First report of the Horse nettle virus infecting tomato (Solanum lycopersicum L.) in the United States.

Tomato (Solanum lycopersicum L.), a member of the Solanaceae family, represents one of the most extensively cultivated vegetable species worldwide and traces its origin to western South America (Caruso et al. 2022). In a field survey conducted in 2023 in Bixby, Tulsa County, Oklahoma, distinct symptoms were noted in two plants: one exhibited mottling and cupping of leaves and brown discoloration on leaves, petioles, and stems, while the other displayed a downward curling of leaves. Leaf samples from both symptomatic tomato plants (labelled as K4 and K5) were collected, and total RNA was extracted individually via the TRI Reagent® method (Molecular Research Center Inc., Cincinnati, OH, USA). Subsequently, the RNA samples were pooled and subjected to high-throughput sequencing (HTS) on the NextSeq 500/550 high-output kit v2.5 (Illumina, U.S.A.) at the genomic facility, Oklahoma State University (Stillwater, OK). Total read count of 8,227,020 (average length =150.5 bp) was obtained, trimmed, and de novo assembled using CLC Genomics Workbench v22.0.1 (QIAGEN) and used for BLASTn and BLASTx analysis. Two contigs: 6,375 bp (average coverage 2,915.92, read count 142,538) and 3,564 bp (average coverage 3,035.91, read count 82,370) from the pooled sample showed 88.6% and 96.7% nucleotide identities with RNA 1 (OP292294) and RNA 2 (OP292295) of Horse nettle virus A (HNA-A) isolate MD-1, respectively. Sequences of both partial contigs (RNA 1, accession no. PP063196) and RNA 2, accession no. PP063197) were submitted to GenBank. The HTS data did not reveal any other viral or viroid sequences in these two tomato samples. To further confirm the presence of HNV-A, total RNA from K4 and K5 samples was tested individually by RT-PCR using HNV specific primers (Supplementary Table 1) based on the two partial contig sequences. The expected PCR products (491 bp and 451 bp) were obtained only from the K4 sample and none from the K5 sample. PCR products were extracted from an agarose gel, cloned into the pGEM®-T Easy vector (Promega), and transformed into Escherichia coli DH5α cells (New England Bio Labs). Two clones for each PCR product were sequenced by Sanger sequencing. Nucleotide sequence comparisons and BLASTn analysis of 491 bp and 451 bp showed 86% and 97% nucleotide identity with RNA 1 and RNA 2 of HNV-A isolate MD-1 (OP292294 and OP292295), respectively. Additionally, eight more leaf samples from eight different symptomatic tomato plants were collected in the same field and tested by RT-PCR as described above. All eight samples were positive by RT-PCR, but no PCR band was obtained in the total RNA from a healthy tomato leaf used as a control. Sequences from the PCR products were identical to the obtained HTS sequences. Our results confirmed for the first time that HNV-A can infect tomatoes. Currently, HNV-A has been reported to only infect a single weed (Horse nettle, Solanum carolinense) (Zhou et al. 2023). The identification of HNV-A in tomatoes in Oklahoma suggests a potential host shift is of concern for local growers as well as tomato growers worldwide. This shift underscores the urgency for an in-depth investigation into the transmission and host specificity of HNV-A. This is the first report in the United States and the world that HNV-A could infect tomatoes naturally in a grower field.

Optimized Extraction of Polyphenols from Unconventional Edible Plants: LC-MS/MS Profiling of Polyphenols, Biological Functions, Molecular Docking, and Pharmacokinetics Study.

Plant bioactive phenolic metabolites have recently attracted the attention of researchers due to their numerous health advantages. Therefore, this study aimed to investigate with advanced techniques the bioactive metabolites and antioxidant and antidiabetic capacity of four unconventional edible plant leaves: lemongrass (Cymbopogon citratus (DC.) Stapf), chicory (Cichorium intybus L.), moringa (Moringa oleifera Lam.), and ryegrass (Lolium perenne L.). The extraction process was optimized using different solvents. These plants' phenolic composition, identification, and characterization have been determined herein using LCESI-QTOF-MS/MS. This research identified 85 phenolic compounds, including 24 phenolic acids, 31 flavonoids, 7 stilbenes and lignans, and 17 other metabolites. Moreover, the study determined that moringa has the highest total phenolic content (TPC; 18.5 ± 1.01 mg GAE/g), whereas ryegrass has the lowest (3.54 ± 0.08 mg GAE/g) among the selected plants. It seems that, compared to other plants, moringa was found to have the highest antioxidant potential and antidiabetic potential. In addition, twenty-two phenolic compounds were quantified in these chosen edible plants. Rosmarinic acid, chlorogenic acid, chicoric acid, ferulic acid, protocatechuic acid, and caffeic acid were the most abundant phenolic acids. In silico molecular docking was also conducted to investigate the structure-function relationship of phenolic compounds to inhibit the alpha-glucosidase. Finally, the simulated pharmacokinetic characteristics of the most common substances were also predicted. In short, this investigation opens the way for further study into these plants' pharmaceutical and dietary potential.

