Weeks Of Maturation Research Articles

Constitutive and Synaptic Activation of GIRK Channels Differentiates Mature and Newborn Dentate Granule Cells.

Sparse neural activity in the dentate gyrus is enforced by powerful networks of inhibitory GABAergic interneurons in combination with low intrinsic excitability of the principal neurons, the dentate granule cells (GCs). Although the cellular and circuit properties that dictate synaptic inhibition are well studied, less is known about mechanisms that confer low GC intrinsic excitability. Here we demonstrate that intact G protein-mediated signaling contributes to the characteristic low resting membrane potential that differentiates mature dentate GCs from CA1 pyramidal cells and developing adult-born GCs. In mature GCs from male and female mice, intact G protein signaling robustly reduces intrinsic excitability, whereas deletion of G protein-activated inwardly rectifying potassium channel 2 (GIRK2) increases excitability and blocks the effects of G protein signaling on intrinsic properties. Similarly, pharmacological manipulation of GABAB receptors (GABABRs) or GIRK channels alters intrinsic excitability and GC spiking behavior. However, adult-born new GCs lack functional GIRK activity, with phasic and constitutive GABABR-mediated GIRK signaling appearing after several weeks of maturation. Phasic activation is interneuron specific, arising primarily from nNOS-expressing interneurons rather than parvalbumin- or somatostatin-expressing interneurons. Together, these results demonstrate that G protein signaling contributes to the intrinsic excitability that differentiates mature and developing dentate GCs and further suggest that late maturation of GIRK channel activity is poised to convert early developmental functions of GABAB receptor signaling into GABABR-mediated inhibition.SIGNIFICANCE STATEMENT The dentate gyrus exhibits sparse neural activity that is essential for the computational function of pattern separation. Sparse activity is ascribed to strong local inhibitory circuits in combination with low intrinsic excitability of the principal neurons, the granule cells. Here we show that constitutive activity of G protein-coupled inwardly rectifying potassium channels (GIRKs) underlies to the hallmark low resting membrane potential and input resistance of mature dentate neurons. Adult-born neurons initially lack functional GIRK channels, with constitutive and phasic GABAB receptor-mediated GIRK inhibition developing in tandem after several weeks of maturation. Our results reveal that GABAB/GIRK activity is an important determinant of low excitability of mature dentate granule cells that may contribute to sparse DG activity in vivo.

Increasing Cell Seeding Density Improves Elastin Expression and Mechanical Properties in Collagen Gel-Based Scaffolds Cellularized with Smooth Muscle Cells.

Vascular tissue engineering combines cells with scaffold materials in vitro aiming the development of physiologically relevant vascular models. For natural scaffolds such as collagen gels, where cells can be mixed with the material solution before gelation, cell seeding density is a key parameter that can affect extracellular matrix deposition and remodeling. Nonetheless, this parameter is often overlooked and densities sensitively lower than those of native tissues, are usually employed. Herein, the effect of seeding density on the maturation of tubular collagen gel-based scaffolds cellularized with smooth muscle cells is investigated. The compaction, the expression, and deposition of key vascular proteins and the resulting mechanical properties of the constructs are evaluated up to 1 week of maturation. Results show that increasing cell seeding density accelerates cell-mediated gel compaction, enhances elastin expression (more than sevenfold increase at the highest density, Day 7) and finally improves the overall mechanical properties of constructs. Of note, the tensile equilibrium elastic modulus, evaluated by stress-relaxation tests, reach values comparable to native arteries for the highest cell density, after a 7-day maturation. Altogether, these results show that higher cell seeding densities promote the rapid maturation of collagen gel-based vascular constructs toward structural and mechanical properties better mimicking native arteries.

