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Ectoine as a promising protective agent in humans and animals
Abstract
Ectoine is a compatible water molecule-binding solute (osmoprotectant) produced by several bacterial species in response to osmotic stress and unfavourable environmental conditions. This amino acid derivative can accumulate inside cells at high concentrations without interfering with natural processes and can protect the cell against radiation or osmotic stress. This brief review presents the current state of knowledge about the effects of ectoine on animals and focuses on its practical use for enzyme stabilisation, human skin protection, anti-inflammatory treatment, inhibitory effects in neurodegenerative diseases, and other therapeutic potential in human or veterinary medicine
... Ectoine (1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid) is a compatible water molecule-binding solute, and a natural osmoprotectant produced by several bacterial species that survive well in extreme conditions of salinity, drought, irradiation, pH and temperature [13]. Ectoine is a strong cell protectant with a unique effect of preferential hydration [14]. Through its reorganization of water molecules, ectoine works to exclude osmolytes from interacting with proteins and membranes, which stabilizes their native structure and morphology and protects cells from environmental stress [15]. ...
... Ectoine is a natural cell-protective and inflammatory-reducing molecule with an excellent tolerability and safety profile [14,30,31]. Ectoine has a strong binding capacity to water molecules and can form a protective hydration shield around biomolecules [32]. ...
... An important clinical study has reported that changes in tear osmolarity do not correlate significantly with changes in patient symptoms or corneal fluorescein staining in dry eye disease [40]. We hypothesize that the protective effects of ectoine, an amino acid derivative, might contribute to its unique effect of strong preferential hydration [14], but not to its function as a lubricant. To clarify how this works, further studies are required to investigate the cellular pathway and molecular mechanism, such as Th1 and Th17 responses, which are well established as playing an important role in the pathogenesis of dry eye disease [41,42]. ...
Ectoine, a novel natural osmoprotectant, protects bacteria living in extreme environments. This study aimed to explore the therapeutic effect of ectoine for dry eye disease. An experimental dry eye model was created in C57BL/6 mice exposed to desiccating stress (DS) with untreated mice as controls (UT). DS mice were dosed topically with 0.5–2.0% of ectoine or a vehicle control. Corneal epithelial defects were detected via corneal smoothness and Oregon Green dextran (OGD) fluorescent staining. Pro-inflammatory cytokines and chemokines were evaluated using RT-qPCR and immunofluorescent staining. Compared with UT mice, corneal epithelial defects were observed as corneal smoothness irregularities and strong punctate OGD fluorescent staining in DS mice with vehicle. Ectoine treatment protected DS mice from corneal damage in a concentration-dependent manner, and ectoine at 1.0 and 2.0% significantly restored the corneal smoothness and reduced OGD staining to near normal levels. Expression of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) and chemokines CCL3 and CXCL11 was significantly elevated in the corneas and conjunctivas of DS mice, whereas 1.0 and 2.0% ectoine suppressed these inflammatory mediators to near normal levels. Our findings demonstrate that ectoine can significantly reduce the hallmark pathologies associated with dry eye and may be a promising candidate for treating human disease.
... Emerging evidence increasingly suggests that dysbiosis of the gut microbiota is associated with a wide range of diseases, including inflammatory bowel disease (IBD), cardiovascular disease, allergies, diabetes, and obesity (Bownik and Stępniewska, 2016;Koh et al., 2022). Previous studies have demonstrated a decrease in the abundance of genera such as Megamonas, Anaerostipes, and Bifidobacterium in patients with IBD (Arboleya et al., 2016;Yachida et al., 2019;Mager et al., 2020;Parker et al., 2020;Ren et al., 2020). ...
... These metabolites suggest the presence of intestinal inflammation and may also imply adverse effects on the digestive system following weaning (Bownik and Stępniewska, 2016;Koh et al., 2022). Furthermore, the downregulation of taurine and hypotaurine pathways after weaning may be linked to the inflammatory response, as studies have suggested that under certain disease conditions or severe stress, the demand for these compounds increases while their synthesis may be impaired (Singh et al., 2023). ...
Introduction
There are complex interactions between host and gut microbes during weaning, many of the mechanisms are not yet fully understood. Previous research mainly focuses on commercial pigs, whereas limited information has been known about the host and gut microbe interactions in miniature pigs.
Methods
To address the issue in Bama miniature piglets that were weaned 30 days after birth, we collected samples on days 25 and 36 for metabolomics, transcriptomics, and microgenomics analysis.
Results and discussion
The average daily weight gain of piglets during weaning was only 58.1% and 40.6% of that during 0-25 days and 36-60 days. Metabolomic results identified 61 significantly different metabolites (SDMs), of which, the most significantly increased and decreased SDMs after weaning were ectoine and taurocholate, respectively, indicating the occurrence of inflammation. Metagenomic analysis identified 30 significantly different microbes before and after weaning. Bacteria related to decreasing intestinal inflammation, such as Megasphaera, Alistipes and Bifidobacterium, were enriched before weaning. While bacteria related to infection such as Chlamydia, Clostridium, Clostridioides, and Blautia were enriched after weaning. The carbohydrate enzymes CBM91, CBM13, GH51_1, and GH94 increase after weaning, which may contribute to the digestion of complex plant fibers. Furthermore, we found the composition of antibiotic resistance genes (ARGs) changed during weaning. Transcriptomic analysis identified 147 significantly differentially expressed genes (DEGs). The upregulated genes after weaning were enriched in immune response categories, whereas downregulated genes were enriched in protein degradation. Combining multi-omics data, we identified significant positive correlations between gene MZB1, genera Alistipes and metabolite stachydrine, which involve anti-inflammatory functions. The reduced abundance of bacteria Dialister after weaning had strong correlations with the decreased 2-AGPE metabolite and the downregulated expression of RHBDF1 gene. Altogether, the multi-omics study reflects dietary changes and gut inflammation during weaning, highlighting complex interactions between gut microbes, host genes and metabolites.”
... Nowadays, ECT gained a significant interest in the healthcare sector due to its promising therapeutic benefits in a variety of human and animal disorders [5,6], such as GIT inflammation disorders [7,8], Alzheimer's disease [9], lung syndromes [10,11], and rhino-conjunctivitis symptoms [12,13]. In preclinical studies, ECT was shown to guard cells against the damage induced by various stressors and prevent the subsequent activation of the inflammatory cascades, leading to reducing the inflammatory process at the membrane level [14]. ...
... LLOQ, low, medium, and high QC samples (1,5,350, 750 ng/mL) were prepared and treated by the mentioned extraction procedure. For within-run accuracy and precision (n = 6), samples were measured continuously, while for between-run accuracy and precision (n = 18), they were measured on three consecutive days. ...
Ectoine (ECT) has recently gained considerable interest in the healthcare sector due to its promising therapeutic benefits in a variety of human disorders. This research aimed to quantify the ECT plasma level in rats by creating and optimizing a sensitive and validated UPLC-MS/MS method. Prior to analysis, ECT extraction from the plasma samples was conducted via a protein precipitation procedure, using hydroxyectoine as an internal standard (IS). A 1.7 μm UPLC C8 column (100 mm × 2.1 mm) was selected for the chromatographic separation, using a gradient mobile phase consisting of acetonitrile and 0.05% formic acid. The electrospray ionization mass spectrometry (ESI-MS) was used to detect ECT in the positive ion mode. To determine the specific precursor and the product ions of ECT, multiple reaction monitoring (MRM) methods were carried out. The selected ion pair of ECT was 143.1 > 97 and 159.1 > 113.13 for the IS. The ECT’s linearity range in rat plasma was found to be 1-1000 ng/mL, with a recovery rate of 96.48–97.37%. Consistent with FDA guidelines for bio-analytical method validation, the suggested method was validated. The method was efficiently employed to quantify the studied drug in spiked rat plasma with good accuracy and precision with no significant matrix effects. Furthermore, it was effectively used to investigate the pharmacokinetic behavior of ECT in rats after a single oral dose of 30 mg/kg.
... With its ability to resist high temperature and osmotic pressure, ectoine has gained significant commercial value in various industries including biopharmaceuticals, biotechnology, and fine chemicals [13]. Additionally, ectoine has been shown to exhibit anti-aging and cytoprotective effects, and may have potential therapeutic applications for diseases such as Alzheimer's disease [14,15]. Ectoine biosynthesis occurs through five sequential enzymatic reactions from L-aspartate, and its production is often triggered by increased environmental salinity [16]. ...
... Ectoine is a compatible solute naturally present in bacterial cells that functions as a protectant against harsh conditions, thus promoting cell growth and metabolism. Prior research has documented the protective effects of ectoine in bacteria, animals, and humans [14]. Ectoine, as a compatible solute and osmoprotectant, can protect halophilic microorganisms from stress factors and also reduce the wrinkled appearance of the spore [25]. ...
Monascus, a key player in fermented food production, is known for generating Monascus pigments (MPs) and monacolin K (MK), possessing bioactive properties. However, the limited stability of MPs and mycotoxin citrinin (CTN) constrain the Monascus industry. Extremolytes like ectoine, derived from bacteria, exhibit cytoprotective potential. Here, we investigated the impact of ectoine on Monascus purpureus ATCC 16365, emphasizing development and secondary metabolism. Exogenous 5 mM ectoine supplementation substantially increased the yields of MPs and MK (105%–150%) and reduced CTN production. Ectoine influenced mycelial growth, spore development, and gene expression in Monascus. Remarkably, ectoine biosynthesis was achieved in Monascus, showing comparable effects to exogenous addition. Notably, endogenous ectoine effectively enhanced the stability of MPs under diverse stress conditions. Our findings propose an innovative strategy for augmenting the production and stability of bioactive compounds while reducing CTN levels, advancing the Monascus industry.
... The reduction of cellular stress by compatible solutes is a highly conserved evolutionary biological process that occurs in living organisms from bacteria up to the highest classes of vertebrates. These molecules strongly bind water molecules and stabilize macromolecules without interfering with cellular processes [14]. ...
... Ectoine behaves as an osmoregulator molecule, attracting water to form a hydrating capsule around the main biological macromolecules, such as lipids and proteins. This mechanism of action is believed to promote the formation of biological macromolecules more thermodynamically stable, thus protecting individual cells and their components from insults mediated by external agents [14]. The ability of ectoine to reduce redness has been demonstrated in several preclinical studies [15][16][17]. ...
... AIDS and AIDS group compared to healthy control group. At the same time, AIDS and pre-AIDS groups had lower levels of phosphonoacetate and anti-inflammatory metabolites, such as vitamin B3 (niacinamide and nicotinamide riboside) (46,47), vitamin B6 [pyridoxine (48) and pyridoxamine (49)], ectoine (50), cinnamaldehyde (51), salidroside (52,53), melatonin (54,55) and fumaric acid (56), among which niacinamide, pyridoxine, ectoine, cinnamaldehyde and salidroside exhibited a decreasing trend with the severity of disease. Furthermore, except for spermidine, levels of other biogenic amines had an upward trend with the disease severity, including polyamines (putrescine, cadaverine, N-acetylputrescine) and phenylethylamine. ...
... Altered microbiota were also correlated to low antiinflammatory metabolites levels, such as vitamin B3 (46,47), vitamin B6 (48,49), ectoine (50), cinnamaldehyde (51), fumaric acid (56), ect. Among the altered microbiota, we observed notably lower abundance of bacteria relevant to the alleviation of inflammation in AIDS group, like Prevotella_9 (39), FIGURE 8 Alteration of KEGG pathways at different stages of HIV infection. ...
Background
The immune activation caused by microbial translocation has been considered to be a major driver of HIV infection progression. The dysbiosis of gut microbiota has been demonstrated in HIV infection, but the interplay between gut microbiota and its metabolites in the pathogenesis of HIV is seldom reported.
Methods
We conducted a case-controlled study including 41 AIDS patients, 39 pre-AIDS patients and 34 healthy controls. Both AIDS group and pre-AIDS group were divided according to clinical manifestations and CD4 + T cell count. We collected stool samples for 16S rDNA sequencing and untargeted metabolomics analysis, and examined immune activation and microbial translocation for blood samples.
Results
The pre-AIDS and AIDS groups had higher levels of microbial translocation and immune activation. There were significant differences in gut microbiota and metabolites at different stages of HIV infection. Higher abundances of pathogenic bacteria or opportunistic pathogen, as well as lower abundances of butyrate-producing bacteria and bacteria with anti-inflammatory potential were associated with HIV severity. The metabolism of tryptophan was disordered after HIV infection. Lower level of anti-inflammatory metabolites and phosphonoacetate, and higher level of phenylethylamine and polyamines were observed in HIV infection. And microbial metabolic pathways related to altered metabolites differed. Moreover, disrupted metabolites contributed by altered microbiota were found to be correlated to microbial translocation and immune activation.
Conclusions
Metabolites caused by dysbiosis of gut microbiota and related metabolic function are correlated to immune activation and microbial translocation, suggesting that the effect of microbiota on metabolites is related to intestinal barrier disruption in HIV infection.