Probiotic Yoghurt Enriched with Mango Peel Powder: Biotransformation of Phenolics and Modulation of Metabolomic Outputs after In Vitro Digestion and Colonic Fermentation.

This study investigated the health-promoting effects and prebiotic functions of mango peel powder (MPP) both as a plain individual ingredient and when incorporated in yoghurt during simulated digestion and fermentation. The treatments included plain MPP, plain yoghurt (YA), yoghurt fortified with MPP (YB), and yoghurt fortified with MPP and lactic acid bacteria (YC), along with a blank (BL). The identification of polyphenols in the extracts of insoluble digesta and phenolic metabolites after the in vitro colonic fermentation were performed employing LC-ESI-QTOF-MS2. These extracts were also subjected to pH, microbial count, production of SCFA, and 16S rRNA analyses. The characterisation of phenolic profiles identified 62 phenolic compounds. Among these compounds, phenolic acids were the major compounds that underwent biotransformation via catabolic pathways such as ring fission, decarboxylation, and dehydroxylation. Changes in pH indicated that YC and MPP reduced the media pH from 6.27 and 6.33 to 4.50 and 4.53, respectively. This decline in pH was associated with significant increases in the LAB counts of these samples. The Bifidobacteria counts were 8.11 ± 0.89 and 8.02 ± 1.01 log CFU/g in YC and MPP, respectively, after 72 h of colonic fermentation. Results also showed that the presence of MPP imparted significant variations in the contents and profiles of individual short chain fatty acids (SCFA) with more predominant production of most SCFA in the MPP and YC treatments. The 16s rRNA sequencing data indicated a highly distinctive microbial population associated with YC in terms of relative abundance. These findings suggested MPP as a promising ingredient for utilisation in functional food formulations aiming to enhance gut health.

Characterization, Antioxidant Potential, and Pharmacokinetics Properties of Phenolic Compounds from Native Australian Herbs and Fruits.

In recent decades, plant bioactive phenolic compounds gained much attention due to their various health benefits. Therefore, this study aimed to analyze native Australian river mint (Mentha australis), bush mint (Mentha satureioides), sea parsley (Apium prostratum), and bush tomatoes (Solanum centrale) for their bioactive metabolites, antioxidant potential, and pharmacokinetics properties. LC-ESI-QTOF-MS/MS was applied to elucidate these plants' composition, identification, and quantification of phenolic metabolites. This study tentatively identified 123 phenolic compounds (thirty-five phenolic acids, sixty-seven flavonoids, seven lignans, three stilbenes, and eleven other compounds). Bush mint was identified with the highest total phenolic content (TPC-57.70 ± 4.57 mg GAE/g), while sea parsley contained the lowest total phenolic content (13.44 ± 0.39 mg GAE/g). Moreover, bush mint was also identified with the highest antioxidant potential compared to other herbs. Thirty-seven phenolic metabolites were semi-quantified, including rosmarinic acid, chlorogenic acid, sagerinic acid, quinic acid, and caffeic acid, which were abundant in these selected plants. The most abundant compounds' pharmacokinetics properties were also predicted. This study will develop further research to identify these plants' nutraceutical and phytopharmaceutical potential.

Identification and characterization of anthocyanins and non-anthocyanin phenolics from Australian native fruits and their antioxidant, antidiabetic, and anti-Alzheimer potential.

Polyphenols are vital bioactive constituents that have beneficial effects on human health. The aim of this study was to characterize the biologically active phenolic metabolites in Australian native commercial fruits (Kakadu plum, Davidson's plum, quandong peach, and muntries) and their antioxidant, α-glucosidase, and acetylcholinesterase inhibition activities. Polyphenols were measured through total phenolic content (TPC), total flavonoid content (TFC), total condensed tannin (TCT), and total monomeric anthocyanin content (TMAC). Moreover, different in-vitro biological assays (DPPH, ABTS, FICA, OH-RSA, α-glucosidase, and acetylcholinesterase inhibition activities) were conducted to measure the antioxidant, anti-diabetic, and anti-Alzheimer's potential of these selected fruits. LC-ESI-QTOF-MS/MS was implied for identification and quantification purposes. In this study, a total of 307 bioactive metabolites (51 phenolic acids, 194 flavonoids, 15 tannins, 23 other polyphenols, 5 stilbenes, 12 lignans, and 7 terpenoids) were putatively identified. A total of 41 phenolic compounds were quantified/semi-quantified. Kakadu plum was identified with a higher concentration of polyphenols and biological activities compared to Davidson plum, quandong peach, and muntries. Molecular docking was also conducted to discover the actual role of the most abundant phenolic metabolites in the α-glucosidase and acetylcholinesterase inhibition activities.

