Secretome from Human Mesenchymal Stem Cells-Derived Endothelial Cells Promotes Wound Healing in a Type-2 Diabetes Mouse Model

Tissue regeneration is often impaired in patients with metabolic disorders such as diabetes mellitus and obesity, exhibiting reduced wound repair and limited regeneration capacity. We and others have demonstrated that wound healing under normal metabolic conditions is potentiated by the secretome of human endothelial cell-differentiated mesenchymal stem cells (hMSC-EC). However, it is unknown whether this effect is sustained under hyperglycemic conditions. In this study, the wound healing effect of secretomes from undifferentiated human mesenchymal stem cells (hMSC) and hMSC-EC in a type-2 diabetes mouse model was analyzed. hMSC were isolated from human Wharton’s jelly and differentiated into hMSC-EC. hMSC and hMSC-EC secretomes were analyzed and their wound healing capacity in C57Bl/6J mice fed with control (CD) or high fat diet (HFD) was evaluated. Our results showed that hMSC-EC secretome enhanced endothelial cell proliferation and wound healing in vivo when compared with hMSC secretome. Five soluble proteins (angiopoietin-1, angiopoietin-2, Factor de crecimiento fibroblástico, Matrix metallopeptidase 9, and Vascular Endothelial Growth Factor) were enriched in hMSC-EC secretome in comparison to hMSC secretome. Thus, the five recombinant proteins were mixed, and their pro-healing property was evaluated in vitro and in vivo. Functional analysis demonstrated that a cocktail of these proteins enhanced the wound healing process similar to hMSC-EC secretome in HFD mice. Overall, our results show that hMSC-EC secretome or a combination of specific proteins enriched in the hMSC-EC secretome enhanced wound healing process under hyperglycemic conditions.     

Character and chlorine reactivity of dissolved organic matter from a mountain pine beetle impacted watershed

Lodgepole pine needle leachates from trees killed by the mountain pine beetle epidemic in Colorado were evaluated for dissolved organic matter (DOM) character, biodegradation, treatability by coagulation and disinfection byproduct (DBP) formation. An average of 8.0 (±0.62) mg-DOC/g-dry weight of litter was leached from three sets of needle samples representing different levels of forest floor degradation. Fluorescence analysis included collection of excitation and emission matrices, examination of peak intensities and development of a 4-component parallel factor (PARAFAC) analysis model. Peak intensity and PARAFAC analyses provided complementary results showing that fresh leachates were initially dominated by polyphenolic/protein-like components (60-70%) and humic-like fluorescence increased (40-70%) after biodegradation. Humic-like components were removed by coagulation (20-64%), while polyphenolic/protein-like components were not, which may create challenges for utilities required to meet OM removal regulations. DBP formation yields after 24 h chlorination were 20.5-26.4 μg/mg-DOC for trihalomethanes and 9.0-14.5 μg/mg-DOC for haloacetic acids for fresh leachates; increased after biodegradation to 19.2-64.2 and 7.1-30.9 μg/mg-DOC, respectively; and decreased after coagulation (fresh: 11.3-17.7;5.7-7.6 μg/mg-DOC, respectively; biodegraded: 12.0-27.3 and 2.9-7.2 μg/mg-DOC, respectively), reflective of changes in concentration of humic material. Humic-like PARAFAC components and peak intensities were positively correlated (R(2) ≥ 0.45) to DBP concentrations, while polyphenolic/protein-like components were not (R(2) ≤ 0.17).     

CD40–CD40L in Neurological Disease

Immune-inflammatory conditions in the central nervous system (CNS) rely on molecular and cellular interactions which are homeostatically maintained to protect neural tissue from harm. The CD40–CD40L interaction upregulates key proinflammatory molecules, a function best understood in the context of infection, during which B-cells are activated via CD40 signaling to produce antibodies. However, the role of CD40 in neurological disease of non-infectious etiology is unclear. We review the role of CD40–CD40L in traumatic brain injury, Alzheimer’s Disease, Parkinson’s Disease, stroke, epilepsy, nerve injury, multiple sclerosis, ALS, myasthenia gravis and brain tumors. We also highlight therapeutic advancements targeting the CD40 system to either attenuate the neuroinflammatory response or leverage the downstream effects of CD40 signaling for direct tumor cell lysis.     

An improved HPLC method for single‐run analysis of the spectrum of hop bittering compounds usually encountered in beers

A single‐run reverse phase‐high performance liquid chromatography method for the quantification of humulinones, α‐acids, iso‐α‐acids and reduced iso‐α‐acids (where present) in commercial beer samples is presented. The method utilizes a binary solvent system consisting of (A) 1% v/v acetic acid and (B) 0.1% v/v orthophosphoric acid in acetonitrile. Separation was achieved on a Purospher® star RP‐18 column (250 × 46 mm, 3 µm) with a flow rate of 0.5 mL/min. The compounds of interest eluted within 32 min. The method was fully validated according to International Conference on Harmonization guidelines and subsequently applied to monitor degradation of hop acids in a storage trial where four lager beers were aged at 28 and 38 °C for 70 and 60 days, respectively. Results confirmed the widely reported degradation through storage of trans‐iso‐α‐acids whilst demonstrating that the HLPC method was sufficiently sensitive to monitor and model this degradation. One beer exhibited a significantly lower (P < 0.05) rate of trans‐iso‐α‐acid degradation than the other conventionally hopped beers in the study, which might have been linked to its higher pH (4.71 vs 4.36). The relative stability of reduced iso‐α‐acids during ageing was also confirmed.     

