Hirami lemon (Citrus reticulata var. depressa) modulates the gut-brain axis in a chronic mild stress-induced depression mouse model.

Citrus reticulata var. depressa, commonly known as Hirami lemon, is a native citrus species found in Taiwan and Okinawa islands of Japan. While several Citrus species are known to possess antidepressant activity by modulating the gut microbiota, the antidepressant effect of Hirami lemon and its underlying mechanisms have not been thoroughly investigated. In this study, we explored the potential antidepressant efficacy of the fruit extract (CD) and the essential oil (CDE) from Hirami lemon peel using a chronic mild stress (CMS)-induced mouse model and analyzed the association of gut microbiome changes. Our findings revealed that mice subjected to CMS exhibited anxiety- and depression-like behaviors as assessed by elevated plus-maze and forced swimming tests, respectively. Significantly, oral administration of CDE and CD notably reversed CMS-induced depression- and anxiety-like behaviors in CMS-induced mice. Moreover, compared to the non-stressed group, CMS significantly altered the gut microbiome, characterized by highly diverse bacterial communities, reduced Bacteroidetes, and increased Firmicutes. However, oral administration of CDE and CD restored gut microbiota dysbiosis. We also performed a qualitative analysis of CD and CDE using UPLC-MS and GC-MS, respectively. The CD contained 25 compounds, of which 3 were polymethoxy flavones and flavanones. Three major compounds, nobiletin, tangeretin and hesperidin, accounted for 56.88% of the total relative peak area. In contrast, the CDE contained 11 terpenoids, of which 8 were identified as major compounds, with D-limonene (45.71%) being the most abundant, followed by γ-terpinene (34.65%), linalool (6.46%), p-cymene (2.57%), α-terpineol (2.04%), α-pinene (1.89%), α-terpinolene (1.46%), and ß-pinene (1.16%), accounting for 95.94% of the total oil. In conclusion, our study demonstrated the potential of Hirami lemon as a source of natural antidepressant agents for the prevention and treatment of major depressive disorders.

Peri-hilar branching pattern and variations of the renal artery among Indian kidney donors using pre-operative computed tomography angiography: an anatomical study and review.

BACKGROUND: The peri-hilar branching pattern of renal arteries and variations such as accessory renal artery and early branching of the renal artery are important factors to be evaluated preoperatively to minimise potential complications during renal transplantation and renal surgeries. The present study was done to assess the peri hilar branching pattern of the renal artery and its variation among the renal donors in India. MATERIALS AND METHODS: One hundred ninety eight kidneys from 99 donors were analysed using pre-operative computed tomography renal angiograms prospectively, over 1 year 2 months. RESULTS: Based on the branching pattern of primary and secondary branches of renal arteries, we identified 8 cardinal and 10 minor peri-hilar branching patterns. Type I (duplicated fork) peri-hilar branching pattern was the most frequently found (70.2%) among 198 kidneys, followed by type II (triplicated fork) (19.2%) and type III (ladder) (10.6%). Duplicated fork pattern was common in both the right renal artery (75.7%) and left renal artery (64.6%), respectively. The prevalence of accessory renal artery was 39.4% with hilar artery at 21.2% and polar artery at 18.2%, and the prevalence of early division of renal artery was 20.2%. No significant association was found between gender and laterality in the prevalence of both early division and accessory renal artery (p > 0.05). Early division of renal artery was more frequent in females (28%) whilst accessory renal arteries were more common in males (40.7%). CONCLUSIONS: The present study showed a consistent peri-hilar branching pattern with high individual variability among Indian renal donors. The knowledge about the peri-hilar branching pattern as well as renal artery variations conferred by this study would greatly compliment urological surgeons during renal transplant surgeries.

Antcins from Antrodia cinnamomea and Antrodia salmonea Inhibit Angiotensin-Converting Enzyme 2 (ACE2) in Epithelial Cells: Can Be Potential Candidates for the Development of SARS-CoV-2 Prophylactic Agents.