Study of Anti-nutritional Compounds, Antioxidant Activity and Fatty Acid Composition of Moringa ( Moringa oleifera Lam.) Foliage

The study was conducted to investigate the effects of cutting intervals on the anti-nutritional factors, antioxidant activity and fatty acid composition of Moringa (Moringa oleifera Lam.) foliage. An established Moringa plot dividing into 12 equal plots was subjected to 3 different maturity stages of harvesting at 4, 6 and 8 weeks in a completely randomized block design experiment. It was found that the level of total phenol and tannin (mg tannic acid equivalent/g dry weight) at 4 (51.86 and 34.90), 6 (43.89, and 27.96) and 8 (29.00 and 16.66) weeks of maturity decreased significantly. Similarly, with the increase of maturity the level of condensed tannin significantly decreased (0.23, 0.17 and 0.14 mg catechin equivalent/g dry weight, respectively). In the case of antioxidant activity, significantly higher DPPH inhibition activity was found after 4 weeks (60.1 %) compared to 6 and 8 weeks of maturity (56.0 and 53.4 %, respectively). However, the fatty acid composition of Moringa foliage was significantly affected with harvesting stage of maturity. It contained the highest level of α-linolenic acid followed by palmitic acid, linoleic acid, stearic acid and oleic acid, respectively (48.71, 21.65, 13.07, 5.89 and 4.63 % of the total identified fatty acids, respectively). The average level of poly unsaturated fatty acid (PUFA) (61.78 %) was about three times higher than the level of saturated fatty acids (31.24 %). In conclusion, harvesting of Moringa foliage at 8 weeks interval would have less anti-nutritional factors with more PUFA and antioxidant activities.

A biomimetic hydrogel functionalized with adipose ECM components as a microenvironment for the 3D culture of human and murine adipocytes.

The lack of relevant in vitro models for adipose tissue makes necessary the development of a more physiological environment providing spatial and chemical cues for the effective maturation of adipocytes. We developed a biofunctionalized hydrogel with components of adipose extracellular matrix: collagen I, collagen VI, and the cell binding domain of fibronectin and we compared it to usual 2D cultures on plastic plates. This scaffold allowed 3D culture of mature adipocytes from the preadipocytes cell lines 3T3-L1 and 3T3-F442A, as well as primary Human White Preadipocytes (HWP), acquiring in vivo-like organization, with spheroid shaped adipocytes forming multicellular aggregates. The size of these aggregates increased with time up to 120 μm in diameter after 4 weeks of maturation, with good viability. Significantly higher lipogenic activity (up to 20-fold at day 28 for HWP cultures) and differentiation rates were also observed compared to 2D. Gene expression analyses highlighted earlier differentiation and complete maturation of 3D HWP compared to 2D, reinforced by the expression of Perilipin protein after 21 days of nutrition. This increase in adipocytes phenotypic and genotypic markers made this scaffold-driven culture as a robust adipose 3D model. Retinoic acid inhibition of lipogenesis in HWP or isoprenalin and caffeine induction of lipolysis performed on mouse 3T3-F442A cells, showed higher doses of molecules than typically used in 2D, underlying the physiologic relevance of this 3D culture system. Biotechnol. Bioeng. 2017;114: 1813-1824. © 2017 Wiley Periodicals, Inc.

Endogenous ABA content in relation to maturation of somatic embryos in Tulipa (L.) ‘Apeldoorn’ cultures

The aim of the present study was to estimate the endogenous abscisic acid (ABA) content in tulip ‘Apeldoorn’ torpedo and mature somatic embryos. Moreover, the effect of exogenous ABA and/or its inhibitor fluridone on somatic embryo maturation and conversion into plantlets was investigated. Torpedo-stage somatic embryos were subcultured on media containing 5 μM of picloram and 1 μM of 6-benzyl-aminopurine (BAP)—control, and combinations of ABA (0 or 10 μM) and/or fluridone (0 or 30 μM) for 1 week. Then, the torpedo embryos were transferred to a maturation medium containing 0.25 μM of α-naphthaleneacetic acid (NAA) and 2.5 μM of BAP, without ABA and fluridone treatment, and cultivated under darkness or light for ten weeks. Endogenous ABA content (first time measured in tulip somatic embryos) was evaluated by ELISA test. The obtained results revealed that the highest level of endogenous ABA, at 17.45 nmol g−1 dry weight (DW), was recorded in torpedo-stage of tulip embryo development, only after 1 week of ABA treatment, and was nearly 10 times higher in comparison with the control. Simultaneous addition of ABA and fluridone to the medium resulted in the lowering of the ABA concentration to 9.58 nmol g−1 DW. During ten weeks of maturation of the embryos, the endogenous ABA content in mature tissue of tulip somatic embryo considerably decreased to an amount 0.87–1.33 nmol g−1 DW (irrespective of ABA and fluridone treatment) and did not differ significantly from control (0.59 nmol g−1 DW). Exogenous ABA and fluridone significantly decreased the growth value of fresh weight (FW) of the tulip torpedo-shaped and mature embryos under light conditions. Percentage of the DW of the torpedo embryos treated with exogenous ABA was significantly higher (15.43–17.02) in comparison with the control (10.87). Three to three and a half times more malformed mature embryos were noted under light conditions than in darkness, irrespective of ABA and fluridone treatment. The highest percentage of mature embryos forming shoots (conversion) was observed under light conditions in the control and after fluridone treatment (26 and 20%, respectively).

