Capillary electrophoresis using triple layer modified capillary facilitating salivary ion analyses: Application to search for potential stress markers induced by cold pressure test.

In this study, we introduce a novel approach for the analysis of salivary ions using capillary electrophoresis (CE) with a triple-layer coated capillary. The capillary is sequentially coated with cationic silylating reagents, poly(vinylsulfonate), and polybrene to form a custom designed surface that effectively inhibits adsorption of protein matrix on the capillary inner wall and allows for reproducible ion analysis. For the CE with capacitively coupled contactless conductivity detection, we used suitable background electrolytes (BGEs) for salivary ion analysis. Anions were separated using a mixture of 2-(N-morpholino)ethanesulfonic acid and l-arginine, and cations were separated using that with 18-crown-6. This setup enabled rapid separation, within 4 min, together with sensitive detection. We quantified nine common anions and five cations typically found in saliva samples using this CE method, both before and after a cold pressure test (CPT, a standard stress test). The CE system demonstrated consistent ion separation across 30 consecutive measurements without requiring capillary replacement. Notably, the salivary ion balance remained predominantly anion-rich, regardless of the CPT. Cold water exposure induced greater variation in the total anion concentration than in the total cation concentration. Further analysis using multiple regression analysis revealed strong relationships between nitrate and nitrite, formate and phosphate, and potassium and nitrate, before and after the CPT. Notably, potassium and nitrate ions exhibited variations in response to stress. These results provided a method for assessing salivary ion composition and insights into the potential of salivary ions as biomarkers for stress.

Acupuncture Relaxation, Vigilance Stage, and Autonomic Nervous System Function: A Comparative Study of Their Interrelationships.

Objective: During acupuncture stimulation, autonomic nervous system (ANS) function changes toward being parasympathetic-dominant, with a transient decrease in heart rate (HR). The aim of this research was to determine the relationships between cortical relaxation and vigilance as observed on background electroencephalograms (EEGs), HR, and ANS function during deep acupuncture. Materials and Methods: This comparative study was conducted at Teikyo Heisei University, in Toshima-ku, Tokyo, Japan, with 27 healthy male volunteers. After resting for 20 minutes, the men received manual acupuncture at LI 10 on the left forearm for 2 minutes at a depth of 15-20 mm at a 1-Hz frequency while undergoing concurrent EEG and electrocardiography (ECG) monitoring. Each subject described his level of comfort during acupuncture. HR; power ratios (normalized units [n.u.]) of EEG alpha, beta, delta, and theta waves; and HR variability (HRV) indices were calculated. Results: In the subgroup who experienced discomfort, delta and theta n.u. were decreased while alpha n.u. were increased, indicating increased vigilance and decreased relaxation. In the subgroup who experienced comfort, there were no significant changes. HRV indices suggested parasympathetic-dominant changes in both subgroups. Weak correlations were observed between a decrease of theta n.u. and sympathetic-dominant changes in HRV indices. Conclusions: Alterations in background EEG activities were not the primary factors changing ANS function to parasympathetic-dominant and decreasing HR, but these alterations related to a weak secondary factor changing ANS function. EEG activity by which cortical relaxation and vigilance were represented was the weak secondary factor changing ANS function during acupuncture; the primary factor might be supraspinal reflection.

Effects of a Standardized Oligomerized-Polyphenol from Litchi chinensis Fruit Extract and Mixed Plant Extract Supplementation on Peripheral Circulation and Cold Sensitivity.

Certain individuals tend to suffer from a cold sensation-particularly in the lower extremities-despite most people not suffering from the same sensation. In Japan, this phenomenon is called "hie-sho" and reduces quality of life for several people, particularly women. A previous study has shown that a standardized oligomerized-polyphenol from Litchi chinensis fruit extract (OPLFE) reportedly causes a significant increase in body surface temperature. The present study aimed to investigate whether supplementation with OPLFE affected peripheral circulation and cold sensitivity. This randomized, double-blind, placebo-controlled trial was performed including 25 participants (age, 45.0±10.4 y; 3 males and 22 females) who were assigned to consume OPLFE, mixed plant extract with OPLFE, or placebo capsules for 14 d. Participants were instructed to relax for 60 min in a temperature-controlled room prior to obtaining measurements. Changes in skin temperature and peripheral blood flow of the middle finger were assessed immediately before and 1, 5, 10, 20, and 30 min after immersion in cold water (10ºC). Participants' height, weight, skin temperature, and blood flow in peripheral tissue were measured; furthermore, their "hie-sho" was measured using the Visual Analog Scale (VAS). Skin temperature and blood flow in peripheral tissue increased in the OPLFE and mixed plant extract with OPLFE groups on day 14 compared with those on day 1. In addition, cold sensitivity in these two groups significantly improved between day 1 and day 14. These findings suggest that OPLFE improves "hie-sho" by increasing peripheral blood flow and skin temperature.

