The Valorisation of Melissa officinalis Distillation By-Products for the Production of Polyphenol-Rich Formulations.

Lemon balm (Melissa officinalis) is an aromatic and medicinal plant, rich in bioactive ingredients and with superior antioxidant activity. The essential oil of this plant is an expensive product, so the use of the by-products of the essential oil industry is particularly useful. The aim of this research was to process Melissa officinalis distillation by-products to develop a series of polyphenol-rich formulations. In the present research, lemon balm was distilled in a laboratory-scale distiller, and the recovered by-product was used for further successive extractions with acetone and water, using a fixed-bed semi-batch extractor. Acetone extract exhibited relatively poor results as far as yield, phenolic composition and antiradical activity are concerned. However, the aqueous extract presented high yield in both total phenolic content (i.e., 111 mg gallic acid equivalents (GAE)/g, on a dry herb basis (dw)), and anti-radical capacity (205 mg trolox equivalents (TE)/g dw). On a dried extract basis, the results were also impressive, with total phenols reaching 322 mg GAE/g dry extract and antiradical capacity at 593 mg TE/g dry extract. The phenolic components of the extract were identified and quantified by HPLC-DAD. Rosmarinic acid was the major component and amounted to 73.5 mg/g dry extract, while the total identified compounds were quantified at 165.9 mg/g dry extract. Finally, formulations with two different wall materials (gum arabic-maltodextrin and maltodextrin) and two different drying methods (spray-drying and freeze-drying) were applied and evaluated to assess their performance, yield, efficiency and shelf-life of total phenolic content and rosmarinic acid concentration. From the present investigation, it is concluded that after one year of storage, rosmarinic acid does not decrease significantly, while total phenolic content shows a similar decrease for all powders. According to the yield and efficiency of microencapsulation, maltodextrin alone was chosen as the wall material and freeze-drying as the preferred drying method.

Melissa officinalis essential oil as an antimicrobial agent against Listeria monocytogenes in watermelon juice.

The burden of foodborne illness has a negative effect on public health, but also in countries' economy. Melissa officinalis is an aromatic plant known for its biological properties, including antioxidant and antimicrobial effects. This work highlighted M. officinalis essential oil's antioxidant potential and antimicrobial activity against L. monocytogenes, presenting a bactericidal action and being able to inhibit some virulence attributes, such as biofilm formation. The pre-exposure of the bacterium to subinhibitory levels of essential oil (0.125 µL/mL) did not induce high tolerance to stresses (such as high temperature, low pH, osmotic stress and desiccation) or cross-resistance with antibiotics, while not modifying the invasion ability to Caco-2 cells. When applied in food model media (lettuce, chicken and milk) and watermelon juice, the essential oil showed to have antimicrobial activity in a lettuce leaf model medium, further diminishing L. monocytogenes contamination and inhibiting the natural microbiota present in watermelon juice. M. officinalis essential oil shows potential to be used as control of L. monocytogenes in watermelon juice, while increasing the food's microbial shelf life.

Melissa officinalis L. nanoethosomal formulation: evaluation of antioxidant, enzyme inhibitory activities and in vitro toxicity.

This work aimed to create an extract of Melissa officinalis L. with strong antiradical efficacy, characterize it, and enhance its long-term efficacy by developing an ethosomal formulation. DPPH and ABTS assays were used to test the antiradical activity of extracts with different ethanol ratios obtained from the aerial part. Phytochemical characterization of the extract with the highest activity, ethyl acetate fraction of 60% ethanol extract, was analyzed by HPLC. The active ethyl acetate fraction was loaded into ethosomes, and characterization and release studies of the formulation were performed. The released extract from the formulation exhibited substantial antiradical action as well as inhibition of collagenase (71.5%) and elastase (75.5%) enzymes. The toxicity of the active extract and the formulation was determined in the mouse fibroblast cell line. This study successfully developed a long-term antioxidant and enzyme inhibitor formulation containing M. officinalis, which stands out for its medicinal properties.

Molecular barcoding of Melissa officinalis L. (badranjboye) in Iran and identification of adulteration in its medicinal services.