... Ectoine (1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid), a cyclic amino acid derivative of L-aspartate, can protect living cells by alleviating the toxic effects on proteins, nucleic acids, and cell membranes aroused from extreme temperature, high osmolarity, dehydration, and radiation [1,2]. Ectoine has been exploited for multiple applications, including the inhibition of neurodegenerative diseases [3], skin protection against cell damage and aging [4], anti-inflammation treatment [5], organ transplantation maintenance [6], and other biomedical applications. However, the high production cost is the primary obstacle to ectoine availability in a broader consumption market [7][8][9]. ...
... In consideration of the significant effects of gene expression dose on metabolic flux and cell growth [39,40], herein, three plasmids pBBR1MCS-2, pCDFDuet-1 and pRSFDuet-1 with different copy numbers (5)(6)(7)(8)(9)(10), and approximate 100, respectively) were applied to reconstruction of the ectoine biosynthesis pathway. We found expression of ectABC gene cluster via the high copy numbered pRSFDuet-1 conferred the significantly increased ectoine accumulation (0.8 g L -1 , Fig. 2b). ...
Ectoine is a natural macromolecule protector and synthesized by some extremophiles. It provides protections against radiation-mediated oxidative damages and is widely used as a bioactive ingredient in pharmaceutics and cosmetics. To meet its growing commercial demands, we engineered Escherichia coli strains for the high-yield production of ectoine. The ectABC gene cluster from the native ectoine producer Halomonas elongata was introduced into different E. coli strains via plasmids and 0.8 g/L of ectoine was produced in flask cultures by engineered E. coli BL21(DE3). Subsequently, we designed the ribosome-binding sites of the gene cluster to fine tune the expressions of genes ectA, ectB, and ectC, which increased the ectoine yield to 1.6 g/L. After further combinatorial overexpression of Corynebacterium glutamicum aspartate kinase mutant (G1A, C932T) and the H. elongate aspartate-semialdehyde dehydrogenase to increase the supply of the precursor, the titer of ectoine reached 5.5 g/L in flask cultures. Finally, the engineered strain produced 60.7 g/L ectoine in fed-batch cultures with a conversion rate of 0.25 g/g glucose.
... Ectoine is a water-binding zwitterionic amino-acid derivative with numerous biotechnological applications and is a common component of cosmetic antiaging and moisturizing creams to improve skin resistance to surfactants in skin-cleansing solutions. Ectoine also alleviates skin inflammation and is currently recommended to treat moderate atopic dermatitis (Bownik and Stȩpniewska 2016). Furthermore, the compound is useful in sunscreens as it strongly absorbs UV radiation and in diverse cell types protects DNA from breaking down. ...
... Furthermore, the compound is useful in sunscreens as it strongly absorbs UV radiation and in diverse cell types protects DNA from breaking down. Ectoine also has other notable applications in medicine (Bownik and Stȩpniewska 2016). ...
Central-Andean Ecosystems (between 2000 and 6000 m above sea level (masl) are typical arid-to-semiarid environments suffering from the highest total solar and ultraviolet-B radiation on the planet but displaying numerous salt flats and shallow lakes. Andean microbial ecosystems isolated from these environments are of exceptional biodiversity enduring multiple severe conditions. Furthermore, the polyextremophilic nature of the microbes in such ecosystems indicates the potential for biotechnological applications. Within this context, the study undertaken used genome mining, physiological and microscopical characterization to reveal the multiresistant profile of Nesterenkonia sp. Act20, an actinobacterium isolated from the soil surrounding Lake Socompa, Salta, Argentina (3570 masl). Ultravioet-B, desiccation, and copper assays revealed the strain’s exceptional resistance to all these conditions. Act20’s genome presented coding sequences involving resistance to antibiotics, low temperatures, ultraviolet radiation, arsenic, nutrient-limiting conditions, osmotic stress, low atmospheric-oxygen pressure, heavy-metal stress, and toxic fluoride and chlorite. Act20 can also synthesize proteins and natural products such as an insecticide, bacterial cellulose, ectoine, bacterial hemoglobin, and even antibiotics like colicin V and aurachin C. We also found numerous enzymes for animal- and vegetal-biomass degradation and applications in other industrial processes. The resilience of Act20 and its biotechnologic potential were thoroughly demonstrated in this work.
... Ectoine is a naturally occurring carboxamidine produced by extremophiles [27] and is compatible with cell metabolism even at high concentrations. Ectoine acts as a kosmotrope and protein stabilizer, boosting hydrophobic contacts and folding, and stabilizing cell membranes [28]. The established beneficial effects of ectoine include anti-inflammatory properties, which result from the deactivation of the pro-inflammatory transcription factor nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB), and anti-oxidant properties, which arise from the elimination of hydroxyl radicals. ...
The most severe form of muscular dystrophy (MD), known as Duchenne MD (DMD), remains an incurable disease, hence the ongoing efforts to develop supportive therapies. The dysregulation of autophagy, a degradative yet protective mechanism activated when tissues are under severe and prolonged stress, is critically involved in DMD. Treatments that harness autophagic capacities therefore represent a promising therapeutic approach. Osmolytes are protective organic molecules that regulate osmotic pressure and cellular homeostasis and may support tissue-repairing autophagy. We therefore explored the effects of the osmolyte ectoine in the standard mouse model of DMD, the mdx, focusing on the autophagy-related proteome. Mice were treated with ectoine in their drinking water (150 mg/kg) or through daily intraperitoneal injection (177 mg/kg) until they were 5.5 weeks old. Hind limb muscles were dissected, and samples were prepared for Western blotting for protein quantification and for immunofluorescence for an evaluation of tissue distribution. We report changes in the protein levels of autophagy-related 5 (ATG5), Ser366-phosphorylated sequestosome 1 (SQSTM1), heat shock protein 70 (HSP70), activated microtubule-associated protein 1A/1B-light chain 3 (LC3 II) and mammalian target of rapamycin (mTOR). Most importantly, ectoine significantly improved the balance between LC3 II and SQSTM1 levels in mdx gastrocnemius muscle, and LC3 II immunostaining was most pronounced in muscle fibers of the tibialis anterior from treated mdx. These findings lend support for the further investigation of ectoine as a potential therapeutic intervention for DMD.
... 27 Ectoine has a unique effect of strong preferential hydration through its reorganization of water molecules. 28 Ectoine was found to inhibit the inflammatory responses in chronic inflammatory diseases. 29 For ocular disease, ectoine has been used in clinical trials to treat dry eye disease 30,31 and seasonal allergic rhinoconjunctivitis. 31,32 However, these clinical trials of ectoine topical application are limited in a few European countries, such as Russia, Ukraine, Switzerland, and Germany, but not in the United States; and the investigations were restricted to clinical efficacy on dry eye symptoms and signs. ...
Purpose
This study aimed to explore protective effects and potential mechanism of ectoine, a natural osmoprotectant, on ocular surface mucin production in dry eye disease.
Methods
A dry eye model was established in C57BL/6 mice exposed to desiccating stress (DS) with untreated (UT) mice as controls. DS mice were topically treated with 2.0% ectoine or PBS vehicle. Corneal epithelial defects were assessed by Oregon Green Dextran (OGD) fluorescent staining. Conjunctival goblet cells, ocular mucins, and T help (Th) cytokines were evaluated by immunofluorescent staining or ELISA, and RT-qPCR.
Results
Compared with UT mice, corneal epithelial defects were detected as strong punctate OGD fluorescent staining in DS mice with vehicle, whereas ectoine treatment largely reduced OGD staining to near-normal levels. Conjunctival goblet cell density and cell size decreased markedly in DS mice, but was significantly recovered by ectoine treatment. The protein production and mRNA expression of two gel-forming secreted MUC5AC and MUC2, and 4 transmembrane mucins, MUC1, MUC4, MUC16, and MUC15, largely decreased in DS mice, but was restored by ectoine. Furthermore, Th2 cytokine IL-13 was inhibited, whereas Th1 cytokine IFN-γ was stimulated at protein and mRNA levels in conjunctiva and draining cervical lymph nodes (CLNs) of DS mice, leading to decreased IL-13/IFN-γ ratio. Interestingly, 2.0% ectoine reversed their alternations and restored IL-13/IFN-γ balance.
Conclusions
Our findings demonstrate that topical ectoine significantly reduces corneal damage, and enhances goblet cell density and mucin production through restoring imbalanced IL-13/IFN-γ signaling in murine dry eye model. This suggests therapeutic potential of natural osmoprotectant ectoine for dry eye disease.
... It is a compatible solute, which means it may accumulate in cells at high concentrations without impairing cellular functions. Ectoine acts as a kosmotrope and protein stabilizer, boosting hydrophobic contacts and folding, and stabilizes cell membranes [26]. Additional beneficial effects include anti-inflammatory properties, via deactivation of the pro-inflammatory transcription factor nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB), and antioxidant properties, through elimination of hydroxyl radicals. ...
The severest form of muscular dystrophy (MD) termed Duchenne MD (DMD) remains to this day an incurable disease, hence the continued effort to further develop supportive therapies. Perturbation of autophagy, a degradative yet protective mechanism activated when tissues face severe and persistent stress, is critically involved in DMD, consequently, treatments harnessing autophagic capacities represent an amenable therapeutic strategy. We studied the effects of the protective osmolyte ectoine in the standard mouse model of DMD, the MDX, zooming in on the autophagy-related proteome. From birth, mice were treated with ectoine in the drinking water (150mg/kg) or through daily intraperitoneal injection (177mg/kg) until 5 weeks old. Hind limb muscles were dissected, performing western blotting for protein quantification and immunofluorescence to investigate tissue distribution. We report significant alterations of total levels of autophagy related 5 (ATG5), Ser366 phosphorylated sequestosome 1 (SQSTM1) and heat shock protein 70 (HSP70), and of activated microtubule-associated protein 1A/1B-light chain 3 (LC3 II) in mitochondrial fractions. Ectoine tends to restore the shifted balance between LC3 II and SQSTM1 levels observed in MDX muscle, and LC3 II upregulation was most pronounced on muscle fibers of MDX treated with ectoine in the drinking water. Findings from this explorative study support a possible beneficial effect of ectoine on the autophagic deficit in the MDX, which warrants its further exploration as a therapeutic supplement for DMD.
... For example, ectoin has been used in anti-aging topicals due to its inhibitory effects on tyrosinase activity, anti-inflammatory effects, and ability to absorb UV radiation, thereby protecting DNA from photodamage. 58 The topical application of astaxanthin has been shown to protect against UV irradiation, 59,60 thereby improving wrinkle severity and skin hydration. 61 Furthermore, THDA has been shown to improve hyperpigmentation and photodamaged skin. ...
Purpose
The topical application of antioxidants has been shown to augment the skin’s innate antioxidant system and enhance photoprotection. A challenge of topical antioxidant formulation is stability and penetrability. The use of a targeted drug delivery system may improve the bioavailability and delivery of antioxidants. In this ex vivo study, we assessed the effects of the topical application of a liposome-encapsulated antioxidant complex versus a free antioxidant complex alone on skin photoaging parameters and penetrability in human skin explants.
Patients and Methods
Human organotypic skin explant cultures (hOSEC) were irradiated to mimic photoaging. The encapsulated antioxidant complex and free antioxidant complex were applied topically onto the irradiated hOSEC daily for 7 days. The two control groups were healthy untreated hOSEC and irradiated hOSEC. Photoprotective efficacy was measured with pro-inflammatory cytokine (IL-6 and IL-8) and matrix metalloproteinase 9 (MMP-9) secretion. Cell viability and metabolic activity were measured via resazurin assay. Tissue damage was evaluated via lactate dehydrogenase (LDH) cytotoxicity assay. Skin penetration of the encapsulated antioxidant complex was assessed via fluorescent dye and confocal microscopy.
Results
Compared to healthy skin, irradiated skin experienced increases in IL-6, IL-8 (p < 0.05), and MMP-9 (p < 0.05) secretion. After treatment with the encapsulated antioxidant complex, there was a 39.3% reduction in IL-6 secretion, 49.8% reduction in IL-8 (p < 0.05), and 38.5% reduction in MMP-9 (p < 0.05). After treatment with the free antioxidant complex, there were no significant differences in IL-6, IL-8, or MMP-9 secretion. Neither treatment group experienced significant LDH leakage or reductions in metabolic activity. Liposomes passed through the stratum corneum and into the epidermis.
Conclusion
The topical application of a liposome-encapsulated antioxidant complex containing ectoin, astaxanthin-rich microalgae Haematococcus pluvialis extract, and THDA improves penetrability and restored IL-6, IL-8, and MMP-9 levels in irradiated human skin explants, which was not seen in the comparator free antioxidant complex group.
... The pathological changes involved in OA are characterized by degeneration and injury of the articular cartilage free radicals. Therefore, ectoine can be widely used in the fine chemical, biomedicine, and biomanufacturing industries [9,10]. Currently, ectoine is used in treating diseases such as upper airway inflammation, inflammatory bowel disease, xerophthalmia, dry rhinitis, Alzheimer's disease, and Parkinson's disease. ...
