Grain-Based Dietary Background Impairs Restoration of Blood Flow and Skeletal Muscle During Hindlimb Ischemia in Comparison With Low-Fat and High-Fat Diets

Background: Among vascular pathologies associated with obesity, peripheral artery disease (PAD) occupies the important position. In clinical practice, nutritional interventions are recommended for patients with PAD. In this work, we investigated how the different dietary backgrounds affect the regeneration rate of ischemic hindlimb in mice.Methods: Male C57BL/6J mice were housed on three types of diet: low-fat (LFD), high-fat (HFD), and grain-based diet (GBD) for 13 weeks. Metabolic parameters including FBG level, ITT, and GTT were evaluated. The blood flow was assessed by laser Doppler scanning on 7, 14, and 21 days after hindlimb ischemia. Necrotic area of m.tibialis, macrophage infiltration, and angiogenesis/arteriogenesis were evaluated by histology. Glucose uptake in recovered skeletal muscle was analyzed using [3H]-2-deoxyglucose, and GLUT1 and GLUT4 expression were assessed by Western blotting.Results: In our work, we developed three experimental groups with different metabolic parameters: LFD with normal glucose metabolism, GBD with mild hyperglycemia, and HFD with impaired glucose tolerance. GBD-fed mice had a tendency to increase necrosis of m. tibialis and significantly higher macrophage infiltration than LFD and HFD groups. Moreover, GBD-fed mice had a trend to decreased blood flow recovery and significantly impaired arteriogenesis. Recovered skeletal muscle of GBD-fed mice had lower glucose uptake and decreased level of GLUT4 expression.Conclusion: Thus, we conclude that dietary background and metabolic status determine the rate of post-ischemic regeneration including angiogenesis, skeletal muscle recovery and metabolic activity. The most effective regeneration is supported by LFD, while the lowest rate of regeneration occurs on GBD.

In Vitro Regeneration of Tea (Camellia sinensis (L). O. Kuntze) By Somatic Embryogenesis from Immature Cotyledon Tissues

Tea (Camellia sinensis) is the world's most popular beverage plant, as well as an important plantation crop with high commercial value. It has been maintained for centuries through conventional vegetative propagation. Tea clonal propagation in vitro has the advantage of producing a large number of elite plants. If an efficient in vitro regeneration technology is available, this technique could be exploited for selection of tea plants for desired trait. The selected plants could be later on multiplied through in vitro or ex vitro techniques. The study aimed to induced somatic embryogenesis from immature embryo explants to genetic variaton. Different concentrations of phenylboronic acid with benzyladenine and phenylboronic acid with kinetin were tested in MS medium with 30 g/L sucrose and 8 g/L agar. MS medium without any plant growth regulators was used as control group. Considering the embryo survival rate, 1.5 mg/ L-1 phenylboronic acid + 1 mg/ L-1 kinetin produced highest result as 87.3% while lowest was in control group as 36.7%. The highest plant regeneration rate was found in 1,5 mg/ L-1 phenylboronic acid + 1 mg/ L-1 kinetin and 1.5 mg/ L-1 phenylboronic acid + 1 mg/ L-1 benzyladenine medium respectively as 58.3% and 55.6%. Kinetin treatment with increasing phenylboronic acid concentrations gave the best results in terms of somatic embryo survival rate. Also, kinetin treatment produced better results when compared to benzyladenine concentrations.