Medicinal plants are an important source for treatment of diseases in many countries. Today, controlling the quality of the products of medicinal plants is an important task. Customer health may be in danger due to the fraud and misconduct by the sales associates in the sales centers. Melissa officinalis L. (Lamiaceae) is an important medicinal plant used for treatment of several diseases. In Iran, the species of Dracocephalum, Hymencrater, Nepeta and Stachys are mistakenly sold under the name of Badranjboye that have different pharmaceutical properties. For avoiding this mistake, this investigation was performed with the following aims: 1) Checking for the cheating and identifying the Badranjboye in the Iran's market of medicinal plants, 2) Providing the molecular barcode for the medicinal species of Melissa. For this purpose, Market-sold plant samples (leaves) and original reference plant species were compared by morphology, odor as well as Internal transcribed spacer (ITS) and chloroplast DNA ((psbA-trnH) and (trnL-trnF)) sequences. Various molecular analyses, such as sequencing, determination of genetic distance, and construction of phylogenetic tree were performed. These reports have shown that internal transcribed spacer (ITS) and chloroplast DNA (psbA-trnH) sequences are an efficient molecular marker to produce barcode gap and differentiating Melissa officinalis from other species.

Synergistic effects of Nepeta menthoides and Melissa officinalis aqueous extracts on reserpine-induced depressive-like behaviors in mice.

Nepeta menthoides Boiss. & Buhse and Melissa officinalis are extensively used in Persian medicine for the treatment of depression. Considering the active ingredients and main phenolic compounds of these plants and possible synergistic effects, this study examined the antidepressant and antioxidant activities of aqueous extract of N. menthoides (NM) and M. officinalis (MO) in reserpinized mice alone and combination. Mice were pretreated orally for 1-week with normal saline (10 ml/kg), fluoxetine (20 mg/kg), imipramine (10 mg/kg), NM (50-100-200-400 mg/kg), MO (150-350-550-750 mg/kg), and combination (NM 50 with MO 150 mg/kg). The behavioral changes were evaluated using forced swim, tail suspension, and open field tests, 24 hr after reserpine injection (4 mg/kg) on eighth day. The amounts of active components in the extracts and catalase (CAT) as a brain oxidative stress were measured with enzyme-linked immunosorbent assay. Data showed that this combination produced a synergistic action on behaviors and a significant increase in CAT activity. High-performance liquid chromatography results showed that rosmarinic acid contents in MO and NM were 6.42 ± 1.1 and 11.03 ± 2.16 mg/g of dried extract, respectively. Total flavonoid and phenolic contents of MO were higher than NM. The findings suggest that the present combination produces an antidepressant-like effect, which is possibly triggered by its antioxidant properties.

Melissa officinalis: Composition, Pharmacological Effects and Derived Release Systems-A Review.

Melissa officinalis is a medicinal plant rich in biologically active compounds which is used worldwide for its therapeutic effects. Chemical studies on its composition have shown that it contains mainly flavonoids, terpenoids, phenolic acids, tannins, and essential oil. The main active constituents of Melissa officinalis are volatile compounds (geranial, neral, citronellal and geraniol), triterpenes (ursolic acid and oleanolic acid), phenolic acids (rosmarinic acid, caffeic acid and chlorogenic acid), and flavonoids (quercetin, rhamnocitrin, and luteolin). According to the biological studies, the essential oil and extracts of Melissa officinalis have active compounds that determine many pharmacological effects with potential medical uses. A new field of research has led to the development of controlled release systems with active substances from plants. Therefore, the essential oil or extract of Melissa officinalis has become a major target to be incorporated into various controlled release systems which allow a sustained delivery.

Rosmarinic Acid, as an NHE1 Activator, Decreases Skin Surface pH and Improves the Skin Barrier Function.