Osteoarthritis (OA) is a chronic degenerative disease that primarily includes articular cartilage destruction and inflammatory reactions, and effective treatments for this disease are still lacking. The present study aimed to explore the protective effects of ectoine, a compatible solute found in nature, on chondrocytes in rats and its possible application in OA treatment. In the in vitro studies, the morphology of the chondrocytes after trypsin digestion for 2 min and the viability of the chondrocytes at 50°C were observed after ectoine treatment. The reactive oxygen species (ROS) levels in chondrocytes pretreated with ectoine and post-stimulated with H2O2 were detected using an ROS assay. Chondrocytes were pretreated with ectoine before IL-1β stimulation. RT‒qPCR was used to measure the mRNA levels of cyclooxygenase-2 (COX-2), metallomatrix proteinase-3, -9 (MMP-3, -9), and collagen type II alpha 1 (Col2A1). In addition, immunofluorescence was used to assess the expression of type II collagen. The in vivo effect of ectoine was evaluated in a rat OA model induced by the modified Hulth method. The findings revealed that ectoine significantly increased the trypsin tolerance of chondrocytes, maintained the viability of the chondrocytes at 50°C, and improved their resistance to oxidation. Compared with IL-1β treatment alone, ectoine pretreatment significantly reduced COX-2, MMP-3, and MMP-9 expression and maintained type II collagen synthesis in chondrocytes. In vivo, the cartilage of ectoine-treated rats exhibited less degeneration and lower Osteoarthritis Research Society International (OARSI) scores. The results of this study suggest that ectoine exerts protective effects on chondrocytes and cartilage and can, therefore, be used as a potential therapeutic agent in the treatment of OA.
... Likewise, another interesting finding was the negative association between VOO consumption and ectoine. Ectoine is a secondary metabolite [41,42] produced by certain bacterial genera, such as Streptomyces spp., and has been identified in small concentrations in the human intestinal microbiota [43]. Its presence in the context of olive oil consumption suggests a potential interaction between the gut microbiome and dietary patterns. ...
Background
Olive oil consumption has been inversely associated with the risk of type 2 diabetes (T2D) and cardiovascular disease (CVD). However, the impact of olive oil consumption on plasma metabolites remains poorly understood. This study aims to identify plasma metabolites related to total and specific types of olive oil consumption, and to assess the prospective associations of the identified multi-metabolite profiles with the risk of T2D and CVD.
Methods
The discovery population included 1837 participants at high cardiovascular risk from the PREvención con DIeta MEDiterránea (PREDIMED) trial with available metabolomics data at baseline. Olive oil consumption was determined through food-frequency questionnaires (FFQ) and adjusted for total energy. A total of 1522 participants also had available metabolomics data at year 1 and were used as the internal validation sample. Plasma metabolomics analyses were performed using LC–MS. Cross-sectional associations between 385 known candidate metabolites and olive oil consumption were assessed using elastic net regression analysis. A 10-cross-validation (CV) procedure was used, and Pearson correlation coefficients were assessed between metabolite-weighted models and FFQ-derived olive oil consumption in each pair of training–validation data sets within the discovery sample. We further estimated the prospective associations of the identified plasma multi-metabolite profile with incident T2D and CVD using multivariable Cox regression models.
Results
We identified a metabolomic signature for the consumption of total olive oil (with 74 metabolites), VOO (with 78 metabolites), and COO (with 17 metabolites), including several lipids, acylcarnitines, and amino acids. 10-CV Pearson correlation coefficients between total olive oil consumption derived from FFQs and the multi-metabolite profile were 0.40 (95% CI 0.37, 0.44) and 0.27 (95% CI 0.22, 0.31) for the discovery and validation sample, respectively. We identified several overlapping and distinct metabolites according to the type of olive oil consumed. The baseline metabolite profiles of total and extra virgin olive oil were inversely associated with CVD incidence (HR per 1SD: 0.79; 95% CI 0.67, 0.92 for total olive oil and 0.70; 0.59, 0.83 for extra virgin olive oil) after adjustment for confounders. However, no significant associations were observed between these metabolite profiles and T2D incidence.
Conclusions
This study reveals a panel of plasma metabolites linked to the consumption of total and specific types of olive oil. The metabolite profiles of total olive oil consumption and extra virgin olive oil were associated with a decreased risk of incident CVD in a high cardiovascular-risk Mediterranean population, though no associations were observed with T2D incidence.
Trial registration: The PREDIMED trial was registered at ISRCTN (http://www.isrctn.com/, ISRCTN35739639).
... Ectoine is a natural component found in extremophilic bacteria that can adapt to extremely harsh environments as well as osmotic stress conditions. This property is related to the prior exclusion of ectoine to the interface of a lipid monolayer of the cell membrane [134], and ectoine can play a protective role. As a bioinspired permeating CPA that does not interfere with normal cellular activities [135] , Choi JK et al. used ectoine as a bioinspired CPA component to replace DMSO and vitrified human oocytes. ...
There have been increasing requirements for women’s fertility preservation due to oncological and nononcological reasons in recent years, and meeting these demands will be a hot topic in the coming years. Oocyte cryopreservation is a workable option for preserving women’s fertility, and great advances have already been made and much progress has been made in mammalian gene banking and human oocyte banks. In this paper, we systematically introduce the history of oocyte cryopreservation and vitrification technology and highlight the vitrification carrier. Furthermore, we summarize the fundamentals of oocyte vitrification and discuss the effects of vitrification on oocyte quality. Strategies to improve the effect of oocyte cryopreservation are also proposed. At the end of this review, we conclude oocyte cryopreservation and outline future perspectives.
... Ectoine is a naturally occurring organic molecule. It functions as a protective agent, preventing damage to biological structures from harsh environmental conditions such as osmotic and thermal stress [34]. Ectoine also has water retention properties, allowing it to maintain hydration levels in cells, which is essential for the survival of organisms [35]. ...
Metabolic engineering is a promising strategy to realize green synthesis of valued chemicals derived from petroleum. According to the literature, cell factories for producing L-aspartate and its derivatives (β-alanine, ectoine, 3-hydroxypropionate, D-pantothenic acid and L-homoserine) have been developed. In this review, we firstly introduced the functions, applications and markets of L-aspartate and its derivatives. Then, the current research progress on microbial production of them was elaborated in detail. Finally, we have discussed the limiting factors and given some suggestions for realizing applications of engineered bacteria in the industry, including metabolic engineering of the bacteria to increase the titer, yield and productivity of the target products, fermentation condition optimization and downstream purification. With the development of novel technologies and increased investments in synthetic biology, it is promising to realize sustainable production of L-aspartate and its derivatives at the industrial scale in the future.
... www.nature.com/scientificreports/ Ectoin and arylpolyene related-BGCs are known to protect cells against oxidative effects 93,94 . We identified ectoin BGCs in Halomonas (LC_1), Halopseudomonas (LC_3) and Cobetia (LC_6), while arylpolyene BGCs were found in Pseudoalteromonas (LC_2), Shewanella (LC_4), Brucella (LC_5) and Cobetia (LC_6). ...
Here we report the oil degradation genetic potential of six oil-degrading bacteria (ODB), previously used as a bioremediation consortium, isolated from the hydrocoral Millepora alcicornis and seawater. The strains were identified as Halomonas sp. (LC_1), Cobetia sp. (LC_6), Pseudoalteromonas shioyasakiensis (LC_2), Halopseudomonas aestusnigri (LC_3), Shewanella algae (LC_4), and Brucella intermedia (LC_5). The taxonomic identification differed from that of the original paper when we used whole genome gene markers instead of just 16S rRNA gene. Genes responsible for the degradation of aromatic hydrocarbons and n-alkanes were found in all genomes, although different (and complementary) steps of the metabolic pathways were unique to each strain. Genes for naphthalene and toluene degradation were found in various strains. We annotated quinate degradation genes in LC_6, while LC_3 and LC_5 presented genes for biosurfactant and rhamnolipid biosynthesis. We also annotated genes related to beneficial mechanisms for corals, such as genes involved in nitrogen and DMSP metabolism, cobalamin biosynthesis and antimicrobial compounds production. Our findings reinforce the importance of using bacterial consortia for bioremediation approaches instead of single strains, due to their complementary genomic arsenals. We also propose a genome-based framework to select complementary ODB that can provide additional benefits to coral health.
... Transgenic plants that can synthesize ectoine because of the introduced bacterial ectoine biosynthesis cluster can resist severe drought stress [17]. Its equal effectiveness in protecting human skin from water loss makes it attractive for use in advanced cosmetics [6,18,19]. Ectoine can protect enzymes [20,21] and DNA damage caused by high doses of UV irradiation [22,23]. It can inhibit the aggregation and neurotoxicity of β-amyloid in neurodegenerative Alzheimer's [24,25]. ...
Ectoine and its derivative 5-hydroxyectoine are compatible solutes initially found in the hyperhalophilic bacterium Ectothiorhodospira halochloris, which inhabits the desert in Egypt. The habitat of ectoine producers implies the primary function of ectoine as a cytoprotectant against harsh conditions such as high salinity, drought, and high radiation. More extensive and in-depth studies have revealed the multiple functions of ectoine in its native producer bacterial cells and other types of cells and its biomolecular components (such as proteins and DNA) as a general protective agent. Its chemical properties as a bio-based amino acid derivative make it attractive for basic scientific research and related industries, such as the food/agricultural industry, cosmetic manufacturing, biologics, and therapeutic agent preparation. This article first discusses the functions and applications of ectoine and 5-hydroxyectoine. Subsequently, more emphasis was placed on advances in bio-based ectoine and/or 5-hydroxyectoine production. Strategies for developing more robust cell factories for highly efficient ectoine and/or 5-hydroxyectoine production are further discussed. We hope this review will provide a valuable reference for studies on the bio-based production of ectoine and 5-hydroxyectoine.
... [15] Due to their safety as showed by animal and human studies, their applications are spread in the medical, cosmetics and biochemistry fields. [16] Recently, they have been used as cryopreservants for mammalian cells, [17,18] and proved to maintain the viability of air dried and freeze dried Escherichia coli. [19] Although the mechanism underlying this cryopreservation ability has not been fully unraveled yet, there are various hypothesis that suggest potential mechanisms of action through biophysical specific features. ...
... Meanwhile, antiSMASH detected the presence of biosynthetic gene clusters accounting for compounds such as ectoine and albaflavenon. Ectoine is an extremolyte naturally found in bacteria that thrive in extreme environmental conditions such as high salinity, irradiation, drought, extreme pH, and temperature [110,111] . Ectoine confers protection to the bacteria by regulating osmotic stress, stabilizing lipid bilayers, preventing DNA and protein damage, and offering hydroxyl radical scavenging activity [112] . ...
A novel strain, Streptomyces griseiviridis MUM 136JT was recovered from a mangrove forest soil in Malaysia. The Gram-positive bacterium forms strong yellow aerial mycelium and moderate yellow substrate mycelium on ISP 2 agar. A polyphasic approachwas used to determine the taxonomy status of strain MUM 136JT. The strain showed a spectrum of phylogenetic and chemotaxonomic properties consistent with those of the members of the genus Streptomyces. The cell wall peptidoglycan was determined to contain LL-diaminopimelic acid. The predominant menaquinones were identified as MK-9(H8) and MK-9(H6), while the identified polar lipids consisted of lipid, aminolipid, phospholipid, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, diphosphatidylglycerol, and phosphatidylinositolmannoside. The cell wall sugars consist of ribose, mannose, and galactose. The predominant cellular fatty acids (>10.0 %) were identified as iso-C16:0 (31.6 %), anteiso-C15:0 (14.8 %), iso-C15:0 (12.0 %), and anteiso-C17:0 (11.1 %). Phylogenetic analysis identified that closely related strains for MUM 136JT are Streptomyces leeuwenhoekii DSM 42122T (98.9 %), Streptomyces erythrogriseus JCM 9650T (98.4 %), Streptomyces griseoincarnatus JCM 4381T (98.5 %). The DNA-DNA relatedness values between MUM 136 JT and closely related type strains ranged from 13.3 ± 1.5 % to 17.4 ± 2.0 %. The name Streptomyces griseiviridis sp. nov. is proposed, and the type strain is MUM 136JT (= NBRC 114249T = MCCC 1K04199T).
... Both molecules provide excellent cosmotropic [1], protein-and DNA-stabilizing [4,6,7], as well as cell-and tissue-protecting and moisturizing properties [4,[6][7][8], which easily explains why they are of increasing commercial interest. Notably, the two ectoines are applied in high-price sectors such as cosmetics [9,10], medicine [11][12][13] and biotechnology [14][15][16][17]. Ectoine is the industrial flagship among the world's extremolytes. ...
Background
Extremolytes enable microbes to withstand even the most extreme conditions in nature. Due to their unique protective properties, the small organic molecules, more and more, become high-value active ingredients for the cosmetics and the pharmaceutical industries. While ectoine, the industrial extremolyte flagship, has been successfully commercialized before, an economically viable route to its highly interesting derivative 5-hydroxyectoine (hydroxyectoine) is not existing.
