Inflammatory Markers in Mother's Own Milk and Infant Stool of Very Low Birthweight Infants.

BACKGROUND: Mother's breastmilk is the gold standard for feeding preterm infants. Preterm delivery may be precipitated by inflammatory maternal states, but little is known about milk cytokine profiles and how they correlate with markers of infant gut inflammation (i.e., stool calprotectin) in this vulnerable population. RESEARCH AIM: To assess cytokines and inflammatory markers in milk from parents of very preterm infants over time as well as correlations between milk and infant's stool calprotectin. METHOD: This is a secondary analysis of milk samples collected during OptiMoM, a triple-blind randomized clinical trial of infants born < 1250 g (NCT02137473). Longitudinally collected samples were analyzed for cytokines, choline, and inflammatory markers (C-reactive protein [CRP], IFN-γ, IL-10, IL-1ß, IL-1ra, IL-6, IL-8, TNF-α). Infant stools were collected for longitudinal calprotectin analysis. Generalized estimating equations quantified longitudinal profiles of milk markers and stool calprotectin, their associations, and the correlation between free choline and C-reactive protein over follow-up. RESULT: Participants included 92 parents and infants (median weeks of gestation 27.3, median birth weight 845 g, and prevalence of male infants 45%). In all, 212 milk samples and 94 corresponding stool calprotectin levels were collected 1-11 weeks postpartum. C-reactive protein was present in much higher concentrations than other markers, and was highest in Week 1 postpartum. It decreased over time. IL-8 and free choline also changed over time while other markers did not. There was no correlation between any milk markers and stool calprotectin. CONCLUSION: Milk from mothers of very preterm infants has detectable inflammatory markers, some of which change over time. Research is needed to determine if infant outcomes are associated with these markers.

Associations of maternal inflammatory states with human milk composition in mothers of preterm infants.

Introduction: Overweight/obesity (ow/ob) is increasing in prevalence in pregnant women, and it is associated with other pro-inflammatory states, such as pre-eclampsia, gestational diabetes, and preterm labor. Data are lacking if mothers experiencing inflammatory states who deliver preterm have mother's own milk (MOM) with differing inflammatory markers or pro-inflammatory fatty acid (FA) profiles. Methods: The aim was to explore associations of maternal pre- and perinatal inflammatory states with levels of inflammatory markers and/or FAs in longitudinal samples of MOM from mothers of preterm infants born <1,250 g. Inflammatory states included pre-pregnancy ow/ob, diabetes, chorioamnionitis (chorio), preterm labor (PTL), premature rupture of membranes (PROM), pre-eclampsia, and cesarian delivery. In MOM, inflammatory markers studied included c-reactive protein (CRP), free choline, IFN-Ɣ, IL-10, IL-1ß, IL-1ra, IL-6, IL-8, and TNF-α, and FAs included omega-6:omega-3 ratio, arachidonic acid, docosahexaenoic acid, linoleic acid, monounsaturated FAs, and saturated FAs. The above inflammatory states were assessed individually, and the healthiest mothers (normal BMI, no chorio, and ± no pre-eclampsia) were grouped. Regression analysis tested associations at baseline (day 5) and over time using generalized estimating equations. Results: A total of 92 infants were included who were delivered to mothers (42% ow/ob) at a median gestational age of 27.7 weeks and birth weight of 850 g. MOM CRP was 116% higher (relative change 2.16) in mothers with ow/ob at baseline than others (p = 0.01), and lower (relative change 0.46, 0.33, respectively) in mothers in the two "healthy groups" at baseline (both p < 0.05) than others. MOM IL-8 levels were lower with chorio and PTL at baseline. No significant associations were found for other individual or grouped inflammatory states nor for other MOM inflammatory markers nor FA profiles at baseline. Discussion: In conclusion, MOM CRP levels are positively associated with inflammatory states, such as ow/ob. Reassuringly, there was no association between FA profiles or most other inflammatory markers and maternal inflammatory states. Further studies are needed to determine potential associations or ramifications of MOM CRP in vulnerable preterm infants.

Formation of bimetallic Ag-Au nanowires by metallization of artificial DNA duplexes.