Electroacupuncture of the Ophthalmic Branch of the Trigeminal Nerve: Effects on Prefrontal Cortex Blood Flow.

Objective: The current authors observed enhanced cerebral blood flow (CBF) in the prefrontal cortex (PFC) in response to 100-Hz electroacupuncture (EA) stimulation of the ophthalmic branch of the trigeminal nerve. However, it is not yet clear if responsiveness to 100-Hz EA depends on stimulus intensity. This study examined the effects of stimulus strength on PFC CBF during 100-Hz EA of the ophthalmic branch of the trigeminal nerve. Materials and Methods: Twelve subjects underwent 3 acupuncture sessions: I, control, no stimulation; II, 0.1 mA EA; and III, 0.2 mA EA). Needles were inserted 1 cm lateral of the head median line; the anterior insertion point was on the front hairline and the posterior insertion point was ∼7 cm behind the hairline. Stimulation frequency was set to 100-Hz. PFC CBF was measured in terms of oxygenated, deoxygenated, and total hemoglobin (OxyHb, DeoxyHb, TotalHb, respectively), using 16-channel (Ch) near-infrared spectroscopy. Results: Stimulation of 0.2 mA was associated with significant elevation of OxyHb levels in the 0.1 mA condition in Chs 6, 10, and 12. Ch 2-6, 10, 12 signals were notably higher than in the control condition. Stimulation of 0.2 mA and 0.1 mA were associated with significant declines in DeoxyHb levels, compared to the control condition in Ch 4. Finally, 0.2 mA stimulation in Chs 12 and 13 was associated with significant elevation of TotalHb levels in the control condition. Conclusions: Using 0.2-mA stimulation, 100-Hz EA of the ophthalmic nerve enhances PFC CBF more strongly than 0.1-mA stimulation.

Effects of Cathode Direct-Current Electroacupuncture Stimulus of the Promixal Anterior Lower Limbs on Heart Rate and Lumbar Blood Flow.

Objective: There has been little research on direct-current acupuncture. This study was conducted to verify the effect of acupuncture current intensity and polarity on heart rate (HR) when applying direct-current electroacupuncture (DCEA) to the proximal anterior part of the lower limbs and to clarify the effect of cathodes (-) on lumbar blood flow. Materials and Methods: This study was conducted at Teikyo Heisei University, Chiba, Japan. The subjects were 144 healthy male volunteers (21.7 ± 0.6 years' old). These participants were divided into 1 control, 1 manual acupuncture (MA), and 3 DECEA groups (-25 µA, +75 µA, and -75 µA). Acupuncture was performed on the lower limbs (at ST 36), and HR and intertissue blood flow (ITBF) were observed as the main outcome measures. Results: For intensity, HR was significantly lower in the -75 µA group than in the control (P < 0.001), MA (P < 0.001), and -25 µA (P < 0.01) groups. For polarity, HR was significantly lower in the -75 µA group than in the +75 µA group (P = 0.0028). For blood flow, the volume of HbO2 in the L-4-L-5 region at 1 minute after stimulation was significantly higher in the -75 µA stimulus group than in the MA group (P < 0.05). There was no change in the T-7-T-8 region. Conclusions: A -75 µA stimulus to ST 36 reduced HR and increased ITBF.

Effects of Acupuncture Sensations on Transient Heart Rate Reduction and Autonomic Nervous System Function During Acupuncture Stimulation.