Antcins are newly identified steroid-like compounds from Taiwan's endemic medicinal mushrooms Antrodia cinnamomea and Antrodia salmonea. Scientific studies of the past two decades confirmed that antcins have various pharmacological activities, including potent anti-oxidant and anti-inflammatory effects. The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused the coronavirus disease-2019 (COVID-19) pandemic and is characterized as a significant threat to global public health. It was recently identified that SARS-CoV-2 required angiotensin converting enzyme 2 (ACE2), a receptor which supports host cell entry and disease onset. Here, we report a novel function of antcins, in which antcins exhibit inhibitory effects on ACE2. Compared to the untreated control group, treatment with various antcins (antcin-A, antcin-B, antcin-C, antcin-H, antcin-I, and antcin-M) significantly inhibited ACE2 activity in cultured human epithelial cells. Indeed, among the investigated antcins, antcin-A, antcin-B, antcin-C, and antcin-I showed a pronounceable inhibition against ACE2. These findings suggest that antcins could be novel anti-ACE2 agents to prevent SARS-CoV-2 host cell entry and the following disease onset.

Impact of COVID-19 outbreak on the mental health status of undergraduate medical students in a COVID-19 treating medical college: a prospective longitudinal study.

BACKGROUND: The COVID-19 pandemic is found to affect the mental health of the population. Undergraduate medical students are especially prone to mental health disorders and hence could be more vulnerable to the impact of the pandemic. METHODS: A prospective longitudinal study was conducted on 217 undergraduate medical students in a medical college at Chennai, India. Depression, anxiety, and stress levels were recorded using Depression Anxiety Stress Scale 21 Items (DASS21) before and during the COVID-19 outbreak in India in December 2019 and June 2020, respectively. In the follow-up survey, in addition to DASS21, the Pittsburgh Sleep Quality Index to assess sleep quality and a self-administered questionnaire to assess the impact of COVID-19 related stressors were used. The self-administered questionnaire assessed the status of COVID-19 testing, interactions with COVID-19 patients, self-perceived levels of concerns and worries related to academics (COVID-19-AA (academic apprehensions)) and those pertaining to the self and family/friends (COVID-19-GA (general apprehensions)). Cross-sectional and longitudinal comparison of overall scores of depression, anxiety, and stress and scores stratified by gender, year of study, place of residence and monthly family income were performed. Predictors for depression, anxiety, and stress during COVID-19 were investigated using adjusted binary logistic regression analysis and results were expressed as adjusted odds ratio with 95% confidence interval (CI). A P value < 0.05 was considered statistically significant. RESULTS: The average scores of depression, anxiety, and stress during the baseline survey were 7.55 ± 7.86, 4.6 ± 6.19 and 7.31 ± 7.34 with the prevalence (95% Cl) of 33.2% [27-39.9%], 21.2% [16-27.2%] and 20.7% [15.5-26.7%]; in follow-up survey, the mean scores were 8.16 ± 8.9, 6.11 ± 7.13 and 9.31 ± 8.18 with the prevalence being 35.5% [29.1-42.2%], 33.2% [27-39.9%] and 24.9% [19.3-31.2%] for depression, anxiety, and stress respectively. There was a significant increase in both the prevalence and levels of anxiety and stress (P < 0.001), with depression remaining unchanged during COVID-19, irrespective of gender, year of study, place of residence and family's monthly income. Poor sleep quality, higher levels of baseline depression, anxiety, and stress, higher COVID-19-GA, COVID-19 patients in family/friends and direct interactions with COVID-19 patients were found to be significant predictors of negative mental health in undergraduate medical students. COVID-19-AA was not significantly associated with depression, anxiety, and stress. CONCLUSION: The COVID-19 pandemic appears to negatively affect the mental health of the undergraduate medical students with the prevalence and levels of anxiety and stress being increased, and depression symptoms remaining unaltered. Addressing and mitigating the negative effect of COVID-19 on the mental health of this population is crucial.

