A cross-sectional study evidences regulations of leukocytes in the colostrum of mothers with obesity.

BACKGROUND: Breastmilk is a dynamic fluid whose initial function is to provide the most adapted nutrition to the neonate. Additional attributes have been recently ascribed to breastmilk, with the evidence of a specific microbiota and the presence of various components of the immune system, such as cytokines and leukocytes. The composition of breastmilk varies through time, according to the health status of mother and child, and altogether contributes to the future health of the infant. Obesity is a rising condition worldwide that creates a state of systemic, chronic inflammation including leukocytosis. Here, we asked whether colostrum, the milk produced within the first 48 h post-partum, would contain a distinct leukocyte composition depending on the body mass index (BMI) of the mother. METHODS: We collected peripheral blood and colostrum paired samples from obese (BMI > 30) and lean (BMI < 25) mothers within 48 h post-partum and applied a panel of 6 antibodies plus a viability marker to characterize 10 major leukocyte subpopulations using flow cytometry. RESULTS: The size, internal complexity, and surface expression of CD45 and CD16 of multiple leukocyte subpopulations were selectively regulated between blood and colostrum irrespective of the study groups, suggesting a generalized cell-specific phenotype alteration. In obesity, the colostrum B lymphocyte compartment was significantly reduced, and CD16+ blood monocytes had an increased CD16 expression compared to the lean group. CONCLUSIONS: This is the first characterization of major leukocyte subsets in colostrum of mothers suffering from obesity and the first report of colostrum leukocyte subpopulations in Latin America. We evidence various significant alterations of most leukocyte populations between blood and colostrum and demonstrate a decreased colostrum B lymphocyte fraction in obesity. This pioneering study is a stepping stone to further investigate active immunity in human breastmilk.

COVID-19 preparedness-a survey among neonatal care providers in low- and middle-income countries.

OBJECTIVE: To evaluate COVID-19 pandemic preparedness, available resources, and guidelines for neonatal care delivery among neonatal health care providers in low- and middle-income countries (LMICs) across all continents. STUDY DESIGN: Cross-sectional, web-based survey administered between May and June, 2020. RESULTS: Of 189 invited participants in 69 LMICs, we received 145 (77%) responses from 58 (84%) countries. The pandemic provides significant challenges to neonatal care, particularly in low-income countries. Respondents noted exacerbations of preexisting shortages in staffing, equipment, and isolation capabilities. In Sub-Saharan Africa, 9/35 (26%) respondents noted increased mortality in non-COVID-19-infected infants. Clinical practices on cord clamping, isolation, and breastfeeding varied widely, often not in line with World Health Organization guidelines. Most respondents noted family access restrictions, and limited shared decision-making. CONCLUSIONS: Many LMICs face an exacerbation of preexisting resource challenges for neonatal care during the pandemic. Variable approaches to care delivery and deviations from guidelines provide opportunities for international collaborative improvement.

HPLC Method for Quantification of Caffeine and Its Three Major Metabolites in Human Plasma Using Fetal Bovine Serum Matrix to Evaluate Prenatal Drug Exposure.

Caffeine is recognized as the first-line therapeutic agent for apnea of prematurity. The dosage regimen is 10 mg/kg loading dose and 2.5 mg/kg maintenance dose. However, the plasma concentration achieved, not always, is therapeutically useful. It makes necessary to increase the doses to reach plasma concentration up to 30 or 35 µg/mL or even higher to attain therapeutic effect. To study why neonates have these differences, and whether these effects are linked to prenatal caffeine exposure, we had to develop an analytical method for an accurate measurement of caffeine and metabolites concentration. The analysis was carried out using fetal bovine serum (FBS) as biological matrix in a high-performance liquid chromatography with an ultraviolet detector method. This method allows acceptable chromatographic resolution between analytes in 15 minutes. It was validated and proved to be linear in the 0.1-40 µg/mL range for caffeine, paraxanthine, theobromine, and theophylline in the same chromatographic analysis. Accuracy for quality control samples for intra- and interday assays was ranged from 96.5 to 105.2% and 97.1 to 106.2%. Precision had CV no more than 10% in all concentration levels for all analytes. No differences were observed between quantification in human and FBS. This method was applied to quantify plasma drug concentration in mothers and their newborns in a Mexican northeast population. In our study, we confirmed self-reported caffeine maternal intake in 85.2% (n=23); meanwhile, in their newborn's plasma, it was detected only in 78% (n=21). Caffeine plasma concentrations in mother and newborn had a linear relationship, and no differences were observed between groups (mothers versus children). These results suggest that our analytical method and substitution of biological matrix was linear, precise, and accurate for caffeine quantification and could be used for measuring prenatal exposure and let us to study, in the future, concentration differences observed during apnea clinical treatment.

