Neonatal intestinal mucus barrier changes in response to maturity, inflammation, and sodium decanoate supplementation.

The integrity of the intestinal mucus barrier is crucial for human health, as it serves as the body's first line of defense against pathogens. However, postnatal development of the mucus barrier and interactions between maturity and its ability to adapt to external challenges in neonatal infants remain unclear. In this study, we unveil a distinct developmental trajectory of the mucus barrier in preterm piglets, leading to enhanced mucus microstructure and reduced mucus diffusivity compared to term piglets. Notably, we found that necrotizing enterocolitis (NEC) is associated with increased mucus diffusivity of our large pathogen model compound, establishing a direct link between the NEC condition and the mucus barrier. Furthermore, we observed that addition of sodium decanoate had varying effects on mucus diffusivity depending on maturity and health state of the piglets. These findings demonstrate that regulatory mechanisms governing the neonatal mucosal barrier are highly complex and are influenced by age, maturity, and health conditions. Therefore, our results highlight the need for specific therapeutic strategies tailored to each neonatal period to ensure optimal gut health.

Effects of prophylactic antibiotics administration on barrier properties of intestinal mucosa and mucus from preterm born piglets.

Early intervention and short-duration treatments with antibiotics in premature infants are reported to reduce the incidence of necrotizing enterocolitis (NEC), a terrible disease with severe inflammation and impaired intestinal barrier properties. Yet, it is unclear how antibiotics exposure, as well as route of administration used for dosing, can minimize the risk of NEC. With this study, we aimed to investigate if and how administration of antibiotics may affect the barrier properties of intestinal mucosa and mucus. We compared how parenteral (PAR) and a combination of enteral and parenteral (ENT+PAR) ampicillin and gentamicin given to preterm born piglets within 48 h after birth affected both barrier and physical properties of ex vivo small intestinal mucosa and mucus. Permeation of the markers mannitol, metoprolol, and fluorescein-isothiocyanate dextran of 4 kDa (FD4) and 70 kDa (FD70) through the mucosa and mucus was evaluated. For all markers, permeation through the mucosa and mucus collected from PAR piglets tended to be reduced when compared to that observed using untreated piglets. In contrast, permeation through the mucosa and mucus collected from ENT+PAR piglets tended to be similar to that observed for untreated piglets. Additionally, rheological measurements on the mucus from PAR piglets and ENT+PAR piglets displayed a decreased G' and G'/G" ratio and decreased viscosity at 0.4 s-1 as well as lower stress stability compared to the mucus from untreated piglets.

Mucoadhesive Dendrons Conjugated to Mesoporous Silica Nanoparticles as a Drug Delivery Approach for Orally Administered Biopharmaceuticals.

Biological drugs are increasingly important for patients and industry due to their application in the treatment of common and potentially life-threatening diseases such as diabetes, cancer, and obesity. While most marketed biopharmaceuticals today are injectables, the potential of mucoadhesive delivery systems based on dendron-coated mesoporous silica nanoparticles for oral delivery of biological drugs is explored in this project. We hypothesize that specifically designed dendrons can be employed as mucoadhesive excipients and used to decorate the surface of nanoparticles with properties to embed a drug molecule. We initially tested a novel synthesis method for the preparation of dendrons, which was successfully validated by the chemical characterization of the compounds. The interaction between dendrons and mucin was studied through isothermal titration calorimetry and quartz crystal microbalance with dissipation monitoring and proved to be spontaneous and thermodynamically favorable. Dendrons were conjugated onto 244.4 nm mesoporous silica nanoparticles and characterized for chemical composition, size, and surface charge, which all showed a successful conjugation. Finally, dynamic light scattering was used to study the interaction between nanoparticles and porcine gastric mucin, whereas the interaction between nanoparticles and porcine intestinal mucus was characterized by rheological measurements. This study shows a deeper biophysical understanding of the interaction between nanoparticles and mucin or native porcine intestinal mucus, further leveraging the current understanding of how dendrons can be used as excipients to interact with mucin. This will provide knowledge for the potential development of a new generation of mucoadhesive nanoformulations for the oral delivery of biopharmaceuticals.