Efficient Plant Regeneration via Indirect Organogenesis in Carnation (Dianthus Caryophyllus Semperflorens flore pleno) Cultivars

ABSTRACT Callus induction and plant regeneration are important steps of in vitro plant breeding of ornamental plants. In this study, the effects of different combinations of plant growth regulators (PGRs), promoters, and minerals on callus induction and plant regeneration in different carnation cultivars were studied in a completely randomized design with three replications. For callus induction, 16 different combinations of 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA), 1-naphthaleneacetic acid (NAA), and casein hydrolysate (CH) were studied using in vitro leaf explants. The Murashige and Skoog (MS) medium supplemented with 0.2 mg·dm-3 of 2,4-D and 200 mg·dm-3 of CH showed the highest frequency of callus induction. Among the cultivars, ‘Noblesse’ showed the highest rate of callus induction (91.67%). Regarding regeneration, BA, NAA, silver nitrate (AgNO3), and adenine hemisulfate (As) were used in ten different combinations. The ‘Cameron’, ‘Tabasco’, and ‘Noblesse’ cultivars with 95.24% regeneration percentage showed the highest rate of plant regeneration. Generally, in most cultivars, the highest regeneration rate and shoot number per explant were found in the MS medium supplemented with 3 mg·dm-3 of BA, 0.6 mg·dm-3 of NAA, 5 mg·dm-3 of AgNO3, and 40 mg·dm-3 of As. According to the results, the highest regeneration frequency was obtained when 40 mg·dm-3 of As was added to the medium. Finally, the flow cytometry analysis indicated that there were no significant differences between in vitro regenerated and control plants in terms of DNA ratios.

Spontaneous Regeneration of Plantlets Derived from Hairy Root Cultures of Lopezia racemosa and the Cytotoxic Activity of Their Organic Extracts

A histological analysis was performed with the aim of elucidating the spontaneous regeneration process of the hairy root lines LRT 2.3 and LRT 6.4, derived from Lopezia racemosa leaf explants and genetically transformed with the Agrobacterium rhizogenes strain ATCC15834/pTDT. The analysis showed both lines regenerate via indirect somatic embryogenesis; LRT 6.4 also regenerated by direct organogenesis. The morphogenic characteristics of the regenerated plantlets from both lines showed the typical characteristics, described previously, including a higher number of axillary shoot formation, short internodes, and plagiotropic roots compared with wild-type seedlings. The regeneration process occurred without the addition of plant growth regulators and was linked to the sucrose concentration in the culture medium. Reducing the sucrose concentration from 3% to 2%, 1%, and 0.5% increased the regeneration rate in LRT 6.4; the effect was less pronounced in LRT 2.3. The cytotoxic activity of different organic extracts obtained from roots and shoots were evaluated in the cancer cell lines HeLa (cervical carcinoma), HCT-15 (colon adenocarcinoma), and OVCAR (ovary carcinoma). The hexane and dichloromethane extracts from roots of both lines showed cytotoxic activity against the HeLa cell line. Only the dichloromethane extract from the roots of PLRT 2.3 showed cytotoxic activity against the OVCAR cell line. None of the methanol extracts showed cytotoxic activity, nor the shoot extracts from any solvent.

Bioindicators of Behavior and Regeneration of a Caatinga Area in the Brazilian Semi-Arid - A Review

Studies that allow the understanding of the nutrient cycling processes and maintenance of soil fertility in the caatinga biome are necessary to assist in the proposal of possible management forms, in view of the sustainability of this ecosystem, to provide data in an area of knowledge the quantification of glomalin can be a good indicator of changes caused by the use of the soil and, in turn, therefore, it can become a good indicator of its recovery because it is correlated with important attributes of the soil. In addition to providing data of this process for a better understanding of the soils of the country, a study on this subject will also contribute with a new database for global monitoring. Research that can follow the regeneration rate of Caatinga forests in the Semi-arid will allow the development of models relating to several variables, not currently available for the semi-arid tropical region.

