The Psychophysiological Profile and Cardiac Autonomic Reactivity in Long-Term Female Yoga Practitioners: A Comparison with Runners and Sedentary Individuals.

Yoga practice, a means of stress management, has been reported to optimize psychophysiological health; however, its underlying mechanisms remain unclear. The purpose of the present study was to examine the psychophysiological profile and cardiac autonomic reactivity in long-term yoga practitioners and compare them to runners and sedentary individuals. Psychological health and aerobic fitness level were evaluated using self-reported questionnaires and a 3-min step test. Blood pressure (BP), heart rate (HR), respiration rate (RR), and heart rate variability (HRV) parameters were recorded at rest, as well as during and following psychological stress, which was elicited by the Stroop color and word test and the mental arithmetic task. The yoga group demonstrated a lower RR (10.35 ± 2.13 bpm) as compared to the other two groups, and a lower HR (66.60 ± 7.55 bpm) and diastolic BP (67.75 ± 8.38 mmHg) at rest when compared to the sedentary group (all p < 0.05). HRV parameters following mental stress returned to the baseline in yoga and running groups, but not in the sedentary group. The anxiety level in the running group was significantly lower than that in the sedentary group (p < 0.05). These findings suggested that yoga practitioners may have a greater homeostatic capacity and autonomic resilience than do sedentary individuals.

Training Benefits and Injury Risks of Standing Yoga Applied in Musculoskeletal Problems: Lower Limb Biomechanical Analysis.

Standing yoga poses strengthen a person's legs and helps to achieve the goal of musculoskeletal rehabilitation, but inadequate exercise planning can cause injuries. This study investigated changes in the electromyogram and joint moments of force (JMOFs) of lower extremities during common standing yoga poses in order to explore the feasibility and possible injury risk in dealing with musculoskeletal problems. Eleven yoga instructors were recruited to execute five yoga poses (Chair, Tree, Warrior 1, 2, and 3). The results revealed significant differences in hip, knee, and ankle JMOFs and varying degrees of muscle activation among the poses. Among these poses, rectus femoris muscle activation during the Chair pose was the highest, Warrior 2 produced the highest muscle activation in the vastus lateralis of the front limb, while Warrior 1 had the highest muscle activation in the vastus medialis of the back limb. Therefore, all three poses can possibly be suggested as a therapeutic intervention for quadriceps strengthening. Warrior 1 was possibly suggested as a therapeutic intervention in order to reduce excessive lateral overload of the patella, but the possible adverse effects of Warrior 2 with the highest knee adductor JMOF in the back limb could raise joint reaction forces across the medial condyles. In single-leg balance postures, Warrior 3 had unique training effects on the hamstring, and is therefore suggested as a part of hamstring rehabilitation exercises. The Tree pose induced low lower-extremity JMOFs and a low level of thigh muscle activations when it was performed by senior instructors with excellent balance control; however, for yoga beginners with insufficient stability, it will be a useful training mode for strengthening the muscles that help to keep one upright. This study quantified the physical demands of yoga poses using biomechanical data and elucidated the structures and principles underlying each yoga movement. This is crucial for yoga practitioners.

Injectable polypeptide hydrogel/inorganic nanoparticle composites for bone tissue engineering.

The general concept of tissue engineering is to restore biological function by replacing defective tissues with implantable, biocompatible, and easily handleable cell-laden scaffolds. In this study, osteoinductive and osteoconductive super paramagnetic Fe3O4 nanoparticles (MNP) and hydroxyapatite (HAP) nanoparticles were incorporated into a di-block copolymer based thermo-responsive hydrogel, methoxy(polyethylene glycol)-polyalanine (mPA), at various concentrations to afford composite, injectable hydrogels. Incorporating nanoparticles into the thermo-responsive hydrogel increased the complex viscosity and decreased the gelation temperature of the starting hydrogel. Functionally, the integration of inorganic nanoparticles modulated bio-markers of bone differentiation and enhanced bone mineralization. Moreover, this study adopted the emerging method of using either a supplementary static magnetic field (SMF) or a moving magnetic field to elicit biological response. These results demonstrate that combining external (magnet) and internal (scaffold) magnetisms is a promising approach for bone regeneration.

