Antioxidant, Anti-Inflammatory, and Anti-Aging Potential of a Kalmia angustifolia Extract and Identification of Some Major Compounds

Skin aging is the most visible element of the aging process, giving rise to a major concern for many people. Plants from the Ericaceae family generally have antioxidant and anti-inflammatory properties, making them potential anti-aging active ingredients. This study aimed to evaluate the safety and anti-aging efficacy of a Kalmia angustifolia extract using reconstructed skin substitutes. The safety evaluation was performed using a 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay, while the efficacy was determined by assessing antioxidant and anti-inflammatory activity and analyzing skin substitutes reconstructed according to the self-assembly method by histology and immunofluorescence staining (elastin, collagen-1, collagen-3, aquaporin-3). The cell viability assay established the safety of the extract at a concentration up to 200 μg/mL. The Oxygen Radical Absorbance Capacity (ORAC) assay and a cell-based assay using 2’,7’-dichlorofluorescein-diacetate (DCFH-DA) revealed a strong antioxidant activity with an ORAC value of 16 µmol Trolox Equivalent/mg and a half-maximal inhibitory concentration (IC50) of 0.37 ± 0.02 μg/mL, while an interesting anti-inflammatory activity was found in the inhibition of NO production, with an inhibition percentage of NO production of 49 ± 2% at 80 µg/mL. The isolation and characterization of the extract allowed the identification of compounds that could be responsible for these biological activities, with two of them being identified for the first time in K. angustifolia: avicularin and epicatechin-(2β-O-7, 4β-6)-ent-epicatechin. Histological analyses of skin substitutes treated with the extract showed an increase in dermal thickness compared with the controls. K. angustifolia extract enhanced the expression of elastin and collagen-1, which are usually decreased with skin aging. These results suggest that K. angustifolia has promising antioxidant efficacy and anti-aging potential.

Twenty-five years post-treatment conifer responses to silviculture on a Kalmia-dominated site in eastern Canada

Research has demonstrated the potential of soil scarification, fertilization, and herbicide application to improve conifer seedling establishment and early growth. However, tree responses to and interactions among silvicultural treatments vary, making it difficult to predict mid- and long-term impacts of silviculture on stand productivity. We thus evaluated the 25-year effects of scarification and herbicide–fertilization combinations on black spruce (Picea mariana), jack pine (Pinus banksiana) and tamarack (Larix laricina) planted on a Kalmia angustifolia-dominated site. Our results show that the effects of scarification and herbicide–fertilization combinations diverged among species. Black spruce was the most responsive species to scarification for height and diameter at breast height. The combination of herbicide and fertilization treatments still had significantly positive effects on the long-term height and diameter growth of all species. Silvicultural treatments resulted in significant reductions in rotation length (based on height) when compared to height in nontreated-plots, depending on the species; reductions in years to attain a given height were greater for black spruce than for the other species. Our results illustrate the need to take species autecology into account when predicting productivity gains associated with early silviculture, and to provide managers with specific guidelines for the reforestation of ericaceous-dominated sites in Canadian boreal ecosystems.

Growth release of stunted black spruce (Picea mariana) in Kalmia heath: the role of ectomycorrhizal fungi and near-ground microclimate

Naturally regenerating and planted black spruce (Picea mariana (Mill.) B.S.P.) in post-fire landscapes in eastern Canada often exhibit stunted growth in the presence of ericaceous shrubs such as Kalmia angustifolia L. After a period of stunted growth, some seedlings experience a growth release, exhibiting growth rates closer to normally growing seedlings. We hypothesized that an increase in colonization of root tips by ectomycorrhizal (ECM) fungi is responsible for this release and that the percentage of root tips colonized by ECM fungi would be higher on seedlings that had a released or normally growing neighbour within close proximity. We quantified ECM fungi diversity and abundance from 255 soil cores from stunted, released, and normally growing black spruce seedlings sampled in two Kalmia-dominated sites in Newfoundland. Growth and microsite characteristics around each seedling were also measured. We found that normal and released seedlings had significantly higher proportions of ECM fungi root tips than stunted seedlings, supporting our final hypothesis; however, there was no significant difference in distance between neighbours. Soil chemical properties are thought to inhibit the vegetative spread of ECM fungi species in this particular system and are identified as an important topic for further research.