Development of An Efficient Protocol for Rapid Propagation of Curculigo orchioides Using Leaf Explant Culture

Curculigo orchioides is one of the most common medicinal plants used by diverse cultures and tribal groups. The roots of the plant are used medicinally in Asian countries. Curculigo orchioides have the ability to regenerate through seeds and tubers, but the regeneration rate is low. Plant tissue culture method was believed to have potential for rapid multiplication of this medicinal plant. An efficient protocol for rapid propagation of Curculigo orchioides, of the family Amaryllidaceae, was developed using leaf explants culture. The leaf explants (1 cm x 1 cm squares) cultured on Murashige and Skoog (MS) basal medium were supplemented with various concentrations and combinations of auxins and cytokinins with temperature 25 ± 2°C, relative humidity 85-90% and photoperiod of 12 hours light (2000-3000 lux). Callus induction was obtained within 4 weeks, 2,4-D at 3 mg/l formed profuse callus and the degree was found to be the highest (+++) among all the treatments. The best response to shoot induction, with maximum shoot number 5.33 (mean number of shoots per explant) was obtained using 1.0 mg/l 6-benzyl aminopurine (BAP) in combination with 1.0 mg/l Kinetin. In vitro shoots were induced for rooting on 0.5 mg/l of NAA supplemented medium. In order for seedlings propagated in vitro to adapt to natural conditions, plants were growned on a substrate coir: husk ash: sand (with the ratio of 0.5: 0.5: 1) in a greenhouse (humidity: 70%, temperature: 28-300C) gave 88.33% survival rate after 8 weeks of culture. With the results received, this is an effective approach to propagating Curculigo orchioides.

Temporary Immersion System Improves Regeneration of in vitro Irradiated Recalcitrant Rice Embryogenic Calli

The development of gamma rays mutant rice lines would be a solution for introducing variability in already farmer using varieties. In vitro gamma (60Co) mutagenesis reduces chimeras and allows a faster selection of desired traits but requires laboratory process optimization. The objective of the present work was the in vitro establishment of a recalcitrant rice embryogenic calli, the determination of its sensitivity to gamma radiation (Co-60), sequencing MATK and Rubisco for identification purposes, as well as generation optimization. The radiosensitivity of embryogenic calli resulted in an LD50 of 110Gy, while the 20% lethal dose was 64Gy. All sequenced genes matched perfectly with already reported MATK and Rubisco O. sativa genes with a clear SNP that identifies the local variety related to the southeast Asia Region. Callus induction improves with an MS with 2mg/L 2,4D, and the regeneration was achieved with an MS medium with 3mg/L BAP and 0,5mg/L NAA. The optimized radiation condition was 60Gy with an 83% regeneration in a semisolid medium, allowing a balance between mutation and regeneration. When increased to 80Gy, the regeneration rate falls to 29%. An immersion system (RITA®) of either 60 or 120 seconds every 8hours allowed a systematic and homogeneous total regeneration of the recalcitrant line, in contrast with the semisolid medium that resulted in positive but irregular regeneration. Other well-known recalcitrant cultivars, CR1821, CR1113 also had an improving regeneration in the immersion system, demonstrating its potential use for recalcitrant materials. To our knowledge, this is the first report on using an immersion system to allow regeneration of gamma-ray mutants from recalcitrant rice materials.

Development of haploids in the winter bread wheat anthers

In the modern world, the use of isolated anther cultivation technology is currently an integral part of the wheat breeding process. The development of haploids in the winter bread wheat anthers will allow obtaining new forms of wheat in the shortest possible time and without large areas. The purpose of the current study was to estimate the F3 winter bread wheat hybrids according to the anthers’ sensitivity to androgenesis and plant regeneration in vitro and to identify the factors affecting the yield of haploid production. There has been studied the ability to androgenesis in vitro in the anthers of four winter bread wheat hybrids of intensive and semi-intensive type of the FSBSI “ARC “Donskoy”. There has been assessed the role of the mineral composition of three induction nutrient media N6, W14 and NPB-99. There has been established a correlation between the main stages of development of haploids and a genotype. The highest regeneration rate of green plants was obtained in the sample F3 623 of intensive type (3.3%). The most suitable medium for androgenesis of the winter bread wheat anthers in vitro is NPB-99. Since the genotype F3 623 of intensive type demonstrated high values of haploid production capacity, it could be successfully used in breeding programs for the rapid production of homozygous wheat anther lines in vitro. Using two-way analysis of variance, there has been identified a correlation between the effects of a genotype, nutrient medium and their interaction with the main parameters of haploid formation in winter wheat. The formation of embryogenic structures is mainly associated with the effect of a genotype (46.52%). The proportion of the nutrient composition of the medium was low (1.82%), and the correlation factor was 2.1%. The genotype had the greatest effect on the indicator of the regenerants’ number. The nutrient medium had little effect. Regarding the regeneration of green plants, which is the main indicator of the haploid production, the share of a genotype effect was the largest (47.32%). The contribution of the medium and the correlation of factors were less important, but statistically significant.