Results
Here , we demonstrate high-level hydroxyectoine production, using metabolically engineered strains of C. glutamicum that express a codon-optimized, heterologous ectD gene, encoding for ectoine hydroxylase, to convert supplemented ectoine in the presence of sucrose as growth substrate into the desired derivative. Fourteen out of sixteen codon-optimized ectD variants from phylogenetically diverse bacterial and archaeal donors enabled hydroxyectoine production, showing the strategy to work almost regardless of the origin of the gene. The genes from Pseudomonas stutzeri (PST) and Mycobacterium smegmatis (MSM) worked best and enabled hydroxyectoine production up to 97% yield. Metabolic analyses revealed high enrichment of the ectoines inside the cells, which, inter alia , reduced the synthesis of other compatible solutes, including proline and trehalose. After further optimization, C. glutamicum Ptuf ectD PST achieved a titre of 74 g L ⁻¹ hydroxyectoine at 70% selectivity within 12 h, using a simple batch process. In a two-step procedure, hydroxyectoine production from ectoine, previously synthesized fermentatively with C. glutamicum ectABC opt , was successfully achieved without intermediate purification.
Conclusions
C. glutamicum is a well-known and industrially proven host, allowing the synthesis of commercial products with granted GRAS status, a great benefit for a safe production of hydroxyectoine as active ingredient for cosmetic and pharmaceutical applications. Because ectoine is already available at commercial scale, its use as precursor appears straightforward. In the future, two-step processes might provide hydroxyectoine de novo from sugar.
... The main plasma components correlating with sphero-echinocytes are lactate and 2,3-diphospho-glycerate (2,3-DPG), associated to increased glycolysis; nicotinamide and tryptophanamide, related to tryptophan metabolism; glutamate, creatine, and hypoxanthine, which was previously found to be positively correlated with creatinine (Thomas et al., 2020b); lactoferrin (LTF), an iron-binding protein with antimicrobial and anti-inflammatory activity (Conneely, 2001); ectoine a compound associated to anti-inflammatory properties (Bownik and Stępniewska, 2016) and found in COVID-19 sera, as well as 2-oxoglutarate (also known as α -ketoglutarate) (Kaur et al., 2021; Figure 4C). The fraction of pathological cells also correlates to hypoxanthine, ectoine, and nicotinamide. ...
Coronavirus disease 2019 (COVID-19) is caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and can affect multiple organs, among which is the circulatory system. Inflammation and mortality risk markers were previously detected in COVID-19 plasma and red blood cells (RBCs) metabolic and proteomic profiles. Additionally, biophysical properties, such as deformability, were found to be changed during the infection. Based on such data, we aim to better characterize RBC functions in COVID-19. We evaluate the flow properties of RBCs in severe COVID-19 patients admitted to the intensive care unit by using microfluidic techniques and automated methods, including artificial neural networks, for an unbiased RBC analysis. We find strong flow and RBC shape impairment in COVID-19 samples and demonstrate that such changes are reversible upon suspension of COVID-19 RBCs in healthy plasma. Vice versa, healthy RBCs resemble COVID-19 RBCs when suspended in COVID-19 plasma. Proteomics and metabolomics analyses allow us to detect the effect of plasma exchanges on both plasma and RBCs and demonstrate a new role of RBCs in maintaining plasma equilibria at the expense of their flow properties. Our findings provide a framework for further investigations of clinical relevance for therapies against COVID-19 and possibly other infectious diseases.
... Unlike the above nine metabolites, ectoine levels were increased in DKD stage III samples but reduced in DKD stage IV samples. Ectoine has been proven to have enzyme-stabilizing and anti-inflammatory properties, exerting protective effects in human skin as well as inhibitory effects in neurodegenerative diseases, while therapeutic potential has been demonstrated in human and veterinary medicine [48]. The change in ectoine content observed during DKD progression suggests that the increase occurring in early DKD may serve as a compensatory mechanism. ...
Objective
Early diagnosis of diabetic kidney disease (DKD) has long been a complex problem. This study aimed to analyze the metabolomic characteristics of plasma extracellular vesicles (EVs) at different stages of DKD in order to evaluate the metabolites of plasma EVs and select new biomarkers for the early diagnosis of DKD.
Patients and methods
A total of 78 plasma samples were collected, including samples from 20 healthy controls, 20 patients with type 2 diabetes mellitus (T2DM), 18 patients with DKD stage III, and 20 patients with DKD stage IV. In addition, EVs were isolated for metabolomics analysis.
Results
The results identified differences in EV metabolomic characteristics in DKD patients at different stages, as well as significant differences in EV metabolomics between T2DM patients without DKD and patients with DKD. Ten Significantly differential metabolites were associated with the occurrence and progression of DKD. Uracil, LPC(O-18:1/0:0), sphingosine 1-phosphate, and 4-acetamidobutyric acid were identified as potential early biomarkers for DKD, showing excellent predictive performance.
Conclusion
Uracil, LPC(O-18:1/0:0), sphingosine 1-phosphate, and 4-acetamidobutyric acid exhibited potential as suitable biomarkers for early DKD diagnosis. Unexpectedly, combining these four candidate metabolites resulted in enhanced predictive ability for DKD.
... Widderich et al., 2014). The ectoine have ABC enzymes, which includes diaminobutyric acid (DABA), it has enzymatic steps that are catalyzed by L-2,4-diaminobutyrate transaminase (EctB), 2,4-diaminobutyrate acetyltransferase (EctA), and ectoine synthase (EctC)(Bownik and Stępniewska, 2016) to yield the cyclic ectoine molecule [(4S)-2-methyl-3,4,5,6-tetrahydropyrimidine-4-carboxylic acid]. These are the genes that are encoded by the enzymes and are organized as (EctABC) from Halomonas elongata. ...
... A key adaptation strategy of piezophiles is the accumulation of organic molecules, known as "compatible solutes", that stabilise biomolecular structures 1,16,17 . In particular, those compatible solutes that stabilise proteins against pressure denaturation are known as "piezolytes" 1,[18][19][20][21][22][23][24][25] . In this work, we focus on the piezolyte trimethylamine N-oxide (TMAO). ...
Trimethylamine N-oxide (TMAO) protects organisms from the damaging effects of high pressure. At the molecular level both TMAO and pressure perturb water structure but it is not understood how they act in combination. Here, we use neutron scattering coupled with computational modelling to provide atomistic insight into the structure of water under pressure at 4 kbar in the presence and absence of TMAO. The data reveal that TMAO resists pressure-induced perturbation to water structure, particularly in retaining a clear second solvation shell, enhanced hydrogen bonding between water molecules and strong TMAO – water hydrogen bonds. We calculate an ‘osmolyte protection’ ratio at which pressure and TMAO-induced energy changes effectively cancel out. Remarkably this ratio translates across scales to the organism level, matching the observed concentration dependence of TMAO in the muscle tissue of organisms as a function of depth. Osmolyte protection may therefore offer a molecular mechanism for the macroscale survival of life in extreme environments.
... The ß-and ε-carotene genes maintain a very high density in layer 1 and then decrease by half in the second layer (Table S3). Bacterioruberin, ß-carotenes, and ectoine could protect the microbes present in inner layers from near-UV and high visible-light damage by sun-screening or by quenching triplet-state photosensitizers and reactive oxygen species [54][55][56]. This is especially convenient to protect the primary producers-cyanobacteria in layer 2-feeding oxygen to the rest of the mat. ...
Modern non-lithifying stromatolites on the shore of the volcanic lake Socompa (SST) in the Puna are affected by several extreme conditions. The present study assesses for the first time light utilization and functional metabolic stratification of SST on a millimeter scale through shotgun metagenomics. In addition, a scanning-electron-microscopy approach was used to explore the community. The analysis on SST unveiled the profile of a photosynthetic mat, with cyanobacteria not directly exposed to light, but placed just below a high-UV-resistant community. Calvin–Benson and 3-hydroxypropinate cycles for carbon fixation were abundant in upper, oxic layers, while the Wood–Ljungdahl pathway was dominant in the deeper anoxic strata. The high abundance of genes for UV-screening and oxidant-quenching pigments and CPF (photoreactivation) in the UV-stressed layers could indicate that the zone itself works as a UV shield. There is a remarkable density of sequences associated with photoreceptors in the first two layers. Also, genetic evidence of photosynthesis split in eukaryotic (layer 1) and prokaryotic (layer 2). Photoheterotrophic bacteria, aerobic photoautotrophic bacteria, and anaerobic photoautotrophic bacteria coexist by selectively absorbing different parts of the light spectrum (blue, red, and IR respectively) at different positions of the mat. Genes for oxygen, nitrogen, and sulfur metabolism account for the microelectrode chemical data and pigment measurements performed in previous publications. We also provide here an explanation for the vertical microbial mobility within the SST described previously. Finally, our study points to SST as ideal modern analogues of ancient ST.
... The effect of ectoine has already been assessed in a wide variety of in vitro and in vivo inflammatory disease models with potential applications in Alzheimer's disease, chronic obstructive pulmonary disease and inflammatory bowel disease [20,23,24]. Moreover, a high tolerability and benign safety profile have been attributed to ectoine [25][26][27][28]. To our knowledge, the compound has not been tested in DMD before. ...
Duchenne Muscular Dystrophy (DMD) is a debilitating muscle disorder that condemns patients to year-long dependency on glucocorticoids. Chronic glucocorticoid use elicits many unfavourable side-effects without offering satisfying clinical improvement, thus, the search for alternative treatments to alleviate muscle inflammation persists. Taurine, an osmolyte with anti-inflammatory effects, mitigated pathological features in the mdx mouse model for DMD but interfered with murine development. In this study, ectoine is evaluated as an alternative for taurine in vitro in CCL-136 cells and in vivo in the mdx mouse. Pre-treating CCL-136 cells with 0.1 mM taurine and 0.1 mM ectoine prior to exposure with 300 U/mL IFN-γ and 20 ng/mL IL-1β partially attenuated cell death, whilst 100 mM taurine reduced MHC-I protein levels. In vivo, histopathological features of the tibialis anterior in mdx mice were mitigated by ectoine, but not by taurine. Osmolyte treatment significantly reduced mRNA levels of inflammatory disease biomarkers, respectively, CCL2 and SPP1 in ectoine-treated mdx mice, and CCL2, HSPA1A, TNF-α and IL-1β in taurine-treated mdx mice. Functional performance was not improved by osmolyte treatment. Furthermore, ectoine-treated mdx mice exhibited reduced body weight. Our results confirmed beneficial effects of taurine in mdx mice and, for the first time, demonstrated similar and differential effects of ectoine.
... It was found to have many uses especially in cosmetics due to its hydration effect. Ectoine has the ability to stabilize enzyme, protect human skin, can be used as anti-inflammatory, and can be utilized in neurodegenerative diseases because of its inhibitory effect (1) . ...
... The main plasma components correlating with sphero-echino cytes are lactate and 2,3-diphosphoglycerate (2,3-DPG), as-sociated to increased glycolysis; nicotinamide and tryptophanamide, related to tryptophan metabolism; glutamate, creatine, and hypoxanthine, which was found to be positively correlated with creatinine (31); lactoferrin (LTF), an iron-binding protein with antimicrobial and anti-inflammatory activity (35); ectoine a compound associated to anti-inflammatory properties (36) and found in COVID-19 sera, as well as 2-oxoglutarate (also known as -ketoglutarate) (37). The shape ratio also correlates to hypoxanthine, ectoine, and nicotinamide. ...
Coronavirus disease 2019 (COVID-19) is caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and can affect multiple organs, among which is the circulatory system. Inflammation and mortality risk markers were previously detected in COVID-19 plasma and red blood cells (RBCs) metabolic and proteomic profiles. Additionally, biophysical properties, such as deformability, were found to be changed during the infection. Based on such data, we aim to better characterize RBC functions in COVID-19. We evaluate the flow properties of RBCs in severe COVID-19 patients admitted to the intensive care unit by using in vitro microfluidic techniques and automated methods, including artificial neural networks, for an unbiased RBC analysis. We find strong flow and RBC shape impairment in COVID-19 samples and demonstrate that such changes are reversible upon suspension of COVID-19 RBCs in healthy plasma. Vice versa, healthy RBCs immediately resemble COVID-19 RBCs when suspended in COVID-19 plasma. Proteomics and metabolomics analyses allow us to detect the effect of plasma exchanges on both plasma and RBCs and demonstrate a new role of RBCs in maintaining plasma equilibria at the expense of their flow properties. Our findings provide a framework for further investigations of clinical relevance for therapies against COVID-19 and possibly other infectious diseases.
... The ß-and ε -carotene genes maintain a very high density in layer 1 and then decrease by half in the second layer (Table S3). According to previous reports, bacterioruberin, ß-carotenes, and ectoine could protect microorganisms from near-UV and high visible-light damage by sunscreening or by quenching triplet-state photosensitizers and reactive oxygen species (Sandmann et al., 1998;Shahmohammadi et al., 1998;Bownik and St pniewska, 2016). The concentration of these pigments, as well as those of the CPF effecting photoreactivation (Fig. 4, Panel B), was significantly higher in the first layer of the microbial mat, which depth faces continuous light exposure during the day. ...