Aquaporine-5 and epithelial sodium channel ß-subunit gene expression in gastric aspirates in human term newborns.

Both transient tachypnea of the newborn and neonatal respiratory distress syndrome have been associated with changes in gene expression of aquaporine-5 (AQP5) and the ß subunit of the epithelial sodium channel (ß-ENaC) in the respiratory epithelium. Gastric aspirate (GA) obtained immediately after birth could represent a new source for gene expression analysis for these respiratory diseases. The aims of this study were to determine the feasibility of estimating AQP5 and ß-ENaC gene expression in exfoliated respiratory epithelial cells from the GA of term neonates, and to compare the values with those found in scraped nasal epithelial cells, previously validated as a surrogate for distal lung epithelium in terms of ionic channel activity. The study had a cross-sectional, proof-of-concept design. Immediately after birth, we obtained GA and nasal mucous membrane scrapings from term newborns, in which total RNA and RT-qPCR assays for AQP5 and ß-ENaC genes were performed. AQP5 gene expression was greater in GA than in nasal scrapings, and ß-ENaC gene expression was at least as great in GA as that obtained in nasal scrapings. Amplification of samples from the two sites was comparable. AQP5 gene expression was greater in babies delivered by cesarean section; ß-ENaC gene expression was greater in babies delivered vaginally, but only in the nasal samples. Quantitation of the expression of AQP5 and of ß-ENaC genes in GA, obtained shortly after birth from term newborns is feasible. If confirmed in preterm neonates, this approach could aid in the differential diagnosis of neonatal respiratory diseases.

Can sTREM-1 predict septic shock & death in late-onset neonatal sepsis? A pilot study.

BACKGROUND AND OBJECTIVES: The transmembrane glycoprotein TREM-1 triggers an inflammatory response. Its soluble fraction (sTREM-1) has been shown to have diagnostic accuracy for late-onset neonatal sepsis (LONS). Until now, the potential of sTREM-1 to predict septic shock and/or death in septic neonates has not been explored. This study obtained estimates of the incidence and prevalence of septic shock and/or death in septic neonates for future sample size calculations for confirmatory studies and evaluated the feasibility of using sTREM-1 as a predictor of septic shock and/or death in neonates with LONS criteria. STUDY DESIGN: A pilot study with a cross-sectional design was performed from May 1(st) to October 31(st), 2012. The participants were hospitalized neonates who, after three days of life, were diagnosed as having LONS. Plasma sTREM-1 was quantified by ELISA. The main outcome measurement was the development of septic shock and/or death. RESULTS: Of 71 eligible subjects, nine (12.7%) progressed to septic shock and/or death. In the LONS-Non-Shock group, the sTREM-1 median and interquartile range (IQR) plasma value were 10 (10 to 70) pg/mL. In the LONS & Shock/Death group, the values were 567 (260 to 649) pg/mL. These values were significantly different (Mann-Whitney's U test, p=0.001). A ROC curve for a proposed sTREM-1 cut-off value of 300 pg/mL exhibited an area under the curve of 0.884 (95% CI=0.73 to 1.0; p<0.0001), with a sensitivity of 0.78 (95% CI=0.46 to 0.94) and specificity of 0.97 (95% CI=0.92 to 0.99); PPV would be 0.78 (95% CI=0.46 to 0.94) and NPV 0.97 (95% CI=0.92 to 0.99). CONCLUSIONS: In neonates with LONS, sTREM-1 has the potential to provide an excellent predictive value for septic shock/death. Larger sample sizes are needed to identify the optimal cut-off value of plasma sTREM-1 for this diagnosis and to provide diagnostic accuracy measures.