Comparison of vibration rolling, nonvibration rolling, and static stretching as a warm-up exercise on flexibility, joint proprioception, muscle strength, and balance in young adults.

Warm-up is an essential component for optimizing performance before an exercise session. This study investigated that the immediate effects of vibration rolling (VR), nonvibration rolling (NVR), and static stretching as a part of a warm-up regimen on the flexibility, knee joint proprioception, muscle strength, and dynamic balance of the lower extremity in young adults. Compared with the preintervention, VR induced the range of motion of knee flexion and extension significantly increased by 2.5% and 6%, respectively, and isokinetic peak torque and dynamic balance for muscle strength and dynamic balance increased by 33%-35% and 1.5%, respectively. In the three conditions, most outcomes between VR and NVR were comparable; however, the participants had a significantly higher knee joint reposition error after NVR than after VR, indicating that NVR would have a hampering knee joint proprioception effect. In particular, compared with static stretching, VR significantly increased the quadriceps muscle strength by 2-fold and dynamic balance by 1.8-fold. These findings suggest that athletic professionals may take VR into account for designing more efficient and effective preperformance routine to improve exercise performances. VR has high potential to translate into an on-field practical application.

Physical fitness characteristics of adolescent wushu athletes.

BACKGROUND: The purpose of this study was to examine the physical fitness of wushu athletes in junior high schools and to compare their fitness with that of track-and-field and basketball athletes, as well as general junior high school students. METHODS: A total of 120 participants aged 12-15 years were recruited for this study (wushu N.=30; track-and-field N.=30; basketball N.=30; general students N.=30). Participants' height, weight, flexibility, muscle endurance, cardiorespiratory endurance, agility, power, and static and dynamic balance were assessed and compared among sports groups and general students. RESULTS: The results revealed that athletes in the track-and-field, basketball, and wushu groups had significantly better physical fitness than general students. Additionally, the wushu group had significantly better flexibility and static balance than the other three groups. The track-and-field group had better cardiorespiratory endurance than the other groups. The basketball group had the best agility performance. CONCLUSIONS: These findings suggest that flexibility and balance may be most impacted by the practice of wushu in adolescents. Moreover, adolescents who engage in school sports had better physical fitness than that of general students.

Effects of Taping on Achilles Tendon Protection and Kendo Performance.

CONTEXT: It has been reported that there is a high rate of Achilles tendon injury among kendo athletes. For protection and to support the area, kendo athletes habitually use taping during practice or games. OBJECTIVE: To investigate the effect of various taping techniques on injury prevention and functional performance in kendo athletes. DESIGN: Case-control study. SETTING: Laboratory. PARTICIPANTS: 15 University Kendo Team athletes with at least 2 y kendo experience. MAIN OUTCOME MEASURES: Athletes completed 5 stepping backwards and striking cycles under 4 taping conditions: no taping, athletic taping of ankle joint (AT-Ankle), athletic taping of Achilles tendon (AT-Achilles), and Kinesio-Tex taping of Achilles tendon (KT-Achilles). Jump distance, lower limb angular motion, left foot-ground contact time, Achilles tendon force (ATF), and soleus and medial gastrocnemius muscle activities were measured. RESULTS: Lowest peak ATF was found in AT-Achilles during heel-down phase, with statistically significant difference from KT-Achilles peak force. Significant decline of soleus muscle electromyography amplitude was also found when compared to no taping during heel-down phase and other conditions during pushing phase. Conversely, KT-Achilles showed significant decrease in foot-ground contact time compared with no taping and greater ankle range of motion than in AT-Ankle. CONCLUSION: To protect the Achilles tendon, AT-Achilles taping is recommended since it tends to decrease ATF. Conversely, to enhance athlete performance, we recommend KT-Achilles taping to speed up kendo striking motion. However, the Achilles tendon must withstand greatest forces concurrently. This finding implies that AT-Achilles taping can protect the injured Achilles tendon and KT-Achilles taping can enhance performance on the kendo striking motion.

Antimelanogenic, Antioxidant and Antiproliferative Effects of Antrodia camphorata Fruiting Bodies on B16-F0 Melanoma Cells.

