Enzymatic pre-treatment of grape seeds for an oil with higher antioxidant activity

The paper investigates the effect of the enzymatic pre-treatment of grape seeds from six Romanian cultivars on the oil extracted. The grape seeds of some white and red Romanian grape varieties were separated from winery waste, washed, dried and ground, with the oil then obtained by extraction with petroleum ether. The extraction was performed directly or after a preliminary treatment with a commercial pectin lyase. The enzymatic procedure applied was more cost effective compared to other treatments previously described in which a cocktail of enzymes was used. The quantity of the extracted oil was measured in both types of processing, with an increase being observed for pre-treated samples. The fatty acid profiles (FAPs) of the oils resulted for the treated and untreated seeds were determined. No change in the composition was noticed. The reductive power of these oils was also investigated. Compared to the untreated samples for the same variety, the enzyme pre-treatment resulted in a superior antioxidant capacity.

Radular Morphology of Thiarid from Raja Ampat

Radula in gastropods is considered as a constant and conservative character that can be used in classification and phylogenetic at certain taxa level. However, character of radula is also ecophenotypic, so that the morphology of the radula can also indicates intraspecific variations that able to detect differences among species. The insights of the radular morphology specific to Thiaridae is important for classifying species within this family. Through an enzymatic procedure using the proteinase-K, five radula of Thiarid from Raja Ampat were extracted. The results of research on radula showed that the five species studied had the taeniglossan radula type. The band of the radula is 1.8 - 4.8 mm long and consists of marginal teeth, lateral teeth, and rachidian. The radula formula is 2/1/R/1/2 with a 3-4/1/3-4 rachidian pattern and a 2-3/1/2-3 lateral teeth pattern. The mesocone shape of the rachidian and lateral teeth varies between rounded and pointed.

Simple and Rapid Non-Enzymatic Procedure Allows the Isolation of Structurally Preserved Connective Tissue Micro-Fragments Enriched with SVF

The stromal vascular fraction (SVF) consists of a heterogeneous population of stem and stromal cells, generally obtained from adipose tissue by enzymatic digestion. For human cell-based therapies, mechanical process methods to obtain SVF represent an advantageous approach because they have fewer regulatory restrictions for their clinical use. The aim of this study was to characterize a novel commercial system for obtaining SVF from adipose tissue by a mechanical approach without substantial manipulations. Lipoaspirate samples collected from 27 informed patients were processed by a simple and fast mechanical system (by means of Hy-Tissue SVF). The Hy-Tissue SVF product contained a free cell fraction and micro-fragments of stromal connective tissue. The enzymatic digestion of the micro-fragments increased the yield of free cells (3.2 times) and CFU-F (2.4 times). Additionally, 10% of free cells from SVF were positive for CD34+, suggesting the presence of endothelial cells, pericytes, and potential adipose-derived stem cells (ADSC). Moreover, the SVF cells were able to proliferate and differentiate in vitro toward adipocytes, osteocytes, and chondrocytes. The immunophenotypic analysis of expanded cells showed positivity for typical mesenchymal stem cell markers. The Hy-Tissue SVF system allows the isolation of stromal vascular fraction, making this product of potential interest in regenerative medicine.

Establishment of a protocol for the isolation of ovarian preantral follicles derived from collared peccaries (Pecari tajacu)

SummaryWe compare the efficiency of mechanical or enzymatic methods, and their combination, for the isolation of ovarian preantral follicles (PFs) from collared peccaries. The ovaries from six females were subjected to the different methods investigated here. For the enzymatic method, ovary fragments were exposed to collagenase type IV in TCM-HEPES medium; the mechanical procedure was based on ovarian cortex dissociation by using a scalpel blade. The residual solution obtained after the mechanical isolation was subjected to the enzymatic procedure. The number of isolated PFs was quantified and classified as primordial, primary, or secondary; their viability was assessed using trypan blue dye assay. To confirm the results, PFs derived from the most efficient method were evaluated for integrity using scanning electron microscopy (SEM) and subjected to a 24 h in vitro culture for subsequent evaluation of viability by using fluorescent probes. A higher number of PFs (P < 0.05) was obtained from the enzymatic method (961.7 ± 132.9) in comparison with the mechanical method (434.3 ± 88.9), but no difference was observed between the two methods and their combination (743.2 ± 92.8). The trypan blue assay showed that the enzymatic method (98.7 ± 0.6%) provided the highest percentage of viable follicles (P < 0.05). Furthermore, SEM confirmed the ultrastructural integrity of the surface architecture of peccary PFs isolated by the enzymatic procedure; epifluorescence microscopy was used to confirm their viability (86.0%). In conclusion, we suggest that the enzymatic method investigated here is useful for the isolation of viable ovarian PFs from collared peccaries.

Synthesis of Ester-linked Taxol-oligosaccharide Conjugate and Its Drug Delivery System Using Bio-nanocapsules and Hybrid-bio-nanocapsules

Synthesis of ester-linked oligosaccharide conjugate of taxol, i.e., 7-glycolyltaxol 2″- O-α-maltotrioside, was investigated by enzymatic procedure. The starting material, 7-glycolyltaxol 2″- O-α-D-glucoside, was glycosylated by cyclodextrin glucanotransferase to 7-glycolyltaxol 2″- O-α-maltooligosides [α-maltooligosaccharides; α-Glc-1→(4-α-Glc-1→)n-14-α-glucosides (n = 1–4)]. The enzymatic hydrolysis of 7-glycolyltaxol 2″- O-α-maltooligosides gave 7-glycolyltaxol 2″- O-α-maltotrioside. Both bio-nanocapsules and hybrid-bio-nanocapsules containing 7-glycolyltaxol 2″- O-α-maltotrioside exhibited high in vitro anti-tumor activities.

Formal Synthesis of (+)-Madindoline A a Potent IL-6 Inhibitor Utilizing Enzymatic Discrimination of Quaternary Carbon

A formal synthesis of (+)-madindoline A was achieved. The Sunazuka's key intermediate (4 R,5 S)-(–)-3-butyl-4-( tert-butyldimethylsiloxy)-5-methoxycarbonyl-2,5-dimethyl-2-cyclopentenone was synthesized from easily available (2 S,3 S)-3-acetoxy-2-ethenyl-2-methylcyclopentanone. The starting material was reliably supplied by a chemo-enzymatic procedure in enantiomerically pure form. The synthesis was performed by straightforward transformations involving enone formation and regioselective introduction of the two alkyl side chains.