Regulation of circulating thyroid hormone levels by hypothalamic tanycytes and hypophysial pars tuberalis-specific cells and their morphological and gene- and protein-expression changes under different photoperiods.

Thyroid hormone in the hypothalamus acts as a key determinant of seasonal transitions. Thyroid hormone-levels in the brain are mainly regulated by the hypothalamic tanycytes and pituitary pars tuberalis (PT)-specific cells. TSHß produced by the PT-specific cells stimulates Dio2 expression and decreases Dio3 expression of the tanycytes. Both tanycytes and PT-specific cells in photosensitive animals exhibit remarkable changes of morphological appearance and expressions of genes and proteins under different photoperiods. Long photoperiods induce increased gene- and protein-expressions and active features. Short photoperiods cause the decreased gene- and protein-expressions and inactive features. In the PT, expressions of TSHß, common α-subunit of glycoprotein hormones (α-GSU), and MT1 receptor of melatonin receptors and eyes absent 3 change under different photoperiods. Diurnal rhythms of α-GSU mRNA expression are observed in the PT of Djungarian hamsters. Hes1, Nkx2.1, and LIM homeodomain gene 2 (Lhx2) are involved in the differentiation of PT. In the hypothalamic tanycytes, expressions of Dio2, Dio3, vimentin, serine/threonine kinase 33, GPR50, Nestin, Retinoid signaling genes (retinaldehyde dehydrogenase 1, cellular retinol binding protein 1, and Stra6), monocarboxylate transporter 8, and neural cell adhesion molecule change under different photoperiods. Rax, Lhx2, Nfia/b/x, and fibroblast growth factor 10 are involved in the differentiation of tanycytes.

Ultrasonic-assisted immersion of parboiled treatment improves head rice yield and nutrition of black rice and provides a softer texture of cooked black rice.

Parboiling is gaining increasing attention as it can enhance the head rice yield (HRY) and nutritional quality of non-pigmented rice. The traditional parboiling process with high-temperature immersion requires a long immersion period and results in hard texture of cooked parboiled black rice (PBR), which may be addressed by ultrasound-assisted immersion. In this study, we evaluated the effect of power, time and temperature of ultrasonic immersion on the HRY, texture profile and nutritional quality of PBR. Proper ultrasound-assisted immersion could increase the HRY by about 20% and the GABA content by up to 133%, as well as reduce the arsenic and cadmium content by up to 61% and 79% relative to untreated black rice (UBR), respectively. Moreover, it could increase the content of essential minerals such as calcium, iron and zinc to some extent, and free and bound polyphenols, despite of a certain loss of anthocyanins. It could also improve the palatability of cooked rice. Furthermore, response surface experiments based on the Box-Behnken design were performed to obtain and validate the optimal conditions of ultrasound-assisted immersion (540 W, 45 min, 57 °C). On this basis, morphological changes might be one reason for the improved HRY, nutrition and texture of PBR compared with those of UBR, namely the disappearance of cracks near the aleurone layer and formation of new cracks in the interior of rice.

The sub-acute toxicity of kavalactone in rats: a study of the effect of oral doses and the mechanism of toxicity in combination with ethanol.

Kava is a herbal supplement and beverage made from the Piper methysticum plant, which is known for its recreational use as a mood enhancer, relaxation, as well as pain relief for centuries. Kava is widely used among alcoholics, but it is dangerous and potentially fatal. The objectives of this study were to examine the sub-acute toxicity effects of different doses of 70% kavalactone (KL) in rats by oral application, as well as to elucidate the mechanisms of toxicity alone and in combination with ethanol (EtOH). The most common side effects observed were abnormal breathing, ataxia, lethargy, loss of appetite, indigestion, and loss of coordination, especially in the 800 mg/kg bw, po bodyweight dosage of kava treatment group alone, and in combination with EtOH. In the sub-acute study, there were dose-related decreases in body weight, feed intake, and water consumption rates. Gross and histopathological findings revealed that the liver was abnormal in color, size, consistency, and the weight significantly increased at a dose of 800 mg/kg bw, po, with KL alone and a greater increase in combination with EtOH. Hepatocellular hypertrophy (HP) and necrosis with Kupffer cells hyperplasia were observed in the periacinar zone of all rats dosed with KL (800 mg/kg bw, po) alone, and extensive changes were observed in combination with EtOH. The periportal (Z1) and mid-zonal (Z2) areas of hepatocytes were less affected as compared to the periacinar zone. These results demonstrate that EtOH exacerbated the sedative and hypnotic activity of KL, and markedly increased toxicity. The histopathological results supported the clinical and biochemical findings and the severity of hepatic damage in a dose-dependent manner.

