Effect-Directed Profiling of Monofloral Honeys from Ethiopia by High-Performance Thin-Layer Chromatography and High-Resolution Mass Spectrometry.

Ethiopian honey is used not only as food but also for treatment in traditional medicine. For its valorization, bioactive compounds were analyzed in nine types of monofloral Ethiopian honey. Therefore, a non-target effect-directed profiling was developed via high-performance thin-layer chromatography combined with multi-imaging and planar effect-directed assays. Characteristic bioactivity profiles of the different honeys were determined in terms of antibacterial, free-radical scavenging, and various enzyme inhibitory activities. Honeys from Hypoestes spp. and Leucas abyssinica showed low activity in all assays. In contrast, others from Acacia spp., Becium grandiflorum, Croton macrostachyus, Eucalyptus globulus, Schefflera abyssinica, Vernonia amygdalina, and Coffea arabica showed more intense activity profiles, but these differed depending on the assay. In particular, the radical scavenging activity of Croton macrostachyus and Coffea arabica honeys, the acetylcholinesterase-inhibiting activity of Eucalyptus globulus and Coffea arabica honeys, and the antibacterial activity of Schefflera abyssinica honey are highlighted. Bioactive compounds of interest were further characterized by high-resolution mass spectrometry. Identifying differences in bioactivity between mono-floral honey types affects quality designation and branding. Effect-directed profiling provides new insights that are valuable for food science and nutrition as well as for the market, and contributes to honey differentiation, categorization, and authentication.

Enzyme activity, amino acid profiles and hydroxymethylfurfural content in Ethiopian monofloral honey.

The enzymes activity, hydroxymethylfurfural (HMF) and amino acids in honeys are relatively low. However, they play very significant role for honey quality. In this study, enzymes, amino acids and HMF contents of Ethiopian monofloral honeys were investigated. Diastase, invertase and HMF were analyzed based on the Harmonized International Honey Commission method and amino acids using amino acids analyzer (HPLC). Diastase activity ranged from 3.91 ± 0.730 (Schefflera abyssinica) to 13.6 ± 2.30 [Becium grandiflorum (L: Lalibella)]; invertase 36.5 ± 1.93 (Leucas abyssinica) to 4.85 ± 2.36 (Schefflera abyssinica); and HMF 0 ± 0 (Hypoestes and Leucas abyssinica) to 3.37 ± 1.73 (Croton macrostachyus). Significant variations were observed among Schefflera abyssinica honeys in diastase content, despite being from the same botanical origin. Significant variations were also observed among Becium grandiflorum honeys in invertase and diastase contents. Bees' geographical race and location affected enzymes activities. Lower level of enzymes could be an intrinsic characteristic of Ethiopian honey. Thus, enzymes activity alone cannot be a worthwhile indicator of quality for Ethiopian honey; besides diastase and invertase activity, the quality control of Ethiopian honeys should be supported by HMF parameters.

STAT1 phosphorylation and cleavage is regulated by the histamine (H4) receptor in human atopic and non-atopic lymphocytes.

Histamine can modulate the balance between T helper lymphocytes 1 and 2 (Th1 and Th2), and there is evidence that allergic reactions can be associated with excessive histamine production causing shifts toward Th2 responses. As signal transduction in Th-cells is specifically correlated to signal transducer and activator of transcription (STAT) activation and expression, the question arises whether histamine acting through histamine receptors (HR) induces modulation of the Janus kinase (JAK)-STAT pathway. Peripheral blood mononuclear cells (PBMC) from atopic and non-atopic donors were stimulated with phytohemagglutinin (PHA). Initial interleukin-4 (IL-4) levels were higher in the atopic group compared to the non-atopics and interferon-gamma (IFN-gamma) levels were lower. This was correlated to lower levels of STAT1 expression and phosphorylation. Furthermore, Western blots showed a 118-kDa STAT1 band at the start of the PHA stimulation that was apparently cleaved to STAT1alpha (91 kDa) and a 28 kDa-fragment with further stimulation. Histamine or the H4R agonist, clobenpropit, led to a significant suppression of the formation and phosphorylation of STAT1alpha in the non-atopic group after 48 h of PHA stimulation, but had no effect in the atopic group where STAT1alpha levels were already reduced. The STAT1alpha levels in the non-atopic group were enhanced by the H4R antagonist JNJ7777120. The phosphorylation of STAT1 could also be potentiated by the H4R antagonist, mimicking the precursor responses of STAT1alpha. JNJ7777120 also increased the binding of STAT1 to DNA and this effect could be reversed by histamine. As for histamine, the effects of the H4R antagonist were only seen in the non-atopic group. These results suggest that, in non-atopic individuals, histamine acting via the H4R can influence STAT1 regulation, but that this pathway is not present in atopics perhaps due to constitutive suppression of STAT1 activity.