Phytochemicals, Antioxidant Activities, and Toxicological Screening of Native Australian Fruits Using Zebrafish Embryonic Model.

Phytochemicals play a pivotal role in human health and drug discovery. The safety evaluation of plant extracts is a prerequisite to ensure that all phytochemicals are safe before translational development and human exposure. As phytochemicals are natural, they are generally considered safe, although this is not always true. The objective of this study was to investigate and compare the phytochemical composition, antioxidant potential, and safety evaluation of native Australian Muntries (Kunzea pomifera), Kakadu plum (Terminalia ferdinandiana), Davidson plum (Davidsonia) and Quandong peach (Santalum acuminatum) through the in vivo vertebrate zebrafish embryonic model. The highest total phenolic content (TPC; 793.89 ± 22.27 µg GAE/mg) was quantified in Kakadu plum, while the lowest TPC (614.44 ± 31.80 µg GAE/mg) was quantified in Muntries. Developmental alterations, mortality, and morbidity were assessed for toxicological screening of these selected native Australian fruit extracts. In this study, muntries were quantified as having the least LC50 value (169 mg/L) compared to Davidson plum (376 mg/L), Kakadu plum (>480 mg/L), and Quandong peach (>480 mg/L), which indicates that muntries extract was more toxic than other fruit extracts. Importantly, we found that adverse effects were not correlated to the total phenolic content and antioxidant potential of these native Australian fruits and cannot simply be predicted from the in vitro analysis. Conclusively, these selected native Australian fruit extracts are categorized as safe. This study could explore the use of these native Australian fruits in cosmetics, pharmaceuticals, and drug discovery.

LC-MS/MS Characterization of Phenolic Metabolites and Their Antioxidant Activities from Australian Native Plants.

Polyphenols are considered vital bioactive compounds beneficial for human health. The Australian flora is enriched with polyphenols which are not fully characterized yet. Thus, the main objective of this study was to identify and characterize the Australian native sandalwood nuts, wattle seeds, lemongrass, and old man saltbush for phenolic compounds and their antioxidant activities. In this study, we tentatively identified a total of 155 phenolic compounds including 25 phenolic acids, 55 flavonoids, 22 isoflavonoids, 22 tannins, 22 lignans, 33 stilbenes, 33 coumarins and derivatives, 12 tyrosols and derivatives, and 6 phenolic terpenes. The highest total phenolic content (TPC) (15.09 ± 0.88 mg GAE/g) was quantified in lemongrass, while the lowest TPC (4.17 ± 0.33 mg GAE/g) was measured in wattle seeds. The highest total flavonoid content (TFC) and total condensed tannins (TCT) were measured in lemongrass and wattle seeds, respectively. A total of 18 phenolic metabolites were quantified/semi-quantified in this experiment. Lemongrass contains a vast number of phenolic metabolites.

A Comparative Study for Nutritional and Phytochemical Profiling of Coffea arabica (C. arabica) from Different Origins and Their Antioxidant Potential and Molecular Docking.

Coffee is the most widely used beverage globally and contains many bioactive compounds, including phenolic compounds, alkaloids, triterpenes, organic acids, amino acids, hormones, and fatty acids. The main objective of this study was the comparative profiling of Australian, Colombian, Ethiopian, and Peruvian C. arabica using LC-ESI-QTOF-MS/MS. In this study, we tentatively identified 136 bioactive metabolites, including five (05) organic acids, six (06) alkaloids, three (03) amino acids (l-phenylalanine, l-tyrosine, and l-pyroglutamic acid), two (02) hormones (melatonin and serotonin), two fatty acids, one (01) furopyrans (goniothalenol), one (01) carotenoid (crocetin), three (03) terpenoids, thirty-eight (38) phenolic acids, forty-one (41) flavonoids, five (05) stilbenes, three (03) lignans and twenty-three (23) other polyphenols in C. arabica. The highest TPC value (17.74 ± 0.32 mg GAE/g) was measured in Colombian coffee while the lowest TPC value (10.24 ± 0.73 mg GAE/g) was in Peruvian coffee. Colombian coffee has a higher antioxidant potential than other studied coffee samples. A total of nineteen phenolic metabolites were mapped through LC-MS/MS. Quinic acid derivatives were quantified in higher concentrations than other metabolites. Furthermore, molecular docking predicted that chlorogenic acid is a main bioactive compound that contributes to anti-Alzheimer and anti-diabetic activities of C. arabica. The obtained results indicate that C. arabica contains a vast number of bioactive compounds which have potential health benefits. Furthermore, research could be conducted to validate the effect of these metabolites on the flavor profile of coffee beverages.

