Influence of Grape Flesh on the Retention and Composition of Polyphenols from Skins and Seeds.

This study examined the impact of grape flesh polysaccharide, protein, and amino acid contents on polyphenol retention from skins and seeds in Pinot noir (Vitis vinifera) and cold-hardy interspecific cultivars Marquette and Frontenac (Vitis spp.). After isolating grape tissues (skin, seed, and flesh), they were soaked either individually or combined with other tissues in a wine-like solution for up to 7 days. Findings revealed that flesh significantly reduces the concentration of condensed tannin, and mono- and diglucoside forms of anthocyanins in the supernatants, due to its rich content in polysaccharides and proteins. Frontenac skin and flesh tissues were the main sources of soluble proteins, amino acids, and soluble polysaccharides. Surprisingly, Marquette exhibited a higher retention of skin tannin than Pinot noir, likely due to its smaller tannin molecular mass, and a potential competitive effect with anthocyanins for the binding sites of flesh.

A CD36 transmembrane domain peptide interrupts CD36 interactions with membrane partners on macrophages and inhibits atherogenic functions.

CD36 is a transmembrane glycoprotein receptor for oxidized low density lipoprotein (LDL) and other endogenous danger signals and promotes athero-thrombotic processes. CD36 has been shown to associate physically with other transmembrane proteins, including integrins, tetraspanins, and toll-like receptors, which modulate CD36-mediated cell signaling. The CD36 N-terminal transmembrane domain (nTMD) contains a GXXXG sequence motif that mediates protein-protein interactions in many membrane proteins. We thus hypothesized that the nTMD is involved in CD36 interactions with other membrane proteins. CD36 interactions with partner cell surface proteins on murine peritoneal macrophages were detected with an immunofluorescence-based proximity ligation cross linking assay (PLA) and confirmed by immunoprecipitation/immunoblot. Prior to performing these assays, cells were incubated with a synthetic 29 amino acid peptide containing the 22 amino acid of CD36 nTMD or a control peptide in which the glycine residues in GXXXG motif were replaced by valines. In functional experiments, macrophages were preincubated with peptides and then treated with oxLDL to assess LDL uptake, foam cell formation, ROS formation and cell migration. CD36 nTMD peptide treated cells compared to untreated or control peptide treated cells showed decreased CD36 surface associations with tetraspanin CD9 and ameliorated pathologically important CD36 mediated responses to oxLDL, including uptake of DiI-labeled oxLDL, foam cell formation, ROS generation, and inhibition of migration.

Effects of Saignée and Bentonite Treatment on Phenolic Compounds of Marquette Red Wines.

To improve the phenolic extraction and color stability of red wine made from cold-hardy grapes, two winemaking practices, saignée and bentonite, were applied separately and in combination on Marquette grapes at crushing. The effects of these winemaking strategies on Marquette wine's basic chemical properties, monomeric and polymeric phenolic compounds were studied, as well as the development of color characteristics from crushing to 5 months of aging. The saignée (9% juice run-off) treatment showed little impact on the phenolic content of the finished wine, but showed an increase in color intensity. A hue shift towards an orange-yellow tone was observed in the bentonite-treated wines, which was associated with a loss of monomeric anthocyanins. The combination of saignée and bentonite showed less impact on removing anthocyanins and wine color, and increased phenolics content, therefore improving the extraction of non-anthocyanins monomeric phenolics. Although this combination treatment led to the highest concentration of tannin content after pressing, this difference between the control and other treatments disappeared over time. These results suggested that the interactions between tannins and other wine compounds still occur after removing proteins in Marquette wines.

Effect of the Application Time of Accentuated Cut Edges (ACE) on Marquette Wine Phenolic Compounds.

Cold-hardy interspecific hybrid grape varieties (Vitis spp.) have distinctive chemical compositions such as high acidity, a high content of anthocyanin diglucoside and a low condensed tannins content, compared to Vitis vinifera varieties. Considering the importance of phenolic compounds on the quality of red wine, a mechanical maceration technique, accentuated cut edges (ACE), has been evaluated when applied directly to crushed grapes (ACE-C), and 24 h before pressing (ACE-P), to improve the extraction of phenolic compounds. Samples were collected at crushing, bottling, and after five months of aging. Phenolic compounds and color characteristics of the wines were analyzed by high-performance liquid chromatography (HPLC) with diode array and fluorescence detectors and UV-Visible spectrophotometry. The color intensity, non-anthocyanin monomeric compounds and total iron-reactive phenolics content increased after applying ACE, compared to the control (CTL) after aging, and was significantly higher (37%) after ACE-C, compared to ACE-P. However, the concentration of condensed tannins was below the limit of detection in all the samples, indicating that ACE did not help their extraction or further interactions occurred with disrupted cell wall material. Applying ACE at crushing was considered as the optimum time to achieve a higher color stability in Marquette red wines.

Effects of Lyophilization on the Release Profiles of 3D Printed Delivery Systems Fabricated with Carboxymethyl Cellulose Hydrogel.