Crystallization of nanostructured cobalt hydroxide carbonate at ambient conditions: a key precursor of Co3O4

AbstractCrystals of Co2CO3(OH)2 have been synthesized under ambient conditions, in contrast to hydrothermal methods reported previously. We have developed a simple but efficient methodology to obtain an initial amorphous phase that evolves to a crystalline cobalt hydroxide carbonate after one week of maturation. X-ray diffraction analysis indicates that this phase crystallizes in the space group P21/a (a = 12.886(6), b = 9.346(3), c = 3.156(1) Å, β = 110.358(6)°). The platelet morphology of Co2CO3(OH)2 agrees with its lamellar crystal structure. High-resolution transmission electron microscopy (HRTEM) reveals that each individual platelet is comprised of nanodomains disoriented with respect to their neighbours. The kinetics and the activation energy (Ea = 6.26 kJ mol–1) of the transformation process have been estimated using the rate constant method. The precipitation of solids leads to a decrease in the cobalt concentration in the solution (∼88%) reaching values of ∼150 ppm, which can be considered a successful reduction from the perspective of water quality. The calcination in air of the synthetized platelets produced exclusively Co3O4. The thermo-X-ray difraction results confirm that Co2CO3(OH)2 is transformed over a small range of temperatures (225–235°C) into pure Co3O4. HRTEM images show that the lamellar nanomorphology is preserved in the Co3O4 phase. Therefore, understanding the crystallization behaviour of Co2CO3(OH)2 can help to minimize environmental problems caused by cobalt pollution and may facilitate the management of methods to obtain phases with specific nanomorphologies used widely in material sciences.

Effect of Different Harvesting Regimes on the Pasting Properties of Selected Varieties of Banana – A Response Surface Study

Banana fruits from three banana varieties – M. Cavendish AAA, M. paradisiaca AAB, and M. acuminata AAA were harvested at 11, 13 and 15 weeks of maturity to determine the effect of different harvesting regimes on the pasting properties of the banana varieties. Flour was prepared from the banana fruits at the different harvesting regimes and the pasting properties of the flour samples were determined. A response surface, central composite for faced central composite design (K=2), was used to study the effect of the experimental variables - banana variety and time of harvesting on the pasting properties of banana flour. Regression models developed showed that the experimental variables had significant (p ≤ 0.05) effect on the peak, trough, breakdown, final and setback viscosities of flour processed from banana varieties. They also had significant (p ≤ 0.05) effect on the peak time and pasting temperature of the flour samples. The large range of values obtained in the pasting profile of the cassava roots studied is indicative of the wide genetic variations existing in the cassava clones analyzed and hence, the flour from the various clones will find wide applications for household and industrial uses.

Quantity of selected probiotic cultures in semi-hard cheese with low-cooking curd during the maturation process.

