Exogenous melatonin's effect on salivary cortisol and amylase: A randomized controlled trial.

This study aimed to examine the effect of acute exogenous melatonin administration on salivary cortisol and alpha-amylase (sCort and sAA) as representatives of the HPA axis and the sympathetic nervous system, respectively. A single-dose prolonged-release melatonin (2 mg) or a placebo tablet was given to healthy volunteers (n = 64) at 20:00 h in a crossover design. The saliva was collected at six time points (20:00, 21:00, awakening, 30 min after awakening, 10:00, and 12:00 h) and was measured for sCort, sAA, and salivary melatonin (sMT) levels. Pulse rates and sleep parameters were also collected. Melatonin was effective in improving sleep onset latency by 7:04 min (p = .037) and increasing total sleep time by 24 min (p = .006). Participants with poor baseline sleep quality responded more strongly to melatonin than participants with normal baseline sleep quality as they reported more satisfaction in having adequate sleep (p = .017). Melatonin administration resulted in higher sCort levels at awakening time point (p = .023) and a tendency of lower sAA levels but these were not significant. Melatonin ingestion at 20:00 h resulted in a marked increase in sMT levels at 21:00 h and remained higher than baseline up to at least 10:00 h (p < .001). Melatonin increases sCort levels at certain time point with a tendency to lower sAA levels. These opposing effects of melatonin suggested a complex interplay between melatonin and these biomarkers. Also, the results confirmed the positive acute effect of a single-dose melatonin on sleep quality.

HPLC analysis and in vitro antioxidant mediated through cell migration effect of C.hystrix water extract on human keratinocytes and fibroblasts.

Citrus hystrix or kaffir lime is a native tropical plant containing a high level of phenolic and flavonoid compounds. Its fruits are used as a food ingredient to enhance the sour-sweet scent and flavor in many dishes. Due to its polyphenol-containing, it has also been used as traditional medicine for health benefits including oral and gum health, stress relief, hair care, and skincare. In this study, we demonstrated the antioxidant activity of C. hystrix water extract and its effect on human keratinocyte and fibroblast migration. The extract showed a high amount of phenolic and flavonoid contents. The HPLC analysis indicated the presence of gallic acid, catechin, caffeic acid, rutin, and quercetin. We showed that C. hystrix water extract exhibited free radical scavenging capacity, determined by DPPH assay, with IC50 of 14.91 mg/mL, and nitrite radical scavenging capacity, determined by NO assay, with IC50 of 4.46 mg/mL. The C. hystrix water extract displayed unnoticeable toxicity at all tested doses. We showed that the treatment of water extracts as low as 50 µg/mL decreased the reactive oxygen species (ROS) from H2O2-induced ROS formation in both cell lines. Besides, C. hystrix water extract promoted cell migration in a dose-dependent manner. Together, these results demonstrated the positive benefit of C. hystrix water extract as a wound-healing accelerator. Its health benefits may be due to the antioxidant capability of its phytochemical compounds contained in C. hystrix water extract that enhances the migration of two major cell types: fibroblast and keratinocytes, responsible for the proliferation and remodeling phase of wound healing.

Photo-Protective and Anti-Inflammatory Effects of Antidesma thwaitesianum Müll. Arg. Fruit Extract against UVB-Induced Keratinocyte Cell Damage.

The main cause of most skin cancers is damage from UVB from sunlight, which penetrate the skin surface and induce inflammation. For this reason, this study aims to identify natural products with photo-protection properties and their mode of action by using the UVB-irradiated HaCaT keratinocyte model. Antidesma thwaitesianum fruit extracts at 25, 50, and 100 µg/mL recovered cell viability following UVB exposure in a dose-dependent manner. Cell survival was associated with the reduction in intracellular ROS and NO. In addition, we showed that the pre-treatment with the fruit extract lowered the phosphorylation level of two MAPK-signaling pathways: p38 MAPKs and JNKs. The resulting lower MAPK activation decreased their downstream pro-inflammatory cascade through COX-2 expression and subsequently reduced the PGE2 proinflammatory mediator level. The photoprotective effects of the fruit extract were correlated with the presence of polyphenolic compounds, including cyanidin, ferulic acid, caffeic acid, vanillic acid, and protocatechuic acid, which have been previously described as antioxidant and anti-inflammation. Together, we demonstrated that the pre-treatment with the fruit extract had photo-protection by inhibiting oxidative stress and subsequently lowered stress-induced MAPK responses. Therefore, this fresh fruit is worthy of investigation to be utilized as a skincare ingredient for preventing UVB-induced skin damage.

