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.

Alpha-mangostin attenuation of hyperglycemia-induced ocular hypoperfusion and blood retinal barrier leakage in the early stage of type 2 diabetes rats.

The present study examined effects of alpha-mangostin (α-MG) supplementation on the retinal microvasculature, including ocular blood flow (OBF) and blood-retinal barrier (BRB) permeability in a type 2 diabetic animal model. Male Sprague-Dawley rats were divided into four groups: normal control and diabetes with or without α-MG supplementation. Alpha-mangostin (200 mg/Kg/day) was administered by gavage feeding for 8 weeks. The effects of α-MG on biochemical and physiological parameters including mean arterial pressure (MAP), OBF, and BRB leakage were investigated. Additionally, levels of retinal malondialdehyde (MDA), advance glycation end products (AGEs), receptor of advance glycation end products (RAGE), tumour necrosis factor alpha (TNF-α), and vascular endothelial growth factor (VEGF) were evaluated. The elevated blood glucose, HbA1c, cholesterol, triglyceride, serum insulin, and HOMA-IR were observed in DM2 rats. Moreover, DM2 rats had significantly decreased OBF but statistically increased MAP and leakage of the BRB. The α-MG-treated DM2 rats showed significantly lower levels of retinal MDA, AGEs, RAGE, TNF-α, and VEGF than the untreated group. Interestingly, α-MG supplementation significantly increased OBF while it decreased MAP and leakage of BRB. In conclusion, α-MG supplementation could restore OBF and improve the BRB integrity, indicating its properties closely associated with antihyperglycemic, antioxidant, anti-inflammatory, and antiglycation activities.

Effects of Long-Term Alpha-mangostin Supplementation on Hyperglycemia and Insulin Resistance in Type 2 Diabetic Rats Induced by High Fat Diet and Low Dose Streptozotocin.

OBJECTIVE: The present study investigated the effects of long-term supplementation of alpha-mangostin (MG; a xanthone isolated from mangosteen fruit) on hyperglycemia, and insulin resistance in type 2 diabetic rats. MATERIAL AND METHOD: Type 2 diabetes (DM2) was induced in male Sprague-Dawley rats by feeding high fat diet for three weeks followed by an IP injection of low dose streptozotocin. The rats were divided into four groups: control and diabetes without or with alpha-MG supplementation (CON, DM2, CON-MG and DM2-MG group, respectively). Alpha-MG was administered by gavage feeding in the amount of 200 mg/kg BW/day for 8 or 40 weeks. Fasting blood glucose, plasma HbA1c, cholesterol, and triglyceride were determined in all groups of rats. Serum insulin, calculated HOMA-IR and Oral glucose tolerance test were also carried out. RESULTS: The results showed that both 8 and 40 weeks DM2 groups had a significant increase in fasting blood glucose, HbA1c, plasma cholesterol and triglyceride compared with their aged-match control groups. Furthermore, the serum insulin and HOMA-IR were significantly elevated in 8 weeks DM2 whereas these two parameters were significantly decreased in 40 weeks DM2 group compared with their aged-match CON groups (p < 0.001). The OGTT showed impaired glucose tolerance in DM2 groups. Interestingly, alpha-MG supplemented DM2-MG group had significantly decreased levels of fasting blood glucose, HbA1c, plasma cholesterol, triglyceride when compared with the untreated DM2 groups. Supplementation of alpha-MG for 40 weeks in DM2-MG group showed significantly increase serum insulin levels compared with that of DM2 group (p < 0.001). Moreover alpha-MG supplemented DM-MG group demonstrated a better glucose tolerance pattern which was different from that of DM2 group at both 8 weeks and 40 weeks experimental periods. CONCLUSION: Long-term alpha-mangostin supplementation has anti-hyperglycemic, anti-hyperlipidemic effects and increase insulin sensitivity by improving beta-cell functions in type 2 diabetes mellitus.

The Pilot Study of the Effect of Meditation to the Serum Brain-Derived Neurotrophic Factor (BDNF) of Medical Students, Srinakharinvirot University.

BACKGROUND: Mindfulness meditation is a method to decrease stress and increase memory. So, mindfulness meditation should increase serum brain-derived neurotrophic factor (BDNF). OBJECTIVE: To study the effect of mindfulness meditation on the serum BDNF of medical students. MATERIAL AND METHOD: The study group consisted of 30 male and female second-year medical students that volunteered to participate in the study, aged 19.1 ± 0.55 year olds (range 18-20) from Srinakharinwirot University. Their blood was drawn to measure BDNF before and after a four-day mindfulness meditation programme. The comparison of serum BDNF levels before and after meditation were analysed by paired t-test. RESULTS: The subjects were 66.77%female and 33.33% male. The average serum BDNF level before the meditation was 17.67 ng/ml (SD 3.58). After meditation, there was a decrease in serum BDNF to 17.34 ng/ml, which was however not statistically significant (SD 4.04, p > 0.05). CONCLUSION: The levels of blood BDNF decreases slightly after practising meditation. We plan to investigate the reason in the future.