The clonal growth in Aconitum carmichaelii Debx.

Aconitum carmichaelii Debx. is used as traditional herbal medicine in China, Japan, and other Asian countries. A. carmichaelii has two modes for reproduction: sexual reproduction with seed and vegetative reproduction with vegetative propagules. The vegetative propagules are belowground and invisible. To date, only a handful of studies for the clonal growth are available. In this study, we investigated the clonal growth by anatomical and morphological changes. Results revealed that the axillary bud appeared on the rhizome. Furthermore, the axillary meristem in the axillary bud differentiated a bud upwards and an adventitious root (AR) downwards. The AR expanded to a tuberous root in order to provide the bud nutrients for the new plant. The AR branched LRs. In addition, some lateral roots (LRs) on the AR also swelled. Both the AR and LR were found to follow a similar pattern of development. However, high lignification in the stele region of LRs inhibited further expansion. AR development was attributed to activities of the cambium and meristem cell, starch accumulation, stele lignification, and a polyarch stele. Our study not only provides a better understanding of clonal growth but also provides clues to explore the regulatory mechanisms underlying AR development in A. carmichaelii.

THE PECULIARITIES OF MORPOLOGICAL CHANGES OF RATS' OVARY AND BIOCHEMICAL STATE UNDER THE DAMAGE WITH DIFFERENT DOSES OF LEAD ACETATE.

OBJECTIVE: The aim of the study was to study the effect of low and high doses of lead acetate on biochemical parameters and morphological status of rat ovaries in the experiment. PATIENTS AND METHODS: Materials and methods: The study was performed on 36 nonlinear female rats weighing 180-210 g, aged 4 months, divided into 3 experimental groups: I - control (C), II - rats, which were given 30 days to drink a solution of lead acetate with at the rate of 0,05 mg / kg of animal weight, group III - rats, which were given for 30 days to drink a solution of lead acetate at the rate of 60 mg/kg of animal weight. Biochemical research methods were included determination of diene conjugate concentration in animals' blood, concentration of TBA-active products, study of oxidative modification of proteins in blood plasma, determination of superoxide dismutase and catalase activities. Endogenous intoxication was assessed by the definition of medium-mass molecules, the content was expressed in units of extinction. The material for light microscopy investigation from the ovary was performed according to the generally accepted method. RESULTS: Results: Lead acetate causes activation of peroxidation of lipids and proteins in the body of female rats, which is directly dependent on the dose of lead. In response to the activation of free radical oxidation there are changes in the antioxidant system, which depend on the dose of lead acetate: at a dose of 0.05 mg / kg superoxide dismutase and catalase activity increase, at a dose of 60 mg / kg superoxide dismutase and catalase activity. Small doses of lead do not cause endogenous intoxication. Lead acetate causes the development of endogenous intoxication in animals only in large doses: increases the formation of toxic compounds, cell apoptosis, decreased excretory function of the kidneys, which is associated with multiorgan disorders. As a result of the action of lead acetate, morphological changes of the ovaries were observed, which increased with increasing dose of lead acetate. There was a dose-dependent decrease in massometric parameters, the number of follicles and changes in the thickness of the surface structures of the ovary, which is more pronounced at 60 mg/kg. CONCLUSION: Conclusions: Under the influence of small and large doses of lead acetate on biochemical changes in blood and morphological changes in the ovaries in male rats the oxidative stress is developed. Under the influence of small doses, the changes are adaptive, and under the influence of large doses - damaging.