Validation of the PTSD screening cartoon test "Darryl" in a Danish clinical sample of children and adolescents.

The current study examined the reliability and validity of a Danish adaptation of "Darryl", a cartoon-based measure of DSM-IV symptoms of post-traumatic stress disorder (PTSD) in a sample of Danish children and adolescents. Exposure to potentially traumatizing events, PTSD symptoms and diagnostic status were assessed in 65 children between the ages of 7 and 17 years old. Participants were recruited from a crisis centre for battered women and a school for children with emotional and behavioural problems. The study revealed that Darryl has good internal consistency for the overall scale and adequate reliability for each DSM-IV symptom cluster. Scores from Darryl were significantly correlated with the Clinician Administered PTSD Scale for Children and Adolescents (CAPS-CA) scores and receiver operating characteristic (ROC) analysis showed that it could significantly predict the presence of a PTSD diagnosis according to the CAPS-CA. In comparison to the CAPS-CA, Darryl has comparable psychometric properties and assesses PTSD symptoms in a developmentally appropriate manner. Furthermore, Darryl is quick to administer and was easier for the children in this sample to comprehend than the more traditional measure of PTSD. Overall, Darryl is an effective instrument for screening children at risk of a PTSD diagnosis. The relatively high specificity suggests that screening positive for PTSD using Darryl merits further diagnostic assessment. Copyright © 2016 John Wiley & Sons, Ltd.

Cell-Penetrating Peptides as Carriers for Transepithelial Drug Delivery In Vitro.

There is a growing interest in the use of cell-penetrating peptides (CPPs) as carriers for transepithelial drug delivery. This chapter gives an introduction to and discussion of the commonly used production and characterization methods for CPP-cargo samples including high-throughput cell viability screening. Moreover, we describe methods for permeation and cell viability assessment in the Caco-2 cell culture model with and without implementation of biosimilar mucus. Last, a method to assess metabolic degradation in vitro is described.

The effect of butter grains on physical properties of butter-like emulsions.

Milk fat exists as globules in its natural state in milk. The potential of using globular fat to modulate the rheological properties and crystallization behavior in butter-like emulsions was studied in the present work. We conducted a comparative study of butter-like emulsions, with a fat phase consisting of 0, 10, 25, 50, or 100% anhydrous milk fat (AMF), the remaining fat being butter grains, and all samples containing 20% water, to obtain systematic variation in the ratio of globular fat. All emulsions were studied over 4wk of storage at 5°C. By combining small and large deformation rheology, we conducted a detailed characterization of the rheological behavior of butter-like emulsions. We applied differential scanning calorimetry to monitor thermal behavior, confocal laser scanning microscopy for microstructural analysis, and low-field pulsed nuclear magnetic resonance spectrometry to measure solid fat content. By combining these techniques, we determined that increasing the fraction of globular fat (by mixing with butter grains) decreases the hardness of butter-like emulsions up to an order of magnitude at d 1. However, no difference was observed in thermal behavior as a function of butter grain content, as all emulsions containing butter grains revealed 2 endothermal peaks corresponding to the high (32.7°C ± 0.6) and medium (14.6°C ± 0.1) melting fractions of fatty acids. In terms of microstructure, decreasing the amount of butter grains in the emulsions resulted in formation of a denser fat crystal network, corresponding to increased hardness. Moreover, microstructural analysis revealed that the presence of butter grains resulted in faster formation of a continuous fat crystal network compared with the 100% AMF sample, which was dominated by crystal clusters surrounded by liquid oil. During storage, hardness remained stable and no changes in thermal behavior were observed, despite an increase in solid fat content of up to 5%. After 28d of storage, we observed no difference in either microstructural or rheological properties, indicating that formation of primary bonds occurs primarily within the first day of storage. The rheological behavior of butter-like emulsions is not determined solely by hardness, but also by stiffness related to secondary bonds within the fat crystal network. The complex rheological behavior of milk fat-based emulsions is better characterized using multiple parameters.