Efficient Plant Regeneration System for New Guinea Impatiens (Impatiens Hawkeri W. Bull) CV. ‘Violet’and ‘Scarlet Bronze Leaf’

New Guinea Impatiens (Impatiens hawkeri W. Bull) is an eye-popping landscaping plant which is of bright and colorful blooms. A highly efficient in vitro plant regeneration system through direct shoot organogenesis was established for the first time from hypocotyl with partial cotyledons of New Guinea Impatiens. The results showed that Explant sterilization method, basic medium type,AgNO3,sucrose and plant growth regulators (PGRs) have greatly influences on in vitro morphogenesis.The regeneration rate in regeneration media that MS supplemented with 0.5mg·L−1 TDZ and 0.1mg·L−1 NAA was acceptable ,the induction rate of 'Violet' was 86.67%, and its proliferation coefficient was 5.27, while the induction rate of 'Scarlet Bronze Leaf' was 83.33%, and its proliferation coefficient was 5.13.PIC was unable to induce clumped sprouts, but it had a better effect on callus induction.We also included a shoot multiplication stage using regeneration New Guinea Impatiens medium that MS supplemented with 0.8mg·L−1 6-BA,0.5mg·L−1 TDZ and 0.05mg ·L−1 NAA.Reducing sucrose concentration to 20g·L−1 or adding 1mg·L−1AgNO3 could alleviate the vitrification phenomenon in the process of tufted bud proliferation.The optimal root culture medium for the regenerated seedlings of 'violet' and 'scarlet bronze leaf' of New Guinea Impatiens was MS supplemented with 0.05mg·L−1IBA, the rooting rate reached 100%.The study examined the micropropagation responses of New Guinea Impatiens in the presence of various growth regulators and provided a simple and more suitable protocol adapted for the mass propagation of clones.

The Role of Farnesoid X Receptor in Accelerated Liver Regeneration in Rats Subjected to ALPPS

Background: the role of bile acid (BA)-induced farnesoid X receptor (Fxr) signaling in liver regeneration following associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) was investigated in a rat model. Methods: Male Wistar rats underwent portal vein ligation (PVL) (n = 30) or ALPPS (n = 30). Animals were sacrificed pre-operatively and at 24, 48, 72, or 168 h after intervention. Regeneration rate, Ki67 index, hemodynamic changes in the hepatic circulation, and BA levels were assessed. Transcriptome analysis of molecular regulators involved in the Fxr signaling pathway, BA transport, and BA production was performed. Results: ALLPS induced more extensive liver regeneration (p < 0.001) and elevation of systemic and portal BA levels (p < 0.05) than PVL. The mRNA levels of proteins participating in hepatic Fxr signaling were comparable between the intervention groups. More profound activation of the intestinal Fxr pathway was observed 24 h after ALPPS compared to PVL. Conclusion: Our study elaborates on a possible linkage between BA-induced Fxr signaling and accelerated liver regeneration induced by ALPPS in rats. ALPPS could trigger liver regeneration via intestinal Fxr signaling cascades instead of hepatic Fxr signaling, thereby deviating from the mechanism of BA-mediated regeneration following one-stage hepatectomy.