The work studies the survival of added selected probiotic bacteria Lactobacillus acidophilus (S1), Lactobacillus casei (S2), and Lactobacillus plantarum96 (S3) in semi-hard cheese with low-cooking curd during the maturation process. Cheeses were made according to the standard procedure (Polyfood SI 050 device). Probiotic lactobacilli strains Lactobacillus acidophilus (S1), Lactobacillus casei (S2), and Lactobacillus plantarum96 (S3) used in this study were added into the milk before the renneting process. The manufactured cheeses were matured for 6months at the temperature of 10°C. Cheese samples were taken for pH and titratable acidity measurements, lactobacilli enumeration, and chemical analysis at 30, 60, 90, 120, 150 and 180days of maturation. At the end of the experiment (180days) the cheese samples were analyzed also for the amount of lactic acid and protein contents. Initial numbers of lactobacilli inoculated into the milk (10(8)CFUmL(-1)) decreased during the first 2weeks of maturation and reached from 2.15 10(7)CFUg(-1) in S1 cheese to 4.32 10(7)CFUg(-1) in S3 cheese. The number of Lactobacillus acidophilus strain bacteria at the beginning of the maturation period was 2.47.10(7)CFUg(-1) and declined until day 120 of maturation to the number of 0.45 10(6)CFUg(-1). In the last month of the experiment day 180 the viable cell numbers started to rise up to the final number of 0.41 10(7)CFUg(-1). The numbers of Lactobacillus plantarum96 varied around 10(8)CFUg(-1) during the whole period of the experiment. According to our results it was detected that in all experimental cheeses, the used probiotic lactobacilli reached the values above 10(6)CFUg(-1). Thus the legislated and therapeutic minimum limits set for the products containing probiotic bacteria for human diet were fulfilled.

Biology of soft tissue wound healing and regeneration – Consensus Report of Group 1 of the 10th European Workshop on Periodontology

The scope of this consensus was to review the biological processes of soft tissue wound healing in the oral cavity and to histologically evaluate soft tissue healing in clinical and pre-clinical models. To review the current knowledge regarding the biological processes of soft tissue wound healing at teeth, implants and on the edentulous ridge. Furthermore, to review soft tissue wound healing at these sites, when using barrier membranes, growth and differentiation factors and soft tissue substitutes. Searches of the literature with respect to recessions at teeth and soft tissue deficiencies at implants, augmentation of the area of keratinized tissue and soft tissue volume were conducted. The available evidence was collected, categorized and summarized. Oral mucosal and skin wound healing follow a similar pattern of the four phases of haemostasis, inflammation, proliferation and maturation/matrix remodelling. The soft connective tissue determines the characteristics of the overlaying oral epithelium. Within 7-14 days, epithelial healing of surgical wounds at teeth is completed. Soft tissue healing following surgery at implants requires 6-8 weeks for maturation. The resulting tissue resembles scar tissue. Well-designed pre-clinical studies providing histological data have been reported describing soft tissue wound healing, when using barrier membranes, growth and differentiation factors and soft tissue substitutes. Few controlled clinical studies with low numbers of patients are available for some of the treatments reviewed at teeth. Whereas, histological new attachment has been demonstrated in pre-clinical studies resulting from some of the treatments reviewed, human histological data commonly report a lack of new attachment but rather long junctional epithelial attachment and connective tissue adhesion. Regarding soft tissue healing at implants human data are very scarce. Oral soft tissue healing at teeth, implants and the edentulous ridge follows the same phases as skin wound healing. Histological studies in humans have not reported new attachment formation at teeth for the indications studied. Human histological data of soft tissue wound healing at implants are limited. The use of barriers membranes, growth and differentiation factors and soft tissue substitutes for the treatment of localized gingival/mucosal recessions, insufficient amount of keratinized tissue and insufficient soft tissue volume is at a developing stage.