Modern non-lithifying stromatolites (STs) on the shore of the volcanic lake Socompa in the Puna are affected by several extreme conditions. Although STs were proposed as ecologic models for understanding stress response and resilience in microbial ecosystems constituting a window into the past, our knowledge of ST function is still nascent. The present study assesses for the first time light utilization and functional metabolic stratification of STs on a millimeter scale through shotgun metagenomics. In addition, a scanning-electron-microscopy approach was used to explore the community. Our results demonstrated that Bacteroidetes and Cyanobacteria play major roles as ST builders and primary producers to sustain a diverse community of heterotrophs. STs manifest a high occurrence of genes for the synthesis of UV-protecting pigments, the cryptochrome-photolyase family (CPF), and rhodopsins in the surface layers. Three different ecologic niches involving the use of light in energy production were defined. Calvin-Benson and Wood-Ljungdahl pathways were proposed as the main mechanisms for carbon fixation. Several genes account for the microelectrode chemical data and pigment measurements performed in previous publications. We also provide here an explanation for the vertical microbial mobility within the ST described previously. Finally, our study points to STs as ideal modern analogues of ancient STs.
... Ectoine is one of the compatible solutes able to accumulate in cells in response to external increases in osmotic pressure without compromising cell physiology. Other than serving as osmotic balancing agents, compatible solutes also show direct protective effects against freezing, high UV irradiation, high temperature, and macromolecule denaturation under adverse conditions [61][62][63][64]. Not all of them are present in all organisms; in fact, their occurrence seems be spread out and sometimes species specific, with some functional specialization. ...
Diatoms are a successful group of microalgae at the base of the marine food web. For hundreds of millions of years, they have shared common habitats with bacteria, which favored the onset of interactions at different levels, potentially driving the synthesis of biologically active molecules. To unveil their presence, we sequenced the genomes of bacteria associated with the centric diatom Thalassiosira rotula from the Gulf of Naples. Annotation of the metagenome and its analysis allowed the reconstruction of three bacterial genomes that belong to currently undescribed species. Their investigation showed the existence of novel gene clusters coding for new polyketide molecules, antibiotics, antibiotic-resistance genes and an ectoine production pathway. Real-time PCR was used to investigate the association of these bacteria with three different diatom clones and revealed their preference for T. rotula FE80 and Skeletonema marinoi FE7, but not S. marinoi FE60 from the North Adriatic Sea. Additionally, we demonstrate that although all three bacteria could be detected in the culture supernatant (free-living), their number is up to 45 times higher in the cell associated fraction, suggesting a close association between these bacteria and their host. We demonstrate that axenic cultures of T. rotula are unable to grow in medium with low salinity (<28 ppt NaCl) whereas xenic cultures can tolerate up to 40 ppt NaCl with concomitant ectoine production, likely by the associated bacteria.
... Other important compounds produced by halophiles are ectoine and hydroxyectoine. Compatible solutes such as ectoine and hydroxyectoine are used as protective factors for cells, DNA, and proteins, which makes them highly valuable for cosmetics and medicine sectors [140,141]. Ectoine is commonly used in the cosmetics industry, but clinical trials are also conducted to indicate its importance in the treatment of eye and respiratory diseases [142][143][144][145]. In silico studies for new microorganisms producing ectoine and hydroxyectoine can be improved by using BGC identification methods like antiSMASH [146][147][148]. ...
Halophiles, the salt-loving organisms, have been investigated for at least a hundred years. They are found in all three domains of life, namely Archaea, Bacteria, and Eukarya, and occur in saline and hypersaline environments worldwide. They are already a valuable source of various biomolecules for biotechnological, pharmaceutical, cosmetological and industrial applications. In the present era of multidrug-resistant bacteria, cancer expansion, and extreme environmental pollution, the demand for new, effective compounds is higher and more urgent than ever before. Thus, the unique metabolism of halophilic microorganisms, their low nutritional requirements and their ability to adapt to harsh conditions (high salinity, high pressure and UV radiation, low oxygen concentration, hydrophobic conditions, extreme temperatures and pH, toxic compounds and heavy metals) make them promising candidates as a fruitful source of bioactive compounds. The main aim of this review is to highlight the nucleic acid sequencing experimental strategies used in halophile studies in concert with the presentation of recent examples of bioproducts and functions discovered in silico in the halophile’s genomes. We point out methodological gaps and solutions based on in silico methods that are helpful in the identification of valuable bioproducts synthesized by halophiles. We also show the potential of an increasing number of publicly available genomic and metagenomic data for halophilic organisms that can be analysed to identify such new bioproducts and their producers.
... It is classified as a compatible solute allowing bacteria to counteract negative effects of high osmolarity or other extreme environments [8,9]. Ectoine works via a mechanism called "preferential exclusion", resulting in the preferential hydration of macromolecules, thereby protecting those from negative influences [10]. The industrial-scale production of ectoine has provided the opportunity to employ its protective and moisturizing effects into a range of medical devices. ...
Introduction
Symptomatic relief of acute rhinosinusitis is commonly achieved with nasal decongestants. The current observational study investigated the efficacy and safety of treatment of acute rhinosinusitis with Ectoin ® Rhinitis Spray compared to or in combination with Xylometazoline-containing decongesting nasal spray.
Methods
Patients with acute rhinosinusitis applied either Ectoin ® Rhinitis Spray, Xylometazoline nasal spray or a combination of both products. Rhinosinusitis symptoms were assessed, and nasal oedema and endonasal redness were determined by rhinoscopy. Patient diaries based on the validated SNOT (Sino Nasal Outcome Test) questionnaire evaluated rhinosinusitis parameters over time and influences of the disease on quality of life. Following treatment, investigators and patients judged the efficacy and tolerability.
Results
Ectoin ® Rhinitis Spray diminished common rhinosinusitis symptoms such as nasal obstruction, nasal secretion, facial pain/headache, and smell/taste impairment. Upon treatment over 7 days, rhinosinusitis sum scores decreased statistically significantly ( p < 0.001) by − 64.25%, which was comparable to that achieved with Xylometazoline-containing decongesting nasal spray (− 67.60%). No side effects were observed during treatment with Ectoin ® Rhinitis Spray, whereas treatment with Xylometazoline-containing nasal spray resulted in nasal mucosa dryness. Concomitant treatment with both products diminished the development of nasal dryness and required fewer applications of Xylometazoline-containing nasal spray.
Conclusion
Ectoin ® Rhinitis Spray is an effective, natural treatment option for acute rhinosinusitis, which may be used as monotherapy or as add-on treatment with a Xylometazoline-containing nasal spray. The concomitant use of Ectoin ® Rhinitis Spray might reduce the needed dose of decongestant nasal spray and counteract bothersome side effects such as dry nasal mucosa.
Trial registration
The current study was registered in the ClinicalTrials.gov database under the identifier: NCT03693976 (date of registration: Oct 3, 2018).
As the global population continues to grow, so does the demand for longer, healthier lives and environmentally responsible choices. Consumers are increasingly drawn to naturally sourced products with proven health and wellbeing benefits. The marine environment presents a promising yet underexplored resource for the cosmetics industry, offering bioactive compounds with the potential for safe and biocompatible ingredients. This manuscript provides a comprehensive overview of the potential of marine organisms for cosmetics production, highlighting marine-derived compounds and their applications in skin/hair/oral-care products, cosmeceuticals and more. It also lays down critical safety considerations and addresses the methodologies for sourcing marine compounds, including harvesting, the biorefinery concept, use of systems biology for enhanced product development, and the relevant regulatory landscape. The review is enriched by three case studies: design of macroalgal skincare products in Iceland, establishment of a microalgal cosmetics spin-off in Italy, and the utilization of marine proteins for cosmeceutical applications.
Tissue engineered constructs prepared using conventional scaffold-based approaches have the potential to repair or regenerate damaged tissues and organs. Various scaffold fabrication strategies such as electrospinning, solvent casting, particulate leaching, gas foaming, hydrogels, freeze-drying, and 3D bioprinting have been used to fabricate artificial tissues. In recent times, 3D bioprinting has been predominantly used in various biomedical fields, including healthcare and pharmaceutical applications due to precision in 3D geometry. However, there are no viable strategies to preserve bioprinted constructs for on-demand applications because of the lack of specialized techniques or cryopreservation agents to maintain the cell viability and functionality of the bioprinted tissues. To solve this issue, cryopreservation of bioprinted tissues has emerged in recent years to develop methods to create and cryopreserve bioprinted constructs for on-demand applications. This review discusses various techniques used for producing ready-to-use tissue engineered products such as electrospinning, hydrogels, 3D bioprinting, and other bioprinting approaches. Further, the factors influencing the bioprinted tissues, such as cryoprotectants, polymer types and crosslinker concentrations, crosslinking approaches, viscoelastic properties, storage facilities, etc, were also discussed in detail. The potential of cryopreservable bioprinted tissues in various healthcare applications are elaborated with lucid examples. Finally, the conclusions and possible future directions for the fabrication and cryopreservation of tissue engineered products are highlighted.
This book delves into the world of natural sources from medicinal plants, microbes, and fungi, to lichen, algae, and clay minerals that have been used for centuries in traditional medicine. These sources are rich in bioactive secondary metabolites that have a wide range of applications in various industries, including cosmetics and personal care products.
This book provides a comprehensive guide to secondary metabolites for cosmeceutical purposes, regulatory perspectives for cosmeceuticals in different countries, and allergic responses from these secondary metabolites. Additionally, this book discusses the impact of nanotechnology on cosmetic products such as skin and hair care.
Bioprospecting of Natural Sources for Cosmeceuticals is a valuable resource for researchers and graduate students in chemistry, botany, biotechnology, microbiology, cosmetic science, and the pharmaceutical sciences. It is also useful for those researching traditional medicine systems and those in the microbiology, biotechnology, pharmaceutical, and nanoscience industries.
Ectoine is an important natural secondary metabolite in halophilic microorganisms. It protects cells against environmental stressors, such as salinity, freezing, drying, and high temperatures. Ectoine is widely used in medical, cosmetic, and other industries. Due to the commercial market demand of ectoine, halophilic microorganisms are the primary method for producing ectoine, which is produced using the industrial fermentation process “bacterial milking.” The method has some limitations, such as the high salt concentration fermentation, which is highly corrosive to the equipment, and this also increases the difficulty of downstream purification and causes high production costs. The ectoine synthesis gene cluster has been successfully heterologously expressed in industrial microorganisms, and the yield of ectoine was significantly increased and the cost was reduced. This review aims to summarize and update microbial production of ectoine using different microorganisms, environments, and metabolic engineering and fermentation strategies and provides important reference for the development and application of ectoine.
Ectoine has gained considerable attention as a high-value chemical with significant application potential and market demand. This study aimed to increase ectoine yields by blocking the metabolic shunt pathway of l-aspartate-4-semialdehyde, the precursor substrate in ectoine synthesis. The homoserine dehydrogenase encoded by hom in H. campaniensis strain XH26 is responsible for the metabolic shunt of l-aspartate-4-semialdehyde to glycine. CRISPR/Cas9 technology was used to seamlessly knockout hom, blocking the metabolic shunt pathway to increase ectoine yields. The ectoine yield of XH26/Δhom was 351.13 mg (g CDW)⁻¹ after 48 h of incubation in 500 mL shake flasks using optimal medium with 1.5 mol L⁻¹ NaCl, which was significantly higher than the 239.18 mg (g CDW)⁻¹ of the wild-type strain. Additionally, the absence of the ectoine metabolic shunt pathway affects betaine synthesis, and thus the betaine yields of XH26/Δhom was 19.98 mg (g CDW)⁻¹, considerably lower than the 69.58 mg (g CDW)⁻¹ of the wild-type strain. Batch fermentation parameters were optimized, and the wild-type strain and XH26/Δhom were fermented in 3 L fermenters, resulting in a high ectoine yield of 587.09 mg (g CDW)⁻¹ for the defective strain, which was significantly greater than the ectoine yield of 385.03 mg (g CDW)⁻¹ of the wild-type strain. This study showed that blocking the metabolic shunt of synthetic substrates effectively increases ectoine production, and a reduction in the competitively compatible solute betaine appears to promote increased ectoine synthesis.
A 7%w/w Ectoine formula, from natural source, formulated to reduce melanomagenesis, enhance penetration by 3D printed microneedles (MN), with specified length, diameter and tip to ensure painless effect. Ectoine Gel formulations were prepared using Carbapol 940 and Pluronic (F127). The effect of the polymers on pH, viscosity, spread-ability, and the in-vitro, ex-vivo release profiles were obtained. The physiochemical investigation showed uniform gel formulations. The formulations' in-vitro and ex-vivo drug release displayed a controllable drug release pattern, reaching 63.7-96% and 73-94.7% after 24 hours. The permeation study of the in vitro and ex vivo release revealed that the drug release from gels followed diffusion mechanism. The selected formula was used, apply 3D printed MN array to treat melanoma. Male rats were used for induction of melanoma using 0.5% of 7,12-Dimethylbenz[a]anthracene three times weekly for 12 weeks, histopathology was applied to ensure development of carcinoma then rats were treated using the selected formula. Following treatment for continuous 6 weeks, histopathology showed a change in anatomy of skin, which started to return to its normal structure. The anti-melanogenesis activity of optimum formula of Ectoine gel, enhanced by 3D printed MN, was found to be effective in reducing the severity of skin cancer reinforcing the efficacy of the promising treatment.