Localization of (+)-Catechin in Picea abies Phloem: Responses to Wounding and Fungal Inoculation.

To understand the positional and temporal defense mechanisms of coniferous tree bark at the tissue and cellular levels, the phloem topochemistry and structural properties were examined after artificially induced bark defense reactions. Wounding and fungal inoculation with Endoconidiophora polonica of spruce bark were carried out, and phloem tissues were frequently collected to follow the temporal and spatial progress of chemical and structural responses. The changes in (+)-catechin, (-)-epicatechin, stilbene glucoside, and resin acid distribution, and accumulation patterns within the phloem, were mapped using time-of-flight secondary ion mass spectrometry (cryo-ToF-SIMS), alongside detailed structural (LM, TEM, SEM) and quantitative chemical microanalyses of the tissues. Our results show that axial phloem parenchyma cells of Norway spruce contain (+)-catechins, the amount of which locally increases in response to fungal inoculation. The preformed, constitutive distribution and accumulation patterns of (+)-catechins closely follow those of stilbene glucosides. Phloem phenolics are not translocated but form a layered defense barrier with oleoresin compounds in response to pathogen attack. Our results suggest that axial phloem parenchyma cells are the primary location for (+)-catechin storage and synthesis in Norway spruce phloem. Chemical mapping of bark defensive metabolites by cryo-ToF-SIMS, in addition to structural and chemical microanalyses of the defense reactions, can provide novel information on the local amplitudes and localizations of chemical and structural defense mechanisms and pathogen-host interactions of trees.

[The development and experimental evaluation of the effectiveness of a new specialized food based on dried mare's milk during exercise].

The development and implementation of effective means to improve performance, endurance, rapid recovery of the body after physical exertion and, ultimately, improve athletic performance are still relevant. The aim of the work was to develop a new specialized product based on dry mare's milk, as well as to evaluate its effectiveness on an experimental model of physical activity. Material and methods. A specialized product has been developed, including powdered mare's milk, skimmed cow's milk, vegetable cream, crushed sea buckthorn fruits, wheat germ, vitamins A, E, C, PP, folic acid, mineral substances (selenium, magnesium, zinc, iron), inulin, dry bacterial starter culture (Lactobacillus acidophilus, Streptococcus lactis, Bifidobacterium bifidum in a 1:1:1 ratio) and fucoidan. Experimental studies were carried out on 70 white male Wistar rats with an initial body weight of 207-226 g. Animals were fed complete semi-synthetic diet with free access to food and water. Animals of the experimental group additionally received 10 g of a specialized product daily. The control group of animals additionally received glucose in an amount corresponding to the calorie content of 10 g of the specialized product (45 kcal). The animals were subjected to physical exertion - forced swimming until they were completely tired. The swimming test was carried out every seven days during the 21-day experimental period with a load of 10% of the animal's body weight. In hemolysates of erythrocytes, liver microsomes, and in the mitochondrial fraction of the femoral muscle, the activity of catalase and superoxide dismutase was assessed using kits, the concentration of malondialdehyde (MDA) and diene conjugates was determined by spectrophotometry. The level of lactic and pyruvic acids in the blood serum and femoral muscle of rats was assessed by spectrophotometry. The liver and heart were histologically examined. Results. Feeding animals the specialized protein product for 21 days resulted in a statistically significant increase in endurance, as evidenced by data on the time of swimming with a load. So, in the experimental group, in comparison with the initial data, the swimming time increased by 223%. In the control group, the time of swimming with a load increased in comparison with the initial data by only 71.4%, which was 3.1 fold lower than the values in the experimental group. The time of swimming with a load of animals from both groups did not change statistically significantly in the next 7 days of feeding exclusively semi-synthetic diet. The consumption of the specialized product was accompanied by a positive trend in the change in the antioxidant status indicators. Thus, in the membranes of erythrocytes, there was a decrease in the concentration of malondialdehyde by 55.2% and an increase in the activity of catalase and superoxide dismutase by 19.6 and 37.9%, respectively, compared with data in the control group. In the microsomal fraction of the liver, the level of MDA decreased by 40.0% and catalase activity increased by 59.6%. In the mitochondrial fraction of the femoral muscle, a decrease in the level of MDA and diene conjugates was noted, respectively, by 46.8 and 40.8%. In rats of the experimental group, the concentration of lactic acid in the blood serum was reduced by 40.6%, and in the femoral muscle - by 24.7% compared with animals of the control group. Histological studies of the hepatic and cardiac tissues confirmed positive changes in the structure of the studied organs. Conclusion. The results obtained indicate a favorable effect of the protein mixture on the state of the antioxidant system, the general physiological state of rats, their endurance in relation to physical activity, which is largely associated with the set of food ingredients included in the composition, and, first of all, complete protein, vitamins-antioxidants (A, E, C), as well as energy sources, pre- and probiotics, minerals and trace elements, immune defense factors that favorably affect the state of the membranes of erythrocytes, myocytes and hepatocytes and increase not only the body's endurance, but also its metabolic functions, which is confirmed by the data of biochemical and morphological studies.