First Report of Tomato Yellow Leaf Curl Virus Infecting Pepper and Tomato in Oklahoma.

Tomato yellow leaf curl virus (TYLCV), (genus Begomovirus and family Geminiviridae) is one of the devastating viruses infecting tomatoes (Solanum Lycopersicon) and could have a significant impact on tomato production worldwide (Moriones and Navas-Castillo, 2000; Rybicki, 2015; Perez-Padilla et al. 2019). TYLCV was first reported in the United States (US) in 1997 in Florida (Polston et al. 1999) and since then it has been reported in several other states but not in Oklahoma (de Sa et al. 2008). Both pepper (Capsicum spp.) and tomato are grown in >50 counties of Oklahoma for fresh produce in the local market, and for processing industries. During a survey in 2021, pepper (bell pepper cv SV3964) and tomato (cv grand marshall) plants grown in a commercial field in Bixby, Oklahoma, showed typical virus-like symptoms including yellowing, leaf curling, cupping, twisting, and mottling (Fig. 1). Estimated disease incidence was 10% in both crops. In addition, whiteflies (Bemisia tabaci) were also observed on these plants. Twenty symptomatic samples from pepper (n=16) and tomato (n=4) plants, and four asymptomatic from pepper (n=2) and tomato (n=2) plants were collected and brought to the Plant Virology Lab at the University of Tulsa for further analysis Total RNA was extracted from all samples using the Spectrum Plant Total RNA Kit (Sigma-Aldrich, LOT: SLCG7913). RNA from two symptomatic samples, one each from pepper and tomato (named BX6 and BX11, respectively) were subjected to high-throughput sequencing (HTS) on the NextSeq 500/550 high-output kit v2.5 (Illumina, USA) at the genomic facility, Oklahoma State University (Stillwater, OK, USA). A total of 22,385,404 (average length 73.6 bp) and 19,012,605 (average length 73.7 bp) trim pair-end for both samples (BX6 and BX11) were assembled using CLC Genomics Workbench (v12.0.3) (Qiagen, Inc) and subjected to BLASTn analysis. The three contigs: 393 bp (coverage 363.24X), 670 bp (coverage 489.92X), and 990 bp (coverage 112.96X) for BX6 isolate, and four contigs: 393 bp (coverage 341.64X), 668 bp (coverage 457.23X), 814 bp (coverage 107.86X), and 990 bp (coverage 93.71X), for BX11 isolate, showed from 96 to 99% nucleotide (nt) identities with movement protein, capsid protein, C3, V2, C4, AC1, AC2, AC3 protein genes of the TYLCV isolates from Australia, China, Iran, Mexico, South Korea, and the US. The HTS data did not reveal any other viral sequence in these two samples. To further confirm the presence of TYLCV in these samples, three overlapping TYLCV-specific primer pairs (Supplementary Table 1) were designed based on the alignment of complete genome sequences of TYLCV isolates present in the Genbank. Total DNA was extracted from fresh leaf tissues of BX6 and BX11 samples using the Plant DNA Kit (Omega BIO-TEK,) and used in the PCR assay. The expected PCR products (814 bp, 1,085 bp and 1,068 bp) were generated from both samples providing further evidence of TYLCV infection. To obtain the complete genome sequence, these PCR amplified DNA fragments of both isolates were gel extracted and cloned using the pGEM-T® Easy Vector system. Three independent clones of each fragment were sequenced and analyzed by Sanger sequencing. The resulting consensus sequences were used in a BLASTn search against the GenBank and had the highest identities (99-99.2%) with TYLCV sequences. The overlapping consensus nt sequence of the BX6 isolate was 2,782 nt (Accession no ON321843), whereas that of BX11 was 2,777 nt (Accession no ON785706) showed 99.2% and 99.0% nt identities with the corresponding sequence of TYLCV isolates (KX347141 and KX347142) respectively from Australia. The nt identity between BX6 and BX11 was 99.7%. Further screening by PCR of the remaining 18 symptomatic field samples (pepper, n=15 and tomatoes, n=3) revealed TYLCV infection in two pepper and all three tomato samples consistent with the greater susceptibility of tomato. None of the asymptomatic pepper and tomato samples were positive for TYLCV. DNA from both BX6 and BX11 samples was also used in the rolling circle amplification (RCA) assay (TempliPhi Amplification Kit, Cytiva) and the presence of circular TYLCV DNA was detected. Our results demonstrated the presence of TYLCV infection in these symptomatic pepper and tomato plants. This is the first report of TYLCV infecting pepper or tomato in open fields in Oklahoma. Further surveys are needed to determine the incidence of TYLCV and the presence of its vector in other counties throughout Oklahoma.