Stratum corneum (SC) pH regulates skin barrier functions and elevated SC pH is an important factor in various inflammatory skin diseases. Acidic topical formulas have emerged as treatments for impaired skin barriers. Sodium proton exchanger 1 (NHE1) is an important factor in SC acidification. We investigated whether topical applications containing an NHE1 activator could improve skin barrier functions. We screened plant extracts to identify NHE1 activators in vitro and found Melissa officinalis leaf extract. Rosmarinic acid, a component of Melissa officinalis leaf extract, significantly increased NHE1 mRNA expression levels and NHE1 production. Immunofluorescence staining of NHE1 in 3D-cultured skin revealed greater upregulation of NHE1 expression by NHE1 activator cream, compared to vehicle cream. Epidermal lipid analysis revealed that the ceramide level was significantly higher upon application of the NHE1 activator cream on 3D-cultured skin, compared to application of a vehicle cream. In a clinical study of 50-60-year-old adult females (n = 21), application of the NHE1 activator-containing cream significantly improved skin barrier functions by reducing skin surface pH and transepidermal water loss and increasing skin hydration, compared to patients who applied vehicle cream and those receiving no treatment. Thus, creams containing NHE1 activators, such as rosmarinic acid, could help maintain or recover skin barrier functions.

Quantitative Proteome Analysis Reveals Melissa officinalis Extract Targets Mitochondrial Respiration in Colon Cancer Cells.

Melissa officinalis (MO), known as lemon balm, is a popular ingredient blended in herbal tea. In recent decades, the bioactivities of MO have been studied in sub-health and pathological status, highlighting MO possesses multiple pharmacological effects. We previously showed that hot water MO extract exhibited anticancer activity in colorectal cancer (CRC). However, the detailed mechanisms underlying MO-induced cell death remain elusive. To elucidate the anticancer regulation of MO extract in colon cancer, a data-driven analysis by proteomics approaches and bioinformatics analysis was applied. An isobaric tandem mass tags-based quantitative proteome analysis using liquid chromatography-coupled tandem mass spectrometry was performed to acquire proteome-wide expression data. The over-representation analysis and functional class scoring method were implemented to interpret the MO-induced biological regulations. In total, 3465 quantifiable proteoforms were identified from 24,348 peptides, with 67 upregulated and 54 downregulated proteins in the MO-treated group. Mechanistically, MO impeded mitochondrial respiratory electron transport by triggering a reactive oxygen species (ROS)-mediated oxidative stress response. MO hindered the mitochondrial membrane potential by reducing the protein expression in the electron transport chain, specifically the complex I and II, which could be restored by ROS scavenger. The findings comprehensively elucidate how MO hot water extract activates antitumor effects in colorectal cancer (CRC) cells.

Effect of graphene / metal nanocomposites on the key genes involved in rosmarinic acid biosynthesis pathway and its accumulation in Melissa officinalis.

BACKGROUND: Recently, numerous investigations have been done to study graphene and silver nanoparticle in the fields of agriculture and medicine. In the present study, the green synthesis of nanoparticles with two concentrations (0, 40, 60 mM) and their effect on the molecular and biochemical biosynthesis pathway of rosmarinic acid in a new method, low cost, and safe for the environment has been investigated. The transcript levels of key genes in the rosmarinic acid biosynthesis pathway (Tyrosine aminotransferase, rosmarinic acid synthase, and phenylalanine-ammonia lyase) were studied using real-time quantitative polymerase chain reaction. Then, the rosmarinic acid content was evaluated using HPLC. RESULTS: The results showed that a concentration-dependent manner was observed in treated plants. At the biochemical level, the use of nanocomposites at concentration of 40 mM showed higher soluble carbohydrate (37%), flavonoids (21%), total phenol (35%) as well as total protein (47%) compared to the control plants. HPLC results showed that rosmarinic acid content in the treated plants with a low concentration of nanocomposite (40 mM) was more affected than plants treated with a high concentration of nanocomposite (60 mM) (26%) and also compared to other treatments. At the molecular level, the result showed that Tyrosine aminotransferase and rosmarinic acid synthase gene expression was positively correlated with both silver nanoparticle concentrations and nanocomposite treatments, but phenylalanine-ammonia lyase gene expression was positively correlated only with nanocomposite at 40 mM concentration. CONCLUDE: It can conclude that the nanocomposite at low concentration is more likely to induce molecular and biochemical parameters. And also, in the rosmarinic acid biosynthesis pathway, the Tyrosine aminotransferase -derived pathway is more efficient than the phenylalanine-ammonia lyase -derived pathway by causing a nano-elicitor. Therefore, it was concluded that studied elicitor at low concentration, can create plants with higher production capacity.

The effects of lemon balm (Melissa officinalis L.) on depression and anxiety in clinical trials: A systematic review and meta-analysis.