Objective: During acupuncture stimulation, heart rate (HR) transiently decreases and autonomic nervous system (ANS) function becomes parasympathetic-dominant. To clarify the effect of acupuncture sensations (pain, De Qi), the effects of deep acupuncture sensations on HR and ANS functions were determined. Materials and Methods: In this comparative study at Teikyo Heisei University, Tokyo, Japan, 40 healthy, male student volunteers rested for 20 minutes before undergoing manual acupuncture to the Shousanli (LI 10) acupoint on the left forearm for 2 minutes at a frequency of 1 Hz, with concurrent electrocardiography. Depth of stimulation was 15-20 mm. These subjects described their subjective acupuncture sensations. Calculations were performed, using HR variability analysis to find HR and low-frequency (LF) normalized units (nu), the ratio of LF components to total components (as an index of sympathetic nervous system function), high-frequency (HF)nu, the ratio of HF components to total components (as an index of parasympathetic nervous system function), and LF/HF (as an index of sympathetic and parasympathetic balance). Results: For the final analysis, data were available for 32 subjects. Compared to before acupuncture, HR decreased during acupuncture. HR decreased when no acupuncture sensations or when weak De Qi sensations were perceived, and remained unchanged when acupuncture sensations without De Qi or strong De Qi were perceived. LFnu decreased, HFnu increased, and LF/HF decreased, regardless of pain or De Qi. Conclusions: Acupuncture stimulation reduced HR even without De Qi sensations and caused ANS function to be parasympathetic-dominant, irrespective of the perception of acupuncture sensations.

Evaluation of Autonomic Nervous System Function Using Heart Rate Variability Analysis During Transient Heart Rate Reduction Caused by Acupuncture.

Objective: Human studies have demonstrated that heart rate (HR) decreases during acupuncture stimulation, and pharmacologic studies have shown that this autonomic nervous system (ANS) response is parasympathetic-dominant. It has become clear that significant changes occur in the ANS after acupuncture, based on HR variability (HRV). However, it is inconclusive, according to HRV analysis, if acupuncture induces a significant change in autonomic function during stimulation. The aim of this study was to investigate ANS function using HRV analysis during HR reduction induced by manual acupuncture stimulation to the muscles. Materials and Methods: In this study, electrocardiograms of 25 adult men were analyzed. After resting for 20 minutes, participants underwent 15-20-mm deep acupuncture stimulation at the Shousanli (LI 10) point at 1 Hz for 2 minutes. Instantaneous HR was recorded. The index of parasympathetic nervous activity high-frequency (HF) normalized units (HFnu) and the ratio of sympathovagal balance (low frequency [LF]/HF) were calculated by HRV analysis. Results: HR during acupuncture was significantly lower, compared to HR both before and after acupuncture. HFnu during acupuncture were significantly higher, compared to HFnu both before and after acupuncture. The LF/HF ratio during acupuncture was significantly lower, compared to the ratio before acupuncture, and remained low after acupuncture, compared to before acupuncture. Conclusions: Acupuncture stimulation to the muscle can effectively reduce HR, increase HFnu, and decrease LF/HF that depends on autonomic regulation of both sympathovagal balances.

Effects of AHCC® on Immune and Stress Responses in Healthy Individuals.

AHCC® is a functional food from the basidiomycete Lentinula edodes. We evaluated the effects of AHCC® on subjects under different kinds of stress and at rest. Physical stress was imposed using an active standing test, known as Schellong's test. Sympathetic nervous activity in the standing position was significantly greater in AHCC®-treated subjects than in a placebo group. In contrast, AHCC® significantly increased parasympathetic nervous activity at rest. Under mental stress, AHCC® increased sympathetic nervous activity, with no difference in the parasympathetic nervous system. In subjects with chronic mental stress, self-reported "initiation and maintenance of sleep" was significantly greater in the AHCC®-intake period than in the placebo intake period, and natural killer cell activity also increased after AHCC® intake, suggesting a possible mechanism of action of AHCC®. Our findings indicate that AHCC® is potentially effective in stress management and may be useful in the treatment of depression.

Effects of electroacupuncture to the trigeminal nerve area on the autonomic nervous system and cerebral blood flow in the prefrontal cortex.