Phytochemical and Safety Evaluations of Finger Lime, Mountain Pepper, and Tamarind in Zebrafish Embryos.

Plants play a pivotal role in drug discovery, constituting 50% of modern pharmacopeia. Many human diseases, including age-related degenerative diseases, converge onto common cellular oxidative stress pathways. This provides an opportunity to develop broad treatments to treat a wide range of diseases in the ageing population. Here, we characterize and assess the toxicological effects of finger lime (Citrus australasica), mountain pepper (Tasmannia lanceolata), and small-leaved tamarind (Diploglottis australis) extracts. The characterization demonstrates that these Australian native plants have antioxidant potential and, importantly, they have high concentrations of distinct combinations of different antioxidant classes. Using zebrafish larvae as a high-throughput pre-clinical in vivo toxicology screening model, our experiment effectively discriminates which of these extracts (and at what exposure levels) are suitable for development towards future therapies. The LC50-96h for finger lime and tamarind were >480 mg/L, and 1.70 mg/L for mountain pepper. Critically, this work shows that adverse effects are not correlated to the properties of these antioxidants, thus highlighting the need for combining characterization and in vivo screening to identify the most promising plant extracts for further development. Thus, we present a high-throughput pre-clinical screening that robustly tests natural plant products to utilize the diversity of antioxidant compounds for drug development.

LC-MS/MS-QTOF Screening and Identification of Phenolic Compounds from Australian Grown Herbs and Their Antioxidant Potential.

Culinary spices and herbs have been used to impart a characteristic flavour and aroma in food due to their appealing fragrance. Recently, bioactive compounds from herbs, especially phenolics, have gained much attention due to their potential health outcomes. The aim of this study was to characterize and quantify the phenolic compounds from 10 widely used Australian-grown herbs (oregano, rosemary, bay, basil, sage, fenugreek, dill, parsley, mint and thyme). For this purpose, liquid chromatography mass spectrometry (LC-MS) was used for the complete profiling of polyphenolic compounds and quantification of abundant phenolic compounds was completed with high-performance liquid chromatography-photodiode array detection (HPLC-PDA). Polyphenols from Australian-grown herbs were estimated through total phenolic content (TP), total flavonoids (TF) and total tannins (TT) along with their in-vitro antioxidant activities. Oregano and mint were estimated with the highest value of TP (140.59 ± 9.52 and 103.28 ± 8.08 mg GAE/g, milligram gallic acid equivalent/gram) while rosemary and mint had the highest TF (8.19 ± 0.74 and 7.05 ± 0.43 mg QE (quercetin equivalent)/g). In this study, eighty-four (84) phenolic compounds were screened and confirmed through LC-MS/MS by comparing their masses and fragmentation pattern with published libraries. The results of this study validate the use of these herbs as bioactives and their positive impact on human health.

LC-ESI-QTOF-MS/MS characterization of phenolic compounds from Pyracantha coccinea M.Roem. and their antioxidant capacity.

Pyracantha coccinea M.Roem. is considered as an important medicinal plant contributing remarkably to health and medicinal benefits. This is attributed to the presence of abundant polyphenols with powerful antioxidant properties. However, little research has been studied on the comprehensive identification and characterization of the phenolic compounds in areal parts of P. coccinea.  This study aimed to investigate, characterize, and quantify the phenolic profiles of P. coccinea through liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS/MS) and high-performance liquid chromatography-photodiode array (HPLC-PDA. Further, it showed a significantly higher value in total phenolic content (TPC) than that of total flavonoids (TFC) and tannins (TTC). As for antioxidant capacities, P. coccinea presented the highest activity in ABTS (7.12 ± 0.25 mg AAE/g dw) compared with DPPH, FRAP, and TAC assays. The LC-ESI-QTOF-MS/MS analysis detected 28 phenolic compounds, including phenolic acids (12), flavonoids (13), other polyphenols (2), and lignans (1) in P. coccinea samples. The results from HPLC-PDA indicated the chlorogenic acid (11.49 ± 1.89 mg/g) was the most abundant phenolic acid, while kaempferol (14.67 ± 2.17 mg/g) was the predominant flavonoid in P. coccinea. This research confirms the benefits of the P. coccinea plant as a potential source of natural antioxidants for the food and pharmaceutical industries.

