mostraron diferencias estadísticamente significativas para el cambio de la HbA1c, en comparación con la terapia periodontal sola. Conclusión: En general se evidencia una discreta reducción de HbA1c con la terapia periodontal usando antibióticos tópicos.

Combined predictors of neurodevelopment in very low birth weight preterm infants.

OBJECTIVE: To evaluate the combined prognostic value of neurological examination, head circumference and cranial ultrasound for neurodevelopmental delay (NDD) in very low birth weight (VLBW, <1500 g) preterm infants. METHODS: Prospective follow-up study. Preterm infants with VLWB were assessed for NDD using the Mullen Scales of Early Learning test at 24 months of corrected age. Abnormal neurological examination (≥2 deviant items of Hammersmith neurological examination), microcephaly and major ultrasound abnormalities, each performed at term age, were evaluated as predictors of NDD in a multivariable Poisson model. RESULTS: 35/132 infants (26.5%) had NDD. In the multivariable analysis, microcephaly (RR, 3.2; 95% CI, 1.6-6.7) and major ultrasound abnormalities (RR, 2.7; 95% CI, 1.3-5.7) were associated to NDD. The combination of the two tests showed the highest positive predictive value (100%; 95% CI, 51%-100%), while the combination of normal neurological examination, no major US findings and normal head size at term showed the highest negative predictive value (89%; 95% CI, 78%-95%). The maximum under receiver operating characteristic curve area was for microcephaly or major ultrasound abnormalities (AUC 0.74 (0.65-0.83)). CONCLUSION: The combination of head circumference, cranial ultrasound and neurological examination at term age is useful to predict NDD in VLBW preterm infants.

Antibiotic Overuse in Premature Low Birth Weight Infants in a Developing Country.

BACKGROUND: Neonatal sepsis is a leading cause of child morbidity and mortality, especially in premature and low birth weight infants. Prompt antibiotic therapy is warranted, but its inappropriate use leads to bacterial resistance and adverse outcomes. Our objective is to describe the antibiotic use for late-onset sepsis in Peruvian premature infants. METHODS: This study is a prospective study as a secondary analysis of a clinical trial in 3 neonatal care units in Peru. We included infants in the first 72 hours of life, with birth weight (BW) <2000 g. We described the antibiotic use as length of therapy (LOT) per 1000 patient days (PD) and antibiotic courses. RESULTS: We included 408 neonates, with 12,204 PD of follow-up; 253 infants (62%) had a BW ≤1500 g. Total antibiotic use for late-onset sepsis was 2395 LOT (196 LOT/1000 PD). Two-hundred and seventy-one patients (66.4%) did not receive antibiotics for late-onset sepsis during their hospitalization. In total, 204 antibiotic courses were administered; 92 infants (22.5%) received 1 course, and 45 (11.0%) received 2-5 antibiotic courses. Mean duration of antibiotic course was 10.8 days (standard deviation: ±7.3). We found a significant association between a lower BW and increased antibiotic use per day (P < 0.001). The most commonly used antibiotics were vancomycin (143 LOT/1000 PD), carbapenems (115 LOT/1000 PD), aminoglycosides (72 LOT/1000 PD) and ampicillin (41 LOT/1000 PD). CONCLUSIONS: Premature infants receive antibiotics for longer than recommended periods of time. Antibiotic overuse is greater in neonates with lower BW. Vancomycin is the most used antibiotic. There is an urgent need to develop antimicrobial stewardship programs in our setting.

Assessment of glycosylated hemoglobin (HbA1c) in type 2 diabetics before and after non-surgical periodontal treatment. A short-term follow-up study

Aim: To evaluate glycosylated hemoglobin (HbA1c) before and after non-surgical periodontal treatment in patients with type 2 diabetes mellitus (DM-2). Methods: Twenty subjects diagnosed with periodontitis and DM-2 were treated using an NSPT protocol. Periodontal examination and blood measurements were performed at baseline and after three months. Patients with DM-2 treated for at least a year, with at least 10 teeth and with probing depths between 4-6 mm in more than three regions were included. The variables evaluated were HbA1c in blood and periodontal measures (probing depths, insertion level, gingival bleeding on probing, dental plaque, calculus, inflammation, clinical attachment and mobility). All patients were informed of the conditions of the therapy used. Scaling and root planning (SRP) of the full mouth was performed using an ultrasonic scaler and hand instrument under local anesthesia, supragingival prophylaxis and oral hygiene instruction. Also, 0.12% chlorhexidine digluconate was formulated twice a day for two weeks. Statistical analyses were performed using StataIC 14. The values are shown as the mean, median and standard deviation (SD) or interquartile rank (IR), and p<0.05 was considered significant. Results: Twenty subjects completed the three-month follow-up and were included in the analysis. Three months after the non-surgical periodontal treatment, most periodontal parameters had a meaningful reduction (p< 0.05) and a large effect size >0.8. Clinical attachment level showed no improvement. The HbA1c values were not significantly decreased (p=0.94). Conclusions: Although non-surgical periodontal therapy eliminates local inflammation, it is insufficient to significantly reduce HbA1c levels in a short time period

[Use of lumbar puncture in the evaluation of late-onset sepsis in low birth weight neonates]. / Uso de la punción lumbar en la evaluación de sepsis neonatal tardía en recién nacidos de bajo peso.