[Regulating effect of electroacupuncture on left ventricular function and autophagy-specific proteins in spontaneously hypertension rats].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) on cardiac hypertrophy and expression of myocardial autophagy-specific proteins in spontaneously hypertensive rats (SHRs). METHODS: Twelve male Wistar Kyoto (WKY) rats were selected as the normal control group, and 24 male SHRs were randomized into model and EA groups (n＝12 in each). EA (2 Hz /15 Hz, 1 mA) was applied to "Taichong" (LR3) and "Baihui" (GV20) for 20 min, once a day for successive 30 days. The other two groups only received the same grasping and fixation procedures. The systolic blood pressure (SBP) was measured using a non-invasive blood detector, and the left ventricular function including the left ventricular anterior wall diameter (LVAWd), left ventricular posterior wall diameter (LVPWd) and left ventricular internal diameter at end-diastolic dimension (LVIDd) was detected using a real-time echocardiography imaging system, and the left ventricular mass index (left ventricle weight / body weight, LVMI) was calculated. The expression of Beclin-1 and LC3 proteins in the left ventricle tissue was detected by Western blot. Pathological changes of the myocardial tissue were observed by Masson staining and transmission electron microscope (TEM). RESULTS: After the intervention, the elevated SBP was significantly lowered in the EA group relevant to the model group (P<0.01) and the increased LVAWd, LVPWd and LVMI were markedly decreased in the EA group (P<0.01, P<0.05), while the reduced LVIDd was significantly ameliorated in the EA group (P<0.01). Masson staining showed that in SHRs, disorder of arrangement of cardiac myofibers, swelling of cardiomyocytes, widened space among myocardial cells with a large number of collagenous fibers were observed, and under TEM, aggregation and unequal size of myocardial mitochondria with vacuolation and rupture, autophagosomes and autophagic lysosomes were seen. These changes were relatively milder in the EA group. Western blot showed that hypertension induced up-regulation of Beclin-1 and LC3-II and ratio of LC3-II/LC3-I, and down-regulation of LC3-I were significantly suppressed in the EA group (P<0.01, P<0.05). CONCLUSION: EA at LR3 and GV20 may improve the left cardiac function, myocardial morphological changes and the degree of autophagy in SHRs, which is related to its effects in down-regulating the expression of myocardial Beclin-1 and LC3-II and in up-regulating expression of LC3-I.

Regulating effect of electroacupuncture on left ventricular function and autophagy-specific proteins in spontaneously hypertension rats / 针刺研究