Geranium and Lemon Essential Oils and Their Active Compounds Downregulate Angiotensin-Converting Enzyme 2 (ACE2), a SARS-CoV-2 Spike Receptor-Binding Domain, in Epithelial Cells.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), also known as coronavirus disease-2019 (COVID-19), is a pandemic disease that has been declared as modern history's gravest health emergency worldwide. Until now, no precise treatment modality has been developed. The angiotensin-converting enzyme 2 (ACE2) receptor, a host cell receptor, has been found to play a crucial role in virus cell entry; therefore, ACE2 blockers can be a potential target for anti-viral intervention. In this study, we evaluated the ACE2 inhibitory effects of 10 essential oils. Among them, geranium and lemon oils displayed significant ACE2 inhibitory effects in epithelial cells. In addition, immunoblotting and qPCR analysis also confirmed that geranium and lemon oils possess potent ACE2 inhibitory effects. Furthermore, the gas chromatography-mass spectrometry (GC-MS) analysis displayed 22 compounds in geranium oil and 9 compounds in lemon oil. Citronellol, geraniol, and neryl acetate were the major compounds of geranium oil and limonene that represented major compound of lemon oil. Next, we found that treatment with citronellol and limonene significantly downregulated ACE2 expression in epithelial cells. The results suggest that geranium and lemon essential oils and their derivative compounds are valuable natural anti-viral agents that may contribute to the prevention of the invasion of SARS-CoV-2/COVID-19 into the human body.

Regulation of pro and anti-inflammatory signaling molecules in effect of Withania Somnifera root extract treated sleep deprivation induced Wistar rats.

Sleep plays an imperative role in maintaining good health. Sleep along with circadian cycle wields strong regulatory control over immunity. Sleep deprivation (SD) is a threat to health developing several immunological disorders. The medicinal plant Withania somnifera (WS) root extract is widely used for its immuno-modulatory properties. Therefore, it is of interest to assess the effect of WS root extract on pro and anti-inflammatory signalling in SD rats. 24 male Wistar rats (120-150g) were divided into 4 groups with 6 animals in each. The groups were divided such that Group I - cage control, Group II - large platform control, Group III - sleep deprived & Group IV - WS treated SD rats. RT-PCR based mRNA expression analysis of pro inflammatory (IL-1ß, IL-6, MCP-1, TNF-α) and anti-inflammatory marker (IL-10) in the cortex of control and SD rats were completed. Concurrent protein expression analysis was completed using western blot. Data was analyzed using one-way ANOVA and Duncan's multiple range test in SPSS software version 20. Data showed elevation of pro-inflammatory markers and depression of IL-10. Thus, WS down regulated the pro-inflammatory and up-regulated the anti-inflammatory molecules, which can be further considered towards the treatment of sleep deprivation induced inflammatory diseases.

A mechanistic and empirical review of antcins, a new class of phytosterols of formosan fungi origin.

Antcins are unique phytosterols isolated from A. cinnamomea and A. salmomea, which are the endemic fungus of Taiwan. A. cinnamomea has long been highly valued medicinal mushroom in Taiwan and traditionally used as a folk remedy for various human illness. Recent scientific explorations claimed that the pharmacological activities of A. cinnamomea and A. salmomonea are gone beyond their original usage. The therapeutic efficacy of these medicinal mushrooms was attributed to their high content of unique bioactive secondary metabolites, including terpenoids, benzenoids, ubiquinol derivatives, polysaccharides, lignans, nucleic acids, steroids, and maleic/succinic acid derivatives. Antcins is a group of steroids in Antrodia spp. with ergostane skeleton received much attention in Taiwan's academic circle due to their broad-spectrum of biological activities. At present, twelve antcins, i.e. antcin A, B, C, D, E, F, G, H, I, K, M, and N along with twelve derivatives/epimers (25R/S-antcin A, B, C, H, I and K) and seven analogs (methyl antcinate A, B, G, H, K, L and N) were identified. Several studies have demonstrated that antcins possessed anti-cancer, anti-inflammation, anti-oxidant, anti-diabetic, anti-aging, immunomodulation, hepatoprotection, and hypolipedimic activities. The main goal of this review is to define the chemistry, isolation, advances in production, and biological activities of antcins and their derivatives/analogs. Special attention has been given to a detail view of their biological activities in vitro and in vivo and their pharmacological potentials.

Lung Cancer Detection Using Image Segmentation by means of Various Evolutionary Algorithms.