Uniform bimetallic nanowires, tunable in size, have been grown on artificial DNA templates via a two-step metallization process. Alkyne-modified cytosines were incorporated into 900-base-pair polymerase-chain-reaction fragments. The alkyne modifications serve as addressable metal-binding sites after conversion to a sugar triazole derivative via click chemistry. Reaction of the Tollens reagent with these sugar-coated DNA duplexes generates Ag0 metallization centers around the sugar modification sites of the DNA. After a subsequent enhancement step using gold, nanowires < or = 10 nm in diameter with a homogeneous surface profile were obtained. Furthermore, the advantage of this two-step procedure lies in the high selectivity of the process, due to the exact spatial control of modified DNA base incorporation and hence the confinement of metallization centers at addressable sites. Besides experiments on a membrane as a proof for the selectivity of the method, atomic force microscopy (AFM) studies of the wires produced on Si-SiO2 surfaces are discussed. Furthermore, we demonstrate time-dependent metallization experiments, monitored by AFM.

Directed DNA metallization.

Genes of interest can be selectively metallized via the incorporation of modified triphosphates. These triphosphates bear functions that can be further derivatized with aldehyde groups via the use of click chemistry. Treatment of the aldehyde-labeled gene mixture with the Tollens reagent, followed by a development process, results in the selective metallization of the gene of interest in the presence of natural DNA strands.

The construction of DNA molecules of figure-eight structure.

Using DNA molecules to construct a structural scaffold for nanotechnology is largely accepted. In this article, we report on two methods for constructing a figure-eight structure of DNA molecules having a relatively high yield that could be used further as a scaffold for nanotechnology applications. In the first method, two plasmids were constructed that, on digestion with a restriction endonuclease producing nicks in the corresponding sites and after heating, produced complementary single-stranded sequences, enabling the plasmids to hybridize to each other and forming a figure-eight structure. The formation of the figure-eight structure was analyzed by restriction analysis and gel electrophoresis as well as by atomic force microscopy. The second method makes use of the bacteriophage M13 that is obtained as either a single- or double-stranded circular DNA molecule. Two M13 molecules harboring complementary sequences were constructed and produced a figure-eight structure on hybridization. The methods described here could be used further for the construction of nanoelectronic devices.

Delayed immediate implants: alveolar bone changes during the healing period.

Delaying the placement of immediate fixtures by 6-8 weeks after extraction of the natural dentition allows for the elimination of associated infective processes, the achievement of maximum osteoblastic activity that theoretically could help the osseointegration process and complete wound covering that simplifies the placement of grafts or membranes. This study examines the healing associated with 21 fixtures in 14 patients. The fixtures were placed into sockets 6-8 weeks after tooth extraction without the use of barrier membranes or bone substitutes. Measurements were taken immediately prior to fixture placement and 3-6 months later at the abutment placement. Alveolar bone height, the remaining socket depth and diameter and the depth to which a 3.75 mm fixture could be inserted into the socket were measured. After fixture placement the vertical and horizontal measurements from the cover screw to the surrounding alveolar bone and the distance from the cover screw to the CEJ of the adjacent tooth were recorded. All fixtures were integrated at exposure with 1 failure during the follow-up period. The distance from the cover screw to the buccal plate decreased by a mean of 2.17 mm. There was an increase in the mean vertical bone height at all 4 surfaces. When horizontal defects were present, the mean vertical distance decreased from 2.5 +/- 0.37 mm to 0.36 +/- 0.64 mm. When horizontal defects were absent, the mean vertical distance decreased from 3.86 +/- 0.58 mm to 0.48 +/- 0.25 mm. There was also a marked decrease in the horizontal distance between the bone margin and the surface of the fixture from 1.6 +/- 1.73 mm to 0.02 +/- 0.02 mm. These results indicate a strong tendency for the defects to fill-in the horizontal plane and for bone growth to occur in the vertical plane to the height of the cover screw. In conclusion the delayed immediate placement of fixtures has a good short-term prognosis with bone regeneration occurring around the defect without the use of barrier membranes or bone substitutes.