BACKGROUND: The autonomic nervous system and trigeminal nerve are involved in adjusting flow through diverging cerebral arteries in the prefrontal cortex. The purpose of this study was to examine the effect of 100 Hz electroacupuncture (EA) to the trigeminal nerve area on cerebral blood flow and autonomic nervous system function. METHOD: This was a randomised crossover study of 16 healthy volunteers who were assigned to an EA or control group. Stimulation (in the EA group) was performed five times, each after 1 min of rest. Needles were inserted at the inner edge of the eyebrows and 1 cm from the front hairline midpoint. We used high-frequency (HF) and low-frequency (LF) components of heart rate (HR) variability to assess autonomic nervous system function. HF and LF/HF ratio were taken as indicators of parasympathetic and sympathetic nervous system activity, respectively. We measured cerebral blood flow using a two-channel near-infrared spectroscope. RESULTS: In the EA group, HR significantly decreased (p=0.004) and HF significantly increased (p=0.006) relative to baseline. By contrast, there were no significant changes in HR or HF within the control group (p>0.05). Accordingly, HR tended to be lower (p=0.087) and HF greater (p=0.071) in the EA group versus the control group. There were no significant differences in LF/HF ratio within/between groups. Compared with the control group, cerebral blood flow was significantly greater in the left (p=0.048) and right (p=0.016) prefrontal cortex in the EA group. CONCLUSIONS: Delivery of 100 Hz EA to the trigeminal nerve area reduces HR and increases parasympathetic nervous activity and cerebral blood flow.

Effect of Enzyme-Treated Asparagus Extract (ETAS) on Psychological Stress in Healthy Individuals.

The aim of this study was to examine the effectiveness of Enzyme-Treated Asparagus Extract (ETAS) on improving stress response. A randomized, double-blind, placebo-controlled cross-over trial was undertaken in healthy volunteers. ETAS (150 mg/d) or a placebo was consumed for 28 d, with a washout period. Psychological parameters were examined using a self-report scale questionnaire and psychological stress was applied using the Uchida-Kraepelin (U-K) test. During the stress load, autonomic nervous function was analyzed. After the stress load, a profile of mood states (POMS) psychological rating was performed, and serum cortisol, plasma catecholamine, salivary secretory immunoglobulin A (sIgA), and salivary cortisol were analyzed. ETAS intake improved the self-reported rating for the items "Feel tired," "Hard to get up," and "Feel heavy" in the psychological questionnaire; ameliorated the self-reported rating for the items "Depression-Dejection" and "Fatigue" in the POMS questionnaire; and increased salivary sIgA levels after the U-K test. In contrast, serum and salivary cortisol levels, and plasma catecholamine did not change. During the U-K test, ETAS significantly upregulated the sympathetic nerve activity. Furthermore, ETAS intake significantly increased the number of answers and the number of correct answers in the U-K test, suggesting that it might improve office work performance with swiftness and accuracy under stressful conditions. In conclusion, ETAS supplementation reduced feelings of dysphoria and fatigue, ameliorated quality of sleep, and enhanced stress-load performance as well as promoted stress response by increasing salivary sIgA levels. These data suggest ETAS intake may exert beneficial effects, resulting from well-controlled stress management, in healthy individuals.

Effects of a Gentle, Self-Administered Stimulation of Perineal Skin for Nocturia in Elderly Women: A Randomized, Placebo-Controlled, Double-Blind Crossover Trial.

BACKGROUND: Somatic afferent nerve stimuli are used for treating an overactive bladder (OAB), a major cause of nocturia in the elderly. Clinical evidence for this treatment is insufficient because of the lack of appropriate control stimuli. Recent studies on anesthetized animals show that gentle stimuli applied to perineal skin with a roller could inhibit micturition contractions depending on the roller's surface material. We examined the efficacy of gentle skin stimuli for treating nocturia. METHODS: The study was a cross-over, placebo-controlled, double-blind randomized clinical study using two rollers with different effects on micturition contractions. Participants were elderly women (79-89 years) with nocturia. Active (soft elastomer roller) or placebo (hard polystyrene roller) stimuli were applied to perineal skin by participants for 1 min at bedtime. A 3-day baseline assessment period was followed by 3-day stimulation and 4-day resting periods, after which the participants were subjected to other stimuli for another 3 days. The primary outcome was change in the frequency of nighttime urination, for which charts were maintained during each 3-day period. RESULTS: Twenty-four participants were randomized, of which 22 completed all study protocols. One participant discontinued treatment because of an adverse event (abdominal discomfort). In participants with OAB (n = 9), change from baseline in the mean frequency of urination per night during the active stimuli period (mean ± standard deviation, -0.74 ± 0.7 times) was significantly greater than that during placebo stimuli periods (-0.15 ± 0.8 times [p < 0.05]). In contrast, this difference was not observed in participants without OAB (n = 13). CONCLUSIONS: These results suggest that gentle perineal stimulation with an elastomer roller is effective for treating OAB-associated nocturia in elderly women. Here the limitation was a study period too short to assess changes in the quality of sleep and life. TRIAL REGISTRATION: UMIN Clinical Trial Registry (CTR) UMIN000015809.

