Explant type and natural ventilation systems influence growth and content of carvacrol and thymol of Lippia gracilis Schauer

The objective of this study was to evaluate the effects of different natural ventilation systems and explant types on the growth and volatile compound content of Lippia gracilis cultured in vitro. The treatments consisted of four membrane systems (without membrane, with one, two, and four porous membranes) and two explant types (nodal segments with and without a pair of leaves). The evaluation of growth, photosynthetic pigments and chemical analysis of the volatile fraction were performed at 35 days of cultivation in half strength MS basal medium. Natural ventilation systems significantly influenced the in vitro growth and volatile fraction of L. gracilis. Explants with a pair of leaves obtained the best experimental responses. The natural ventilation system with four membranes provided the best growth parameters and leaf area response of L. gracilis explants with leaves. The photosynthetic pigments decreased with an increase in the number of porous membranes in the culture flask. Variations in the number, content, and profile of volatile compounds under the influence of natural ventilation systems were observed. Major constituents such as ρ-cymene, γ-terpinene, thymol, carvacrol, and E-caryophyllene, regardless of experimental conditions, were identified. The highest carvacrol and thymol contents were observed in plantlets grown in culture flasks with four porous membranes. To maximize the content of carvacrol and thymol from the in vitro culture of L. gracilis, explants with a pair of leaves and four porous membranes in culture flasks are recommended for use.

     

In Vitro Regeneration of Achillea millefolium L from Shoot-tips and Root Segments of Seedlings

This report describes, for the first time, an efficient plant regeneration system for Achillea millefolium L (yarrow), a medicinal plant, via shoot multiplication from shoot-tips and adventitious shoot regeneration from root segments. Higher numbers of shoots were obtained when shoot-tips were cultured on MSMO medium supplemented with 3.0 mg l^?1 BA and 0.5 mg l^?1 IAA, or 5.0 mg l^?1 KIN and 1.0 mg l^?1 IBA, producing 17.3 and 17.0 shoots per explant at 100% frequency, respectively. For adventitous shoot regeneration, only root segments developed shoots when cultured on medium containing a combination of 1 mg l^?1 TDZ, 0.5 mg l^?1 IAA and 0.5 mg l^?1 GA3 (18.9 shoots per explant at 100% frequency), while other types of explants (i.e., cotyledons, leaf lamina and petiole segments) or hormonal combinations tested were found ineffective. Regenerated shoots rooted readily on MSMO medium containing different concentrations of IAA, IBA, NAA or 2,4-D, however, NAA at 0.5 mg l^?1, or IBA at 0.5 or 1.0 mg l^?1 were found to be the most productive. Nearly all of the regenerated plants (98%) survived through the hardening process when the rooted plantlets were kept at 55–65% relative humidity for 2 weeks, which were then planted in pots containing potting soil and kept at 25–35% humidity.     

CuO nanoparticles significantly influence in vitro culture,steviol glycosides,and antioxidant activities of <Emphasis Type="Italic">Stevia rebaudiana</Emphasis> Bertoni

Copper oxide (CuO) nanoparticles (NPs) synthesized through co-precipitation method were employed in MS media during in vitro culture of Stevia rebaudiana. Physiological characteristics, production of steviol glycosides, and antioxidative parameters were investigated in regenerated plants. CuO NPs had crystalline monoclinic cubic cuprous oxides with average size 47 nm. The NPs were applied at 0, 0.1, 1.0, 10, 100 and 1000 mg/L in MS media for direct organogenesis of S. rebaudiana from nodal segments. Shoot organogenesis was found highest (88.5%) at 10 mg/L CuO and average shoot length, mean number of shoot per explant, and fresh weight were also found significantly higher at the same concentration. High performance liquid chromatography (HPLC) illustrated significant rise of bioactive major steviol glycosides (rebaudioside A and stevioside) at 10 mg/L CuO NPs in MS media. The oxidative stress produced by CuO nanoparticles on S. rebaudiana was affirmed by antioxidant activities i.e. total antioxidant activity (TAC), total reducing power (TRP) and 2,2-diphenyl-1-picryl hydrazyl (DPPH)-free radical scavenging activity. The oxidative stress generated by NPs involved production of antioxidative molecules total phenolic content (TPC), total flavonoid content (TFC) depending on NPs concentration. The study concludes that copper oxide nanoparticles functions as a stimulator of bioactive components productions, and can be employed in in vitro batch cultures.     

GSPE is superior to NAC in the prevention of contrast-induced nephropathy: Might this superiority be related to caspase 1 and calpain 1?

Aims

Our study was intended to evaluate the role of inducible nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS), caspases 1 and 3 and calpain 1 in the pathogenesis of contrast-induced nephropathy (CIN) and to compare the protective effects of N acetyl cysteine (NAC) and grape seed proanthocyanidin extract (GSPE) against the development of CIN.

Main methods

32 rats were divided into four groups; control, contrast media (CM), CM + NAC and CM + GSPE. CIN was induced by administration of 7 ml/kg diatrizoate. The experiment was discontinued on the ninth day. Blood was collected for blood urea nitrogen (BUN) and creatinine measurement. Rat kidney tissues were removed for histopathological evaluation and the investigation of caspases 1 and 3, iNOS, eNOS, TUNEL and calpain 1.

Key findings

A significant increase in BUN, creatinine, renal histopathological injury, TUNEL, caspases 1, 3, calpain 1, iNOS and eNOS was observed in the CM group compared to the control group. There was amelioration in all these parameters in the CM + GSPE group, while there was no significant amelioration in BUN, creatinine and renal histopathological injury in the CM + NAC group. In addition, calpain 1 staining and creatinine were significantly lower in the CM + GSPE group compared to the CM + NAC group.