Comparison of salivary cortisol, amylase, and chromogranin A diurnal profiles in healthy volunteers.

OBJECTIVE: This study aimed to compare the diurnal patterns of three salivary biomarkers (cortisol, amylase, and chromogranin A), as well as the factors affecting their levels. DESIGN: A total of 110 participants with mean age of 26.93 ± 6.05 years old took part in the study. The saliva was collected at awakening, 30 min after awakening, 10:00, 12:00, 16:00, and 19:00 h. Cortisol, amylase, chromogranin A, and total protein levels were determined. RESULTS: The diurnal patterns of three biomarkers were different with cortisol showing the least variance and chromogranin A showing the highest variance among individuals. Participants with lower BMI exhibited higher cortisol levels (p = 0.044). Age oppositely affected amylase and chromogranin A, as being older was associated with higher amylase (p = 0.029) and lower chromogranin A levels (p < 0.01) even though both markers represent the sympathetic activity. Male participants also showed lower chromogranin A levels than females (p = 0.045). Total protein concentration affected chromogranin A levels only around awakening period but not at other time points suggesting that protein adjustment may not be necessary if the experiment is performed during the day. The afternoon was the period where all three biomarkers showed rather stable levels. CONCLUSIONS: These results illustrate the nature of cortisol, amylase, and chromogranin A patterns throughout the day in a normal physiological state and help in choosing the right condition to perform the experiment with these biomarkers.

The Leaf Extract of Coccinia grandis (L.) Voigt Accelerated In Vitro Wound Healing by Reducing Oxidative Stress Injury.

The impairment in the regulation of the physiological process in the inflammatory phase of wound healing results in oxidative stress damage, which increases the severity and extends the healing time. In this study, we aimed to evaluate the radical scavenging properties of Coccinia leaf extract and its ability to ameliorate a migration process in vitro. Coccinia is a medicinal plant that was used in ancient times for relieving insect bite itching and swelling. However, the role of Coccinia leaf extract as an antioxidant related to the process of wound healing has never been studied. In this study, we demonstrated that the leaf extract possessed antioxidant properties that acted as a proton donor to neutralize reactive oxygen species with the IC50 value of 4.85 mg/mL of the extract. It could chelate iron with the IC50 value of 21.39 mg/mL of the extract. The leaf extract protected the human fibroblasts and keratinocytes from hydrogen peroxide-induced oxidative stress by increasing cell survival rate by more than 20% in all test doses. The protective property was dose-dependently correlated with the decrease in reactive oxygen species formation. In addition, the leaf extract enhanced the cell migration rate of fibroblasts and keratinocytes up to 23% compared with vehicle control. The results suggested that Coccinia leaf extract may be a potential herb for increasing the wound healing process with its antioxidant capacity and can be used as an herbal ingredient for the utilization of skincare products.

Antioxidant and anti-inflammatory effects of piperine on UV-B-irradiated human HaCaT keratinocyte cells.

