Emergence of Lasiodiplodia theobromae induced leaf necrosis in tea (Camellia sinensis [L.] O. Kuntze) from India.

The tea plant, Camellia sinensis [L.] O. Kuntze, is a vital global agricultural commodity, yet faces challenges from fungal infections, which affects its production. To reduce the loss in the tea production, the fungal infections must be removed which is managed with fungicides, which are harmful to the environment. Leaf necrosis, which decreases tea quality and quantity, was investigated across Assam, revealing Lasiodiplodia theobromae as the causative agent. Pathogenicity tests, alongside morphological and molecular analyses, confirmed its role in leaf necrosis. Genome and gene analysis of L. theobromae showed multiple genes related to its pathogenicity. The study also assessed the impact of chemical pesticides on this pathogen. Additionally, the findings in this study highlight the significance of re-assessing management approaches in considering the fungal infection in tea.

Quantitative 1H NMR with global spectral deconvolution approach for quality assessment of natural and cultured Cordyceps sinensis.

Cordyceps sinensis is a precious medicinal food which has been successfully cultivated indoors. It remains to be investigated for a simultaneous comparison on aqueous components of natural and cultivated samples. Herein, an approach of quantitative nuclear magnetic resonance (qNMR) analysis combined with global spectral deconvolution (GSD) was established for simultaneous quantification of 26 aqueous components in C. sinensis. Processed by GSD, the distorted baselines of 1H NMR spectra were greatly improved, and overlapped signals were also well separated so as to achieve accurate identification and quantitation of components in C. sinensis. Method validation by UHPLC-QTOF-MS and TOF-SIMS analysis revealed that qNMR combined with GSD is a reliable approach for simultaneous quantification of multiple components including characteristic markers of glutamine, GABA and trehalose in authentic and fake C. sinensis. The well-established qNMR approach can be used for quality assessment of natural and cultivated C. sinensis as well as differentiation from fake ones.

Pollution and health risk assessment of rare earth elements in Citrus sinensis growing soil in mining area of southern China.

Background: Analyzing the pollution and health risk of rare earth elements (REEs) in crop-growing soils around rare earth deposits can facilitate the improvement of REE mining-influenced area. In this study, pollution status, fraction and anomaly, plant accumulation characteristics, and potential risks of REEs (including heavy and light rare earth elements, HREEs and LREEs) in C. sinensis planting soil near ion-adsorption deposits in southern Ganzhou were analyzed. The influence of the soil environment on REEs in soil and fruit of C. sinensis was also explored. Methods: The geo-accumulation index (Igeo) and ecological risk index(RI) were used to analyze the pollution potential and ecological risks of REEs in soils, respectively. Health risk index and translocation factor (TF) were applied to analyze the accumulation and health risks of REEs in fruit of C. sinensis. The influence of soil factors on REEs in soil and fruit of C. sinensis were determined via correlation and redundancy analysis. Results: Comparison with background values and assessment of Igeo and RI indicated that the soil was polluted by REEs, albeit at varying degrees. Fractionation between LREEs and HREEs occurred, along with significant positive Ce anomaly and negative Eu anomaly. With TF values < 1, our results suggest that C. sinensis has a weak ability to accumulate REEs in its fruit. The concentrations of REEs in fruit differed between LREEs and HREEs, with content of HREE in fruit ordered as Jiading > Anxi > Wuyang and of LREE in fruit higher in Wuyang. Correlation and redundancy analysis indicated that K2O, Fe2O3 and TOC are important soil factors influencing REE accumulation by C. sinensis, with K2O positively related and Fe2O3 and TOC negatively related to the accumulation process.

Essential and Potentially Toxic Elements (PTEs) Content in European Tea (Camellia sinensis) Leaves: Risk Assessment for Consumers.

Tea (Camellia sinensis) is the second most consumed beverage worldwide, playing a key role in the human diet. Tea is considered a healthy drink, as its consumption has been linked to a lower risk of cardiovascular disease-related events and death, stroke, metabolic syndrome and obesity. However, several studies have shown that C. sinensis is a hyperaccumulator of Al and other elements that are considered potentially toxic. In the present study, the contents of 15 elements (both essential and toxic) were determined for the first time in tea leaves collected in tea gardens located in six different European countries and processed to provide black and green tea. The results showed that Al was the major toxic element detected, followed by Ni, Cr, Pb, As, Cd, Ag, and Hg. Essential elements were detected in the order of Mn, Fe, Zn, Cu, Co, and Se. Statistically significant correlations (p < 0.05) were found in the distribution of some elements, highlighting mechanisms of synergic or antagonist interaction. Multivariate analysis revealed that geographical origin was the main driver in clustering the samples, while the different treatment processes (black or green) did not significantly affect the contents of elements in the leaves. The estimation of potential non-carcinogenic risk revealed no risk for the consumption of European teas for consumers in terms of potentially toxic elements.