Significance

Our study showed, for the first time in the literature, that GSPE has a greater renoprotective effect compared with NAC and that this effective protection may be related to decrease in calpain 1 levels.     

Embryogenesis,plant regeneration and cardiac glycoside determination in Digitalis ferruginea subsp. ferruginea L

The present study reports, for the first time, an efficient in vitro plant regeneration protocol for Digitalis ferruginea subsp. ferruginea L. (rusty foxglove). We have used different concentrations of gibberellic acid (GA₃) on Murashige and Skoog (MS) medium to assess the germination frequency of seeds. High frequency of germination was achieved on MS medium with 1.0 mg l^?1 GA₃. 6-Benzylaminopurine (BAP) combined with α-naphtaleneacetic acid (NAA) or 2, 4-dichlorophenoxy acetic acid (2, 4-D) in the induction MS medium induced both somatic embryogensis and shoot organogenesis. The highest percentage of callus growth (85 %) was obtained when hypocotyl explants were cultured on MS medium containing 0.5 mg l^?1 2, 4-D plus 1.0 mg l^?1 BAP. The maximum mean number of somatic embryos (7.3 ± 1.3 embryos) or shoots (12.0 ± 1.1 shoots) per callus was obtained when medium contained 0.25 mg l^?1 NAA plus 1.0 mg l^?1 BAP or 0.5 mg l^?1 NAA plus 2.0 mg l^?1 BAP. The regenerated shoots easily rooted on MS medium. Higher amounts of lanatoside C [13.2 ± 0.5 mg 100 g^?1 dry weight (dw)] and digoxin (2.93 ± 0.31 mg 100 g^?1 dw) accumulation were obtained when shoots were obtained by indirect regeneration. We also investigated derivatives of cardenolides, i.e., digitoxigenin (730 ± 180 mg 100 g^?1 dw), gitoxigenin (50 ± 20 mg 100 g^?1 dw) and digoxigenin (490 ± 170 mg 100 g^?1 dw) from natural samples.     

An efficient in vitro regeneration system for Lythrum salicaria

This report describes an efficient plant regeneration system for the medicinal plant Lythrum salicaria via direct adventitious shoot development from leaf and stem explants. Leaf explants were much more responsive to regeneration than stem segments. Of the hormonal combinations tested, those involving thidiazuron (TDZ; 0.1, 0.3 or 0.5 mg dm⁻³) were more effective than the combinations of other hormones and 0.1 mg dm-3 TDZ combined with either indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) was the most productive. Rooting was readily achieved when multiple shoots were singled out and cultured on medium containing different auxins. IAA was the most effective on root development in terms of both the number of roots per shoot and the frequency of rooted shoots. More than 90 % of the regenerants survived after hardening for four weeks at gradually decreased air humidity.     

Regeneration and cardiotonic glycoside production in <Emphasis Type="Italic">Digitalis davisiana</Emphasis> Heywood (Alanya Foxglove)

A high-frequency in vitro regeneration of Digitalis davisiana Heywood (Alanya foxglove) and cardiotonic glycoside production from both in vitro produced materials (regenerated plantlets or germinated seedlings) and leaves of natural populations were obtained. Cardiac glycosides regulate heart rhythm and are effective in cancer chemotherapy, in particular for prostate and breast cancer treatments. Testing six different types of culture media revealed that Linsmaier and Skoog (LS) was the most effective for shoot production. Shoot regeneration efficiency was higher when flamingo-bill or hypocotyl explants were cultured on LS medium containing 0.5 mg/l thidiazuron (TDZ) and 0.25 mg/l indoleacetic acid (IAA). Rooting of all shoots that regenerated was readily achieved, even in the absence of plant growth regulators (PGRs). Production of cardenolides (lanatoside C and digoxin) in the materials grown in vitro seemed to correlate with several parameters, such as nutritional and hormonal compositions of the culture medium as well as the duration of culture on the initial regeneration and/or final growth medium. Higher amounts of digoxin accumulation were obtained when shoots were regenerated on LS or Gamborg’s B5 medium containing 0.5 mg/l TDZ and 0.25 mg/l IAA, producing 12.59 and 11.93 mg/kg dry weight (dw) digoxin, respectively. For natural populations, seasonal variations seemed to affect the production of digoxin in the leaves. The highest amount of digoxin (246.58 mg/kg dw) was in leaf samples collected in July, which coincides with the flowering stage of the plant in the region of collection.     

Solid state coordination chemistry of organodiphosphonates with copper(II) and auxilliary aromatic nitrogen heterocyclic ligands

The hydrothermal reactions of CuSO₄ · 5H₂O, a phosphonate ligand and an appropriate aromatic nitrogen heterocycle yield a series of materials of the Cu(II)/phosphonate/nitrogen ligand donor family. Two of the materials [Cu(2,2′-bpy)(HO₃PCH₂CH₂CH₂PO₃H)] (1) and [Cu(4,4′-bpy)(H₂O)₂(HO₃PCH₂CH₂CH₂CH₂PO₃H)] (2) are two-dimensional, while [{Cu(2,2′-bpy)}₂(O₃PC₆H₄PO₃)] · 8H₂O (3 · 8H₂O) is three-dimensional. When 1,3,5-benzenetricarboxylic acid is introduced as a reactant, the one-dimensional material [Cu(2,2′-bpy){OC₆H₂(CO₂H)₃}(HOPC₆H₅)] (4) is isolated. This is an example of an in situ hydrothermal ligand transformation in which the 1,3,5-benzene tricarboxylic acid is hydroxylated to give 1,3,5-benzene tricarboxylic acid-2-hydroxide. Compound 5 [Cu(terpy)(HO₃PC₆H₄PO₃H] is molecular.