The increase in intracellular reactive oxygen and nitrogen species plays a key role in ultraviolet B (UV-B)-induced inflammatory responses in the human skin. Piperine exhibits many pharmacological benefits. In the present study, the photoprotective effects and the possible underlying mechanisms of the anti-inflammatory effects of piperine on UV-B-irradiated keratinocytes were investigated. Piperine exerted strong, direct scavenging effects on DPPH radicals and exhibited free radical scavenging capabilities as demonstrated by the DCFH-DA and Griess assays. Consistent with these results, 10, 20, and 40 µM piperine pretreatments attenuated UV-B irradiation-induced keratinocyte cytotoxicity as reported by the resazurin assay. The highest concentration of piperine inhibited UV-B irradiation-induced cell apoptosis, as revealed by Hoechst 33342 staining. Moreover, we demonstrated the anti-inflammatory effects of piperine using western blot analysis, real-time PCR, and ELISA. Pretreatment with piperine suppressed the activation of phosphorylated p38, JNK, and AP-1 as well as the levels of COX-2/PGE2 and iNOS synthesis, while UV-B-irradiated cells triggered the induction of these signaling molecules. These results indicated that the inhibition of these inflammatory signaling pathways might play a key role in the regulation of the anti-inflammatory effects of piperine. In addition, piperine showed stronger anti-inflammatory effects than celecoxib which served as a positive control at the same concentration. All these results suggested that the anti-inflammatory properties of piperine protected keratinocytes from UV-B-induced damage, which might be due to its antioxidant properties. Therefore, piperine may be an effective therapeutic candidate compound for the treatment of UV irradiation-induced skin inflammation.

Daily White kwao krua dietary supplement alleviates LDL oxidative susceptibility, plasma LDL level and improves vasculature in a hypercholesterolemia rabbit model.

BACKGROUND AND AIM: White kwao krua is an edible plant that grows in Southeast Asia. It is very rich in natural phytoestrogens. Previous clinical studies revealed that the use of White kwao krua as a hormone replacement therapy has beneficial effects on the lipid profile of menopause women. In this present study, we utilized the hypercholesterolemia rabbit model to demonstrate the effect of White kwao krua on the daily intake of high-fat diet. EXPERIMENTAL PROCEDURE: We induced hypercholesterolemia in rabbits by feeding with high-fat diet (1% cholesterol-containing diet). The animals were maintained 12 weeks for the experimentation. The White kwao krua supplement was administered 100 mg/kg/day, and the effects were monitored comparing with Statins and turmeric. Blood was collected periodically to monitor the plasma cholesterol level and the oxidative susceptibility of isolated LDL-cholesterol. At the end of the experiment, the aorta was collected from the animal and performed endothelial-dependent relaxation and endothelial-independent relaxation assays. The relative ratio of intima to media layer was microscopically evaluated from hematoxylin/eosin-stained tissues. RESULTS AND CONCLUSION: We showed that the White kwao krua supplement reduced LDL-cholesterol about 40% compared with high-fat diet consumption alone. Administration of White kwao krua had significantly prolonged the susceptibility of LDL-cholesterol to oxidation. Besides, it led to the improvement of vascular function by recovering endothelium-dependent relaxation and alleviating vascular structure impairment induced by high-fat dietary intake. Together, we suggest that White kwao krua should be used as a dietary supplement to reduce the atherogenesis in high-fat dietary consumption. SECTION: Dietary therapy/nutrients supplements. TAXONOMY: Inflammation, Disease.

Protective effects of γ-mangostin on 6-OHDA-induced toxicity in SH-SY5Y cells.

γ-Mangostin is a xanthone with hydroxyl groups that confer the substance-free radical scavenging effects. As opposed to the other more extensively studied mangostins, scarce research has been conducted on neuroprotective effects of γ-mangostin on models of Parkinson's disease (PD). Therefore, this investigation aimed to elucidate its antioxidant and neuroprotective effects on 6-OHDA-induced toxicity in SH-SY5Y cells. 6-OHDA treatment, an inducer of PD pathology in vitro studies, decreased cell viability and increased the level of intracellular ROS production. Furthermore, the substance-induced the expression of phosphorylated p38 MAPK, negatively affected the Bax/Bcl-2 ratio and increased caspase-3 activity; all of which were factors that are associated with apoptosis. Pretreatment of cells with γ-mangostin at concentrations of 0.5, 1, and 2.5µM markedly increased cell survival and reduced the level of intracellular ROS formation as shown by DPPH radical scavenging activity of the compound. Furthermore, a significant suppression of p-p38, improved Bax/Bcl-2 ratio expression, and reduced caspase-3 activity was exhibited in the cells after γ-mangostin pretreatment. The reduction of apoptosis was further supported by the reduction of pyknotic nuclei indicated by Hoescht 33342 staining. These findings indicate that γ-mangostin could attenuate 6-OHDA-induced neuronal cell death and that the protective effect of γ-mangostin is associated with its antioxidative potential and through the modulation of the apoptotic signalling pathway. Therefore, γ-mangostin may be an effective xanthone among other mangostins for preventing neurodegeneration in PD caused by oxidative stress.