Ectoine is a compatible solutes that is diffusely dispersed in bacteria and archaea. It plays a significant role as protectant against various external pressures, such as high temperature, high osmolarity, dryness and radiation, in cells. Ectoine can be utilized in cosmetics due to its properties of moisturizing and antiultraviolet. It can also be used in the pharmaceutical industry for treating various diseases. Therefore, strong protection of ectoine creates a high commercial value. Its current market value is approximately US$1000 kg−1. However, traditional ectoine production in high-salinity media causes high costs of equipment loss and wastewater treatment. There is a growing attention to reduce the salinity of the fermentation broth without sacrificing the production of ectoine. Thus, heterologous production of ectoine in nonhalophilic microorganisms may represent the new generation of the industrial production of ectoine. In this review, we summarized and discussed the biological activities of ectoine on cell and human health protection and its heterologous production.
The term "zwitterionic polymers" refers to polymers that bear a pair of oppositely charged groups in their repeating units. When these oppositely charged groups are equally distributed at the molecular level, the molecules exhibit an overall neutral charge with a strong hydration effect via ionic solvation. The strong hydration effect constitutes the foundation of a series of exceptional properties of zwitterionic materials, including resistance to protein adsorption, lubrication at interfaces, promotion of protein stabilities, antifreezing in solutions, etc. As a result, zwitterionic materials have drawn great attention in biomedical and engineering applications in recent years. In this review, we give a comprehensive and panoramic overview of zwitterionic materials, covering the fundamentals of hydration and nonfouling behaviors, different types of zwitterionic surfaces and polymers, and their biomedical applications.
Introduction:
Polymorphic light eruption (PLE) is the most common idiopathic, acquired photodermatosis. The pathophysiology of PLE is not yet fully understood but seems to involve immunological mechanisms, UVA-induced oxidative stress, and the subsequent elicitation of a cellular stress response affecting keratinocyte gene expression and skin immune function. In the present study, a high broad-spectrum sunscreen medical device (MD), containing a very high protection complex of UVB and UVA filters and ectoin, was investigated for its ability to protect against UVA-induced PLE.
Methods:
The study was carried out as a monocentric, double-blinded, randomized, untreated controlled design. The test MD was applied (2 mg/cm2) on one side of the chest according to a randomization list of 15 patients with a typical history of PLE, and the contralateral area remained untreated. After product application, the test areas were exposed daily to increasing doses of UVA radiation (from 40 to 60 J/cm2) until a PLE reaction was detected or for a maximum of five consecutive days. Evaluations of induced PLE included clinical scoring and chromametry for erythema and pigmentation.
Results:
Overall, no positive PLE reaction was observed on the side of the chest treated by the test MD, whereas positive PLE reactions were triggered on the untreated side of 13 subjects. Subjective sensations were very rare on the MD-treated side but were numerous and more severe on the untreated side. Chromametry and clinical visual inspection indicated that the skin color was unchanged on the MD-protected side, whereas high increased values of erythema and pigmentation were observed on the untreated chest side.
Conclusion:
This MD sunscreen based on a complex of UVA-UVB filters and 1% of ectoin may be effective in preventing UVA-induced PLE. New studies comparing this MD sunscreen versus the same product without ectoin should be conducted.
Clinicaltrials:
gov identifier: NCT05320315 (retrospectively registered 09/17/2021).
The marine ecosystem is a unique and complicated system that is characterized primarily by the great biodiversity, representing the richness of diverse habitats and the variations of life forms. It involves various marine organisms with great chemical diversity, and they are rich sources of secondary metabolites relative to terrestrial organisms. The oceans thus always offer excellent opportunities for discovering valuable materials from various organisms. Such compounds contain a wide range of chemical structures and functions, which for more specific potentialities are wider. The produced compounds have a vital role in the human and animal life and are widely used as antimicrobial, antioxidants, antiviral, anticancer, and food and feed and in food and pharmaceuticals industry. However, in many fields such as food, cosmetics, dietary supplements, animal feed, bioactive packaging, and industrial products, as well as in high-tech biomedical sectors, applications of novel marine molecules are now found.
Understanding adaptation to extreme environments remains a challenge of high biotechnological potential for fundamental molecular biology. The cytosol of many microorganisms, isolated from saline environments, reversibly accumulates molar concentrations of the osmolyte ectoine to counterbalance fluctuating external salt concentrations. Although they have been studied extensively by thermodynamic and spectroscopic methods, direct experimental structural data have, so far, been lacking on ectoine-water-protein interactions. In this paper, in vivo deuterium labeling, small angle neutron scattering, neutron membrane diffraction and inelastic scattering are combined with neutron liquids diffraction to characterize the extreme ectoine-containing solvent and its effects on purple membrane of H. salinarum and E. coli maltose binding protein. The data reveal that ectoine is excluded from the hydration layer at the membrane surface and does not affect membrane molecular dynamics, and prove a previous hypothesis that ectoine is excluded from a monolayer of dense hydration water around the soluble protein. Neutron liquids diffraction to atomic resolution shows how ectoine enhances the remarkable properties of H-bonds in water—properties that are essential for the proper organization, stabilization and dynamics of biological structures.
The aim of this observational trial was to evaluate the efficacy and tolerability of a mouth and throat spray containing ectoine in the treatment of acute pharyngitis and/or laryngitis. The outcome was compared with control treatment using saline lozenges. This study was designed as a prospective, controlled, non-randomized, observational multicenter clinical trial and was conducted in Germany. The study population consisted of 95 patients. The decision for treatment with either spray or lozenges was based on the patients’ preference for pharyngeal or oral application. Investigators assessed symptoms specific to acute pharyngitis/laryngitis and determined the pharyngitis symptom score. Both patients and investigators evaluated the tolerability and efficacy of the treatment applied. Treatment with the spray showed higher efficacy, 1.95 ± 0.81 versus 1.68 ± 0.67 (investigators) and 1.97 ± 0.88 versus 1.57 ± 0.69 (patients, p < 0.05). Treatment with the spray resulted in significantly greater reduction of cervical lymph node swelling (p < 0.05), ∆ spray = 0.44 ± 0.62, ∆ lozenges = 0.21 ± 0.62. The lozenges showed some advantage in relieving cough, ∆ lozenges = 0.62 ± 0.94 versus ∆ spray = 0.44 ± 0.85. Both patients and investigators rated the tolerability of both medical devices as “good” to “very good”. Adverse events of mild to moderate severity were either possibly related or not related to the medical devices used. No serious adverse events occurred. Taken together, while the tolerability was consistent in both treatment groups, the ectoine-based spray showed superior efficacy in treating acute pharyngitis and/or laryngitis.
Electronic supplementary material
The online version of this article (doi:10.1007/s00405-016-4060-z) contains supplementary material, which is available to authorized users.
Ectoine and hydroxyectoine are compatible solutes widely synthesized by members of the Bacteria to cope with high osmolarity surroundings. Inspection of 557 archaeal genomes revealed that only 12 strains affiliated with the Nitrosopumilus, Methanotrix, or Methanobacterium genera harbor ectoine/hydroxyectoine gene clusters. Phylogenetic considerations suggest that these Archaea have acquired these genes through horizontal gene transfer events. Using the thaumarchaeon "Candidatus Nitrosopumilus maritimus" as an example, we demonstrate that the transcription of its ectABCD genes is osmotically induced and functional since it leads to the production of both ectoine and hydroxyectoine. The ectoine synthase and the ectoine hydroxylase were biochemically characterized and their properties resemble those of their counterparts from Bacteria. Transcriptional analysis of osmotically stressed "Ca. N. maritimus" cells demonstrated that they possess an ectoine/hydroxyectoine gene cluster (hyp-ectABCD-mscS) different form those recognized previously since it contains a gene for an MscS-type mechanosensitive channel. Complementation experiments with an Escherichia coli mutant lacking all known mechanosensitive channel proteins demonstrated that the (Nm)MscS protein is functional. Hence, "Ca. N. maritimus" cells cope with high salinity not only through enhanced synthesis of osmostress-protective ectoines but they already prepare themselves simultaneously for an eventually occurring osmotic down-shock by enhancing the production of a safety-valve (NmMscS).
Exposure of the airways to carbonaceous nanoparticles can contribute to the development of immune diseases both via the aggravation of the allergic immune response in sensitized individuals and by adjuvant mechanisms during the sensitization against allergens. The cellular and molecular mechanisms involved in these adverse pathways are not completely understood. We recently described that the reduction of carbon nanoparticle-induced lung inflammation by the application of the compatible solute ectoine reduced the aggravation of the allergic response in an animal system. In the current study we investigated the influence of carbon nanoparticles on the sensitization of animals to ovalbumin via the airways. Ectoine was used as a preventive strategy against nanoparticle-induced neutrophilic lung inflammation.
Balb/c mice were repetitively exposed to the antigen ovalbumin after induction of airway inflammation by carbon nanoparticles, either in the presence or in the absence of ectoine. Allergic sensitization was monitored by measurement of immunoglobulin levels and immune responses in lung and lung draining lymph nodes after challenge. Furthermore the role of dendritic cells in the effect of carbon nanoparticles was studied in vivo in the lymph nodes but also in vitro using bone marrow derived dendritic cells.
Animals exposed to antigen in the presence of carbon nanoparticles showed increased effects with respect to ovalbumin sensitization, to the allergic airway inflammation after challenge, and to the specific TH2 response in the lymph nodes. The presence of ectoine during the sensitization significantly reduced these parameters. The number of antigen-loaded dendritic cells in the draining lymph nodes was identified as a possible cause for the adjuvant effect of the nanoparticles. In vitro assays indicate that the direct interaction of the particles with dendritic cells is not able to trigger CCR7 expression, while this endpoint is achieved by lung lavage fluid from nanoparticle-exposed animals.
Using the intervention strategy of applying ectoine into the airways of animals we were able to demonstrate the relevance of neutrophilic lung inflammation for the adjuvant effect of carbon nanoparticles on allergic sensitization.
Ectoine (ECT) is an amino acid produced and accumulated by halophilic bacteria in stressful conditions in order to prevent the loss of water from the cell. There is a lack of knowledge on the effects of ECT in heat-stressed aquatic animals. The purpose of our study was to determine the influence of ECT on Daphnia magna subjected to heat stress with two temperature gradients: 1 and 0.1 °C/min in the range of 23-42 °C. Time to immobilisation, survival during recovery, swimming performance, heart rate, thoracic limb movement and the levels of heat shock protein 70 kDa 1A (HSP70 1A), catalase (CAT) and nitric oxide species (NOx) were determined in ECT-exposed and unexposed daphnids; we showed protective effects of ECT on Daphnia magna subjected to heat stress. Time to immobilisation of daphnids exposed to ECT was longer when compared to the unexposed animals. Also, survival rate during the recovery of daphnids previously treated with ECT was higher. ECT significantly attenuated a rapid increase of mean swimming velocity which was elevated in the unexposed daphnids. Moreover, we observed elevation of thoracic limb movement and modulation of heart rate in ECT-exposed animals. HSP70 1A and CAT levels were reduced in the presence of ECT. On the other hand, NOx level was slightly elevated in both ECT-treated and unexposed daphnids, however slightly higher NOx level was found in ECT-treated animals. We conclude that the exposure to ectoine has thermoprotective effects on Daphnia magna, however their mechanisms are not associated with the induction of HSP70 1A.
Objectives. Allergic rhinitis is a common disease with increasing prevalence and high impact on economic burden and comorbidities. As treatment with pharmacological drugs is not always satisfactory due to side effects and incomplete efficacy, alternative treatment strategies are needed. Ectoine is an osmolyte with membrane stabilizing and inflammation reducing capacities. Nasal spray and eye drops containing ectoine are promising new treatment regimens for allergic rhinitis sufferers. Design and Methods. The current two noninterventional trials evaluated the efficacy and safety of ectoine containing nasal spray and eye drops for treating allergic rhinitis in comparison with either azelastine or cromoglycic acid containing products. Nasal and ocular symptom developments as well as judgment of tolerability and efficacy were assessed both by investigators and patients over a time period of one to two weeks. Results. Both trials confirmed that ectoine containing products reduced nasal and ocular symptoms in allergic rhinitis patients. Results clearly demonstrated good safety profiles of the ectoine products comparable to those of azelastine and even better to those of cromoglycate products. Conclusion. Ectoine containing nasal spray and eye drops are interesting new treatment strategies for sufferers of allergic rhinitis, combining both good efficacy and absence of side effects.
Objectives. The current study aimed to compare the efficacy and safety of a classical anti-inflammatory beclomethasone nasal spray in comparison to a physic-chemical stabilizing ectoine containing nasal spray in the treatment of allergic rhinitis. Design and Methods. This was a noninterventional, open-label, observational trial investigating the effects of beclomethasone or ectoine nasal spray on nasal symptoms and quality of life. Over a period of 14 days, patients were asked to daily document their symptoms. Efficacy and tolerability were assessed by both physicians and patients. Results. Both treatments resulted in a significant decrease of TNSS values. An equivalence test could not confirm the noninferiority of ectoine treatment in comparison with beclomethasone treatment. Although clear symptom reduction was achieved with the ectoine products, the efficacy judgment showed possible advantages for the beclomethasone group. Importantly, tolerability results were comparably good in both groups, and a very low number of adverse events supported this observation. Both treatments resulted in a clear improvement in the quality of life as assessed by a questionnaire answered at the beginning and at the end of the trial. Conclusion. Taken together, it was shown that allergic rhinitis can be safely and successfully treated with beclomethasone and also efficacy and safety were shown for ectoine nasal spray.