Antrodia camphorata is a fungus that is endemic to Taiwan, and its fruiting body has been used as a folk medicine for the prevention or treatment of diverse diseases. The present study is aimed at investigating the antimelanogenesis and antioxidation effect of the ethanolic extract of Antrodia camphorata fruiting body (EE-AC), as well as its antiproliferation effects in B16-F0 melanoma cells. Regarding antimelanogenic effects, EE-AC had effective cupric ions reducing capacity and expressed more potent inhibitory effect than kojic acid on mushroom tyrosinase activity. Moreover, EE-AC significantly inhibited cellular tyrosinase activity and the melanin content in B16-F0 cells at 12.5 µg/mL concentration without cell toxicities. Regarding antioxidant effects, EE-AC exhibited potent DPPH radical- and SOD-like-scavenging activities. Regarding antiproliferative effects, EE-AC exhibited a selective cytotoxic effect and markedly inhibited the migration ability of B16-F0 cells. EE-AC increased the population of B16-F0 cells at sub-G1 phase of the cell cycle. EE-AC also caused the increase of early apoptotic cells and chromatin condensation, which indicated the apoptotic effects in B16-F0 cells. We demonstrated that EE-AC possessed antimelanogenic, antioxidant and anti-skin cancer actions. The results would contribute to the development and application of cosmetics, healthy food and pharmaceuticals.

Effects of Yoga on Heart Rate Variability and Depressive Symptoms in Women: A Randomized Controlled Trial.

OBJECTIVES: The purpose of the study was to investigate the effects of a 12-week yoga program on heart rate variability (HRV) and depressive symptoms in depressed women. METHODS: This was a randomized controlled trial. Twenty-six sedentary women scoring ≥14 on the Beck Depression Inventory-II were randomized to either the yoga or the control group. The yoga group completed a 12-week yoga program, which took place twice a week for 60 min per session and consisted of breathing exercises, yoga pose practice, and supine meditation/relaxation. The control group was instructed not to engage in any yoga practice and to maintain their usual level of physical activity during the course of the study. Participants' HRV, depressive symptoms, and perceived stress were assessed at baseline and post-test. RESULTS: The yoga group had a significant increase in high-frequency HRV and decreases in low-frequency HRV and low frequency/high frequency ratio after the intervention. The yoga group also reported significantly reduced depressive symptoms and perceived stress. No change was found in the control group. CONCLUSIONS: A 12-week yoga program was effective in increasing parasympathetic tone and reducing depressive symptoms and perceived stress in women with elevated depressive symptoms. Regular yoga practice may be recommended for women to cope with their depressive symptoms and stress and to improve their HRV.

Acute Effects of Foam Rolling, Static Stretching, and Dynamic Stretching During Warm-ups on Muscular Flexibility and Strength in Young Adults.

CONTEXT: Foam rolling has been proposed to improve muscle function, performance, and joint range of motion (ROM). However, whether a foam rolling protocol can be adopted as a warm-up to improve flexibility and muscle strength is unclear. OBJECTIVES: To examine and compare the acute effects of foam rolling, static stretching, and dynamic stretching used as part of a warm-up on flexibility and muscle strength of knee flexion and extension. DESIGN: Crossover study. SETTING: University research laboratory. PARTICIPANTS: 15 male and 15 female college students (age 21.43 ± 1.48 y, weight 65.13 ± 12.29 kg, height 166.90 ± 6.99 cm). MAIN OUTCOME MEASURES: Isokinetic peak torque was measured during knee extension and flexion at an angular velocity of 60°/second. Flexibility of the quadriceps was assessed by the modified Thomas test, while flexibility of the hamstrings was assessed using the sit-and-reach test. The 3 interventions were performed by all participants in random order on 3 days separated by 48-72 hours. RESULTS: The flexibility test scores improved significantly more after foam rolling as compared with static and dynamic stretching. With regard to muscle strength, only knee extension peak torque (pre vs. postintervention) improved significantly after the dynamic stretching and foam rolling, but not after static stretching. Knee flexion peak torque remained unchanged. CONCLUSIONS: Foam rolling is more effective than static and dynamic stretching in acutely increasing flexibility of the quadriceps and hamstrings without hampering muscle strength, and may be recommended as part of a warm-up in healthy young adults.

Effects of Yoga on Heart Rate Variability and Mood in Women: A Randomized Controlled Trial.