Wnt signaling inhibitor FH535 selectively inhibits cell proliferation and potentiates imatinib-induced apoptosis in myeloid leukemia cell lines.

Wnt signaling pathway plays a major role in leukemogenesis of myeloid leukemia. Aberrancy in its regulation results in hyperactivity of the pathway contributing to leukemia propagation and maintenance. To investigate effects of Wnt pathway inhibition in leukemia, we used human leukemia cell lines (i.e., K562, HL60, THP1, and Jurkat) and several Wnt inhibitors, including XAV939, IWP2 and FH535. Our results showed that leukemia cell lines (>95 % cells) had increased endogenous levels of ß-catenin as compared to mononuclear cells from healthy donors (0 %). Among the tested inhibitors, FH535 demonstrated a markedly suppressive effect (IC50 = 358 nM) on mRNA levels of ß-catenin target genes (LEF1, CCND1, and cMYC). In addition, FH535 significantly potentiated imatinib-induced apoptosis. Evaluation of erythrocyte and megakaryocyte lineage using flow cytometry demonstrated that the potentiation mechanism is independent of the developmental stage, and is more likely due to crosstalk between other pathways and ß-catenin. FH535 also displayed antiproliferative properties in other cell lines used in this study. In summary, FH535 showed significantly high antiproliferative effects at submicromolar dosages, and additionally enhanced imatinib-induced apoptosis in human leukemia cell lines. Our results highlight its potential antileukemic promise when used in conjunction with other conventional therapeutic regimens.

Thermal Steam Aerosolization Protects Against Lipopolysaccharide-Induced Sepsis in Rats.

Background: Sepsis is a severe systemic inflammatory state in response to infection. The induction of heat shock protein 70 (HSP70) by heat stress has been reported to protect against lethal effects of sepsis. In clinical situation, inhalation of thermal water has been used empirically in the treatment of chronic diseases of respiratory tract. Thus, thermal steam aerosolization may have beneficial effects on sepsis via HPS70 induction. Objective: The present study tested the hypothesis that thermal steam aerosolization could protect against lipopolysaccharideinduced sepsis in rats. Material and Method: Male Spraque-Dawley rats were subjected to steam aerosolization at 40°C for 1 hour before intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS). Blood pressure, heart rate and core temperature were recorded in two hours interval. At the end of the study period, vascular response to vasoconstrictor of isolated aortic rings ex vivo was assessed. Serum proinflammatory cytokines, IL-1beta and IL-6, were analyzed using ELISA technique. Plasma nitric oxide was determined using nitrate/nitrite fluorometric assay Kit. HSP70 expression, heat shock factor-1 (HSF-1) mRNA level and nuclear factor-kappa B (NF-kappa B) activity in the lungs of rats were investigated using western blot analysis, real-time quantitative PCR and transcription factor kits for NF-kappa B p65, respectively. Results: Thermal steam aerosolization treatment prevented the fall in systolic, diastolic, mean arterial blood pressures induced by LPS and restored the vascular response to adrenaline. LPS significantly increased plasma nitrate/nitrite concentration, serum IL-1 beta and IL-6 levels, and NF-kappa B activity in rat lung lysate which were reduced by thermal steam aerosolization. Thermal steam aerosolization induced both HSP70 and HSF-1 mRNA expression. Conclusion: The present study suggests that thermal steam aerosolization can delay the stage of shock in LPS-induced septic rats. It shows a beneficial therapeutic effect and may be applied to the clinical approach for septic shock patient.