The objective of this paper is to explore an expedient image segmentation algorithm for medical images to curtail the physicians' interpretation of computer tomography (CT) scan images. Modern medical imaging modalities generate large images that are extremely grim to analyze manually. The consequences of segmentation algorithms rely on the exactitude and convergence time. At this moment, there is a compelling necessity to explore and implement new evolutionary algorithms to solve the problems associated with medical image segmentation. Lung cancer is the frequently diagnosed cancer across the world among men. Early detection of lung cancer navigates towards apposite treatment to save human lives. CT is one of the modest medical imaging methods to diagnose the lung cancer. In the present study, the performance of five optimization algorithms, namely, k-means clustering, k-median clustering, particle swarm optimization, inertia-weighted particle swarm optimization, and guaranteed convergence particle swarm optimization (GCPSO), to extract the tumor from the lung image has been implemented and analyzed. The performance of median, adaptive median, and average filters in the preprocessing stage was compared, and it was proved that the adaptive median filter is most suitable for medical CT images. Furthermore, the image contrast is enhanced by using adaptive histogram equalization. The preprocessed image with improved quality is subject to four algorithms. The practical results are verified for 20 sample images of the lung using MATLAB, and it was observed that the GCPSO has the highest accuracy of 95.89%.

MicroRNA-708 activation by glucocorticoid receptor agonists regulate breast cancer tumorigenesis and metastasis via downregulation of NF-κB signaling.

Therapeutic administration of glucocorticoids (GCs) is frequently used as add-on chemotherapy for palliative purposes during breast cancer treatment. Recent studies have shown that GC treatment induces microRNA-708 in ovarian cancer cells, resulting in impaired tumor cell proliferation and metastasis. However, the regulatory functions of GCs on miR-708 and its downstream target genes in human breast cancer cells (BCCs) are poorly understood. In this study, we found that treatment with either the synthetic GC dexamethasone (DEX) or the natural GC mimic, antcin A (ATA) significantly increased miR-708 expression by transactivation of glucocorticoid receptor alpha (GRα) in MCF-7 and MDA-MB-231 human BCCs. Induction of miR-708 by GR agonists resulted in inhibition of cell proliferation, cell-cycle progression, cancer stem cell (CSC)-like phenotype and metastasis of BCCs. In addition, GR agonist treatment or miR-708 mimic transfection remarkably inhibited IKKß expression and suppressed nuclear factor-kappaB (NF-κB) activity and its downstream target genes, including COX-2, cMYC, cyclin D1, Matrix metalloproteinase (MMP)-2, MMP-9, CD24, CD44 and increased p21CIP1 and p27KIP1 that are known to be involved in proliferation, cell-cycle progression, metastasis and CSC marker protein. BCCs xenograft models indicate that treatment with GR agonists significantly reduced tumor growth, weight and volume. Overall, our data strongly suggest that GR agonists induced miR-708 and downstream suppression of NF-κB signaling, which may be applicable as a novel therapeutic intervention in breast cancer treatment.

Severe Respiratory Illness Associated with Human Metapneumovirus in Nursing Home, New Mexico, USA.

Human metapneumovirus is an emerging pathogen that causes upper and lower respiratory illness. Nursing home outbreaks of infection with this virus can cause severe illness and lead to poor patient outcomes. We report an outbreak investigation in a nursing home during 2018 and infection control guidelines to assist in disease control.

Receive diversity based transmission data rate optimization for improved network lifetime and delay efficiency of Wireless Body Area Networks.

Wireless Body Area Network (WBAN) has become the emerging technology due to its ability to provide intelligent and cost-effective healthcare monitoring solution. The biological sensors used in WBAN are energy-constrained and required to be functional for a longer duration. Also, the sensed data should be communicated in reasonable time. Therefore, network lifetime and delay have become the primary concerns in the design of WBAN. In this paper, Receive Diversity based Transmission Data Rate Optimization (RDTDRO) scheme is proposed to improve the network lifetime and delay efficiency of Multi level-Quadrature Amplitude Modulation (M-QAM) based WBAN. In the proposed RDTDRO scheme, minimum energy consumption is ensured by optimizing the transmission data rate with respect to a given transmission distance and number of receive antennas while satisfying the Bit Error Rate (BER) requirements. The performance of proposed RDTDRO is analyzed in terms of network lifetime and delay difference and is compared with conventional Baseline and Rate optimized schemes. The results show that at a transmission distance of 0.3 m, the proposed RDTDRO scheme with a receive diversity order of 4 achieves 1.30 times and 1.27 times improvement in network lifetime over conventional Baseline and Rate optimized schemes respectively. From the results, it is also evident that at a transmission distance of 0.3 m, the proposed RDTDRO scheme with a receive diversity order of 4 is delay efficient as it achieves delay difference of 0.75 µs and 0.29 µs over conventional Baseline and Rate optimized schemes respectively.