Diagnostic accuracy of buffy coat culture compared to total blood culture in late-onset sepsis of the newborn.

OBJECTIVES: To study the potential of buffy coat culture as a diagnostic tool for neonatal late-onset sepsis. METHODS: This was a study of diagnostic accuracy in newborn infants born at 28-41 weeks of gestation, weighing >800g, with ≥8 points on the NOSEP-1 scale. Paired samples for total blood culture (TBC) and buffy coat culture were drawn. We established the positivity rate, sensitivity, specificity, predictive values, and likelihood ratios, and compared time to positivity and contamination rates. RESULTS: Fifty-two newborns were included in the study. Twenty-one TBC and 22 buffy coat cultures were positive. The positivity rate for TBC was 40.4% and for buffy coat culture was 42.3% (p=not significant). Three TBC were positive with negative buffy coat culture. Four buffy coat cultures were positive with negative TBC; Kappa agreement was 0.723, p <0.001. Buffy coat culture sensitivity was 86% (95% confidence interval (CI) 68.5-95.4%), specificity 87% (75.4-93.7%), positive predictive value 82% (65.4-91.1%), negative predictive value 90% (77.9-96.8%), positive likelihood ratio 6.64 (2.79-15.05), and negative likelihood ratio 0.16 (0.05-0.42). We found no difference in time to positivity in hours; Wilcoxon Z=1224, p=0.22. The contamination rate was 1.9% for both methods. CONCLUSIONS: Buffy coat culture is as good as TBC for the microbiological diagnosis of late-onset sepsis of the newborn. Buffy coat culture allows the use of remaining plasma for further analysis.

Ectopic liver within the umbilical cord in a very preterm infant from a multiple gestation.

A 29 weeks' gestational age newborn, the product of a multiple gestation, was found to have a round mass in the umbilical cord; the resected lesion was an ectopic liver tissue (3 × 2 cm). She also had an imperforated hymen; otherwise, no other abnormalities were seen. The infant survived. Some possible mechanisms that may be associated with an ectopic liver tissue are presented.

Microbiological and toxicological effects of Perla black bean (Phaseolus vulgaris L.) extracts: in vitro and in vivo studies.

We investigated the microbiological and toxicological effects of three Perla black bean extracts on the growth and culture of selected pathogenic microorganisms, the toxicity over Vero cell lines and an in vivo rat model. Three different solvents were used to obtain Perla black bean extracts. All three Perla black bean extracts were tested for antibacterial and antiparasitic activity and further analysed for intrinsic cytotoxicity (IC(50)). Methanol Perla black bean extract was used for acute toxicity test in rats, with the up-and-down doping method. All Perla black bean extracts inhibited bacterial growth. Pseudomonas aeruginosa, Proteus vulgaris, Klebsiella oxytoca, Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis and Listeria monocytogenes showed inhibition, while Escherichia coli and Enterobacter aerogenes did not. Acidified water and acetic acid Perla black bean extract were tested in parasites. The best IC(50) was observed for Giardia lamblia, while higher concentrations were active against Entamoeba histolytica and Trichomonas vaginalis. The Vero cells toxicity levels (IC(50)) for methanol, acidified water and acetic acid Perla black bean extract were [mean +/- S.D. (95% CI)]: 275 +/- 6.2 (267.9-282.0), 390 +/- 4.6 (384.8-395.2) and 209 +/- 3.39 (205.6-212.4) microg/ml, respectively. In vivo acute toxicity assays did not show changes in absolute organ weights, gross and histological examinations of selected tissues or functional tests. The acetic acid and methanol Perla black bean extract proved to exhibit strong antibacterial activity and the acidified water Perla black bean extract exerted parasiticidal effects against Giardia lamblia, Entamoeba hystolitica and Trichomonas vaginalis. The three Perla black bean extracts assayed over Vero cells showed very low toxicity and the methanol Perla black bean extract in vivo did not cause toxicity.