Objectives. The meta-analysis aims to investigate the efficacy of ectoine nasal spray and eye drops in the treatment of allergic rhinitis and rhinoconjunctivitis symptoms. Design and Methods. This meta-analysis is based on yet unpublished data of four studies. Both nasal and eye symptoms were documented in patient diary cards. All scales were transformed into a 4-point scale: 0 = no, 1 = mild, 2 = moderate, and 3 = severe symptoms. Each symptom was analysed individually in a meta-analysis of the area under the curve values as well as in a meta-analysis of pre- and posttreatment comparison. Results. After seven days of treatment with ectoine nasal spray both nasal and ocular symptoms decreased significantly. A strong reduction of symptom severity was shown for the parameters rhinorrhoea (31.76% reduction) and nasal obstruction (29.94% reduction). Furthermore, the meta-analyses of individual symptoms to investigate the strength of effect after seven days of medication intake showed significant improvement for nasal obstruction, rhinorrhoea, nasal itching, sneezing, itching of eyes, and redness of eyes. The improvement of the symptom nasal obstruction was associated with a strong effect 0.53 (±0.26). Conclusions. The ectoine nasal spray and eye drops seem to be equally effective as guideline-recommended medication in the treatment of rhinoconjunctivitis symptoms.
Objectives. The safety and efficacy of ectoine nasal spray and ectoine nasal spray with dexpanthenol in the treatment of rhinitis sicca were evaluated in two studies. Design and Methods. Two noninterventional observational studies were performed to evaluate the efficacy and safety of a nasal spray containing ectoine (study 1) and ectoine/dexpanthenol (study 2) over a period of two weeks including comparable numbers of patients suffering from rhinitis sicca anterior. Patients and physicians were asked to rate the efficacy in reducing symptoms and the tolerability over the treatment phase. Results. The treatment in both studies resulted in a clinical and statistical significant reduction of the main diagnosis parameters, nasal airway obstruction, and crust formation. There was also a significant reduction in the secondary diagnosis parameters in both studies. Importantly, the tolerability was very good. During the whole observational study, neither patients nor doctors stopped the medication due to unwanted effects. Conclusion. Rhinitis sicca could be successfully treated with a nasal spray containing ectoine and a nasal spray combining ectoine with dexpanthenol. The combination of both substances led to slight advantages.
We were able to demonstrate that hydroxyectoine, in contrast to ectoine, is a good glass-forming compound. Fourier transform infrared and spin label electron spin resonance studies of dry ectoine and hydroxyectoine have shown that the superior glass-forming properties of hydroxyectoine result from stronger intermolecular H-bonds with the OH group of hydroxyectoine. Spin probe experiments have also shown that better molecular immobilization in dry hydroxyectoine provides better redox stability of the molecules embedded in this dry matrix. With a glass transition temperature of 87°C (vs. 47°C for ectoine) hydroxyectoine displays remarkable desiccation protection properties, on a par with sucrose and trehalose. This explains its accumulation in response to increased salinity and elevated temperature by halophiles such as Halomonas elongata and its successful application in ``anhydrobiotic engineering'' of both enzymes and whole cells.
The stabilizing and function-preserving effects of ectoines have attracted considerable biotechnological interest up to industrial scale processes for their production. These rely on the release of ectoines from high-salinity-cultivated microbial producer cells upon an osmotic down-shock in rather complex processor configurations. There is growing interest in uncoupling the production of ectoines from the typical conditions required for their synthesis, and instead design strains that naturally release ectoines into the medium without the need for osmotic changes, since the use of high-salinity media in the fermentation process imposes notable constraints on the costs, design, and durability of fermenter systems.
Here, we used a Corynebacterium glutamicum strain as a cellular chassis to establish a microbial cell factory for the biotechnological production of ectoines. The implementation of a mutant aspartokinase enzyme ensured efficient supply of L-aspartate-beta-semialdehyde, the precursor for ectoine biosynthesis. We further engineered the genome of the basic C. glutamicum strain by integrating a codon-optimized synthetic ectABCD gene cluster under expressional control of the strong and constitutive C. glutamicum tuf promoter. The resulting recombinant strain produced ectoine and excreted it into the medium; however, lysine was still found as a by-product. Subsequent inactivation of the L-lysine exporter prevented the undesired excretion of lysine while ectoine was still exported. Using the streamlined cell factory, a fed-batch process was established that allowed the production of ectoine with an overall productivity of 6.7 g L-1 day-1 under growth conditions that did not rely on the use of high-salinity media.
The present study describes the construction of a stable microbial cell factory for recombinant production of ectoine. We successfully applied metabolic engineering strategies to optimize its synthetic production in the industrial workhorse C. glutamicum and thereby paved the way for further improvements in ectoine yield and biotechnological process optimization.
Background
Particulate air pollution in lung epithelial cells induces pathogenic endpoints like proliferation, apoptosis, and pro-inflammatory reactions. The activation of the epidermal growth factor receptor (EGFR) is a key event responsible for signalling events involving mitogen activated protein kinases specific for these endpoints. The molecular events leading to receptor activation however are not well understood. These events are relevant for the toxicological evaluation of inhalable particles as well as for potential preventive strategies in situations when particulate air pollution cannot be avoided. The current study therefore had the objective to elucidate membrane-coupled events leading to EGFR activation and the subsequent signalling cascade in lung epithelial cells. Furthermore, we aimed to identify the molecular target of ectoine, a biophysical active substance which we described to prevent carbon nanoparticle-induced lung inflammation.
Methods
Membrane signalling events were investigated in isolated lipid rafts from lung epithelial cells with regard to lipid and protein content of the signalling platforms. Using positive and negative intervention approaches, lipid raft changes, subsequent signalling events, and lung inflammation were investigated in vitro in lung epithelial cells (RLE-6TN) and in vivo in exposed animals.
Results
Carbon nanoparticle treatment specifically led to an accumulation of ceramides in lipid rafts. Detailed analyses demonstrated a causal link of ceramides and subsequent EGFR activation coupled with a loss of the receptor in the lipid raft fractions. In vitro and in vivo investigations demonstrate the relevance of these events for carbon nanoparticle-induced lung inflammation. Moreover, the compatible solute ectoine was able to prevent ceramide-mediated EGFR phosphorylation and subsequent signalling as well as lung inflammation in vivo.
Conclusion
The data identify a so far unknown event in pro-inflammatory signalling and contribute to the understanding of particle cell interaction and therefore to risk identification and risk assessment of inhalable xenobiotics. Moreover, as this cellular reaction can be prevented by the well tolerated substance ectoine, a molecular preventive strategy for susceptible persons against airway inflammation is proposed.
The protective properties of ectoine, formerly described for only extremophilic microorganisms, can be transferred to human skin. Our present data show that the compatible solute ectoine protects the cellular membrane from damage caused by surfactants. Transepidermal water loss measurements in vivo suggest that the barrier function of the skin is strengthened after the topical application of an oil in water emulsion containing ectoine. Ectoine functions as a superior moisturizer with long-term efficacy. These findings indicating that ectoine is a strong water structure-forming solute are explained in silico by means of molecular dynamic simulations. Spherical clusters containing (1) water, (2) water with ectoine, and (3) water with glycerol are created as model systems. The stronger the water-binding activity of the solute, the greater the quantity of water molecules remaining in the cluster at high temperatures. Water clusters around ectoine molecules remain stable for a long period of time, whereas mixtures of water and glycerol break down and water molecules diffuse out of the spheres. On the basis of these findings, we suggest that the hydrogen bond properties of solutes are not solely responsible for maintaining the water structure form. Moreover, the particular electrostatic potential of ectoine as an amphoteric molecule with zwitterionic character is the major cause for its strong affinity to water. Because of its outstanding water-binding activity, ectoine might be especially useful in preventing water loss in dry atopic skin and in recovering skin viability and preventing skin aging.
All microorganisms possess a positive turgor, and maintenance of this outward-directed pressure is essential since it is generally considered as the driving force for cell expansion. Exposure of microorganisms to high-osmolality environments triggers rapid fluxes of cell water along the osmotic gradient out of the cell, thus causing a reduction in turgor and dehydration of the cytoplasm. To counteract the outflow of water, microorganisms increase their intracellular solute pool by amassing large amounts of organic osmolytes, the so-called compatible solutes. These osmoprotectants are highly congruous with the physiology of the cell and comprise a limited number of substances including the disaccharide trehalose, the amino acid proline, and the trimethylammonium compound glycine betaine. The intracellular amassing of compatible solutes as an adaptive strategy to high-osmolality environments is evolutionarily well-conserved in Bacteria, Archaea, and Eukarya. Furthermore, the nature of the osmolytes that are accumulated during water stress is maintained across the kingdoms, reflecting fundamental constraints on the kind of solutes that are compatible with macromolecular and cellular functions. Generally, compatible solutes can be amassed by microorganisms through uptake and synthesis. Here we summarise the molecular mechanisms of compatible solute accumulation in Escherichia coli and Bacillus subtilis, model organisms for the gram-negative and gram-positive branches of bacteria.
The heat shock proteins (Hsps) have an important role in the cytoprotection and repair of cells and tissues. One potential mechanism of protection is the ability of Hsp to inhibit genetic expression of proinflammatory cytokines, the transcription of which is dependent on nuclear factor-kappa B (NF-kappaB) activation. In this study, we evaluated the ability of ectoine, a novel natural biomolecule produced by halophilic microorganisms, to activate the hsp70 and hsp70B'. By reverse transcriptase-polymerase chain reaction and Western blot analysis, we demonstrated increased hsp70B' gene expression in human keratinocytes treated with ectoine and heat stressed. In contrast, in the absence of heat shock, ectoine was unable to induce hsp70B' but had the ability to induce another member of the Hsp family, the hsp70. The latter is not only elevated in response to stress but is also present at basal level in unstressed cells. In addition, ectoine had no effect on proinflammatory cytokines interleukin (IL)-1alpha, IL-6, IL-8, and tumor necrosis factor-alpha and on NF-kappaB and IkappaB-alpha pathway, whereas it downregulated the expression of cited proinflammatory cytokines, in lipopolysaccharides-treated keratinocytes. These results highlighted the ability of ectoine to protect cells from stress conditions and to prevent cell damage by maintaining an elevated level of the Hsp70. Overall, these data might suggest the use of this compatible solute in cosmetic and even pharmaceutical preparations aiming to activate a cytoprotective heat shock response in human cells.
Ectoine is a widespread osmolyte enabling halophilic bacteria to withstand high osmotic stress which has many potential applications ranging from cosmetics to its use as a therapeutic agent. In this contribution, combining experiment and theory, the hydration and ion-binding of this zwitterionic compound was studied to gain information on the functioning of ectoine in particular and of osmolytes in general. Dielectric relaxation spectroscopy was used to determine the effective hydration number of ectoine and its effective dipole moment in aqueous solutions with and without added NaCl. The obtained experimental data were compared with structural results from 1D-RISM and 3D-RISM calculations. It was found that ectoine is strongly hydrated, even in the presence of high salt concentrations. Upon addition of NaCl, ions are bound to ectoine but the formed complexes are not very stable. Interestingly, this osmolyte strongly rises the static relative permittivity of its solutions, shielding thus effectively long-range Coulomb interactions among ions in ectoine-containing solutions. We believe that via this effect, which should be common to all zwitterionic osmolytes, ectoine protects against excessive ions within the cell, in addition to its strong osmotic activity protecting against ions outside.
Ectoine (ECT) is produced by halophilic microorganisms in response to various stressful factors. Its protective properties in bacteria and some populations of isolated cells are known; however, no data are available on its protective influence on aquatic invertebrates subjected to a common pollutant, formaldehyde (FA). The purpose of this study was to determine the effects of FA alone (at 20 and 60 mg/L) and in the combination with various concentrations of ECT (5, 10 and 25 mg/L) at various times of exposure on behavioural, physiological and biochemical parameters of Daphnia magna. Specifically, mortality, heart rate, thoracic limb movement, reduced glutathione (GSH)/oxidised glutathione (GSSG) ratio, catalase (CAT) activity and nitric oxide (NOx) levels were determined. The results showed that both concentrations of FA when administered alone induced significant alterations of the determined parameters. On the other hand, animals treated with the combinations of FA + ECT showed decreased mortalities, attenuated inhibition of heart rates and thoracic limb activities, less decreased GSH/GSSG ratios, lower stimulation of CAT activities and NOx levels when compared to the crustaceans subjected to FA alone. The most distinct attenuation of toxic effects was observed in the combinations in which the highest concentrations of ECT were used. The results suggest that oxidative stress induced by FA in daphnids is likely to be alleviated by the antioxidative action of ECT.