Detection of Pathogenic Leptospires by Loop-Mediated Isothermal Amplification Targeting LipL32 Gene.

BACKGROUND: Leptospirosis is a worldwide re-emerging infectious disease caused by pathogenic leptospires including Leptospira interrogans. OBJECTIVE: In the present study, a loop-mediated isothermal amplification (LAMP) was developed to detect L. interrogans using lipL32 as a gene target. MATERIAL AND METHOD: Four specific primers were designed based on the conserved region of lipL32 gene of various serovars ofpathogenic leptospires. LAMP reaction was performed at 65 °C for 1 hour The LAMP products were detected by agarose gel electrophoresis andfluorescence dye. RESULTS: The lipL32 LAMP assay showed highly specificity to the reference stains of L. interrogans serovar Autumnalis, Bataviae, Javanica, Pyrogenes, Icterohaemorrhagiae, and Saigon. No product was produced from non-pathogenic leptospire (L. biflexa), human, or Escherichia coli. The lower limit of detection analyzed by agarose gel electrophoresis andfluorescence dye visualization was 0.02 pg/µl which equivalent to 4 genomic equivalents/reaction. Moreover the clinical strain of leptospires including pathogenic and intermediate group of L. interrogans were detected by lipL32 LAMP CONCLUSION: The developed lipL32 LAMP is high specificity and sensitivity that can be applied to detect pathogenic leptospires in clinical samples.

Effect of Inhaling Bergamot Oil on Depression-Related Behaviors in Chronic Stressed Rats.

BACKGROUND: Bergamot essential oil (BEO) possesses sedation and anxiolytic properties similar to diazepam. After long period of exposure to stressors, including restrained stress, depressive-like behavior can be produced. BEO has been suggested to reduce depression. However, there is no scientific evidence supporting this property. OBJECTIVE: To investigate the effect of BEO in chronic stressed rats on: 1) behavior related depressive disorder, 2) hypothalamic pituitary adrenal (HPA) axis response, and iii) brain-derived neurotrophic factor (BDNF) protein levels in hippocampus. MATERIAL AND METHOD: Male Wistar rats, weighing 200 to 250 g, were induced chronic restrained stress 15 minutes dailyfor two weeks. For the next two weeks, these rats were divided intofour groups, control-i.p., fluoxetine-i.p., control-inhale, and BEO-inhale. Fluoxetine (10 mg/kg i.p.) or saline was intraperitoneally administered daily while 2.5% BEO or saline was inhaled daily. At the end of the treatment, rats were assessed for depressive-like behavior using the forced swimming test (FST). After the behavioral test, the animals were immediately decapitated and trunk blood samples were collected for the measurement ofcorticosterone and adrenocorticotropic hormone (ACTH) levels and hippocampus was dissected and stored in afreezer at -80 °C until assay for BDNF protein. RESULTS: BEO andfluoxetine significantly decreased the immobility time in the FST (p < 0.05). Fluoxetine tended to decrease serum corticosterone and significantly (p < 0.05) decreased serum ACTH while BEO had no effect on these two stress hormones. For BDNF protein determination, neither BEO norfluoxetine had any effect on BDNF protein levels in hippocampus compared to their controls. CONCLUSION: The inhalation ofBEO decrease behavior related depressive disorder similar tofluoxetine but has no effect on HPA axis response and BDNF protein levels in chronic restrained stress.

Antileptospiral activity of xanthones from Garcinia mangostana and synergy of gamma-mangostin with penicillin G.