Risk Communication Through Health Beliefs for Preventing Opisthorchiasis-Linked Cholangiocarcinoma: A Community- Based Intervention in Multicultural Areas of Thailand.

OBJECTIVE: This research aimed to study the effects of the risk communication program through the Cambodian folk song to prevent Opisthorchiasis-linked cholangiocarcinoma (OV-CCA). METHODS: We conducted the quasi-experimental research between August and December 2017 in the Cambodian communities, one-fourth of ethnic minorities residing in multicultural areas of Sisaket Province, Thailand. The samples consisted of 94 equally people divided into experimental group and control group. The experimental group included 47 people at-risk of OV-CCA who received the program for 12 weeks, while the control group received regular services. We collected data by using a questionnaire with a reliability of 0.93. Descriptive and inferential statistics were used for data analysis. RESULTS: The study indicated that the socioeconomic information of both groups was not different. The mean scores of all issues (health beliefs, social support, and prevention behavior in the experimental group were higher than those of the control group with statistical significance. Closer inspection showed that the mean difference of the health beliefs was 55.61 points (95%CI: 52.39-57.42, p<0.001), social support was 9.09 points (95%CI: 8.12-10.05, p<0.001), and prevention behavior was 6.38 points (95%CI: 5.43-7.33, p<0.001). CONCLUSION: Through the Cambodian folk song, the risk communication program by applying the health beliefs and social support to prevent OV-CCA is beneficial for behavior modification in areas with similar cultures.

Anti-Inflammatory Effects of Heat-Processed Artemisia capillaris Thunberg by Regulating IκBα/NF-κB Complex and 15-PGDH in Mouse Macrophage Cells.

Growing evidence suggests that dietary nutrients in herbs and plants are beneficial in improving inflammatory disorders. Artemisia capillaris Thunberg (AC) is a traditional herbal medicine widely used in East Asia to treat pain, hepatotoxicity, and inflammatory disorders. Heat processing is a unique pharmaceutical method used in traditional herbal medicine to enhance the pharmacological effects and safety of medicinal plants. This study demonstrates the anti-inflammatory effects of heat-processed AC (HPAC) in lipopolysaccharide- (LPS-) treated mouse macrophage cells. HPAC reduced LPS-induced inflammatory mediators such as IL-6, IL-1ß, TNF-α, NO, and PGE2 in RAW 264.7 cells. Interestingly, 15-PGDH appears to play a pivotal role rather than COX-2 and mPGES-1 when HPAC regulated PGE2 levels. Meanwhile, HPAC showed anti-inflammatory effects by blocking IκBα phosphorylation and NF-κB nuclear translocalization. Also, we found that HO-1 upregulation was mediated by the mitogen-activated protein kinase (MAPK) pathways in HPAC-treated RAW 264.7 cells. And, in RAW 264.7 cells challenged with LPS, HPAC restored HO-1 expression, leading to NF-κB inhibition. Through further experiments using specific MAPK inhibitors, we found that, in response to LPS, the phosphorylated IκBα and activated NF-κB were attenuated by p38 MAPK/HO-1 pathway. Therefore, HPAC targeting both the IκBα/NF-κB complex and 15-PGDH may be considered as a potential novel anti-inflammatory agent derived from a natural source.

Comprehensive Profiling of Most Widely Used Spices for Their Phenolic Compounds through LC-ESI-QTOF-MS2 and Their Antioxidant Potential.