Activation of Nrf2-mediated anti-oxidant genes by antrodin C prevents hyperglycemia-induced senescence and apoptosis in human endothelial cells.

In the present study, we investigated the effects of antrodin C (ADC), a maleimide derivative isolated from mycelia of Antrodia cinnamomea, on high glucose (HG, 30 mM)-accelerated endothelial dysfunction in vitro. HG-induced cytotoxicity in human umbilical vein endothelial cells (HUVECs) was significantly ameliorated by ADC. In addition, treatment with ADC significantly prevented HG-induced senescence, growth arrest at the G1-S transition phase and apoptosis in HUVECs. Moreover, the increased level of intracellular reactive oxygen species (ROS) under HG condition was significantly ameliorated by ADC. Further analysis revealed that ADC-mediated anti-oxidant effects were due to up-regulation of cellular anti-oxidant genes, such as HO-1 and NQO-1 via promotion of the transcriptional activity of Nrf2, which was further confirmed by the failure of ADC to protect HUVECs from HG-induced dysfunction under HO-1 inhibition or Nrf2 silencing. Furthermore, hyperosmotic glucose (HOG, 60 mM)-induced uncontrolled production of ROS, rapid apoptotic cell death and HUVEC injury were significantly prevented by ADC, whereas these preventive effects were barely observed in HO-1 inhibited or Nrf2 silenced cells. Taken together, these results suggest that ADC may represent a promising intervention in diabetic-associated cardiovascular diseases by activating the Nrf2-dependent cellular anti-oxidant defense system.

A steroid like phytochemical Antcin M is an anti-aging reagent that eliminates hyperglycemia-accelerated premature senescence in dermal fibroblasts by direct activation of Nrf2 and SIRT-1.

The present study revealed the anti-aging properties of antcin M (ANM) and elucidated the molecular mechanism underlying the effects. We found that exposure of human normal dermal fibroblasts (HNDFs) to high-glucose (HG, 30 mM) for 3 days, accelerated G0/G1 phase arrest and senescence. Indeed, co-treatment with ANM (10 µM) significantly attenuated HG-induced growth arrest and promoted cell proliferation. Further molecular analysis revealed that ANM blocked the HG-induced reduction in G1-S transition regulatory proteins such as cyclin D, cyclin E, CDK4, CDK6, CDK2 and protein retinoblastoma (pRb). In addition, treatment with ANM eliminated HG-induced reactive oxygen species (ROS) through the induction of anti-oxidant genes, HO-1 and NQO-1 via transcriptional activation of Nrf2. Moreover, treatment with ANM abolished HG-induced SIPS as evidenced by reduced senescence-associated ß-galactosidase (SA-ß-gal) activity. This effect was further confirmed by reduction in senescence-associated marker proteins including, p21CIP1, p16INK4A, and p53/FoxO1 acetylation. Also, the HG-induced decline in aging-related marker protein SMP30 was rescued by ANM. Furthermore, treatment with ANM increased SIRT-1 expression, and prevented SIRT-1 depletion. This protection was consistent with inhibition of SIRT-1 phosphorylation at Ser47 followed by blocking its upstream kinases, p38 MAPK and JNK/SAPK. Further analysis revealed that ANM partially protected HG-induced senescence in SIRT-1 silenced cells. A similar effect was also observed in Nrf2 silenced cells. However, a complete loss of protection was observed in both Nrf2 and SIRT-1 knockdown cells suggesting that both induction of Nrf2-mediated anti-oxidant defense and SIRT-1-mediated deacetylation activity contribute to the anti-aging properties of ANM in vitro. Result of in vivo studies shows that ANM-treated C. elegens exhibits an increased survival rate during HG-induced oxidative stress insult. Furthermore, ANM significantly extended the life span of C. elegans. Taken together, our results suggest the potential application of ANM in age-related diseases or as a preventive reagent against aging process.

Upper and lower pharyngeal airway space in West-Tamil Nadu population.