OBJECTIVE: To investigate the effect of electroacupuncture (EA) on cardiac hypertrophy and expression of myocardial autophagy-specific proteins in spontaneously hypertensive rats (SHRs). METHODS: Twelve male Wistar Kyoto (WKY) rats were selected as the normal control group, and 24 male SHRs were randomized into model and EA groups (n＝12 in each). EA (2 Hz /15 Hz, 1 mA) was applied to "Taichong" (LR3) and "Baihui" (GV20) for 20 min, once a day for successive 30 days. The other two groups only received the same grasping and fixation procedures. The systolic blood pressure (SBP) was measured using a non-invasive blood detector, and the left ventricular function including the left ventricular anterior wall diameter (LVAWd), left ventricular posterior wall diameter (LVPWd) and left ventricular internal diameter at end-diastolic dimension (LVIDd) was detected using a real-time echocardiography imaging system, and the left ventricular mass index (left ventricle weight / body weight, LVMI) was calculated. The expression of Beclin-1 and LC3 proteins in the left ventricle tissue was detected by Western blot. Pathological changes of the myocardial tissue were observed by Masson staining and transmission electron microscope (TEM). RESULTS: After the intervention, the elevated SBP was significantly lowered in the EA group relevant to the model group (P<0.01) and the increased LVAWd, LVPWd and LVMI were markedly decreased in the EA group (P<0.01, P<0.05), while the reduced LVIDd was significantly ameliorated in the EA group (P<0.01). Masson staining showed that in SHRs, disorder of arrangement of cardiac myofibers, swelling of cardiomyocytes, widened space among myocardial cells with a large number of collagenous fibers were observed, and under TEM, aggregation and unequal size of myocardial mitochondria with vacuolation and rupture, autophagosomes and autophagic lysosomes were seen. These changes were relatively milder in the EA group. Western blot showed that hypertension induced up-regulation of Beclin-1 and LC3-II and ratio of LC3-II/LC3-I, and down-regulation of LC3-I were significantly suppressed in the EA group (P<0.01, P<0.05). CONCLUSION: EA at LR3 and GV20 may improve the left cardiac function, myocardial morphological changes and the degree of autophagy in SHRs, which is related to its effects in down-regulating the expression of myocardial Beclin-1 and LC3-II and in up-regulating expression of LC3-I.

Antibacterial activity of polyphenolic fraction of Kombucha against Vibrio cholerae: targeting cell membrane.

The present study was undertaken to determine the mechanism of antibacterial activity of a polyphenolic fraction, composed of mainly catechin and isorhamnetin, previously isolated from Kombucha, a 14-day fermented beverage of sugared black tea, against the enteropathogen Vibrio cholerae N16961. Bacterial growth was found to be seriously impaired by the polyphenolic fraction in a dose-dependent manner. Scanning Electron Microscopy demonstrated morphological alterations in bacterial cells when exposed to the polyphenolic fraction in a concentration-dependent manner. Permeabilization assays confirmed that the fraction disrupted bacterial membrane integrity in both time- and dose-dependent manners, which were proportional to the production of intracellular reactive oxygen species (ROS). Furthermore, each of the polyphenols catechin and isorhamnetin showed the ability to permeate bacterial cell membranes by generating oxidative stress, thereby suggesting their role in the antibacterial potential of Kombucha. Thus, the basic mechanism of antibacterial activity of the Kombucha polyphenolic fraction against V. cholerae involved bacterial membrane permeabilization and morphological changes, which might be due to the generation of intracellular ROS. To the best of our knowledge, this is the first report on the investigation of antibacterial mechanism of Kombucha, which is mostly attributed to its polyphenolic content. SIGNIFICANCE AND IMPACT OF THE STUDY: The emergence of multidrug-resistant Vibrio cholerae strains has hindered an efficient anti-Vibrio therapy. This study has demonstrated the membrane damage-mediated antibacterial mechanism of Kombucha, a popular fermented beverage of sugared tea, which is mostly attributed to its polyphenolic content. This study also implies the exploitation of Kombucha as a potential new source of bioactive polyphenols against V. cholerae.

Activity of the pterophyllins 2 and 4 against postharvest fruit pathogenic fungi. Comparison with a synthetic analog and related intermediates.