OBJECTIVE: To examine the effects of an 8-week yoga program on heart rate variability and mood in generally healthy women. DESIGN: Randomized controlled trial. PARTICIPANTS: Fifty-two healthy women were randomly assigned to a yoga group or a control group. INTERVENTIONS: Participants in the yoga group completed an 8-week yoga program, which comprised a 60-minute session twice a week. Each session consisted of breathing exercises, yoga pose practice, and supine meditation/relaxation. The control group was instructed not to engage in any yoga practice and to maintain their usual level of physical activity during the study. OUTCOME MEASURES: Participants' heart rate variability, perceived stress, depressive symptoms, and state and trait anxiety were assessed at baseline (week 0) and after the intervention (week 9). RESULTS: No measures of heart rate variability changed significantly in either the yoga or control group after intervention. State anxiety was reduced significantly in the yoga group but not in the control group. No significant changes were noted in perceived stress, depression, or trait anxiety in either group. CONCLUSIONS: An 8-week yoga program was not sufficient to improve heart rate variability. However, such a program appears to be effective in reducing state anxiety in generally healthy women. Future research should involve longer periods of yoga training, include heart rate variability measures both at rest and during yoga practice, and enroll women with higher levels of stress and trait anxiety.

The acute effects of yoga on cognitive measures for women with premenstrual syndrome.

OBJECTIVES: Recently, yoga classes specifically for women with premenstrual syndrome (PMS) have increased, but there is little research about the efficacy of these classes. The primary aims of this study were to evaluate the effect of yoga exercise on women with PMS and to evaluate the immediate change of attention performance after yoga classes. METHODS: This study examined the attention task results of women with PMS. Eleven women with PMS and 9 women without PMS were recruited. The PMS group took the tests before and immediately after the yoga class both in the luteal and follicular phase of one menstrual cycle, while the control group took the tests only twice: once in the luteal phase and once in the follicular phase. Both groups were required to finish resting electroencephalography (EEG) and cognitive task of the 2-back task with EEG recording. RESULTS: The alpha brain wave percentage was higher immediately after yoga exercise in the PMS group. This suggests that the participants felt more relaxed or were in a more peaceful mental condition after yoga exercise. In the 2-back task, the PMS group needed a longer reaction time to respond to the target stimulus in the luteal phase and performed better with higher accuracy and shorter reaction time after yoga exercise. The event-related potentials of the EEG recording displayed a significant variability at the P3 amplitude throughout the menstrual cycle in the PMS group, but the P3 amplitude was unchanged throughout the menstrual cycle in the control group. CONCLUSION: The results of this study suggest that women with PMS could attend short-term yoga exercise in the luteal phase to make themselves feel better and maintain a better attention level.

Monascus-fermented red mold rice exhibits cytotoxic effect and induces apoptosis on human breast cancer cells.

Red mold rice (RMR) is a traditional food and folk medicine to Asian people and has recently become a popular health supplement. RMR has been shown to have some anticancer activities, although the mechanism for inducing cell death of human breast cancer cells is still not fully understood. In this study, bioactive extracts of RMR fermented by Monascus purpureus NTU 803 were analyzed for effects on apoptosis induction in human breast cancer cells. The RMR ethanol extract and ethyl acetate extract contain monacolin K, total phenols, and flavonoids, the three components that have been reported to have anticancer activity. Red mold rice extracts (RMRE) exhibited selective cytotoxic effect on MCF-7 cells. RMRE treatment induced apoptosis and cell cycle arrest at G2/M phase. Apoptosis was confirmed by annexin V-fluorescein isothiocyanate (FITC)/propidium iodide staining, the observation of characteristic chromatin condensation, nuclear DNA fragmentation, and poly(ADP-ribose) polymerase cleavage. Furthermore, the RMRE-induced apoptosis in MCF-7 cells may occur through a mitochondria-dependent pathway while triggering an appropriate balance of bax/bcl-2 and activation of caspase-9 and caspase-3 in a time-dependent manner. To conclude, RMRE exhibits direct cytotoxic and proapoptotic effects on MCF-7 cells and could be considered as a potential functional food for breast cancer prevention.

(-)-N-Formylanonaine from Michelia alba as a human tyrosinase inhibitor and antioxidant.