A systematic review and a meta-analytic approach were considered to investigate the effects of lemon balm as a medicinal herb on anxiety and depression in clinical trials and its side effects. All randomized clinical trials published up to October 30, 2020 that examined lemon balm in patients with symptoms of depression or anxiety, with acute or chronic manifestations, were searched in 12 online databases. Statistical analysis was performed using RevMan software. Continuous data were analyzed using standardized mean differences. Statistical heterogeneity was assessed using Chi2 , I2 , and p value tests. Based on meta-analysis results, lemon balm significantly improved mean anxiety and depression scores compared with the placebo (SMD: -0.98; 95% CI: -1.63 to -0.33; p = 0.003), (SMD: -0.47; 95% CI: -0.73 to -0.21; p = 0.0005) respectively, without serious side effects. Current evidence suggests that lemon balm may be effective in improving anxiety and depressive symptoms, particularly in the acute setting. Due to the high level of heterogeneity between studies, results should be interpreted with caution. The small number of clinical trials and differences between their methods were the limitations of the present study. Further high-quality studies are needed to firmly establish the clinical efficacy of the lemon balm.

Effect of Melissa officinalis on systolic and diastolic blood pressures in essential hypertension: A double-blind crossover clinical trial.

For a long time, Melissa officinalis (M. officinalis) has been used to treat cardiovascular diseases. Therefore, this study aimed at evaluating the effects of M. officinalis on systolic and diastolic blood pressures in hypertensive patients. We conducted a double-blind, controlled, randomized crossover clinical trial on 49 patients who received either M. officinalis capsules (400 mg/d; n = 23) or the placebo (n = 26) three times per day for a 4-week period. After a 2-week washout period, the M. officinalis group received placebo and the other group received M. officinalis for another 4-week period. The systolic and diastolic blood pressures were measured once at baseline and then every 2 weeks for 10 weeks. The statistical analysis of the obtained data revealed that the chronology of the consumption of M. officinalis and placebo had no effect on the systolic and diastolic blood pressures in these two studied groups. Moreover, it was found that systolic and diastolic blood pressures significantly decreased after the consumption of M. officinalis, compared to placebo. Systolic and diastolic blood pressures in group A at the beginning of the study were 152.30 ± 5.312 mmHg and 95.52 ± 1.988 mmHg, respectively, and, after the first phase (drug use), reached 129.88 ± 9.009 mmHg and 80.13 ± 5.488 mmHg, respectively. Systolic and diastolic blood pressures in group B at the beginning of the study was 152.26 ± 5.640 mmHg and 94.44 ± 2.607 mmHg, respectively, and after the second phase (drug use), reached 131.77 ± 8.091 mmHg and 81.46 ± 7.426 mmHg, (p = .005), respectively. Also, no significant side effects were observed during the study. According to the results, M. officinalis can reduce systolic and diastolic blood pressures of the patients with essential hypertension.

Melissa officinalis L. Aqueous Extract Exerts Antioxidant and Antiangiogenic Effects and Improves Physiological Skin Parameters.

Melissa officinalis (MO) is a medicinal plant well-known for its multiple pharmacological effects, including anti-inflammatory, anticancer and beneficial effects on skin recovery. In this context, the present study was aimed to investigate the in vitro and in vivo safety profile of an MO aqueous extract by assessing cell viability on normal (HaCaT-human keratinocytes) and tumor (A375-human melanoma) cells and its impact on physiological skin parameters by a non-invasive method. In addition, the antioxidant activity and the antiangiogenic potential of the extract were verified. A selective cytotoxic effect was noted in A375 cells, while no toxicity was noticed in healthy cells. The MO aqueous extract safety profile after topical application was investigated on SKH-1 mice, and an enhanced skin hydration and decreased erythema and transepidermal water loss levels were observed. The in ovo CAM assay, performed to investigate the potential modulating effect on the angiogenesis process and the blood vessels impact, indicated that at concentrations of 100 and 500 µg/mL, MO aqueous extract induced a reduction of thin capillaries. No signs of vascular toxicity were recorded at concentrations as high as 1000 µg/mL. The aqueous extract of MO leaves can be considered a promising candidate for skin disorders with impaired physiological skin parameters.