Genome-Wide Identification of 2-Oxoglutarate and Fe (II)-Dependent Dioxygenase (2ODD-C) Family Genes and Expression Profiles under Different Abiotic Stresses in Camellia sinensis (L.).

The 2-oxoglutarate and Fe (II)-dependent dioxygenase (2ODD-C) family of 2-oxoglutarate-dependent dioxygenases potentially participates in the biosynthesis of various metabolites under various abiotic stresses. However, there is scarce information on the expression profiles and roles of 2ODD-C genes in Camellia sinensis. We identified 153 Cs2ODD-C genes from C. sinensis, and they were distributed unevenly on 15 chromosomes. According to the phylogenetic tree topology, these genes were divided into 21 groups distinguished by conserved motifs and an intron/exon structure. Gene-duplication analyses revealed that 75 Cs2ODD-C genes were expanded and retained after WGD/segmental and tandem duplications. The expression profiles of Cs2ODD-C genes were explored under methyl jasmonate (MeJA), polyethylene glycol (PEG), and salt (NaCl) stress treatments. The expression analysis showed that 14, 13, and 49 Cs2ODD-C genes displayed the same expression pattern under MeJA and PEG treatments, MeJA and NaCl treatments, and PEG and NaCl treatments, respectively. A further analysis showed that two genes, Cs2ODD-C36 and Cs2ODD-C21, were significantly upregulated and downregulated after MeJA, PEG, and NaCl treatments, indicating that these two genes played positive and negative roles in enhancing the multi-stress tolerance. These results provide candidate genes for the use of genetic engineering technology to modify plants by enhancing multi-stress tolerance to promote phytoremediation efficiency.

Comprehensive Genomic Analysis of Trihelix Family in Tea Plant (Camellia sinensis) and Their Putative Roles in Osmotic Stress.

In plants, Trihelix transcription factors are responsible for regulating growth, development, and reaction to various abiotic stresses. However, their functions in tea plants are not yet fully understood. This study identified a total of 40 complete Trihelix genes in the tea plant genome, which are classified into five clades: GT-1 (5 genes), GT-2 (8 genes), GTγ (2 genes), SH4 (7 genes), and SIP1 (18 genes). The same subfamily exhibits similar gene structures and functional domains. Chromosomal mapping analysis revealed that chromosome 2 has the most significant number of trihelix family members. Promoter analysis identified cis-acting elements in C. sinensis trihelix (CsTH), indicating their potential to respond to various phytohormones and stresses. The expression analysis of eight representative CsTH genes from four subfamilies showed that all CsTHs were expressed in more tissues, and three CsTHs were significantly induced under ABA, NaCl, and drought stress. This suggests that CsTHs plays an essential role in tea plant growth, development, and response to osmotic stress. Furthermore, yeast strains have preliminarily proven that CsTH28, CsTH36, and CsTH39 can confer salt and drought tolerance. Our study provides insights into the phylogenetic relationships and functions of the trihelix transcription factors in tea plants. It also presents new candidate genes for stress-tolerance breeding.

Coinfection of Clonorchis sinensis and hepatitis B virus: clinical liver indices and interaction in hepatic cell models.

BACKGROUND: In China, people infected with hepatitis B virus (HBV) are commonly found in areas with a high prevalence of Clonorchis sinensis, a trematode worm. Published studies have reported that the progression of hepatitis B is affected by coinfection C. sinensis. METHODS: Clinical data from a total of 72 patients with C. sinensis and HBV (as sole infection or with coinfections) and 29 healthy individuals were analysed. We also incubated the hepatic stellate cell line LX-2 with total proteins from C. sinensis adult worms (CsTPs) and HBV-positive sera. In addition, the human hepatoblastoma cell line HepG2.2.15 was treated with the antiviral drug entecavir (ETV), CsTPs and the anti-C. sinensis drug praziquantel (PZQ). RESULTS: Our clinical data indicated that the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TB) and hyaluronic acid (HA) were significantly higher in patients with coinfection than in those infected with HBV only. In cell models, compared with the model in which LX-2 cells were incubated with HBV-positive sera (HBV group), transcripts of alpha-smooth muscle actin and types I and III collagen were significantly elevated in the models of LX-2 cells treated with CsTPs and HBV-positive sera (CsTP+HBV group), while the messenger RNA levels of tumour necrosis factor-α, interleukin (IL)-1ß and IL-6 in the CsTP+HBV group were clearly lower. The HBV surface antigen and hepatitis B e-antigen levels were higher in the HepG2.2.15 cells treated with ETV and CsTPs than in those in the ETV group and in the cells administered a mixture of ETV, CsTPs and PZQ. CONCLUSIONS: These results confirmed that C. sinensis and HBV coinfection could aggravate the progression of liver fibrosis. CsTPs might promote chronic inflammation of the liver in individuals with HBV infection, resulting in the development of hepatic fibrosis.