Genome Sizes and Ploidy Levels in the Genus Kalmia

Kalmia is a highly ornamental genus of shrubs native to North America and Cuba and grown as a valuable nursery crop throughout much of the temperate world. Although most species of Kalmia have previously been found to be diploid with 2n = 2x = 24, one species, Kalmia polifolia, has been found to be tetraploid. However, sampling within the genus has been limited, and information on the ploidy levels of specific cultivars is lacking. The objective of this study was to determine the relative genome sizes and ploidy levels of species, hybrids, and cultivars of Kalmia. Flow cytometry was used to determine the relative genome sizes of 67 accessions representing species, interspecific hybrids, cultivars, and chemically induced polyploids. Traditional cytology was used to calibrate genome sizes with ploidy levels. Results showed that relative genome sizes were conserved with 1Cx values ranging from 0.57 pg for Kalmia carolina to 0.70 pg for Kalmia latifolia. Most species of Kalmia were diploid including K. buxifolia (Leiophyllum buxifolium), K. carolina, K. cuneata, K. hirsuta, K. latifolia, and K. microphylla. Although plants of K. carolina (Kalmia angustifolia var. carolina) were uniformly diploid, the closely related, but more northerly distributed, K. angustifolia was primarily tetraploid, providing additional justification for treating these as separate species. An unusual triploid of K. angustifolia f. candida was also documented. Kalmia polifolia included both tetraploid and potentially pentaploid individuals, indicating a ploidy series within this species. Kalmia latifolia cultivars also included one triploid, two cytochimeras, and two chemically induced tetraploids. Overall, polyploidy was more prevalent in Kalmia than previously reported and varied both within and among species. This broader survey of relative genome sizes and ploidy levels in Kalmia provides valuable information for plant breeders and new insights into the systematics and cytogenetics of the genus.

Chromobacterium vaccinii sp. nov., isolated from native and cultivated cranberry (Vaccinium macrocarpon Ait.) bogs and irrigation ponds

A large number of Gram-negative, motile, mesophilic, violacein-producing bacteria were isolated from the soils and roots of Vaccinium macrocarpon Ait. and Kalmia angustifolia L. plants and from irrigation ponds associated with wild and cultivated cranberry bogs in Massachusetts, USA. Phylogenetic analyses of 16S rRNA gene sequences placed these isolates in a clade with Chromobacterium species, but the specialized environment from which they were isolated, their low genomic DNA relatedness with Chromobacterium violaceum ATCC 12472T and C. subtsugae PRAA4-1T, significant differences in fatty acid composition and colony morphology indicate that the cranberry and Kalmia isolates comprise a separate species of Chromobacterium , for which the name Chromobacterium vaccinii sp. nov. is proposed. Strain MWU205T ( = ATCC BAA-2314T = DSM 25150T) is proposed as the type strain for the novel species. Phenotypic analysis of 26 independent isolates of C. vaccinii sp. nov. indicates that, despite close geographical and biological proximity, there is considerable metabolic diversity among individuals within the population.

Cree antidiabetic plant extracts display mechanism-based inactivation of CYP3A4

Seventeen Cree antidiabetic medicinal plants were studied to determine their potential to inhibit cytochrome P450 3A4 (CYP3A4) through mechanism-based inactivation (MBI). The ethanolic extracts of the medicinal plants were studied for their inhibition of CYP3A4 using the substrates testosterone and dibenzylfluorescein (DBF) in high pressure liquid chromatography (HPLC) and microtiter fluorometric assays, respectively. Using testosterone as a substrate, extracts of Alnus incana , Sarracenia purpurea , and Lycopodium clavatum were identified as potent CYP3A4 MBIs, while those from Abies balsamea , Picea mariana , Pinus banksiana , Rhododendron tomentosum , Kalmia angustifolia , and Picea glauca were identified as less potent inactivators. Not unexpectedly, the other substrate, DBF, showed a different profile of inhibition. Only A. balsamea was identified as a CYP3A4 MBI using DBF. Abies balsamea displayed both NADPH- and time-dependence of CYP3A4 inhibition using both substrates. Overall, several of the medicinal plants may markedly deplete CYP3A4 through MBI and, consequently, decrease the metabolism of CYP3A4 substrates including numerous medications used by diabetics.