Melissa officinalis L. hydro-alcoholic extract inhibits anxiety and depression through prevention of central oxidative stress and apoptosis.

NEW FINDINGS: What is the central question of this study? How does an extract of Melissa officinalis L. ameliorate anxiety- and depressive-like behaviour of mice? What is the main finding and its importance? An extract of Melissa officinalis L. possessed anxiolytic and anti-depressant effects, which could mainly be mediated through its antioxidant and anti-apoptotic properties. ABSTRACT: This study evaluated the effects of a hydro-alcoholic extract of Melissa officinalis (HAEMO) on anxiety- and depressive-like behaviours, oxidative stress and apoptosis markers in restraint stress-exposed mice. In order to induce a depression-like model, mice were subjected to restraint stress (3 h day-1 for 14 days) and received normal saline or HAEMO (50, 75 and 150 mg kg-1  day-1 ) for 14 days. The administered doses of HAEMO were designated based on the concentration of one of the main phenolic compounds present in the extract, rosmarinic acid (2.55 mg kg-1 at lowest dose); other phytochemical analyses including assays for antioxidant activity, total phenols and flavonoids were also carried out. The behavioural changes in an open field task, elevated plus maze, tail suspension and forced swimming tests were evaluated. Also, malondialdehyde (MDA) levels and enzyme activities of superoxide dismutase and glutathione peroxidase, and total antioxidant capacity were assessed in the prefrontal cortex and hippocampus. Moreover, levels of Bcl-2, Bax and caspase 3 in the brain as well as serum concentration of corticosterone were evaluated. HAEMO (75 and 150 mg kg-1 ) significantly reversed anxiety- and depressive-like behaviours. Also, HAEMO reduced MDA levels, enhanced enzymatic antioxidant activities and restored serum levels of corticosterone. An immunoblotting analysis also demonstrated that HAEMO decreased levels of pro-apoptotic markers and increased anti-apoptotic protein levels in the prefrontal cortex and hippocampus of restraint stress-exposed mice. Our findings suggested that HAEMO reduced inflammation and had anxiolytic and antidepressant effects in mice.

Effects of Melissa officinalis (Lemon Balm) on cardio-metabolic outcomes: A systematic review and meta-analysis.

Recent evidence indicates a beneficial effect of Melissa officinalis (MO) intake on several chronic diseases. However, the effects of MO intake have not yet been systematically reviewed. Therefore, we conducted a systematic review and meta-analysis of randomized controlled trials (RCTs) to evaluate the effect of MO intake and focused on several cardiometabolic outcomes. MEDLINE, Scopus, EMBASE, Web of Science and the Cochrane Central Register of Controlled Trials were searched for MO-RCTs evaluating cardiometabolic outcomes. Random-effects meta-analyses estimated the pooled standardized mean differences (SMD) between intervention and control groups. Risk of bias was assessed with the Cochrane Collaboration's tool for assessing the risk of bias in RCTs. Seven RCTs were finally deemed eligible. MO intake was associated with a reduced total cholesterol (TC) (SMD: -0.26; 95% CI: -0.52, -0.01; I2 = 13.7%; k = 6) and a reduced systolic blood pressure (SBP) (SMD: -0.56; 95% CI: -0.85, -0.27; I2 = 00.0%; k = 3). MO intake was not associated with statistically significant changes in triglycerides, low-density lipoprotein, diastolic blood pressure, high sensitivity c-reactive protein levels, fasting blood sugar, HbA1c, insulin or high-density lipoprotein levels. No serious adverse events were reported. The risk of bias was high in a considerable amount of studies. Our study suggests that MO is a safe supplement with beneficial effects on TC and SBP. However, the findings of our study must be seen in the light of major limitations such as a low number of included studies and a serious risk of bias. High-quality RCTs are needed for firm conclusions concerning the effects of MO on cardiometabolic outcomes.

Dietary Intake of Rosmarinic Acid Increases Serum Inhibitory Activity in Amyloid A Aggregation and Suppresses Deposition in the Organs of Mice.