Black bean extract ameliorates liver fibrosis in rats with CCl4-induced injury.

UNLABELLED: We assessed the anti-fibrotic effects of methanolic black bean extract antioxidants in a carbon tetrachloride (CCl4) liver injury model in rats. Experimentally intoxicated animals received CCl4 for eight weeks, the reference and test groups received daily intragastric quercetin or daily intragastric black bean extract. Liver fibrosis was assessed and quantified using morphometric analysis. Expression of fibrosis related genes was measured by real time RT-PCR. Qualitative and quantitative histological analysis showed that administration of 70 mg/kg b.w. of black bean extract reduced hepatic fibrosis index by 18% compared to positive controls (P 0.006), as a result of a decrease in type I (44.3% less, P 0.03) and type IV (68.9% less, P 0.049) collagen gene expression compared to CCl4-injured and Quercetin treated rats. In conclusion, we provide evidence that this methanol black bean extract ameliorates liver fibrosis and types I and IV collagen gene expression, in the animal model used. PRACTICAL APPLICATIONS: The compounds contained in this black bean extract exhibited strong antifibrotic effects in the CCl4 chronic liver injury model used; considering that this compounds are contained in a leguminous that has been used in human diet for a long time, their toxic potential should be very low, and this characteristic should favor their potential use in some other chronic or degenerative states that include an increase in inflammation and oxidative burst in their pathogenesis. Another possible application of this kind of extract could be its use as an antimicrobial or even antiparasitic therapeutic agent, although it is purely speculative.

Complicaciones por el uso de catéteres percutáneos de silicón en una unidad de cuidados intensivos neonatales / Complications in the use of percutaneus silastic cathethers in a Neonatal Intensive Care Unit

Se reportan las complicaciones observadas con el uso de catéteres percutáneos de silicón en una Unidad de Cuidados Intensivos Neonatales. A 72 recién nacidos se les instalaron un total de 93 catéteres. El promedio de punciones para lograr la instalación fue de dos. El principal sitio de entrada fue la vena basílica. La duración promedio in situ del catéter fue de 17.3 días. La principal compilación observada fue colonización bacteriana (10.06/1000 días/catéter), seguida de oclusión; el germen aislado con mayor frecuencia de la punta del catéter fue Staphylococcus epidermidis. Se encontró asociación estadísticamente significativa entre cultivos positivos y el tiempo de duración in situ del catéter

Traumatismo obstétrico: incidencia, clasificación y factores asociados / Obstetical trauma: Incidence, classification and associated factors

Evaluación prospectiva de una cohorte de recién nacidos, para determinar la incidencia de Traumatismo Obstétrico (TO), la clasificación en sus diferentes tipos, y los factores asociados a la ocurrencia de TO, definido éste como toda lesión producto de fuerzas que intervengan en el periodo comprendido entre el inicio del trabajo de parto y el pinzamiento del cordón umbilical. De agosto 1987 a julio de 1990 (tres años) se diagnosticó TO en 39 de 1155 recién nacidos vivos, lo que da una incidencia de 3.4 por ciento o 33.8 por 1000 nacidos vivos. El TO más frecuente fue cefalohematoma (25/39), seguido de fractura de clavícula (8/39), equimiosis de tejidos blandos (4/39), hematoma de tejidos blandos (2/39), heridas (2/39), lesión de plexo braquial (1/39) y hematoma subgaleal (1/39). Se encontró asociación significativa de la ocurrencia de TO con peso para la edad gestacional: nacimiento vía vaginal;aplicación de fórceps, sobre todo medios, y con el uso de inductoconducción.