The anti-actin drugs latrunculin and cytochalasin affect the maturation of spruce somatic embryos in different ways

The role of the actin cytoskeleton in somatic embryo development was investigated using latrunculin B and cytochalasin D. Brief treatments (1h) with either drug at the start of maturation fragmented the actin in suspensor cells and/or depolymerized actin filaments in meristematic cells. The drugs targeted different cells: latB primarily affected the suspensor cells, but cchD damaged both suspensor and meristematic cells. Lethal damage to the meristematic and suspensor cells was observed when the drugs were applied throughout the maturation period, although the severity of this effect depended on their concentrations.The drugs’ effects on the yield of mature somatic embryos were investigated by applying them to embryo cultures throughout the maturation period or for one week at three different points in the maturation process: immediately prior to the start of maturation, during the first week of maturation, and during the fourth week of maturation. The strongest effects were observed when the drugs were applied at the start of maturation. Under these conditions, latB destroyed the suspensors, eliminating the underdeveloped embryos that depend on them. This accelerated the development of embryos that were capable of separating from the suspensors. Thus, while the total number of embryos at the end of the maturation period was lower than in untreated control cultures, the surviving mature embryos were of high quality. cchD treatment at the start of maturation strongly inhibited embryo development. Drug treatment at the end of the maturation period did not significantly affect embryo development: latB caused no change in the yield of somatic embryos, but cchD treatment increased the number of malformed embryos compared to untreated controls.

Properties of a proline-containing glass ionomer dental cement

Proline-containing glass ionomers are promising fast-set dental restorative materials with superior mechanical properties; however, little information is available on other physical properties of this type of glass ionomer. The objectives of this study were to synthesize and characterize a polyacrylic acid terpolymer containing proline derivative (PD) and to investigate the physical properties of this glass ionomer cement (GIC) and its cytotoxicity invitro. A terpolymer of AA (acrylic acid), IA (itaconic acid), and proline derivative (MP) with an 8:1:1 molar ratio was synthesized and characterized. Experimental GIC specimens were made from the synthetized terpolymer with Fuji IX (GC America, Alsip, Ill) commercial glass ionomer powder as recommended by the manufacturer. Specimens were mixed and fabricated at room temperature and were conditioned in distilled water at 37°C for 1 day and 1 week. Vickers hardness was determined with a microhardness tester. The water sorption characteristics and fluoride releasing properties of the specimens were investigated. The invitro cytotoxicity of the experimental glass ionomer was assessed by evaluating the C2C12 cell metabolism with methyltetrazolium (MTT) assay. Commercial Fuji IX was used as a control for comparison. The data obtained for the experimental GIC (PD) were compared with the control group by using 1- and 2-way ANOVA and the Tukey multiple range test at α=.05. Proline-modified GIC (PD) exhibited significantly higher surface hardness values (Vickers hardness number [VHN] 58 ±6.1) in comparison to Fuji IX GIC (VHN 47 ±5.3) after 1 week of maturation. Statistical analysis of data showed that the water sorption properties of the experimental cement (PD) were significantly greater than those of the control group (P<.05). The experimental GIC showed a significant increase in the amounts of initial fluoride release (P<.05) with continued fluoride release from the bulk of the material. The experimental group showed slightly reduced cell metabolism and cell number in comparison to the control group. However, the results were not statistically different (P>.05). An amino acid-containing GIC had better surface hardness properties than commercial Fuji IX GIC. This formulation of fast-set glass ionomer showed increased water sorption without adversely affecting the amount of fluoride release. Considering its biocompatibility, this material shows promise not only as a dental restorative material but also as a bone cement with low cytotoxicity.

Physicochemical quality of tequila during barrel maturation. A preliminary study

Silver Tequila 100% agave was aged in new oak barrels from four French regions. The evolution of physicochemical parameters regulated by Official Mexican Standard, as well as color, turbidity, total acidity and pH were assayed during 34 weeks of maturation. Analysis of variance (ANOVA) showed significant differences between barrel origin on furan-2-carboxaldehyde, dry extract, color and turbidity; and throughout maturation time on all evaluated parameters, except for total acidity, ethanol, and butan-2-ol. The principal component analysis (PCA) made it possible to separate three different groups corresponding to Silver (S), Aged (A), and tequilas during ripening transition (G), as well as to describe an evolution in two steps. Finally, general discriminant analysis (GDA) made it possible to classify correctly 69% of the samples according to barrel origin and 100% of the samples as G and A tequilas.