The antifungal activity of pterophyllin 2, pterophyllin 4, a 5-desmethyl analog of the latter and some of their synthetic intermediates, against three postharvest phytopathogenic fungi, was evaluated. The target fungi were Rhizopus stolonifer, Botrytis cinerea and Monilinia fructicola, which affect fruits worldwide, causing important economic losses. The tests were carried out with imazalil and carbendazim as positive controls. Minimum inhibitory concentrations and minimum fungicidal concentrations were determined, and the morphology of the colonies was examined microscopically. In liquid medium, it was found that pterophyllin 4 exhibited selective fungicidal activity toward M. fructicola, whereas its congener pterophyllin 2 proved to be less potent and not selective and the 5-desmethyl analog of pterophyllin 4 displayed a different activity profile. Morphological changes were observed in the colonies exposed to pterophyllin 4. The results highlighted the importance of small structural features for the antifungal behavior and also suggested that, in Nature, the pterophyllins may act as plant defenses against pathogens.

A comparative assessment of antiproliferative properties of resveratrol and ethanol leaf extract of Anogeissus leiocarpus (DC) Guill and Perr against HepG2 hepatocarcinoma cells.

BACKGROUND: Epidemiological and experimental evidences have shown cancer as a leading cause of death worldwide. Although the folklore use of plants as a reliable source of health-restoring principles is well-documented, the search for more of such plants that are active against diseases, such as cancer, continues. We report here a laboratory-based evidence of the relevance of an ethanol leaf extract of Anogeissus leiocarpus (A2L) in comparison with resveratrol, a natural polyphenol, in cancer therapy. METHODS: The quantitative assessment of flavonoid and phenolic contents involved quercetin and gallic acid as standards, respectively were determined using spectrophotometry. Cytotoxicity was determined fluorometrically using propidium-iodide-staining method. Antioxidant status, adenosine triphosphate (ATP) levels, caspase activities and mitochondrial integrity were assessed using fluorometry/luminometry. RESULTS: The antioxidant assay demonstrated that A2L possesses a strong antioxidant capacity as compared with the reference compounds, ascorbic acid and butylated hydroxytoluene. This is further buttressed by the significantly high level of phenolics obtained in the quantitative assessment of the extract. A 72-h post-treatment examination indicated that both A2L and resveratrol modulate the proliferation of HepG2 liver carcinoma cells in a time- and concentration-dependent manner. Determination of the total nuclei area, propidium-iodide negative and positive nuclei areas all further buttress the modulation of cell proliferation by A2L and resveratrol with the indication that the observed cell death is due to apoptosis and necrosis at lower and higher concentrations of treatments respectively. At lower concentrations (0.39-3.13 µg/mL), resveratrol possesses higher tendencies to activate caspases 3 and 7. Bioenergetically, both resveratrol and A2L do not adversely affect the cells at lower concentrations (0.39-6.25 µg/mL for resveratrol and 12.5-100.0 µg/mL for A2L) except at higher concentrations (12.5-25.0 µg/mL for resveratrol and 200-800 µg/mL for A2L) which are more pronounced in A2L-treated cells. Furthermore, the antioxidant status of HepG2 cells is not perturbed by resveratrol as compared with A2L. Assessment of 24-h post-treatment mitochondrial function shows that resveratrol is not mitotoxic as compared with A2L which exhibits mitotoxicity at its highest concentration. CONCLUSIONS: Taken together, findings from this study showed that A2L possesses strong antiproliferative activity and its prospect in the management of hepatocellular carcinoma deserves further investigation.

Electroacupuncture reduces apoptotic index and inhibits p38 mitogen-activated protein kinase signaling pathway in the hippocampus of rats with cerebral ischemia/reperfusion injury.