Effect of Forefoot Strike on Lower Extremity Muscle Activity and Knee Joint Angle During Cutting in Female Team Handball Players.

BACKGROUND: The purpose of this study is to examine the effects of different strike forms, during cutting, on knee joint angle and lower limb muscle activity. METHODS: Surface electromyography was used to measure muscle activity in individuals performing cutting manoeuvres involving either rearfoot strikes (RFS) or forefoot strikes (FFS). Three-dimensional motion analysis was used to calculate changes in knee angles, during cutting, and to determine the relationship between muscle activity and knee joint angle. Force plates were synchronized with electromyography measurements to compare muscle activity immediately before and after foot strike. RESULTS: The valgus angle tends to be smaller during FFS cutting than during RFS cutting. Just prior to ground contact, biceps femoris, semitendinosus, and lateral head of the gastrocnemius muscle activities were significantly greater during FFS cutting than during RFS cutting; tibialis anterior muscle activity was greater during RFS cutting. Immediately after ground contact, biceps femoris and lateral head of the gastrocnemius muscle activities were significantly greater during FFS cutting than during RFS cutting; tibialis anterior muscle activity was significantly lower during FFS cutting. CONCLUSIONS: The results of the present study suggest that the hamstrings demonstrate greater activity, immediately after foot strike, during FFS cutting than during RFS cutting. Thus, FFS cutting may involve a lower risk of anterior cruciate ligament injury than does RFS cutting.

Pathological analysis of the Candida albicans-infected tongue tissues of a murine oral candidiasis model in the early infection stage.

OBJECTIVE: The early pathological process of Candida infection and immunological responses in tongues of the mice with experimental oral candidiasis was analysed. METHODS: CD-1 mice, pretreated by prednisolone were orally inoculated with Candida albicans. Symptoms were monitored by measuring the area of white tongue coating and number of viable Candida cells in oral cavity. The histopathological analysis was carried by PAS-stain and immunofluorescent staining. IL-4, IL-12p70, IFN-γ, TNF-α in recovered from the homogenates of the tongues were measured by ELISA. RESULTS: The fungus invaded the tongue surface of the mice and white patches developed within 24h after inoculation. Histopathological examination indicated the presence of local acute inflammation in superficial tissues of tongues covered by mycelium of C. albicans. Pathological exacerbation was observed from 24 to 48 h after the inoculation and from then the symptoms of oral candidiasis appeared to move into the recovery phase. Inflammatory cells mainly consisting of neutrophils was accumulated and located under the lesions covered by Candida-hyphae. An increase in IL-12p70 and IFN-γ in tongue homogenates was observed at 48 h after inoculation. CONCLUSIONS: The worst condition in the pathological process in experimental oral candidiasis was found 48 h after C. albicans inoculation. When the surface of the Candida-inoculated tongues was covered with Candida-hyphae, a dense accumulation of neutrophils was observed under the lesions and homogenates of the tongues contained increased levels of IL-12p70 and IFN-γ. These suggested that local pathological condition of Candida-infected tongues may be affected by neutrophils accumulation and increased levels of some cytokines.

[Suppression of experimental inflammation by anti-fungal agent liranaftate in mice].