In search of markers for somatic embryo maturation in hybrid larch (Larix × eurolepis): global DNA methylation and proteomic analyses

A global DNA methylation and proteomics approach was used to investigate somatic embryo maturation in hybrid larch. Each developmental step during somatic embryogenesis was associated with a distinct and significantly different global DNA methylation level: from 45.8% mC for undifferentiated somatic embryos (1-week proliferation) to 61.5% mC for immature somatic embryos (1-week maturation), while maturation was associated with a decrease in DNA methylation to 53.4% for mature cotyledonary somatic embryos (8-weeks maturation). The presence of 5-azacytidine (hypo-methylating agent) or hydroxyurea (hyper-methylating agent) in the maturation medium altered the global DNA methylation status of the embryogenic cultures, and significantly reduced both their relative growth rate and embryogenic potential, suggesting an important role for DNA methylation in embryogenesis. Maturation was also assessed by examining changes in the total protein profile. Storage proteins, identified as legumin- and vicilin-like, appeared at the precotyledonary stage. In the proteomic study, total soluble proteins were extracted from embryos after 1 and 8 weeks of maturation, and separated by two-dimensional gel electrophoresis. There were 147 spots which showed significant differences between the stages of maturation; they were found to be involved mainly in primary metabolism and the stabilization of the resulting metabolites. This indicated that the somatic embryo was still metabolically active at 8 weeks of maturation. This is the first report of analyses of global DNA methylation (including the effects of hyper- and hypo-treatments) and proteome during somatic embryogenesis in hybrid larch, and thus provides novel insights into maturation of conifer somatic embryos.

Enhanced Osteoblastogenesis of Adipose-Derived Stem Cells on Spermine Delivery via β-Catenin Activation

The molecular mechanisms underlying spermine osteo-inductive activity on human adipose-derived stem cells (ASCs) grown in 3-dimensional (3D) cultures were investigated. Spermine belongs to the polyamine family, naturally occurring, positively charged polycations that are able to control several cellular processes. Spermine was used at a concentration close to that found in platelet-rich plasma (PRP), an autologous blood product containing growth and differentiation factors, which has recently become popular in in vitro and in vivo bone healing and engineering. Adipose tissue was surgically obtained from the hypodermis of patients undergoing hip arthroplasty. Patient age negatively affected both ASC yield and ASC ability to form 3D constructs. ASC 3D cultures were seeded in either non differentiating or chondrogenic conditions, with or without the addition of 5 μM spermine to evaluate its osteogenic potential across 1, 2, and 3 weeks of maturation. Osteogenic medium was used as a reference. The effects of the addition of spermine on molecular markers of osteogenesis, at both gene and protein level, and mineralization were evaluated. The effects of spermine were temporally defined and responsible for the progression from the early to the mature osteoblast differentiation phases. Spermine initially promoted gene and protein expression of Runx-2, an early marker of the osteoblast lineage; then, it increased β-catenin expression and activation, which led to the induction of Osterix gene expression, the mature osteoblast commitment factor. The finding that spermine induces ASC to differentiate toward mature osteoblasts supports the use of these easily accessible mesenchymal stem cells coupled with PRP for orthopedic applications.

High performance liquid chromatography-diode array detector (HPLC-DAD) detection of trans-resveratrol: Evolution during ripening in grape berry skins

Trans-resveratrol (3, 5, 4' – trihydroxy-trans-stilbene) is a stilbene naturally present in a number of plant families. Berry skins of grape (Vitis vinifera L.), Alicante, Black Malvasia, Nerello and Prunesta cultivars were analyzed at weekly intervals for trans-resveratrol production at five ripening stages during the last five weeks of maturation, from August 26th to September 23th. Analysis was carried out using the high performance liquid chromatography-diode array detector (HPLC-DAD) technique after fractioning of trans-resveratrol through a 500 mg C18 column (Solid Phase Isolation – SPI technique). A continuous decrease in trans-resveratrol content in all cultivars was observed during ripening. Alicante had the highest trans-resveratrol content, showing a decrease of 46% from the first to last sampling, while Black Malvasia produced the lowest amount of trans-resveratrol, 4 to 5 times less than Alicante at each sampling date. Key words: Antioxidants, grape, phenols, stilbenes, trans-resveratrol, high performance liquid chromatograph (HPLC).