Time Fractionation of Minor Lipids from Cold-Pressed Rapeseed Cake Using Supercritical CO<Subscript>2</Subscript>

This work explored the possibility of using supercritical carbon dioxide (SC-CO₂) to achieve fractionation of pre-pressed rapeseed (Brassica napus) cake oil at 30–50 MPa, at 40 or 80 °C, and increase the concentration of minor lipids (sterols, tocopherols, carotenoids) in the oil. Minor lipids are partially responsible for desirable antioxidant effects that protect against degradation and impart functional value to the oil. The weight and concentration of minor lipids in oil fractions collected during the first 60 min were analyzed. Cumulative oil yield increased with pressure, and with temperature at ≥40 MPa, but was lower at 80 °C than at 40 °C when working at pressure ≤35 MPa. Differences in solubility between the oil and minor lipids explained fractionation effects that were small for tocopherols. Unlike tocopherols, which are more soluble in SC-CO₂ than the oil, sterols and carotenoids are less soluble than the oil, and their concentration increased in the later stages of extraction, particularly at ≥40 MPa, when there was not enough oil to saturate the CO₂ phase. Because of the fractionating effects on rapeseed oil composition, there was an increase in the antioxidant activity of the oil in the second half as compared to the first half of the extraction. Consequently, this study suggests that SC-CO₂ extraction could be used to isolate vegetable oil fractions with increased functional value.     

Removing low ppb level perchlorate, RDX, and HMX from groundwater with cetyltrimethylammonium chloride (CTAC) pre-loaded activated carbon

Perchlorate contaminates vast amounts of groundwater throughout the United States which could potentially be used as potable water. Activated carbon pre-loaded with cetyltrimethylammonium chloride has been shown in this research to be an effective adsorbent for removing perchlorate from three low conductivity (50-66 microS/cm) groundwaters containing perchlorate (ClO(4)(-)) concentrations of 0.85, 1.0, and 5.6 parts per billion (ppb), respectively. In rapid small-scale column tests (RSSCTs), the virgin granular activated carbon (GAC) (used as a control) treated between 20,000 and 40,000 bed volumes (BV) of water. In contrast, the activated carbon that was pre-loaded with CTAC processed 170,000-270,000 BV before perchlorate was detected above 0.25 ppb in the effluent. Though this pre-loading significantly increased the capacity for perchlorate, it also diminished the GAC's capacity to remove organics. The groundwater containing 1 ppb ClO(4)(-) also contained the nitro-organics HMX (0.6 ppb) and RDX (5.5-6.6 ppb). RDX was detected in the effluent from the CTAC-pre-loaded bed after only 8000 BV had been processed whereas 308,000 BV could be processed through the virgin bed before RDX was detected. Likewise, HMX breakthrough was observed after 116,000 BV in the CTAC-pre-loaded bed while the virgin RSSCT exhibited no breakthrough of HMX during a test that was operated for 309,000 BV. However, by combining a CTAC-pre-loaded bed followed by a virgin GAC bed in series, both perchlorate and RDX could be removed for the same length of time.     

An Expedient Synthesis of Flexible Nucleosides through Enzymatic Glycosylation of Proximal and Distal Fleximer Bases

The structurally unique “fleximer” nucleosides were originally designed to investigate how flexibility in a nucleobase could potentially affect receptor–ligand recognition and function. Recently they have been shown to have low-to-sub-micromolar levels of activity against a number of viruses, including coronaviruses, filoviruses, and flaviviruses. However, the synthesis of distal fleximers in particular has thus far been quite tedious and low yielding. As a potential solution to this issue, a series of proximal fleximer bases (flex-bases) has been successfully coupled to both ribose and 2′-deoxyribose sugars by using the N-deoxyribosyltransferase II of Lactobacillus leichmannii (LlNDT) and Escherichia coli purine nucleoside phosphorylase (PNP). To explore the range of this facile approach, transglycosylation experiments on a thieno-expanded tricyclic heterocyclic base, as well as several distal and proximal flex-bases were performed to determine whether the corresponding fleximer nucleosides could be obtained in this fashion, thus potentially significantly shortening the route to these biologically significant compounds. The results of those studies are reported herein.     

Commercial Milk Enzyme‐Linked Immunosorbent Assay (ELISA) Kit Reactivities to Purified Milk Proteins and Milk‐Derived Ingredients

Numerous commercial enzyme‐linked immunosorbent assay (ELISA) kits exist to quantitatively detect bovine milk residues in foods. Milk contains many proteins that can serve as ELISA targets including caseins (α‐, β‐, or κ‐casein) and whey proteins (α‐lactalbumin or β‐lactoglobulin). Nine commercially‐available milk ELISA kits were selected to compare the specificity and sensitivity with 5 purified milk proteins and 3 milk‐derived ingredients. All of the milk kits were capable of quantifying nonfat dry milk (NFDM), but did not necessarily detect all individual protein fractions. While milk‐derived ingredients were detected by the kits, their quantitation may be inaccurate due to the use of different calibrators, reference materials, and antibodies in kit development. The establishment of a standard reference material for the calibration of milk ELISA kits is increasingly important. The appropriate selection and understanding of milk ELISA kits for food analysis is critical to accurate quantification of milk residues and informed risk management decisions.