The Effective Factors on the Structure of Butter and Other Milk Fat-Based Products.

Butter and other milk fat-based products are valuable products for the dairy industry due to their unique taste, their textural characteristics, and nutritional value. However, an increased consumer demand for low-fat-based products increases the need for an increased essential understanding of the effective factors governing the structure of milk fat-based products. Today, 2 manufacturing techniques are available: the churning method and the emulsification method. The first is typically used for production of butter with a globular structure, which has become increasingly popular to obtain low-fat-based products, typically without presence of milk fat globules. The microstructure of milk fat-based products is strongly related to their structural rheology, hence applications. Structural behavior is not determined by one single parameter, but by the interactions between many. This complexity is reviewed here. Parameters such as thermal treatment of cream prior to butter making, water content, and chemical composition influence not only crystal polymorphism, but also the number and sizes of fat crystals. The number of crystal-crystal interactions formed within the products is related to product hardness. During storage, however, postcrystallization increases the solid fat content and strengthens the fat crystal network. The fat crystal network is strengthened by the formation of more and stronger crystal-crystal interactions due to mechanically interlinking of fat crystals, which occurs during crystal growth. Postcrystallization is directly linked to chemical composition. The initially observed microstructural difference causing different rheological behavior will disappear during storage due to postcrystallization and formation of more crystal-crystal interactions.

Polymorphism, microstructure and rheology of butter. Effects of cream heat treatment.

The effect of cream heat treatment prior to butter manufacturing, fluctuating temperatures during storage and presence of fat globules vs. no fat globules was examined in laboratory scale produced butter. X-ray diffraction and differential scanning calorimetry was used to study crystallization behaviour and nuclear magnetic resonance to measure solid fat content and water droplet size distribution. Furthermore, the crystal structure was linked to the rheological properties and microstructure of the butter using confocal laser scanning microscopy. Butter produced from non-matured cream mainly formed α- and ß'-crystals with minor traces of ß-crystals. Maturing of the cream caused a transition from α- to ß'- and ß-form. The rheological behaviour of slow cooled butter deviated from the matured ones by having a lower elastic modulus, caused by a weaker crystal network. Presence of fat globules did not affect the rheological properties significantly.

Calcipotriol delivery into the skin with PEGylated liposomes.

The D-vitamin analogue calcipotriol is commonly used for topical treatment of psoriasis, but skin penetration is required for calcipotriol to reach its pharmacological target: the keratinocytes in the lower epidermis. Liposomes can enhance the delivery of drugs into the skin, but a major challenge for the development of dosage forms containing liposomes is to maintain the colloidal stability in the formulation. The purpose of this study was to investigate the effect of stabilising liposomes with the lipopolymer poly(ethylene glycol)-distearoylphosphoethanolamine (PEG-DSPE) on the physicochemical properties of the liposomes and the ability to deliver membrane-intercalated calcipotriol into the skin. Inclusion of 0.5, l and 5 mol% PEG-DSPE in the membrane enhanced the colloidal stability of the liposomes without compromising the delivery of calcipotriol from the vehicle into excised pig skin. Calcipotriol-loaded liposomes with 1 mol% PEG-DSPE did even provide for a significantly increased deposition of calcipotriol into the stratum corneum. The size of the liposomes affected the penetration of calcipotriol into the stratum corneum since small unilamellar vesicles enhanced calcipotriol penetration as compared to large multilamellar vesicles, indicating that the liposomes to some extent migrate as intact vesicles into the stratum corneum. However, calcipotriol penetrated the skin better than the lipid component of the liposomes, suggesting that at least a fraction of the drug is released from the liposomes during skin migration. In conclusion, PEGylation is therefore a promising approach for stabilising calcipotriol-containing liposomal dispersions without compromising their favourable skin accumulation properties.