Tyrosinase is the first and rate limiting enzyme in the synthesis of melanin pigments for coloring hair, skin, and eyes. As reported in this study, a natural product, (-)-N-formylanonaine isolated from the leaves of Michelia alba D.C. (Magnolianceae), was found to inhibit mushroom tyrosinase with an IC50 of 74.3 microM and to have tyrosinase and melanin reducing activities in human epidermal melanocytes without apparent cytotoxicity to human cells, superior to the known tyrosinase inhibitors, such as kojic acid and 1-phenyl-2-thiourea (PTU). Based on homology modeling, the compound binds the active site by coordinating with two Cu2+ ions. In addition, the compound had antioxidation activities in tests for scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH), reducing power, and chelating metal ions. To our knowledge, this is the first study to reveal the bioactivities of (-)-N-formylanonaine from this plant species.

Effects of Tai Chi rehabilitation on heart rate responses in patients with coronary artery disease.

The objective of the present study was to evaluate the effect of a six-month Tai Chi (TC) exercise cardiac rehabilitation program on two prognostic factors of cardiac events, rate-pressure product and rate-pressure product reserve, in patients with coronary artery disease (CAD). Patients (N = 54) with CAD were recruited from the clinics of cardiology and cardiovascular surgery at a regional hospital in Taiwan. Twenty-two of them enrolled in the TC rehabilitation program which consisted of weekly 90-min sessions of Yang's style TC for six months in addition to receiving usual care. The remaining 32 patients received usual care only. Modified Bruce treadmill exercise test was performed to evaluate their exercise test responses at baseline and at six months. The change over time was significantly different between the TC and control group in peak rate-pressure product (RPP) (interaction between group and time, p = 0.029) and in RPP reserve (interaction between group and time p = 0.009) over the six-month period, there was a decrease in peak RPP of 32.0 mmHg x bpm x 10(-2) and in RPP reserve of 37.4 mmHg x bpm x 10(-2) in the TC group. In conclusion, participating in a six-month TC exercise-based cardiac rehabilitation program was associated with improved peak RPP and RPP reserve during exercise testing in patients with CAD. TC exercise program may lead to a better prognosis for cardiac events in patients with CAD.

The effect of t'ai chi exercise on autonomic nervous function of patients with coronary artery disease.

OBJECTIVE: The objective of this study was to evaluate the effect of t'ai chi (TC) on heart rate variability (HRV) from baseline to 9 months in patients with coronary artery disease (CAD). DESIGN: A comparative trial was undertaken comparing the acute and long-term effect of TC on HRV in patients with CAD. PARTICIPANTS: Sixty-one (61) subjects with CAD undergoing percutaneous intervention or coronary bypass grafting for a period of more than 1 month were recruited from the clinics of cardiology and cardiovascular surgery at Chia-Yi Christian Hospital, Taiwan. INTERVENTIONS: The experimental group (n = 22) practiced weekly 90-minute Yang's style TC for 9 months and the control group (n = 39) continued their normal daily physical activity. MAIN OUTCOME MEASURES: HRV was recorded at baseline, 3 months, 6 months, and 9 months in the control group. Resting HRV was recorded before TC exercise and recovery HRV was recorded 30 minutes post TC at the same four time points in the experimental group. RESULTS: The change in the normalized low-frequency power, normalized high-frequency power, and the low-/high-frequency power ratio between resting and post-TC was significantly different at 9 months when compared with those at baseline, 3, and 6 months. The mean difference in normalized low-frequency power and the low/high-frequency power ratio changed from positive values at baseline, 3, and 6 months to negative values at 9 months. However, there were no significant differences in resting HRV between the patients in TC and control groups in either time domain or frequency domain HRV indices. CONCLUSIONS: The change in heart rate and HRV between resting and post-TC suggested that TC exercise could enhance vagal modulation. The potential beneficial effect of long-term regular TC exercise in patients with CAD merits further investigation.

Effects of three biodiesels and a low sulfur diesel in male rats--a pilot 4-week oral study.