Serum amyloid A (SAA) is one of the most important precursor amyloid proteins and plays a vital step in AA amyloidosis, although the underlying aggregation mechanism has not been elucidated. Since SAA aggregation is a key step in this pathogenesis, inhibitors are useful to prevent and treat AA amyloidosis, serving as tools to investigate the pathogenic mechanism. In this study, we showed that rosmarinic acid (RA), which is a well-known inhibitor of the aggregation of amyloid ß (Aß), displayed inhibitory activity against SAA aggregation in vitro using a microliter-scale high-throughput screening (MSHTS) system with quantum-dot nanoprobes. Therefore, we evaluated the amyloid aggregation inhibitory activity of blood and the deposition of SAA in organs by feeding mice with Melissa officinalis extract (ME) containing RA as an active substance. Interestingly, the inhibitory activity of ME-fed mice sera for SAA and Aß aggregation, measured with the MSHTS system, was higher than that of the control group. The amount of amyloid deposition in the organs of ME-fed mice was lower than that in the control group, suggesting that the SAA aggregation inhibitory activity of serum is associated with SAA deposition. These results suggest that dietary intake of RA-containing ME enhanced amyloid aggregation inhibitory activity of blood and suppressed SAA deposition in organs. This study also demonstrated that the MSHTS system could be applied to in vitro screening and to monitor comprehensive activity of metabolized foods adsorbed by blood.

Regulation of Obesity by Antiangiogenic Herbal Medicines.

Obesity is the result of an energy imbalance caused by an increased ratio of caloric intake to energy expenditure. In conjunction with obesity, related metabolic disorders, such as dyslipidemia, atherosclerosis, and type 2 diabetes, have become global health problems. Obesity progression is thought to be associated with angiogenesis and extracellular matrix (ECM) remodeling. Angiogenesis occurs in growing adult adipose tissues, which are similar to neoplastic tissues. Adipose tissue is highly vascularized, and each adipocyte is nourished by an extensive capillary network. Adipocytes produce proangiogenic factors, such as vascular endothelial growth factor A and fibroblast growth factor 2, which promote neovascularization within the adipose tissue. Furthermore, matrix metalloproteinases (MMPs), including MMP-2 and MMP-9, play important roles in adipose tissue development and microvessel maturation by modifying the ECM. Thus, modulation of angiogenesis and MMP activity provides a promising therapeutic approach for controlling human obesity and its related disorders. Over the past decade, there has been a great increase in the use of alternative treatments, such as herbal remedies, for these diseases. This review will focus on the role of angiogenesis in adipose tissue growth and the regulation of obesity by antiangiogenic herbal medicines.

Glycerosome of Melissa officinalis L. Essential Oil for Effective Anti-HSV Type 1.

Essential oils are complex mixtures of strongly active compounds, very volatile and sensitive to light, oxygen, moisture and temperature. Loading inside nanocarriers can be a strategy to increase their stability and successfully use them in therapy. In the present study, a commercial Melissa officinalis L. (Lamiaceae) essential oil (MEO) was analyzed by gas chromatography-mass spectrometry, loaded inside glycerosomes (MEO-GS) and evaluated for its anti-herpetic activity against HSV type 1. MEO-GS analyses were prepared by the thin layer evaporation method and they were characterized by light scattering techniques, determining average diameter, polydispersity index and ζ-potential. By transmission electron microscopy, MEO-GS appeared as small nano-sized vesicles with a spherical shape. MEO encapsulation efficiency inside glycerosomes, in terms of citral and ß-caryophyllene, was found to be ca. 63% and 76% respectively, and MEO release from glycerosomes, performed by dialysis bag method, resulted in less than 10% within 24h. In addition, MEO-GS had high chemical and physical stability during 4 months of storage. Finally, MEO-GS were very active in inhibiting HSV type 1 infection of mammalian cells in vitro, without producing cytotoxic effects. Thus, MEO-GS could be a promising tool in order to provide a suitable anti-herpetic formulation.

Effect of lemon balm (Melissa officinalis) extract on growth performance, digestive and antioxidant enzyme activities, and immune responses in rainbow trout (Oncorhynchus mykiss).