ZMIANY BIOCHEMICZNE I MIKROBIOLOGICZNE W SERZE WYPRODUKOWANYM PRZY UDZIALE DROŻDŻY YARROWIA LIPOLYTICA

S u m m a r y A Yarrowia lipolytica JII1c yeast strain, isolated from the Polish ‘Rokpol’ mould cheese, was used as an adjunct culture in the production of a Dutch-type cheese. Its effect on the microbiological and biochemical characteristics of the cheese was evaluated in this research study. Milk used to produce the cheese was inoculated with 10 5 cfu/mL yeast cells. During the ripening process, the yeast population grew systematically to reach a maximum level of 7.9 log cfu/g in the sixth week of maturation, whereas the number of lactic acid bacteria increased until the fourth week of ripening. Thereafter, the number of microorganisms in the both groups decreased. After 8 weeks of ripening, the pH value of cheese inoculated with yeasts was significantly higher than that of the control cheese sample (produced without those microorganisms) and reached the levels of 6.37 and 5.47, respectively. In the experimental cheeses, it was also found that the utilization rate of lactic and citric acids was higher. Additionally, the concentration levels of water-soluble nitrogen (WSN) and free amino groups (FAG) in the experimental cheeses were about twice as high as in the control cheese sample. A more intensive proteolysis in the experimental cheese was accompanied by a higher accumulation of biogenic amines, especially of tyramine, putrescine, and 2-phenylethylamine; in the experimental cheese, after 8 weeks, their contents amounted to: 167.01, 77.90, and 69.54 mg/100 g, respectively. In contrast, the concentration of histamine was similar in both cheeses (9.47 and 9.81 mg/100 g in the control and experimental cheese samples, respectively). Also, the experimental cheese revealed more pronounced lipolysis resulting in a higher accumulation of free fatty acids, especially of butyric, myristic, palmitic, stearic, and oleic acids. It can be concluded that the Y. lipolytica JII1c grew well in the cheese causing the ripening process of the cheese to significantly accelerate.

Bypassing the Patient: Comparison of Biocompatible Models for the Future of Vascular Tissue Engineering

The objective of vascular tissue engineering is to develop tissue-engineered, biocompatible, small-diameter vessels suitable to withstand in vivo systolic pressures as well as be immunologically compatible with the patient, in order to minimize graft rejection. In this study, we present and compare two models of biocompatible, tissue-engineered vascular grafts (TEVG), using chitosan and acellularized rat aortas as options for scaffolds. Human aortic adventitial smooth muscle cells and fibroblasts were seeded onto a fibrin gel and subsequently wrapped around either of the two scaffolds. After several weeks of maturation in standard culturing conditions, the graft models were analyzed and compared by mechanical testing, cell viability, and histology. Histological and viability data showed that both models were viable and developed similarly, with a scaffold surrounded by two layers of cells, the fibroblasts lying on top of the smooth muscle cells. Both models responded to 200 mM potassium chloride (KCl) (tensions of 38 ± 3, 78 ± 13, and 52 ± 7 μN) and 25 mM 8-bromo-cyclic AMP (tensions of -23 ± 4, -39 ± 10, and -31 ± 12 μN) stimulation by vasoconstriction and vasorelaxation (n = 3), respectively; however, the chitosan model was unable to maintain the contracted and relaxed tension. Because the acellularized aorta TEVGs were able to maintain stimulated tension better than chitosan TEVGs, we concluded that the acellularized aorta model was better suited for further development.