Ectoine (ECT) is a compatible solute produced by soil, marine and freshwater bacteria in response to stressful factors. The purpose of our study was to determine the possible toxic influence of ECT on Daphnia magna. We determined the following endpoints: survival rate during exposure and recovery, swimming performance, heart rate, thoracic limb movement determined by image analysis, haemoglobin level by ELISA assay, catalase and nitric oxide species (NOx) by spectrophotometric methods. The results showed 80% survival of daphnids exposed to 50 mg/L of ECT after 24 h and 10% after 90 h, however lower concentrations of ECT were well tolerated. A concentration-dependent reduction of swimming velocity was noted at 24 and 48 h of the exposure. ECT (at 2.5 and 4 mg/L) induced an increase of heart rate and thoracic limb movement (at 2.5, 4 and 20 mg/L) after 24 h. After 10 h of the exposure to ECT daphnids showed a concentration-dependent increase of haemoglobin level synthesized and accumulated in the epipodite epithelia. After 24 h we noted a concentration-dependent decrease of haemoglobin level and its lowest value was found after 48 h of the exposure. ECT at a concentration of 20 and 25 mg/L slightly stimulated catalase activity after 24 h. NOx level was also increased after 10 h of the exposure to 20 and 25 mg/L of ECT reaching maximal activity after 24 h. Our results suggest that ECT possesses some modulatory potential on the behaviour, physiology and biochemical parameters in daphnids.
Current red blood cell cryopreservation methods utilize bulk volumes, causing cryo-injury of cells, which results in irreversible disruption of cell morphology, mechanics, and function. An innovative approach to preserve human red blood cell morphology, mechanics, and function following vitrification in nanoliter volumes is developed using a novel cryo-ink integrated with a bio-printing approach.
The tear fluid lipid layer is present at the outermost part of the tear film which lines the ocular surface and functions to maintain the corneal surface moist by retarding evaporation. Instability in the structure of the tear fluid lipid layer can cause an increased rate of evaporation and thus dry eye syndrome. Ectoine has been previously shown to fluidize lipid monolayers and alter the phase behavior. In the current study we have investigated the effect of ectoine on the artificial tear fluid lipid layer composed of binary and ternary lipid mixtures of dipalmitoyl phosphatidylcholine (DPPC), Cholesteryl esters and tri-acyl-glycerols. The focus of our study was mainly the structural and the biophysical aspects of the artificial tear fluid lipid layer using surface activity studies and topology analysis. The presence of ectoine consistently causes an expansion of the pressure-area isotherm indicating increased intermolecular spacing. The topology studies showed the formation of droplet-like structures due to the addition of ectoine only when tri-acyl-glycerol is present in the mixture of DPPC and chol-palmitate, similar to the natural meibomian lipids. Consequently, the hypothesis of an exclusion of tri/di-acyl-glycerol from the meibomian lipid film in the presence of ectoine in the subphase is confirmed. A model describing the effect of ectoine on meibomian lipid films is further presented which may have an application for use of ectoines in eye drops as a treatment for the dry eye syndrome.
Background:
Due to the drastic shortage of organ donors, clinicians are increasingly considering the use of deceased after cardiac death donors (DCD). Compatible solutes like Ectoine and Hydroxyectoine are produced by extremophilic bacteria as a cell protectant to survive in harsh environments. We hypothesized that the addition of Hydroxyectoine to Histidine-Tryptophan-Ketoglutarate solution (HTK) could ameliorate cold ischemic preservation injury of DCD livers.
Material and methods:
Rat livers were harvested from male Wistar rats weighing 250-300 g. Three experimental groups (n=5 per group) were studied: (1) CONTROLS: cold static storage in HTK for 24 h, (2) DCD: 30-min warm ischemia time and 24-h cold static storage in HTK, and (3) DCD+Hydroxyectoine: like DCD, but with 24-h cold static storage in HTK+Hydroxyectoine. Viability of the livers was assessed after 24 h of preservation by isolated perfusion for 45 min with oxygenated Krebs-Henseleit buffer solution.
Results:
(mean ±SEM, Control vs. DCD vs. DCD+Hydroxyectoine) Parenchymal enzyme release was significantly lower in DCD+Hydroxyectoine compared to DCD (AST: 9±0.54; 56.8±2.05; 32.2±7.25 U/L, ALT: 9.5±0.5; 37.75±9.6; 17.5±4.17 U/L). Bile production at the end of 45 min reperfusion was significantly higher in DCD+Hydroxyectoine (5.16±1.32; 1.36±0.34; 10.75±2.24 µL/g liver weight/45 min). Malondialdehyde values were significantly lower in DCD+Hydroxyectoine (0.8±0.09, 1.14±0.18, 0.77±0.08 nmol/mL). Intercellular adhesion molecule-1 showed significantly lower values in DCD+Hydroxyectoine (219.07±51.79, 431.9±35.70, 205.2±37.71 pg/mL) and the portal venous pressure at 45 min was lower compared to DCD (20.41±0.12, 27.47±0.45, 22.08±0.78 mmHg).
Conclusions:
Our data provide evidence for the beneficial role of Hydroxyectoine-modified HTK solution for the preservation of DCD livers compared to HTK.
The present work investigates the effects of a variety of natural cryoprotectants in combination on post-thaw viability and apoptosis of cryopreserved mononuclear cells (MNCs) derived from umbilical cord blood. The extracellular cryoprotectants (10mM) namely trehalose, hydroxyl ethyl starch, polyvinyl pyrrolidine and intracellular CPAs (5mM) like erythritol, taurine and ectoine were used to prepare different combinations of freezing medium following L9 (34) Taguchi orthogonal array. Catalase, coenzyme Q10 and n-acetyl cystine (100μg/m) were added as antioxidants. Among various combinations, freezing medium consisting of hydroxyl ethyl starch, ectoin and co-enzyme Q10 with 10% FBS is found to be most effective combination achieving maximum cell viability of 93%, 5.6% early apoptotic, 0.7% late apoptotic and 0.1% necrotic cells. SEM and phase contrast microscopy confirmed the normal cell morphology of the post-thaw cultured cells with retaining their membrane integrity. The survival rate of MNCs is higher than the rate achieved using conventional Me2SO.
Abstract Compatible solutes are small, uncharged, zwitter ionic, osmotically active molecules produced and accumulated by microorganisms inside their cell to counteract different kinds of environmental stress. They enhance protein stability without interfering with the metabolic pathways even at molar concentrations. In this paper, we report the stabilizing effects of compatible solutes, ectoine, betaine and taurine on membrane protein Bacteriorhodopsin at different concentrations. Using Atomic Force Microscopy-Single Molecule Force Spectroscopy the impact of the osmolytes was quantified by measuring the forces required to pull the protein out of the membrane and the change in the persistence lengths of the unfolded polypeptide chain. Increasing unfolding forces were observed indicating the strengthening of intra-molecular interactions, which are vital for protein stability. The decrease in persistence lengths was recorded showing increasing tendencies of the polypeptide strand to coil up. Interestingly, it was revealed that these molecules have different stabilizing effects on protein unfolding at different concentrations. The results show that the unfolding of single proteins provides insight to the structure-dynamic relationship between the protein and compatible solute molecules at sub-nanometer scale. This also helps to understand the molecular mechanism involved in protein stabilization by organic osmolytes.
Introduction:
The natural cyclic tetrahydropyrimidine, ectoine, is a low-molecular, water-binding, organic osmolyte. Previously, topical application of ectoine to healthy human skin was shown to improve skin hydration as well as skin barrier function.
Objectives:
We therefore speculated that topical application of ectoine would be beneficial for patients with atopic dermatitis (AD), in which a genetically defined defect in skin barrier function is of major pathogenetic relevance. We assessed the efficacy of an ectoine-containing cream (EHK02-01) in the management of 65 patients with mild to moderate AD in a randomized, intra-individual, double-blind, multi-center trial, in which the efficacy of ectoine was compared to a nonsteroidal anti-inflammatory cream previously found to primarily act on skin barrier function and therefore with a comparable mode of action.
Methods:
Sixty-five patients with mild to moderate AD aged 18-65 years were enrolled. The patients applied EHK02-01 and the control cream on two symmetrical lesions twice daily for 28 days. At the beginning, after 7 and after 28 days, treated skin areas were assessed by modified, objective local SCORAD (Scoring Atopic Dermatitis) and IGA (Investigator's Global Assessment) as well as the patients' judgment of efficacy and their assessment of pruritus.
Results:
EHK02-01 was found to be very well tolerated. Even more important, efficacy of EHK02-01 treatment was equivalent to that achieved with the reference product.
Conclusion:
These results indicate that topical treatment with EHK02-01 may represent a novel option for the treatment of patients with AD.
Dimethyl sulfoxide (DMSO) is till now a widely used cryoprotective agent for cryopreservation of cells. Since high concentrations of DMSO have detrimental effects on cell functioning the aim of this study is to reduce the DMSO concentration by using compatible solutes (CS). These are small organic osmolytes that microorganisms synthesize and accumulate to counteract stress factors. Three CS, hydroxyectoine, ectoine and L-proline were investigated as cryoprotective agents for the cryopreservation of the human endothelial cell line HPMEC-ST1.6R. They were either supplemented to freezing or to cell culture medium. L-proline was the most effective CS, the efficiency of recultivation was improved by more than 100 percent. A combination of L-proline and ectoine in the cell culture medium resulted in an improvement by 63 percent. Our results show that L-proline and ectoine could be used as additional CPAs to improve the cryopreservation of human endothelial cells in the presence of low DMSO concentration.
Background/aims:
Ischemia reperfusion (I/R) injury after small bowel transplantation leads to inflammatory reactions and loss of structural integrity with subsequent graft contractile dysfunction in the early postoperative phase. The natural tetrahydropyrimidine ectoine (1-,4-,5-,6-tetrahydro-2-methyl-4-pyrimidine carboxylic acid; THP) protects the ileal mucosa and muscularis against effects of I/R injury in an experimental model of isolated graft reperfusion. The effects of THP treatment were evaluated in an established experimental intestinal transplant model.
Methods:
Isogenic, orthotopic small bowel transplantation was performed in Lewis rats (6 h cold ischemia time). Perioperative THP treatment (intraluminal/intravascular) groups were compared to vehicle-treated animals (after 3 and 24 h) and non-transplanted controls (n = 5/group). Park's score defined the effects of I/R injury. The infiltration of neutrophils, monocytes and macrophages, mRNA expression of IL-6 and TNF-α, serum levels of IL-6 and NO and smooth muscle contractility were evaluated.
Results:
Improved graft outcome after intraluminal and intravascular THP treatment was defined by considerably ameliorated neutrophil infiltration and less histological signs of I/R injury (p ≤ 0.05). In the presence of THP, mRNA expression of IL-6 and TNF-α and IL-6 and NO serum levels were reduced and smooth muscle function was improved.
Conclusion:
THP treatment offers protection against the effects of I/R injury in intestinal transplantation in vivo, however, only as supplementary treatment option.
The aim of this study was to elucidate the protective effect of the new compatible solutes, ectoine and hydroxyectoine, on two sensitive enzymes (lactic dehydrogenase, phosphofructokinase). The solutes tested also included (for reasons of comparison) other compatible solutes such as glycine betaine and a number of disaccharides (sucrose, trehalose, maltose). All compatible solutes under investigation displayed remarkable stabilizing capabilities. However, the degree of protection depended on both the type of solute chosen and the enzyme used as a test system. The most prominent protectants were trehalose, ectoine and hydroxyectoine, which are very often found in nature (singly or in combinationn) as part of the compatible solute cocktail of moderately halophilic eubacteria.
We have performed Molecular Dynamics simulations of ectoine, hydroxyectoine and urea in explicit solvent. Special attention has been spent on the local surrounding structure of water molecules. Our results indicate that ectoine and hydroxyectoine are able to accumulate more water molecules than urea by a pronounced ordering due to hydrogen bonds. We have validated that the charging of the molecules is of main importance resulting in a well defined hydration sphere. The influence of a varying salt concentration is also investigated. Finally we present experimental results of a DPPC monolayer phase transition that validate our numerical findings.
Ectoine and hydroxyectoine belong to the family of compatible solutes and are among the most abundant osmolytes in nature. These compatible solutes protect biomolecules from extreme conditions and maintain their native function. In the present study, we have investigated the effect of ectoine and hydroxyectoine on the domain structures of artificial lung surfactant films consisting of dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylglycerol (DPPG) and the lung surfactant specific surfactant protein C (SP-C) in a molar ratio of 80:20:0.4. The pressure-area isotherms are found to be almost unchanged by both compatible solutes. The topology of the fluid domains shown by scanning force microscopy, which is thought to be responsible for the biophysical behavior under compression, however, is modified giving rise to the assumption that ectoine and hydroxyectoine are favorable for a proper lung surfactant function. This is further evidenced by the analysis of the insertion kinetics of lipid vesicles into the lipid-peptide monolayer, which is clearly enhanced in the presence of both compatible solutes. Thus, we could show that ectoine and hydroxyectoine enhance the function of lung surfactant in a simple model system, which might provide an additional rationale to inhalative therapy.