Because of the accessible and renewable nature of feedstock and the potential for the reduction of harmful combustion emissions and greenhouse gases, biodiesels have received increasing interest as an alternate fuel. Oral exposure to biodiesels is a concern because of contact during refuelling, accidental ingestion and exposure through ground water contamination. Although biodiesels from various feedstock are in use commercially and experimentally, very little is known about their potential adverse effects and no data is available on their potential for ground water contamination. A study was performed on male rats following oral treatment with experimental biodiesels (dissolved in corn oil) derived from canola oil (Bio-C), soy oil (Bio-S) and fish oil (Bio-F), at 500 mg/kg body weight/day, 5 days per week, for 4 weeks. Separate groups of animals were treated with low sulfur diesel (LSD) for comparison purpose, and with corn oil alone to serve as control. The potential for ground water contamination by biodiesels was investigated by the preparation of water-accommodated fractions (WAF) followed by gas chromatographic analysis. WAF from Bio-F and Bio-S was found to have the highest level of dichloromethane extractable materials. Gas chromatographic analysis indicated that the extractable materials from biodiesels contained much higher proportion of C15-C30 materials than LSD. Increased liver weight was observed in animal treated with Bio-C, Bio-S and LSD and decreased thymus weight was found in those treated with Bio-S. Histopathological changes typical of male-rat specific hyaline-droplet nephropathy were detected in kidney tubules of animals treated with LSD, Bio-S and Bio-C. Mild adaptive changes were observed in thyroids of animals treated with LSD, Bio-S and Bio-F. Clinical chemical and biochemical changes were confined to Bio-S and LSD treated rats and included elevation in some hepatic phase-I and phase-II drug metabolizing enzymes and hepatic palmitoyl Co-A oxidase, and elevated urinary concentrations of ascorbic acid and albumin. At the given dose level of 500 mg/kg bw/day, the overall treatment-related effects of biodiesels and LSD are mild, and the severity of the treatment effects may be ranked as: LSD>Bio-S>Bio-C>Bio-F. Considered together with the presence of a higher level of water extractable materials, Bio-S may be more of a concern for potential human health than Bio-C and Bio-F in an oral exposure scenario. Further studies are needed to identify and characterize the constituents contributing to the treatment-related effects specific to these experimental biodiesels.

Subchronic toxicity of chloral hydrate on rats: a drinking water study.

The subchronic toxicity of chloral hydrate, a disinfection byproduct, was studied in rats following 13 weeks of drinking water exposure. Male (262 +/- 10 g) and female (190 +/- 8 g) Sprague-Dawley rats, ten animals per group, were administered chloral hydrate via drinking water at 0.2, 2, 20 and 200 ppm. Control animals received distilled water only. Gross and microscopic examinations, serum chemistry, hematology, biochemical analysis, neurogenic amine analysis and serum trichloroacetic acid (TCA) analysis were performed at the end of the treatment period. Bronchoalveolar fluids were collected at necropsy and urine specimens were collected at weeks 2, 6 and 12 for biochemical analysis. No treatment-related changes in food and water intakes or body weight gains were observed. There were no significant changes in the weights of major organs. Except for a mild degree of vacuolation within the myelin sheath of the optic nerves in the highest dose males, there were no notable histological changes in the tissues examined. Statistically significant treatment-related effects were biochemical in nature, with the most pronounced being increased liver catalase activity in male rats starting at 2 ppm. Liver aldehyde dehydrogenase (ALDH) was significantly depressed, whereas liver aniline hydroxylase activity was significantly elevated in both males and females receiving the highest dose. A dose-related increase in serum TCA was detected in both males and females starting at 2 ppm. An in vitro study of liver ALDH confirmed that chloral hydrate was a potent inhibitor, with an IC(50) of 8 micro M, whereas TCA was weakly inhibitory and trichloroethanol was without effect. Analysis of brain biogenic amines was conducted on a limited number (n = 5) of male rats in the control and high dose groups, and no significant treatment-related changes were detected. Taking into account the effect on the myelin sheath of male rats and the effects on liver ALDH and aniline hydroxylase of both males and females at the highest dose level, the no-observed-effect level (NOEL) was determined to be 20 ppm or 1.89 mg kg(-1) day(-1) in males and 2.53 mg kg(-1) day(-1) in females. This NOEL is ca. 1000-fold higher than the highest concentration of chloral hydrate reported in the municipal water supply.

Short-term oral toxicity of gasohol in female rats.