Effect of harvest age and thermal processing on poly-γ-glutamate folates and minerals in African cowpea leaves (Vigna unguiculata)

Cowpea leaves (Vigna unguiculata) are highly consumed in Africa because the plant is easy to cultivate and highly disease/drought tolerant. The vegetables are harvested between 4 and 9 weeks of maturity. The objectives of this study were to (i) investigate the effect of vegetable maturity on total folate content, poly-γ-glutamate profile and mineral content and (ii) evaluate the effect of thermal treatments on folate stability and its poly-γ-glutamate profile changes. The total folate content and the folate poly-γ-glutamate chain length increased with increasing maturity of the vegetables. Folate occurred predominantly as tri-γ-glutamates (55%); chain lengths of up to tetra-γ-glutamates (14%) for 4-week-old vegetables. For 6- and 8-week-old vegetables, they occurred mainly as penta-γ-glutamates (78 and 60% respectively); chain lengths of up to hexa and hepta-γ-glutamates respectively. On crushing raw vegetables, at all stages of maturity, there was enzymatic conversion to short chain folate poly-γ-glutamates indicating the presence of endogenous γ-glutamyl hydrolase. Heating (30–70°C for10min) affected the stability and profile of folates. There was a general decline (>60%) in the levels of minerals during maturation of the vegetables. In conclusion, the maturity stage is an important factor in optimizing the type and quantity of these micronutrients.

Surgical Option for the Correction of Peyronie's Disease: An Autologous Tissue‐Engineered Endothelialized Graft

Surgical treatment is indicated in severe cases of Peyronie's disease. Incision of the plaque with subsequent graft material implantation is the option of choice. Ideal graft tissue is not yet available. To evaluate the use of an autologous tissue-engineered endothelialized graft by the self-assembly method, for tunica albuginea (TA) reconstruction in Peyronie's disease. Two TA models were created. Human fibroblasts were isolated from a skin biopsy and cultured in vitro until formation of fibroblast sheets. After 4 weeks of maturation, human umbilical vein endothelial cells (HUVEC) were seeded on fibroblasts sheets and wrapped around a tubular support to form a cylinder of about 10 layers. After 21 days of tube maturation, HUVEC were seeded into the lumen of the fibroblast tubes for the endothelialized tunica albuginea (ETA). No HUVEC were seeded into the lumen for the TA model. Both constructs were placed under perfusion in a bioreactor for 1 week. Histology, immunohistochemistry, and burst pressure were performed to characterize mature tubular graft. Animal manipulations were also performed to demonstrate the impact of endothelial cells in vivo. Histology showed uniform multilayered fibroblasts. Extracellular matrix, produced entirely by fibroblasts, presented a good staining for collagen 1. Some elastin fibers were also present. For the TA model, anti-human von Willebrand antibody revealed the endothelial cells forming capillary-like structures. TA model reached a burst pressure of 584 mm Hg and ETA model obtained a burst pressure of 719 mm Hg. This tissue-engineered endothelialized tubular graft is structurally similar to normal TA and presents an adequate mechanical resistance. The self-assembly method used and the autologous property of this model could represent an advantage comparatively to other available grafts. Further evaluation including functional testing will be necessary to characterize in vivo implantation and behavior of the graft.

The role of auxins in somatic embryogenesis of Abies alba

Abstract The somatic embryogenesis of conifers is a process susceptible to exogenous phytohormonal treatments. We report the effects of the synthetic auxin 2,4-dichlorophenoxyacetic acid (2,4-D) and the auxin inhibitor p-chlorophenoxyisobutyric acid (PCIB) on the endogenous level of the auxin indole-3-acetic acid (IAA) and on the anatomical composition of early somatic embryos of Abies alba (European silver fir). The embryogenic suspensor mass (ESM) of Abies alba proliferated on a medium supplemented by 2,4-D as well as on an auxin-free medium. The endogenous level of IAA was significantly higher in the ESM cultivated on a medium supplemented by 2,4-D. The decrease in the endogenous level of IAA in the first week of maturation is one of the most important stimuli responsible for the subsequent development of embryos. However, suppression of IAA synthesis by an auxin inhibitor did not stimulate the development of embryos. The maturation of somatic embryos from the globular to the cotyledonary stage occurs when the concentration of endogenous auxin in the ESM (including the embryos) increases. Early somatic embryos proliferating on a medium supplemented by auxin had an increased probability of maturing successfully. Exogenous auxin treatment during maturation did not compensate for the auxin deficiency during proliferation.