AIM: To compare the upper and lower pharyngeal airway (LPA) width in Class II malocclusion patients with low, average, and high vertical growth patterns. STUDY DESIGN: Cross-sectional analytical study. MATERIALS AND METHODS: Pretreatment lateral cephalometric films of 90 Class II subjects were used to measure the upper and LPAs. The inclusion criteria were subjects of West-Tamil Nadu, aged between 14 and 25 years, only skeletal Class II subjects of either gender and no pharyngeal pathology at initial visit. The sample comprised a total of 90 Class II subjects divided into three groups according to the vertical facial pattern: Normodivergent (n = 30), hypodivergent (n = 30), and hyperdivergent (n = 30). The assessment of upper and LPAs was done according to McNamara's airway analysis. STATISTICAL ANALYSIS: The intergroup comparison of the upper and LPAs was performed with one-way analysis of variance and the Tukey test was used to compare among the various vertical patterns. RESULTS: Skeletal Class II subjects with hyperdivergent facial pattern showed statistically significant narrow upper pharyngeal width when compared to normodivergent and hypodivergent facial patterns. No statistically significant difference was found in the lower pharyngeal width in all three vertical facial growth patterns. CONCLUSION: Subjects with Class II malocclusions and hyperdivergent growth pattern have significantly narrow upper pharyngeal airway space when compared to other two vertical patterns. Narrow pharyngeal airway space is one of the predisposing factors for mouth breathing and obstructive sleep apnea.

2,3,5-Trimethoxy-4-cresol, an anti-metastatic constituent from the solid-state cultured mycelium of Antrodia cinnamomea and its mechanism.

Antrodia cinnamomea is a valuable and unique edible fungus originating from the forests of Taiwan. In this study, an anti-metastatic compound, 2,3,5-trimethoxy-4-cresol (TMC), was isolated from the solid-state cultured mycelium of A. cinnamomea. According to the results obtained from cell wound healing, cell migration and invasion assays, TMC effectively suppressed movement, migration and invasion of lung cancer cells at the dosage of 5-40 µM, which was non-toxic to A549 cells. In addition, TMC reduced protein expression of Akt, MMP-2 and MMP-9 and enhanced E-cadherin and TIMP-1 protein expression, which are known to regulate cell adhesion, migration and invasion. Taken together, TMC effectively suppresses movement, migration and invasion of lung cancer cells, and achieves an anti-cancer metastasis effect.

Investigation on antidiarrhoeal activity of Aristolochia indica Linn. Root extracts in mice.

BACKGROUND: The present study aimed at investigating the effect of ethanolic extract (EtAI), and aqueous extract (AqAI) of Aristolochia indica Linn roots on castor oil-induced diarrhoea and study on small intestinal transit. Phytochemical analysis of extracts was performed as per standard procedure. MATERIALS AND METHODS: The oral toxicity study using Swiss albino mice was performed in accordance with OECD guidelines. The EtAI and AqAI extracts of Aristolochia indica Linn were studied for antidiarrhoeal property using castor oil-induced diarrhoeal model and charcoal-induced gastrointestinal motility test in Swiss albino mice. RESULTS: Among the tested doses of 200 and 400 mg/kg body weight, the extracts reduced the frequency and severity of diarrhoea in test animals throughout the study period. At the same doses, the extract delayed the intestinal transit of charcoal meal in test animals as compared to the control and the results were statistically significant. CONCLUSION: Experimental findings showed that ethanol extract of Aristolochia indica Linn root possess significant antidiarrheal activity and may be a potent source of anti-diarrhoeal drug in future.

Kalicephalus sp. in a captive Russell's viper: a case report.

The occurrence of Kalicephalus sp. of hookworms in a Russell's viper snake maintained in Chennai snake park trust, Chennai is reported. These worms were found in the stomach and intestine. They were cylindrical, thread like and had an elongate body. Anterior end was obliquely truncated with bivalvular buccal capsule. Short, thick and muscularised oesophagus ending in a rounded bulb was observed. In males, a well developed trilobed bursa was observed and the spicules were short and equal. Typical strongyle type eggs were found in the uterus of female worms as well as in the intestinal contents.

Antrodia salmonea in submerged culture exhibits antioxidant activities in vitro and protects human erythrocytes and low-density lipoproteins from oxidative modification.