Spices have long been used to improve food flavor, due to their appealing fragrance and sensory attributes. Nowadays, spices-based bioactives, particularly phenolic compounds, have gained attention due to their wide range of significant effects in biological systems. The present study was conducted to characterize the 12 widely used spices (allspice, black cardamom, black cumin, black pepper, cardamom, cinnamon, clove, cumin, fennel, nutmeg, star-anise, and turmeric) for their phenolics with the liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS2), polyphenols estimation, and their antioxidant potential. Total phenolics, total flavonoids, and total tannin content and their antioxidant activities were estimated in all spices. Clove and allspice had the highest value of total polyphenol content (215.14 and 40.49 mg gallic acid equivalent (GAE) per g of sample), while clove and turmeric had the highest total flavonoids (5.59 mg quercetin equivalent (QE) per g of sample) and total tannin contents (23.58 mg catechin equivalent (CE) per g of sample), respectively. On the other hand, black cumin and black pepper had the highest phosphomolybdate activity (15.61 and 15.43 mg ascorbic acid equivalent (AAE) per g of sample), while clove was almost identified with highest free radical scavenging capacity. A positive correlation was observed among phenolic compounds and antioxidant activities. In this quest, a total of 79 phenolic compounds were tentatively characterized by using LC-ESI-QTOF-MS2 including 26 phenolic acids, 33 flavonoids, 16 other polyphenols, and 4 lignans. The high-performance liquid chromatography coupled with photodiode array detector (HPLC-PDA) quantification of phenolic compounds exhibited higher phenolic acids. These results provided us some valuable information that spices have powerful antioxidant potential that can be further used in human food and animal feed as a supplement for different health promoting applications.

Synthesis of Silver Nanoparticles using Euphorbia wallichii Extract and Assessment of their Bio-functionalities.

BACKGROUND: Silver nanoparticles synthesized by the bio-green method have been applied to various biomedical applications. These procedures are simple, eco-friendly and serve as an alternative to complex chemical methods for the preparation of nanomaterials. OBJECTIVE: In the present study, phytosynthesis of silver nanoparticles, to examine their antioxidant potential, toxic effects towards bacterial-, fungal-strains, brine shrimp nauplii and cancer cells was focused. METHODS: Methanolic extract of Euphorbia wallichii roots was used for the synthesis of silver nanoparticles. The synthesis was monitored and confirmed by UV-visible spectroscopy, Fourier Transform Infra-Red (FTIR) spectrometric analysis, Field Emission Scanning Electron Microscope (FESEM), Energy Dispersive X-ray (EDX) and X-Ray Powder Diffraction (XRD). RESULTS: The synthesized particles were average 63±8 nm in size. Involvement of phenolic (46.7±2.4 µg GAE/mg) and flavonoid (11.7±1.2 µg QE/mg) compounds as capping agents was also measured. Nanoparticles showed antioxidant properties in terms of free radical scavenging potential (59.63±1.0 %), reducing power (44.52±1.34 µg AAE/mg) and total antioxidant capacity (60.48±2.2 µg AAE/mg). The nanoparticles showed potent cytotoxic effects against brine shrimp nauplii (LD50 66.83 µg/ml), proliferation and cell death of HeLa cells as determined by MTT (LD50 0.3923 µg/ml) and TUNEL assays, respectively. Antimicrobial results revealed that silver nanoparticles were found to be more potent against pathogenic fungal (maximum active against A. fumigatus, MIC 15 µg/disc) and bacterial strains (maximum active against S. aureus, MIC 3.33 µg/disc) than the E. wallichii extract alone. CONCLUSION: These results support the advantages of using an eco-friendly and cost-effective method for synthesis of nanoparticles with antioxidant, cytotoxic and antimicrobial potential.

Phytochemical analysis and antidiabetic potential of Elaeagnus umbellata (Thunb.) in streptozotocin-induced diabetic rats: pharmacological and computational approach.

BACKGROUND: The fruit of Elaeagnus umbellata has high medicinal values and is an excellent source of phytochemicals. This study was aimed to evaluate the antioxidant, enzyme inhibitory and antidiabetic potential of Elaeagnus umbellata. METHODS: The antioxidant potential of the crude extract and subfractions of E. umbellata fruit were determined using DPPH (2, 20-diphenyl-1-picrylhydrazyl) and ABTS (2, 2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid) assays. The enzyme inhibitory potentials of extracts against α-amylase and α-glucosidase enzymes were also determined. The in vivo anti-hyperglycemic effects of the extract in STZ-induced type 2 diabetes were determined using Sprague Dawley adult rats. HPLC system (Agilent 1260) was used for the identification of bioactive compounds present in extracts. Molecular docking was used to identify and compare the interaction between the compounds (active constituents) and standard inhibitor acarbose with the α-amylase and α-glucosidase active sites. RESULTS: The chloroform, ethyl acetate, and butanol fractions showed significant antioxidant potential with IC50 values of 40, 45 and 60 µg/mL against DPPH and 57, 70 and 120 µg/mL against ABTS free radicals respectively. The chloroform and ethyl acetate were highly active against α-amylase and α-glucosidase (IC50 values 58 and 200 µg/ml against α-amylase 60 and 140 µg/ml against α-glucosidase. The crude extract, chloroform, and ethyl acetate fractions were more potent in controlling the hyperglycemia in STZ-induced type 2 diabetes in rats and considerable reduction of glucose level was observed compared to the non-treated group. Furthermore, the extracts were also found useful in controlling the secondary complications associated with type 2 diabetes mellitus which was evident from the observed substantial reduction in the blood level of serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase, alkaline phosphatase, total cholesterol, low-density lipoproteins, and triglycerides. The molecular docking approach indicated the favorable inhibitory interaction between the docked compounds and the active sites of the α-amylase and α-glucosidase. All docked compounds occupied the same binding site as occupied by acarbose. CONCLUSION: It was concluded that E. umbellata can be used in the treatment of type 2 diabetes and oxidative stress. The extracts were also found to be effective in relieving the secondary complications associated with type 2 diabetes.

