Objective

In Western medicine, hie (cold sensation) is thought as a subjective complaint. Consequently, despite women's complaints of health problems due to hie, such as distress and decreased quality of life, it is not diagnosed as a disease or given active treatment. However, hie sometimes requires emergency treatment or is a symptom of other disorders. Thus, diagnosing hie and eliminating distress due to hie are clinically significant. This research aimed to discriminate between hie and hiesho (cold disorder) and quantify hie by using objective assessments in addition to subjective assessments.

Methods

A questionnaire was administered to 159 healthy young women without any underlying disease that could cause hie, followed by assessment of vascular function. These data were examined through exploratory factor analysis, exploratory cluster analysis, and multivariate logistic analysis. Two- and three-group comparisons were made using the Mann-Whitney test and Kruskal-Wallis test, respectively, followed by multiple comparisons using Scheffé's method.

Results

The hie (including hiesho) and normal groups comprised 81 (51%) and 78 (49%) subjects, respectively. The questionnaire was a low reliability screening method for determining the severity of hie symptoms. As an index of discrimination of hie and hiesho, we determined the toe-brachial index (TBI) to be 0.81 and toe systolic blood pressure (SBP) to be 87.73 mmHg with a linear function (TBI=0.007 5×toe SBP+0.152, adjusted R²=0.829, F_(1,79)=388.79, P<0.001).

Conclusion

Toe SBP and TBI were suggested to be capable of discriminating and quantifying symptoms when used in combination with a questionnaire.

Objective

Tephrosia purpurea (Linn.) Pers. is widely used in traditional medicine to treat liver disorders, febrile attacks, enlargement and obstruction of liver, spleen, and kidney. In the present study, investigations were carried out to determine the seasonal impact on the content of flavonoid glycosides and on antioxidant activities so as to identify the optimal time of harvesting.

Methods

The plant materials were collected in different seasons during 2013-2014. Air-dried, powdered plant materials were extracted with 95% ethanol and ethanol:water (1:1) by ultrasound-assisted extraction process. Their chemical composition in terms of total polyphenol and flavonoid contents (TPCs and TFCs) was determined using modified colorimetric Folin-Ciocalteu method and aluminum chloride colorimetric assay respectively. To determine the in vitroantioxidant activity, diphenyl-picryl hydrazyl (DPPH) radical-scavenging assay and total antioxidant capacity by phosphomolybdate antioxidant assay were carried out. High-performance liquid chromatography (HPLC)/photo-diode array (PDA) analysis was used to quantify the flavonoid glycosides in the samples collected in different seasons. Correlation studies were also carried out between antioxidant activities and TPCs.

Results

The highest TPC and TFC were found to be in the 95% ethanolic extract of the August sample and the lowest in the 50% hydro-alcoholic extract of the plant sample collected in winter season. It was observed that in both the assays used to determine the antioxidant activity, the 95% ethanolic extracts in all the seasons showed a higher activity than their respective 50% hydro-alcoholic extracts with an increase in activity as we go from cold to hot to rainy seasons. Based on correlation analysis, DPPH radical-scavenging activities as well as the spectrophotometrically measured phosphomolybdenum complex were also strongly correlated with TPC of the extracts. The most abundant flavonoid glycoside was quercetin-3-O-rhamnoglucoside in all the seasons. The content of all flavonoids was observed highest in the 95% ethanolic extract of the plant collected in August (TP-3). The 50% hydro-alcoholic extract of the plant collected in December (TP-6) showed the lowest amount of flavonoids and antioxidant activity.

Conclusion

The findings of this study confirmed that the metabolism and production of flavonoids in T. purpurea are vigorously affected by seasonal factors. Significant differences were observed in the TPC, TFC and flavonoid glycoside composition in the 95% ethanolic as well as 50% hydro-alcoholic extracts of T. purpurea collected in different seasons. Since the 95% ethanolic extract of the August sample (TP-3), which also happens to be its flowering season, exhibited the highest TPC, TFC and antioxidant activities in both DPPH and phosphomolybdate assays as well as contained the highest content of all flavonoids, it could be recommended as the optimal season of harvesting T. purpurea with respect to its pharmaceutically important constituents, i.e., flavonoids.

Objective

This study investigated the effects of allylpyrocatechol (APC), the major component in ethanolic extract of Piper betle, on key oxidative stress resistance enzymes important for the survival of Staphylococcus aureus, a major pathogen in the human host.

Methods

Effects of APC on expressions of genes encoding catalase (katA), superoxide dismutases (SODs), including sodA and sodM, and alkyl hydroperoxide reductase (ahpC) in S. aureus were quantitated by RT-qPCR in reference to gyrA and 16S rRNA. Corresponding activities of the enzymes were also investigated. The Livak analysis was performed for verification of gene-fold expression data. Effects of APC on intracellular and extracellular reactive oxygen species (ROS) levels were determined using the nitroblue tetrazolium (NBT) reduction assay.