This study was conducted to determine the effects of dietary supplementation of lemon balm (Melissa officinalis) aqueous methanolic extract on growth performance, blood parameters, digestive and antioxidant enzyme activities, and non-specific immune responses in rainbow trout (Oncorhynchus mykiss). Fish with an average weight of 23.03 ± 0.07 g were fed a diet supplemented with an aqueous methanolic extract of lemon balm at a dose of 0 (control), 0.1 (LB0.1), 0.5 (LB0.5), and 1 g kg-1 (LB1) for 75 days. The final weight, weight gain, and specific growth rate were observed to be significantly increased in LB0.5 and LB1 groups compared with that of the control. No differences were observed in feed conversion ratio values. WBC increased at the 45th day of the study in LB0.1 group. No differences were determined in RBC (P > 0.05). At the end of the study, lipase increased significantly in all experimental groups compared with the control. Pepsin was also elevated in LB0.5 and LB1 groups compared with the control. Increased trypsin was determined in LB1 group (P < 0.05). SOD activity increased at the end of the study in LB1 group (P < 0.05). CAT values had no differences compared with control. GR activity increased in all experimental groups compared with control. GPx improved in LB0.5 and LB1 groups significantly (P < 0.05). Lipid peroxidation was decreased in LB1 group compared with that of control, but this decrease was not significant (P < 0.05). Oxidative radical production and lysozyme activity significantly increased in LB1 group (P < 0.05). The highest MPO was determined in control group (P < 0.05). Current results suggest that lemon balm extract stimulates growth promoting antioxidant and immune responses in rainbow trout.

Physiological and molecular mechanisms underlying salicylic acid-mitigated mercury toxicity in lemon balm (Melissa officinalis L.).

Mercury (Hg) is one of the most toxic heavy metals with strong negative effects on the plant growth and functions. Salicylic acid (SA) is an important signaling molecule which confers tolerance to metal toxicities but little is known about the mechanisms of SA-mediated alleviation of Hg stress. Here, physiochemical and molecular responses of Hg-stressed lemon balm (Melissa officinalis L.) to exogenous SA were investigated to reveal SA-induced tolerance mechanisms. The CHLG gene of lemon balm which encodes chlorophyll synthase was also partly isolated and sequenced for the first time. Hg stress markedly decreased growth, relative water content (RWC) and photosynthetic pigments of the plant. However, exogenous SA significantly mitigated the toxic effects of mercury on the growth and RWC and enabled plant to maintain chlorophylls to the similar levels of unstressed plants. Hg-induced oxidative damage was also reduced following treatment with SA and treated plants showed the lower extent of lipid peroxidation which was accompanied with the higher free proline and phenolics contents and elevation of the antioxidant capacity as evidenced by DPPH radical scavenging and FRAP assays. Moreover, SA treatment resulted in up-regulation of CHLG and phenylalanine ammonia-lyase (PAL) genes as key components of chlorophyll and phenylpropanoid routes, respectively. Our results collectively indicate the ameliorative effects of exogenous SA in mercury toxicity through coordinated alternations in plant metabolic processes which provide insights to better understand mechanisms of Hg tolerance in lemon balm plant.

Efficacy of Melissa officinalis L. (lemon balm) extract on glycemic control and cardiovascular risk factors in individuals with type 2 diabetes: A randomized, double-blind, clinical trial.

Melissa officinalis is a plenteous source of antioxidant flavonols and flavonoids that contain health-promoting and antidiabetic properties, so this study was undertaken to provide the first assessment of the antidiabetic properties of hydroalcoholic extract of M. officinalis in type 2 diabetic patients. We did a randomized, placebo-controlled trial which included 62 patients, receiving either M. officinalis capsules (700 mg/d; n = 31) or the placebo (n = 31) twice daily for 12 weeks. There were significant differences in serum FBS (P = 0.007), HbA1c (P = 0.002), ß-cell activity (P = 0.05), TG (P = 0.04), HDL-c (P = 0.05), hs-CRP (P = 0.001), and systolic blood pressure (P = 0.04) between the two groups at the end of the study; but total cholesterol, LDL-c, insulin, and HOMA-IR showed no significant changes between the groups. In M. officinalis group, there was a significant change in HDL-c (P = 0.009) and QUICKI (P = 0.005) compared with baseline values. No adverse effects were observed. It seems that M. officinalis is safe and effective in improvement of lipid profile, glycemic control, and reduction of inflammation.