Association of Kaphaja and Kapha-Pittaja Prakriti and methylenetetrahydrofolate reductase C677T allele with type 2 diabetes.

BACKGROUND AND OBJECTIVES: Type 2 diabetes is a multifactorial disorder that results from the interaction between genetic predisposition and environmental factors. Different Prakriti (body constitution) individuals have different susceptibility for the diseases, and this Prakriti is determined by both genetic and environmental factor. This study was undertaken to determine the association status of Methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C with type 2 diabetes and Prakriti. MATERIALS AND METHODS: After informed consent, 54 patients with type 2 diabetes and 56 individuals as normal controls were analyzed. Their constitution and pathological data were collected and MTHFR C677T and A1298C genotypes were determined. RESULTS: Kapha/Kapha-Pittaja Prakriti were associated and found to be strong risk factors (Chi-square test = 39.67, P < 0.00001, odds ratio [OR] = 16.133, 95% confidence interval [CI] = 6.32-41.20) for type 2 diabetes. MTHFR C677T was associated (Chi-square test = 7.743, P = 0.02) with type 2 diabetes where the major CC genotype was found to be a risk for type 2 diabetes (OR = 3.78, 95% CI = 1.14-12.45). A1298C was not associated with type 2 diabetes (Chi-square test = 2.264, P = 0.322). None of the Prakriti was associated with C677T and A1298C variants. INTERPRETATION AND CONCLUSION: In the present study, an extremely strong association between Prakriti (Kaphaja/Kapha-Pittaja) and type 2 diabetes (P < 0.00001) was detected. The present study gives a strong clue for the association of Prakriti (body constitutional) and clinical phenotype.

Clinical evaluation of unani ear drops herbotic for earache.

An ear infection (acute otitis media) is most often a bacterial or viral infection that affects the middle ear. Children as compared to adults are more are prone to infections. A study has been conducted on 200 patients with herbal medicine Herbotic and allopathic drug Ofloxacin for the treatment of otitis media. The efficacy of test and control drug was monitored before and after treatment and diagnosed on clinical history, clinical presentation and pathological investigation. This study was a case control, multicenter, prospective randomized authentic allopathic controlled, two arm parallel group clinical trial The data on clinical proforma was gathered between April 2014-March 2015 and subjected to statistical analysis. From the statistical results it was concluded that Herbotic and Ofloxacin are equally effective for the treatment of earache, effect being confirmed by physicians and patients alike.

Green tea extract can potentiate acetaminophen-induced hepatotoxicity in mice.

Green tea extract (GTE) has been advocated as a hepatoprotective compound and a possible therapeutic agent for acetaminophen (APAP) overdose. This study was conducted to determine if GTE can provide protection against APAP-induced hepatotoxicity. Three different exposure scenarios were tested. The first involved administering APAP (150 mg/kg, orally) to mice followed 6h later by GTE (500 or 1000 mg/kg). The other two involved administering GTE prior to the APAP dose. GTE (500 or 1000 mg/kg, orally) was administered 3h prior to APAP (200 mg/kg, orally) or for three consecutive days (once-daily) followed by APAP (300 mg/kg) on the fourth day. Indices of hepatotoxicity were assessed 24h after the APAP dose. GTE potentiated APAP-induced hepatotoxicity when administered after the APAP dose. GTE caused significant glutathione depletion and this effect likely contributed to the observed potentiation. In contrast, GTE provided protection against APAP-induced hepatotoxicity when administered prior to the APAP dose. GTE dramatically decreased APAP covalent binding to protein indicating that less reactive metabolite was available to cause hepatocellular injury. These results highlight the potential for drug-dietary supplement interactions and the importance of testing multiple exposure scenarios to adequately model different types of potential interactions.