Antrodia salmonea is well known in Taiwan as a beneficial mushroom. In the present study, we investigated the antioxidant activity of whole fermented broth (AS), filtrate (ASF), and mycelia (ASM) of A. salmonea using different antioxidant models. Furthermore, the effect of A. salmonea on AAPH-induced oxidative hemolysis of human erythrocytes and CuSO4-induced oxidative modification of human low-density lipoproteins (LDLs) was examined. We found that the AS, ASF, and ASM possess effective antioxidant activity against various oxidative systems including superoxide anion scavenging, reducing power, metal chelation, and DPPH radical scavenging. Further, AAPH-induced oxidative hemolysis in erythrocytes was prevented by AS, ASF, and ASM. Notably, AS, ASF, and ASM appear to possess powerful antioxidant activities against CuSO4-induced oxidative modification of LDL as assessed by malondialdehyde (MDA) formation, cholesterol degradation, and the relative electrophoretic mobility of oxidized LDL. It is noteworthy that AS had comparatively strong antioxidant ability compared to ASF or ASM, which is well correlated with the content of their total polyphenols. Thus, A. salmonea may exert antioxidant properties and offer protection from atherogenesis.

Antrodia salmonea inhibits TNF-α-induced angiogenesis and atherogenesis in human endothelial cells through the down-regulation of NF-κB and up-regulation of Nrf2 signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Antrodia salmonea (AS) is known as a traditional Chinese medicine, but very few biological activities have been reported. MATERIALS AND METHODS: The present study was aimed to investigate the anti-angiogenic and anti-atherosclerotic potential of the fermented culture broth of AS against tumor necrosis factor-α (TNF-α)-stimulated human endothelial (EA.hy 926) cells. RESULTS: The non-cytotoxic concentrations of AS significantly inhibited TNF-α-induced migration/invasion and capillary-like tube formation in EA.hy 926 cells. Furthermore, AS suppressed TNF-α-induced activity and expression of matrix metalloproteinase-9 (MMP-9), and cell-surface expression of intercellular adhesion molecule-1 (ICAM-1), which was associated with abridged adhesion of U937 leukocytes to endothelial cells. Moreover, AS significantly down-regulated TNF-α-induced nuclear translocation and transcriptional activation of nuclear factor κB (NF-κB) followed by suppression of I-κB degradation and phosphorylation of I-κB kinase-α (IKKα). Notably, the protective effect of AS was directly correlated with the increased expression of hemeoxygenase-1 (HO-1) and γ-glutamylcysteine synthetase (γ-GCLC), which was reasoned by nuclear translocation and transactivation of NF-E2 related factor-2 (Nrf2)/antioxidant response element (ARE). Furthermore, HO-1 knockdown by HO-1-specific shRNA diminished the protective effects of AS on TNF-α-stimulated invasion, tube formation, and U937 adhesion in EA.hy 926 cells. CONCLUSIONS: Taken together, these results suggest that Antrodia salmonea may be useful for the prevention of angiogenesis and atherosclerosis.

Humic acid in drinking well water induces inflammation through reactive oxygen species generation and activation of nuclear factor-κB/activator protein-1 signaling pathways: a possible role in atherosclerosis.

Humic acid (HA) has been implicated as one of the etiological factors in the peripheral vasculopathy of blackfoot disease (BFD) in Taiwan. However, the underlying pathophysiological mechanisms of BFD are not well defined. In this study, we used an in vitro and in vivo model, in which HA (25-200µg/mL) activated macrophages to produce pro-inflammatory molecules by activating their transcriptional factors. HA exposure induced NO and PGE2 production followed by induction of iNOS and COX-2 through NF-κB/AP-1 transactivation in macrophages. In addition, the production of TNF-α and IL-1ß was significantly increased by HA. Moreover, HA-induced iNOS and COX-2 expression were down-regulated by the NF-κB and AP-1 inhibitors pyrrolidine dithiocarbamate (PDTC) and Tanshinone, respectively. Furthermore, generations of ROS and nitrotyrosine, as well as activation of the AKT and MAPKs signaling cascades were observed after HA exposure. Specifically, HA-induced NF-κB activation was mediated by ROS and AKT, and that HA-induced AP-1 activation was mediated by JNK and ERK. Notably, HA-mediated AKT, JNK, and ERK activation was ROS-independent. The inflammatory potential of HA was correlated with increased expression of HO-1 and Nrf2. Furthermore, an in vivo study confirms that mice exposed to HA, the serum levels of TNF-α and IL-1ß was significantly increased in a dose-dependent manner. This report marks the first confirmation that environmental exposure of HA induces inflammation in macrophages, which may be one of the main causes of early atherogenesis in blackfoot disease.