Recently, increasing numbers of researchers are becoming interested in 3D bioprinting because it provides customizability and structural complexity, which is difficult for traditional subtractive manufacturing to achieve. One of the most critical factors in bioprinting is the material. Depending on the bio-applications, materials should be bio-inert or bio-active, non-toxic, and along with those characteristics, mechanical properties should also meet the applicational or manufacturing requirement. As previously validated for bioprinting, carboxymethyl cellulose (CMC) hydrogel is focused on the printability and release control test in this study. With a differentiated weight percentage of CMC hydrogels were used to 3D print capsules filled with food degradable colorant at designated voids to mimic capsules manufactured for oral delivery. Standard USP (United States Pharmacopeia) dissolution apparatus II (Paddle) evaluations were performed both on lyophilized and non-lyophilized printed capsules. The first-order model was selected due to high linear fitting regression. Upon 24 h dissolution, non-lyophilized capsules showed a different release efficiency when the CMC percentage varied, while lyophilized capsules showed no significant difference. This study signifies the possibility of customizing oral drug delivery by printing capsules with CMC hydrogel. The improved delivery efficiency demonstrated by capsules with post-process lyophilizing proposed potential optimization options for pharmaceutical manufacturing industries.

Development of methylcellulose-based sustained-release dosage by semisolid extrusion additive manufacturing in drug delivery system.

The objective of this study is to fabricate customized dosage forms using extrusion-based 3D printing for the sustained delivery of theophylline. The therapeutic paste was prepared by combining various doses of theophylline (0, 75, 100, and 125 mg) with different concentrations of methylcellulose (MC) A4M (8, 10, and 12%). The paste was then 3D printed into semisolid tablets under optimized printing conditions. The rheological properties of printing pastes were related to the 3D printability. Our results indicated that to be 3D printed using the current platform, the storage modulus (G') of the printing paste should be higher than the loss modulus (G″) during the frequency sweep (0.1-600 rad/s), and the tan δ should fall in the range of 0.25-0.27 at 0.63 rad/s. The printed tablets formulated with 10% MC showed the highest overall quality, considering the aspects of resolution, texture, and shape retention regardless of the dosage. The scanning electron microscopy images indicated that the cross-linked structure of MC A4M formed the microscale porous microstructure, which has the potential to embed the theophylline, thus delayed the release through the barrier effect. The in vitro dissolution test revealed that the 3D printed tablets exhibited a sustained release during the first 12 hr. The findings in this study will support the development of customized, personalized medicine with improved efficacy.

3D printing of extended-release tablets of theophylline using hydroxypropyl methylcellulose (HPMC) hydrogels.

An extrusion based 3D printer was used to prepare the semi-solid tablets with different drug loading dosages (75, 100, 125 mg) under ambient temperature. The active pharmaceutical ingredient, theophylline, was uploaded within the hydrogels prepared of hydroxypropyl methylcellulose (HPMC) K4M or E4M. The HPMC concentrations were adjusted to different levels (10 and 12% w/w) to fulfill the requirements for 3D printing. Rheological and textural properties, as well as release profiles, were significantly affected by the type and concentration of excipient regardless of theophylline doses used. The printing material should exhibit shear-thinning behavior, keeping yield stress less than 4000 Pa and a loss factor (tanδ = G''/G') between 0.2 and 0.7, especially for 3D printing purposes using the current platform. The SEM images demonstrated that the hydrogel matrix exhibited a porous structure, which had the potential to encapsulate the theophylline clusters within its microstructure. The in vitro dissolution test showed that the release of all tablets was extended over 12 h, and the calculation of drug release kinetic models revealed that the 3D printed HPMC matrices release the theophylline by diffusion and erosion mechanisms. The excipient HPMC K4M 12% w/w hydrogel was optimal to load the theophylline with flexible dosage combinations due to the great extrudability and shape retention ability. The exploration of rheological properties was investigated in this study, and the results revealed that it is a feasible method to predict the SSE 3D printability and quality of hydrogel-API blend materials for the drug delivery system.

Determination of specific IgG4 for diagnosis and therapeutic evaluation of cerebral cysticercosis.

OBJECTIVE: To probe the significance of specific IgG4 in sera of patients with cerebral cysticercosis for diagnosis and therapeutic evaluation. METHODS: Specific IgG4 in sera of patients with cerebral cysticercosis was assessed using colloidal gold-labeled mouse-anti-human IgG4 McAb as probe. The results were compared with the CT image manifestation. RESULTS: The specific IgG4 positive rate in sera of patients with cerebral cysticercosis was 97.8%, whereas sera from patients with other kinds of parasitosis or central nerve system disease and the control group were all negative, except for a weak cross-reaction of sera from patients with hepatic echinococoosis. The determination of specific IgG4 in sera of patients with cerebral cysticercosis during different times of treatment showed that along with an increase in treatment time and improvement of clinical symptoms, specific IgG4 level gradually decreased. The positive rate and intensity of specific IgG4 in sera from patients with cerebral cysticercosis were consistent with the number of cysticercus parasites in the brain and pathologic changes, such as survival, disintegration, death and calcification. Survival of cysticercus in the brain was objectively evaluated using this technique. CONCLUSIONS: The determination of specific IgG4 in sera is a practical method for diagnosis and therapeutic evaluation of cerebral cysticercosis.