BACKGROUND: Leptospirosis, one of the most widespread zoonotic infectious diseases worldwide, is caused by spirochetes bacteria of the genus Leptospira. The present study examined inhibitory activity of purified xanthones and crude extracts from Garcinia mangostana against both non-pathogenic and pathogenic leptospira. Synergy between γ-mangostin and penicillin G against leptospires was also determined. METHODS: Minimal inhibitory concentrations (MIC) of crude extracts and purified xanthones from G. mangostana and penicillin G for a non-pathogenic (L. biflexa serovar Patoc) and pathogenic (L. interrogans serovar Bataviae, Autumnalis, Javanica and Saigon) leptospires were determined by using broth microdilution method and alamar blue. The synergy was evaluated by calculating the fractional inhibitory concentration (FIC) index. RESULTS: The results of broth microdilution test demonstrated that the crude extract and purified xanthones from mangosteen possessed antileptospiral activities. The crude extracts were active against all five serovars of test leptospira with MICs ranging from 200 to ≥ 800 µg/ml. Among the crude extracts and purified xanthones, garcinone C was the most active compound against both of pathogenic (MIC =100 µg/ml) and non-pathogenic leptospira (MIC = 200 µg/ml). However, these MIC values were higher than those of traditional antibiotics. Combinations of γ-mangostin with penicillin G generated synergistic effect against L. interrogans serovars Bataviae, Autumnalis and Javanica (FIC = 0.52, 0.50, and 0.04, respectively) and no interaction against L. biflexa serovar Patoc (FIC =0.75). However, antagonistic activity (FIC = 4.03) was observed in L. interrogans serovar Saigon. CONCLUSIONS: Crude extracts and purified xanthones from fruit pericarp of G. mangostana with significant antibacterial activity may be used to control leptospirosis. The combination of xanthone with antibiotic enhances the antileptospiral efficacy.

Effects of mindfulness meditation on serum cortisol of medical students.

BACKGROUND: Mindfulness meditation is a method to relax the mind that decreases stress, which otherwise would increase serum cortisol. So, mindfulness meditation should decrease serum cortisol. OBJECTIVE: To study the effect of mindfulness meditation on mental health by using Thai GHQ28 questionaire and study the effect of mindfulness meditation on stress by using serum cortisol. MATERIAL AND METHOD: Volunteer subjects were 30 second year medical students, aged 19.1 +/- 0.55 year olds (range 18-20) from Srinakharinwirot University. They were screened by Thai GHQ28 and blood was drawn to measure cortisol at 8:00 am before and after a four-day mindfulness meditation programme. The comparison of Thai GHQ28 scores and serum cortisol levels before and after meditation were analysed by paired t-test. RESULTS: The subjects were 66.77% female and 33.33% male. The average score of Thai GHQ28 before and after the mindfulness meditation was 1.50 (SD 2.53) and 0.77 (SD 2.08) respectively. The average serum cortisol levels before mindfulness meditation was 381.93 nmol/L (SD 97.74) becoming significantly lower after mindfulness meditation 306.38 nmol/L (SD 90.95). The difference was statistically significant in cortisol level, but not statistically significant in Thai GHQ28. CONCLUSION: Mindfulness meditation lowers the cortisol levels in the blood suggesting that it can lower stress and may decrease the risk of diseases that arise from stress such as psychiatric disorder, peptic ulcer and migraine. Then, mindfulness meditation should be used in combination with standard treatment.

Antimicrobial and antioxidant activities of traditional Thai herbal remedies for aphthous ulcers.

Four medicinal plants (Quercus infectoria, Kaempferia galanga, Coptis chinensis and Glycyrrhiza uralensis) as well as one traditional Thai treatment for aphthous ulcers based on these four plants were tested for antimicrobial activity. MIC values for a range of bacteria and Candida albicans were determined, with both type strains and clinical isolates being used. Antioxidant activity was determined using the ABTS radical scavenging assay. Among the four plants, Q. infectoria showed antimicrobial activity against Staphylococcus aureus with an MIC of 0.41 mg/mL, while C. chinensis showed antifungal activity against C. albicans with an MIC of 6.25 mg/mL. Activity was also shown against a range of other organisms including Salmonella typhi, Serratia marcescens, Vibrio cholerae, Vibrio parahaemolyticus, Pseudomonas aeruginosa and Enterococcus faecalis. The antimicrobial activity of the traditional aphthous ulcer preparation (a powder) was comparable to that for the individual plant extracts, however, incorporation of the powder into a gel formulation resulted in the loss of almost all activity. All extracts, with the exception of K. galanga, also showed good antioxidant activity. This study supports the traditional use of these plants and suggests that they may also be useful in the treatment of other infections.

Modified GC medium for identification of group B streptococci.

The modified GC medium (MGC) was developed for identification of beta-hemolytic group B streptococci. This medium was developed on the basis of enhancing-pigment production of group B streptococci. Three hundred and thirty isolates were tested including 180 isolates of beta-hemolytic group B streptococci, 102 isolates of beta-hemolytic non-group B streptococci, and 48 isolates of Enterococcus faecalis. All isolates of group B streptococci gave carotenoid pigment by this medium. On the other hand, all of non-group B streptococci and E. faecalis did not show pigment after 72 h incubation. The specificity and sensitivity of MGC was 100%. There were no false positive and false negative in this medium. The MGC may be the alternative of choice for the presumptive identification of group B streptococci.