To evaluate the anti-inflammatory activity of the thiocarbamate antifungal agent liranaftate, the edema and the neutrophil accumulation detected by the activity of neutrophil marker enzyme, myeloperoxidase (MPO), were examined following application of liranaftate to mouse ears with inflammation induced by phorbol 12-myristate 13-acetate (PMA). Topical 20 microl administration of liranaftate in a dose-range between 1-4% suppressed the increase in ear thickness 6 hr after PMA application dose-dependently. Similarly, it decreased the weight increase of an ear section after 24 hr dose-dependently. More than 1% of liranaftate also suppressed augmentation of MPO activity of the ear section. This and histological observation indicate that liranaftate treatment suppressed neutrophil accumulation in PMA-applied ear lesion. From these results, we discussed that liranaftate might suppress inflammatory symptoms caused by trychophytosis in a clinical condition.

Enhanced gene replacements in Ku80 disruption mutants of the dermatophyte, Trichophyton mentagrophytes.

The frequency of targeted gene disruption via homologous recombination is low in the clinically important dermatophyte, Trichophyton mentagrophytes. The Ku genes, Ku70 and Ku80, encode key components of the nonhomologous end-joining pathway involved in DNA double-strand break repair. Their deletion increases the homologous recombination frequency, facilitating targeted gene disruption. To improve the homologous recombination frequency in T. mentagrophytes, the Ku80 ortholog was inactivated. The nucleotide sequence of the Ku80 locus containing a 2788-bp ORF encoding a predicted product of 728 amino acids was identified, and designated as TmKu80. The predicted TmKu80 product showed a high degree of amino acid sequence similarity to known fungal Ku80 proteins. Ku80 disruption mutant strains of T. mentagrophytes were constructed by Agrobacterium tumefaciens-mediated genetic transformation. The average homologous recombination frequency was 73.3 +/- 25.2% for the areA/nit-2-like nitrogen regulatory gene (tnr) in Ku80(-) mutants, about 33-fold higher than that in wild-type controls. A high frequency (c. 67%) was also obtained for the Tri m4 gene encoding a putative serine protease. Ku80(-) mutant strains will be useful for large-scale reverse genetics studies of dermatophytes, including T. mentagrophytes, providing valuable information on the basic mechanisms of host invasion.

Suppression of anti-Candida activity of macrophages by a quorum-sensing molecule, farnesol, through induction of oxidative stress.

Farnesol is well known as a quorum-sensing molecule of Candida albicans. To assess the pathological function of farnesol, its effects on macrophage viability and functions including growth inhibitory activities against C. albicans were examined in vitro. Murine macrophages, when cultured in the presence of 56-112 microM of farnesol for 1-2 hr, decreased their activity inhibiting the mycelial growth of C. albicans and lost their viability. This suppression of macrophage function by farnesol was neutralized by the coexistence of the anti-oxidants probucol and trolox. Macrophages cultured in the presence of farnesol for 2 hr displayed morphological change of nuclei and DNA fragmentation, which suggested apoptosis of the cells. Intracellular production of ROS in the farnesol-treated macrophages was shown by fluorescence of DCFH-DA and increase of peroxidized materials. These effects of farnesol were blocked by probucol or trolox. These results indicate that farnesol lowered viability of the murine macrophages and suppressed their anti-Candida activity, perhaps through induction of ROS.

Pharmaco-physio-psychologic effect of Ayurvedic oil-dripping treatment using an essential oil from Lavendula angustifolia.

Ayurvedic oil-dripping treatment, Shirodhara, involves the use of medicated herbal sesame oils. In our previous reports, we found that Shirodhara with plain sesame oil induced anxiolysis and an altered state of consciousness (ASC) in healthy subjects. We studied the pharmaco-physio-psychologic effect of Shirodhara with medicated sesame oil including an essential oil from Lavendula angustifolia (lavender) in the present study. Sixteen (16) healthy females (38 +/- 8 years old) were assigned at random to three treatments applied by a robotic oil-dripping system: plain sesame oil (plain Shirodhara), medicated sesame oil with a 0.3 volume % of lavender essential oil (lavender Shirodhara), or the control supine position. Psychophysiologic parameters including the heart rate, skin temperature of the dorsum of hands and feet, as well as anxiety and ASC were monitored, and the rates of change of these items were calculated to assess the psychophysiologic changes brought about by Shirodhara. Lavender Shirodhara showed potent anxiolytic and ASC-inducing or promoting effects, and induced the largest increase in foot skin temperature. The correlation between anxiolysis and ASC, as well as the correlation between these psychologic effects and the elevated foot skin temperature were larger in the lavender Shirodhara than in the other two conditions. It was speculated that the psycho-physiologic effects of lavender Shirodhara would be brought about by three mechanisms: (1) the well-known relaxing action of essential oils from L. angustifolia mediated by olfactory nerves, (2) the pharmacologic action of substances absorbed through the skin or mucosa in the sesame oil or lavender essential oil, and (3) the physiologic effect of sesame oil dripped on the forehead induced by the somato-autonomic reflex through thermosensors or pressure sensors in the skin or hair follicles via the trigeminal cranial nerve. The complicated pharmaco-physio-psychologic action of Ayurvedic oil treatment may provide a useful model for future pharmaco-physio-psychotherapy.