Results

APC-treated S. aureus cells had higher sodA and sodM transcripts at 1.5-fold and 0.7-fold expressions respectively with corresponding increase in total SOD activity of 12.24 U/mL compared to untreated cells, 10.85 U/mL (P<0.05). Expression of ahpC was highest in APC-treated cells with 5.5-fold increased expression compared to untreated cells (P<0.05). Correspondingly, ahpC activity was higher in APC-treated cells at 0.672 (A_310nm) compared to untreated cells which was 0.394 (A_310nm). In contrast, katA expression was 1.48-fold and 0.33-fold lower respectively relative to gyrA and 16S rRNA. Further, APC-treated cells showed decreased catalase activity of 1.8 × 10^-4 (U/L or μmol/(min·L)) compared to untreated cells, which was 4.8 × 10^-4 U/L (P<0.05). Absorbance readings (A_575nm) for the NBT reduction assay were 0.709 and 0.695 respectively for untreated and treated cells, which indicated the presence of ROS. APC-treated S. aureuscells had lower ROS levels both extracellularly and intracellularly, but larger amounts remained intracellularly compared to extracellular levels with absorbances of 0.457 and 0.137 respectively (P<0.05).

Conclusion

APC induced expressions of both sodA and sodM, resulting in increased total SOD activity in S. aureus. Higher sodAexpression indicated stress induced intracellularly involving O₂^-, presumably leading to higher intracellular pools of H₂O₂. A concommittant decrease in katA expression and catalase activity possibly induced ahpC expression, which was increased the highest in APC-treated cells. Our findings suggest that in the absence of catalase, cells are propelled to seek an alternate pathway involving ahpC to reduce stress invoked by O₂^- and H₂O₂. Although APC reduced levels of ROS, significant amounts eluded its antioxidative action and remained intracellularly, which adds to oxidative stress in treated cells.

Objective

Ligusticum porteri is a traditional Native American herb. The roots of L. porteri are traditionally used in the treatment of many diseases, however, its cytotoxicity, antioxidative and immune-modulatory effects need to be investigated. In this study, we evaluated the effects of the root extract at different doses on human peripheral blood lymphocytes (PBLs).

Methods

The lymphocytes were incubated with different concentrations of the root extracts (0, 50, 100, 200, and 400 μg/mL) and harvested every 6 h for 2 d (P<0.05). The protective effect of the herb against oxidative damage was determined by inducing oxidative stress with the administration of 50 μmol/L of hydrogen peroxide (H₂O₂).

Results

Treatments with L. porteri at 200 and 400 μg/mL increased the viability of PBLs. The deleterious effect of H₂O₂ was ameliorated by 400 μg/mL L. porteri treatment. Addition of 400 μg/mL L. porteri reduced lipid peroxidation in stressed PBLs by 94% (P<0.05). Treatment with 400 μg/mL of L. porteri resulted in a 26.4% increase of reduced glutathione levels. Activities of superoxide dismutase and catalase increased by 17.5% and 55.2% respectively, when stressed PBLs were treated with 400 μg/mL L. porteri for 2 d (P<0.05). Treatment with 400 μg/mL L. porteri increased interferon-γ and interleukin-2 expressions in H₂O₂-challenged PBLs (P<0.05), however, the root extract did not cause a significant difference in interleukin-10 levels compared to the control (P>0.05).

Conclusion

The findings suggest that L. porteri might be a potential immune-modulating agent involving protective effects against oxidative damage.

Objective

Chronic exposure to ultraviolet (UV) radiation induces skin photoaging (premature skin aging). UV irradiation generates reactive oxygen species (ROS), which are shown to play a pivotal role in skin photoaging. Ayurveda is a holistic traditional medical system, and Chyawanprash is one of the most popular formulations in Ayurveda. Since maintenance of the function and appearance of skin is important, we examined whether Chyawanprash has a protective effect on skin photoaging.

Methods

To examine the effect of Chyawanprash on skin photoaging, hairless mice were administered with Chyawanprash in drinking water for 3 weeks, and then repeatedly exposed to ultraviolet light B (UVB) irradiation (225 or 450 mJ/cm²) to induce skin photoaging. To further examine the function of Chyawanprash, its effects were examined in cells cultured in vitro. Chyawanprash was added in culture medium, and examined for the effect on the growth of human keratinocytes, and for the ability to eliminate ROS which generated by paraquat (50 μmol/L) in HeLa cells.

Results

UVB irradiation caused symptoms such as rough skin, erythema, and edema on the skin in hairless mice, but administration of Chyawanprash relieved these symptoms. Further, Chyawanprash significantly suppressed epidermal thickening, a typical marker of skin photoaging, in mice. We then analyzed the effect of Chyawanprash in human cells in culture, and found that Chyawanprash enhanced the growth of human keratinocytes, and efficiently eliminated ROS, which are causally involved in skin photoaging, in HeLa cells.

Conclusion

These findings suggested that Chyawanprash may have beneficial effects on slowing skin photoaging.