The objective of this study was to analyze the use of lumbar punctures (LP) in the evaluation of late-onset neonatal sepsis. It is recommended to perform an LP as part of the evaluation of late-onset sepsis. We used a cohort of 414 newborns with a birth weight <2000g in three hospitals in Lima. A LP was performed in 45/214 (21.0%) of sepsis evaluations and in 13/48 (27.1%) of culture-proven sepsis. Meningitis was diagnosed in 8/214 (3.7%) of the episodes and 8/45 (17.5%) of the evaluations that included an LP. The duration of treatment of the sepsis episodes without a LP and the episodes with a negative LP was similar, and shorter than the episodes with a positive LP. The use of LP in the evaluation of late-onset sepsis is low and can result in undiagnosed and undertreated meningitis. The use of LP in the evaluation of neonatal sepsis must be encouraged in the neonatal units.

Uso de la punción lumbar en la evaluación de sepsis neonatal tardía en recién nacidos de bajo peso / Use of lumbar puncture in the evaluation of late-onset sepsis in low birth weight neonates

RESUMEN El objetivo del estudio fue analizar el uso de la punción lumbar (PL) en las sospechas de sepsis neonatal tardía. Se utilizó una cohorte de 414 neonatos con peso al nacer <2000 g en tres hospitales de Lima. Se realizó la PL en 45/214 (21,0%) sospechas de sepsis y en 13/48 (27,1%) sepsis confirmadas por hemocultivo. Se diagnosticó meningitis en 8/214 (3,7%) sospechas y en 8/45 (17,5%) episodios en los que se realizó la PL. El tiempo de tratamiento de los episodios sin PL fue similar a los episodios de sepsis con meningitis descartada y menor a los episodios de meningitis. El uso de la PL es bajo, lo que puede resultar en meningitis no diagnosticadas y tratadas inadecuadamente. Es necesario reforzar la importancia de la PL en la evaluación de sepsis neonatal.

ABSTRACT The objective of this study was to analyze the use of lumbar punctures (LP) in the evaluation of late-onset neonatal sepsis. It is recommended to perform an LP as part of the evaluation of late-onset sepsis. We used a cohort of 414 newborns with a birth weight <2000g in three hospitals in Lima. A LP was performed in 45/214 (21.0%) of sepsis evaluations and in 13/48 (27.1%) of culture-proven sepsis. Meningitis was diagnosed in 8/214 (3.7%) of the episodes and 8/45 (17.5%) of the evaluations that included an LP. The duration of treatment of the sepsis episodes without a LP and the episodes with a negative LP was similar, and shorter than the episodes with a positive LP. The use of LP in the evaluation of late-onset sepsis is low and can result in undiagnosed and undertreated meningitis. The use of LP in the evaluation of neonatal sepsis must be encouraged in the neonatal units.

Chemical salivary composition and its relationship with periodontal disease and dental calculus

Aim: To determine the relationship between the chemical composition of saliva, periodontal disease and dental calculus. Methods: An observational analytical cross-sectional study was conducted with patients over 55 years of age. Ethical principles of autonomy and risk protection were applied according to the international standards. Sociodemographic and diagnosis variables (presence of dental calculus and periodontal status) were considered to measure salivary concentrations of glucose (by the glucose oxidase/peroxidase method, amylase (by the colorimetric test), urea (by the amount of indophenol), total protein (by the Bradford method) and albumin (by the nephelometric method). Patients chewed a sterile rubber band and 3 mL of stimulated saliva were collected. The samples were stored at -5 °C, centrifuged at 2,800 rpm for 10 min, and the supernatant was removed and stored at -20 °C. Data were presented as frequencies and proportions for qualitative variables and measures of central tendency and dispersion for quantitative variables. Data were analyzed by either analysis of variance or Kruskal Wallis test . A p value <0.05 was considered statistically significant. Results: Significant relationships were observed between the concentration of salivary urea and periodontal status (p = 0.03) and the presence of dental calculus and urea (p = 0.04) was demonstrated. Conclusions: A relationship between the salivary urea concentration and the presence of periodontal disease and dental calculus is suggested.

Lactoferrin for prevention of neonatal sepsis.

Preterm neonates are at risk to acquire infections. In addition to the high mortality associated with sepsis, these patients are at risk for long-term disabilities, particularly neurodevelopment impairment. Several interventions have been evaluated to reduce rates of infections in neonates but have not proven efficacy. Lactoferrin (LF), a milk glycoprotein with anti-inflammatory, immunomodulatory and anti-microbial properties, has the potential to prevent infections in young children. We performed a review of current and ongoing clinical trials of LF for prevention of neonatal sepsis, and found eleven registered clinical trials that include more than 6,000 subjects. Few of these trials have finished; despite their small sample size, the preliminary results show a trend towards a positive protective effect of LF on neonatal infections. Larger trials are underway to confirm the findings of these initial studies. This information will help to define LF's role in clinical settings and, if proven effective, would profoundly affect the treatment of low birth weight neonates as a cost-effective intervention worldwide.