Protective effects of farnesol against oral candidiasis in mice.

Farnesol is known as a quorum-sensing molecule for Candida albicans and is recognized to play pathogenic roles in Candida infection. To assess the possible role of farnesol in mucosal C. albicans infection, the effects of farnesol treatment against experimental oral candidiasis in mice were examined. Prednisolone-pretreated ICR mice were orally infected with C. albicans and 3, 24 and 30 hr later the animals were orally given farnesol. Forty-eight hr later they were killed for observation. Farnesol treatment in a dose ranging between 1.125 and 9 micromol/mouse showed a protective effect against oral candidiasis in a dose-dependent manner, at least as estimated by symptom scores of tongues. At 9 micromol/mouse it decreased bodyweight loss. Histological studies of 2.25 micromol/mouse farnesol-treated animals indicated that farnesol suppressed mycelial growth of C. albicans on the surface of tongues, but microbiological study did not prevent the change of CFU of C. albicans cells not only on tongues but also in feces, kidneys and livers. These results suggest that farnesol has very characteristic roles in protection against mucosal candidiasis.

Protective activity of geranium oil and its component, geraniol, in combination with vaginal washing against vaginal candidiasis in mice.

In order to evaluate an effective administration method of essential oils for vaginal candidiasis, efficacy of vaginal application of essential oils against murine experimental candidiasis was investigated. The effect on vaginal inflammation and Candida growth form was also studied. Vaginal candidiasis was established by intravaginal infection of C. albicans to estradiol-treated mice. These mice intravaginally received essential oils such as geranium and tea tree singly or in combination with vaginal washing. Vaginal administration of clotrimazole significantly decreased the number of viable C. albicans cells in the vaginal cavity by itself. In contrast, these essential oils did not lower the cell number. When application of geranium oil or geraniol was combined with vaginal washing, the cell number was decreased significantly. The myeloperoxidase activity assay exhibited the possibility that essential oils worked not only to reduce the viable cell number of C. albicans, but also to improve vaginal inflammation. The smear of vaginal washing suspension suggested that more yeast-form cells appeared in vaginal smears of these oil-treated mice than in control mice. In vitro study showed that a very low concentration (25 microg/ml) of geranium oil and geraniol inhibited mycelial growth, but not yeast growth. Based on these findings, it is estimated that vaginal application of geranium oil or its main component, geraniol, suppressed Candida cell growth in the vagina and its local inflammation when combined with vaginal washing.

Oral candidiasis deteriorated by local application of a glucocorticoid-containing film in a mouse model.

In order to estimate predisposing activity of oral application of beclomethasone dipropionate (BDP)-containing mucoadhesive films for oral candidiasis, the effects of BDP on growth of Candida albicans were examined in vivo and in vitro. Murine neutrophils inhibited the mycelial growth of C. albicans in vitro, but this anti-Candida activity was clearly suppressed by the presence of 10(-6) M of BDP. In vitro, a BDP-release test showed that the amount of BDP released from BDP-containing films into the fluid phase increased in a time- and concentration-dependent manner and reached about 10-15% of the total amount of BDP in the film within 30 min. When the BDP-containing film was attached to the tongues of mice orally infected with C. albicans, oral infection by C. albicans deteriorated, but not as severely as in mice systemically immunosuppressed with prednisolone. Based on these findings, we also discuss the problems associated with the clinical application of BDP-film as an anti-inflammatory tool.