Electroacupuncture attenuates cerebral hypoxia and neuronal apoptosis induced by cerebral ischemia/reperfusion injury. To further identify the involved mechanisms, we assumed that electroacupuncture used to treat cerebral ischemia/reperfusion injury was associated with the p38 mitogen-activated protein kinase (MAPK) signaling pathway. We established rat models of cerebral ischemia/reperfusion injury using the modified Zea-Longa's method. At 30 minutes before model establishment, p38 MAPK blocker SB20358 was injected into the left lateral ventricles. At 1.5 hours after model establishment, electroacupuncture was administered at acupoints of Chize (LU5), Hegu (LI4), Zusanli (ST36), and Sanyinjiao (SP6) for 20 minutes in the affected side. Results showed that the combination of EA and SB20358 injection significantly decreased neurologic impairment scores, but no significant differences were determined among different interventional groups. Hematoxylin-eosin staining also showed reduced brain tissue injuries. Compared with the SB20358 group, the cells were regularly arranged, the structures were complete, and the number of viable neurons was higher in the SB20358 + electroacupuncture group. Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling assay showed a decreased apoptotic index in each group, with a significant decrease in the SB20358 + electroacupuncture group. Immunohistochemistry revealed reduced phosphorylated p38 expression at 3 days in the electroacupuncture group and SB20358 + electroacupuncture group compared with the ischemia/reperfusion group. There was no significant difference in phosphorylated p38 expression between the ischemia/reperfusion group and SB20358 group. These findings confirmed that the electroacupuncture effects on mitigating cerebral ischemia/reperfusion injury are possibly associated with the p38 MAPK signaling pathway. A time period of 3 days could promote the repair of ischemic cerebral nerves.

Impact of germination on nutritional and physicochemical properties of adlay seed (Coixlachryma-jobi L.).

Adlay has garnered a great deal of research attentions in recent years as a highly nutritious food material and herbal medicine. This study characterized the changes of nutritional and physicochemical properties of adlay seeds during a 60-h germination. The results showed that the 60-h germination brought about a 3.4-fold increase in γ-aminobutyric acid (GABA) and 3.6-fold increase in coixol compared to ungerminated adlay seeds, while the triolein content slightly decreased. Some high molecular proteins were hydrolyzed into smaller proteins, peptides and amino acids after germination. Scanning electron micrographs (SEM) showed that the germination process destroyed the continuous matrix structure of adlay flour and created pits and holes on the surface of some starch granules. Germination resulted to changes in the pasting and gelatinization properties of adlay flour. The results of present study suggest that germination efficiently enhances the nutritional compounds while altering the physicochemical characteristics of adlay seeds.

Anti-Trichomonas vaginalis properties of the oil of Amomum tsao-ko and its major component, geraniol.

CONTEXT: Trichomonosis, caused by the flagellate protozoan Trichomonas vaginalis, is the most common non-viral sexually transmitted disease (STD) and 5-nitroimidazole drugs are used for the treatment. However, a growing number of T. vaginalis isolates are resistant to these drugs, which make it becomes an urgent issue. OBJECTIVE: The current study was designed to evaluate the anti-T. vaginalis activity of the essential oil from A. tsao-ko used in traditional Chinese medicine and as a spice and its main component, geraniol. MATERIALS AND METHODS: The anti-T. vaginalis activities of A. tsao-ko essential oil and geraniol were evaluated by the minimum lethal concentration (MLC) and 50% inhibitory concentration (IC50) in vitro. The morphological changes of T. vaginalis were observed by transmission electron microscopy (TEM). Additionally, sub-MLC concentration treatment with sub-MLC A. tsao-ko essential oil and geraniol was also performed. RESULTS: This study shows that MLC/IC50 of A. tsao-ko essential oil was 44.97 µg/ml/22.49 µg/ml for T. vaginalis isolate Tv1, and 89.93 µg/ml/44.97 µg/ml for T. vaginalis isolate Tv2. Those of geraniol were 342.96 µg/ml/171.48 µg/ml, respectively. After A. tsao-ko essential oil or geraniol treatment, obvious similar morphological changes of T. vaginalis were observed by TEM: the nuclear membrane was damaged, nuclei were dissolved, and the chromatin was accumulated; in the cytoplasm, numerous vacuoles appeared, rough endoplasmic reticulum dilated, the number of ribosomes were reduced, organelles disintegrated, the cell membrane was partially damaged, with cytoplasmic leakage, and cell disintegration was observed. The action time did not increase the effect of A. tsao-ko essential oil or geraniol against T. vaginalis, as no significant difference was observed after sub-MLC concentration treatment for 1, 3, and 5 h with A. tsao-ko essential oil and geraniol. DISCUSSION AND CONCLUSION: The study describes the first report on the activity and morphological changes of A. tsao-ko essential oil and geraniol against T. vaginalis. The results obtained herein presented new opportunities for antitrichomonal drugs.