Caffeine potentiates methamphetamine-induced toxicity both in vitro and in vivo.

Ya-Ba, a combination of the two potent psychostimulants methamphetamine (METH) and caffeine (CAF), is commonly used by drug abusers in Thailand and neighboring countries. While the neurotoxic effects of METH are well documented, the toxicity of this combination is mostly unknown. This study aimed to elucidate the effects of this particular drug combination using both in vitro and in vivo models. We found that combined treatment of METH and CAF at individually non-toxic concentrations significantly decreased viability of human neuroblastoma SK-N-SH cells. The reduction in cell survival was accompanied by an increase in reactive oxygen species (ROS) formation and the Bax/Bcl-2 ratio. In vivo data showed that combined administration of METH and CAF increased the mortality rate of rats, with an increase in the level of thiobarbituric acid reactive substances (TBARS), the indicator of oxidative stress, in striatal tissues. The results indicate that caffeine potentiates the toxic effects of methamphetamine, possibly via a mechanism involving an increase in dopamine release and excess ROS generation.

Curcumin I protects the dopaminergic cell line SH-SY5Y from 6-hydroxydopamine-induced neurotoxicity through attenuation of p53-mediated apoptosis.

Oxidative stress (OS) plays a pivotal role in the pathogenesis of Parkinson's disease (PD). 6-Hydroxydopamine (6-OHDA) is a neurotoxin used to induce oxidative cell death of dopaminergic neurons in experimental models of PD. Curcumin I, or diferuloylmethane is a pure compound isolated from Curcuma longa Linn. that has been reported to have neuroprotective properties. The precise mechanism, however, remains unclear. This study aims to elucidate the mechanisms by which curcumin I exerts its effects, using 6-OHDA-induced neurotoxicity in the human dopaminergic cell line SH-SY5Y. In our experiments, pretreatment with curcumin I improved cell viability, and significantly reduced reactive oxygen species (ROS). Further investigations revealed a reduction of p53 phosphorylation and decrease of the Bax/Bcl-2 ratio, as measured by mRNA expression and protein level. Taken together, these findings indicate that curcumin I protects dopaminergic neurons from 6-OHDA-induced toxicity via the reduction of ROS production, and subsequent attenuation of p53 phosphorylation and reduction of the Bax/Bcl-2 ratio.

The in vitro and ex vivo antioxidant properties, hypolipidaemic and antiatherosclerotic activities of water extract of Moringa oleifera Lam. leaves.

UNLABELLED: Moringa oleifera is used in Thai traditional medicine as cardiotonic. Recent studies demonstrated its hypocholesterolaemic effect. However, to be clinically useful, more scientific data are needed. AIM OF THE STUDY: We investigated the antioxidant, hypolipidaemic and antiatherosclerotic activities of Moringa oleifera leaf extract. MATERIALS AND METHODS: Scavenging activity of the extract on 1,1-diphenyl-2-picrylhydrazyl radicals (DPPH), and the inhibitory effect on Cu(2+)-induced low-density lipoprotein (LDL) oxidation were determined in in vitro experiment. The effects of the extract on cholesterol levels, conjugated diene (CD) and thiobarbituric acid reactive substances (TBARS) and plaque formations in cholesterol-fed rabbits were investigated. RESULTS: We found that in scavenging DPPH radicals the extract and Trolox had IC(50) of 78.15+/-0.92 and 2.14+/-0.12microg/ml, respectively. The extract significantly (P<0.05) prolonged the lag-time of CD formation and inhibited TBARS formation in both in vitro and ex vivo experiments in a dose-dependent manner. In hypercholesterol-fed rabbits, at 12 weeks of treatment, it significantly (P<0.05) lowered the cholesterol levels and reduced the atherosclerotic plaque formation to about 50 and 86%, respectively. These effects were at degrees comparable to those of simvastatin. CONCLUSIONS: The results indicate that this plant possesses antioxidant, hypolipidaemic and antiatherosclerotic activities and has therapeutic potential for the prevention of cardiovascular diseases.