The systemic toxicity of gasohol (10% ethanol in gasoline by volume) in female rats following 4-week oral administration was studied. Female Sprague-Dawley rats (198+/-14 g) were divided into four groups of ten animals each. The low- and medium-dose groups received by gavage corn oil containing gasoline/ethanol at 16/1.8 and 160/18 (mg kg(-1) body weight), respectively, for 28 consecutive days. The high-dose animals were administered gasoline/ethanol at 1600/180 mg kg(-1) on the first day and the dose was reduced to 800/90 mg kg(-1) for the rest of the study period. Control animals received corn oil only. Urine was obtained from all rats after weeks 1, 2 and 4 for biochemical analysis. At termination of the study, kidneys of four rats from each group were examined by electron microscopy. Body weight gains, organ weights, tissue and organ histopathology, serum biochemistry, hematology, liver enzymes and biochemistry were determined in the remaining six animals of each group. No treatment-related changes were observed in the following endpoints: body weight gain or relative weights of the brain, lungs, liver, kidneys, spleen and thymus. A significant increase in pentoxyresorufin O-deethylase (PROD) and benzoylresorufin O-dealkylase (BROD) activities was detected in the high-dose animals, whereas ethoxyresorufin O-deethylase (EROD) activity was unchanged. Treatment with gasohol did not produce any significant changes in hematology and serum clinical chemistry parameters. Biomarkers of oxidative stress such as serum and liver thiobarbituric acid reactive substances (TBARS) and liver glutathione also were unaffected by treatments. Urinary ascorbic acid was elevated markedly in the medium- and high-dose groups following the first, second and fourth weeks of treatment. Urine hippuric acid was increased significantly in the high-dose groups. A dose-related increase in urinary aldehydes also was observed in animals after the first, second and fourth week of treatment. Interestingly, a separate 1-week dosing study revealed that the increase in urinary aldehydes was associated with gasoline and not with ethanol treatment. In the high-dose animals slight increases in urinary protein and N-acetylglucosaminidase activity were observed after week 1 but not after week 2 or week 4. No histopathological changes were detected in the liver, kidneys, stomach, brain, lungs or other tissues examined. Electron microscopic examination of the kidneys also did not reveal any abnormalities. It was concluded that short-term oral administration of gasoline/ethanol at 800/90 mg kg(-1) produced a biochemical response in the liver but no adverse effects in the kidneys and lungs. The biological significance of elevated urinary aldehydes at gasoline/ethanol concentrations of 160/18 mg kg(-1) and higher remains to be studied.

Factorial designs combined with the steepest ascent method to optimize serum-free media for CHO cells.

A serum free medium for recombinant CHO NTHU 108 cell growth and fusion protein (CD20 linked to a human IgG-Fc gamma4 fragment) synthesis were systematically developed using factorial designs combined with the steepest ascent method. Experimental results indicate that the optimal composition of serum replacement for specific fusion protein production was 1% SITE (selenium, insulin, transferrin, ethanolamine), 0.3 g/L yeast extract, and 0.09% linoleic acid-BSA. Cell growth and fusion protein production of the adapted CHO NTHU 108 cultured in Iscove's modified Dulbecco's medium supplemented with these serum substitutes were comparable to those in the Ex-Cell 301 commercial serum-free medium. These serum substitutes can also promote CHO cell growth and fusion protein production in nine kinds of commercial media. The low protein content of the developed medium facilitates downstream processing and product purification.

Validation of higher-throughput high-performance liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry assays to conduct cytochrome P450s CYP2D6 and CYP3A4 enzyme inhibition studies in human liver microsomes.

In the early stage of drug discovery, thousands of new chemical entities (NCEs) may be screened before a single drug candidate can be identified for development. In order to accelerate the drug discovery process, we have developed higher-throughput enzyme assays to evaluate the inhibition of cytochrome P450 isoforms 2D6 (CYP2D6) and 3A4 (CYP3A4) in human liver microsomes. The assays are based on high-performance liquid chromatography/tandem mass spectrometry (LC/MS/MS) techniques. The analysis time for each sample was reduced from approximately 20 minutes for the conventional HPLC assay to 30 seconds for the LC/MS/MS assay. For both LC/MS/MS assays, the linearity (r(2) > 0.99), precision (%CV < 15%) and accuracy (% bias <15%) for both inter- and intraday validations were satisfactory. Since the implementation of the LC/MS/MS assays, our sample throughput has increased by over 40-fold.