Protective effect of Lepidium sativum seed extract against hydrogen peroxide-induced cytotoxicity and oxidative stress in human liver cells (HepG2).

CONTEXT: Garden cress [Lepidium sativum (Brassicaceae)] has been widely used to treat a number of ailments in traditional medicine. The pharmacological and preventive potential of Lepidium sativum, such as anti-inflammatory, antipyretic, antihypertensive, anti-ashthamatic, anticancer, and anti-oxidant, are well known. OBJECTIVE: The present investigation was designed to study the protective effects of chloroform extract of Lepidium sativum seed (LSE) against oxidative stress and cytotoxicity induced by hydrogen peroxide (H2O2) in human liver cells (HepG2). MATERIALS AND METHODS: Cytotoxicity of LSE and H2O2 was identified by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), neutral red uptake (NRU) assays, and morphological changes in HepG2. The cells were pre-exposed to biologically safe concentrations (5-25 µg/ml) of LSE for 24 h, and then cytotoxic (0.25 mM) concentration of H2O2 was added. After 24 h of the exposures, cell viability by MTT, NRU assays, and morphological changes in HepG2 were evaluated. Further, protective effects of LSE on reactive oxygen species (ROS) generation, mitochondrial membrane potential (MMP), lipid peroxidation (LPO), and reduced glutathione (GSH) levels induced by H2O2 were studied. RESULTS: Pre-exposure of LSE significantly attenuated the loss of cell viability up to 48% at 25 µg/ml concentration against H2O2 (LD50 value = 2.5 mM). Results also showed that LSE at 25 µg/ml concentration significantly inhibited the induction of ROS generation (45%) and LPO (56%), and increases the MMP (55%) and GSH levels (46%). DISCUSSION AND CONCLUSION: The study suggests the cytoprotective effects of LSE against H2O2-induced toxicity in HepG2. The results also demonstrate the anti-oxidative nature of LSE.

Effects of sub-acute exposure to TiO2, ZnO and Al2O3 nanoparticles on oxidative stress and histological changes in mouse liver and brain.

Nanomaterials are at the leading edge of the rapidly developing field of nanotechnology. However the information regarding toxicity of these nanoparticles on humans and environment is still deficient. The present study investigated the toxic effects of three metal oxide nanoparticles, TiO2, ZnO and Al2O3 on mouse erythrocytes, brain and liver. Male mice were administered a single oral dose of 500 mg/kg of each nanoparticles for 21 consecutive days. The results suggest that exposure to these nano metallic particles produced a significant oxidative stress in erythrocyte, liver and brain as evident from enhanced levels of Reactive Oxygen Species (ROS) and altered antioxidant enzymes activities. A significant increase in dopamine and norepinephrine levels in brain cerebral cortex and increased brain oxidative stress suggest neurotoxic potential of these nanoparticles. Transmission electron microscopic (TEM) analysis indicated the presence of these nanoparticles inside the cytoplasm and nucleus. These changes were also supported by the inhibition of CuZnSOD and MnSOD, considered as important biomarkers of oxidative stress. The toxic effects produced by these nanoparticles were more pronounced in the case of zinc oxide, followed by aluminum oxide and titanium dioxide, respectively. The present results further suggest the involvement of oxidative stress as one of the main mechanisms involved in nanoparticles induced toxic manifestations.

Effects of Kampo Medicine Extracts on Morphological Changes in the Small Intestinal Mucosa of Mice with Age / 日本東洋医学雑誌

The effects of Rikkunshi-to (Liu jun zi tang), Hochu-ekki-to (Bu zhong yi qi tang), and Hange-shashin-to (Ban xia xie xin tang) on morphological changes in the small intestinal mucosa of mice with age were observed. The results indicated that Rikkunshi-to and Hochu-ekki-to had a beneficial effect on intestinal mucosal changes in 36-weeks-old mice. In particular, the improvement of the absorptive area of the intestinal villi and microvilli depending on the concentration of medicine was found. Ninjin-yoei-to had a strong effect on aged mice of 50 and 70 weeks. The morphology and absorptive function were significantly improved.