The Role of Calmodulin Binding Transcription Activator in Plants under Different Stressors: Physiological, Biochemical, Molecular Mechanisms of Camellia sinensis and Its Current Progress of CAMTAs.

Low temperatures have a negative effect on plant development. Plants that are exposed to cold temperatures undergo a cascade of physiological, biochemical, and molecular changes that activate several genes, transcription factors, and regulatory pathways. In this review, the physiological, biochemical, and molecular mechanisms of Camellia sinensis have been discussed. Calmodulin binding transcription activator (CAMTAs) by molecular means including transcription is one of the novel genes for plants' adaptation to different abiotic stresses, including low temperatures. Therefore, the role of CAMTAs in different plants has been discussed. The number of CAMTAs genes discussed here are playing a significant role in plants' adaptation to abiotic stress. The illustrated diagrams representing the mode of action of calcium (Ca2+) with CAMTAs have also been discussed. In short, Ca2+ channels or Ca2+ pumps trigger and induce the Ca2+ signatures in plant cells during abiotic stressors, including low temperatures. Ca2+ signatures act with CAMTAs in plant cells and are ultimately decoded by Ca2+sensors. To the best of our knowledge, this is the first review reporting CAMAT's current progress and potential role in C. sinensis, and this study opens a new road for researchers adapting tea plants to abiotic stress.

Sustainable Extraction, Chemical Profile, Cytotoxic and Antileishmanial Activities In-Vitro of Some Citrus Species Leaves Essential Oils.

Anti-leishmanial drugs extracted from natural sources have not been sufficiently explored in the literature. Until now, leishmaniasis treatments have been limited to synthetic and expensive drugs. This study investigated, for the first time, the anti-leishmanial efficacy of essential oils (EOs) from the leaves of Citrus species (C. sinensis, C. limon, and C. clementina). Essential oils were extracted from three species by solvent free microwave extraction (SFME); in addition, lemon oil was also isolated by hydro-distillation (HD). These were investigated using gas chromatography coupled with mass spectrometry (GC-MS) and evaluated against Leishmania species, namely Leishmania major and Leishmania infantum, using a mitochondrial tetrazolium test (MTT) assay. The chemical compositions of Citrus limon EOs obtained by HD and SFME showed some differences. The identified peaks of C. limon (SFME) represented 93.96%, where linalool was the major peak (44.21%), followed by sabinene (14.22%) and ocimene (6.09%). While the hydro-distilled oil of C. limon contained geranial (30.08%), limonene (27.09%), and neral (22.87%) in the identified peaks (96.67%). The identified components of C. clementina leaves oil (68.54%) showed twenty-six compounds, where the predominant compound was geranial (42.40%), followed by neral (26.79%) and limonene (14.48%). However, 89.82% C. sinensis oil was identified, where the major peaks were for neral (27.52%), linalool (25.83%), and geranial (23.44%). HD oil of lemon showed the highest activity against L. major, with moderate toxicity on murine macrophage (RAW 264.7) cells, and possessed the best selectivity index on both Leishmanial species (SI: 3.68; 6.38), followed by C. clementina oil and C. limon using SFME (0.9 ± 0.29, 1.03 ± 0.27, and 1.13 ± 0.3), respectively. C. clementina oil induced the greatest activity on Leishmania infantum, followed by HD lemon and SFME lemon oils (0.32 ± 0.18, 0.52 ± 0.15, and 0.57 ± 0.09, respectively) when compared to Amphotericin B (0.80 ± 0.18 and 0.23 ± 0.13) as a positive control, on both species, respectively. Our study suggests a potent anti-leishmanial activity of lemon oil (HD) on L. major, followed by C. clementina. With the same potency on L. infantum shown by C. clementina oil, followed by HD lemon oil. This effect could be attributed to the major compounds of limonene, citral, and neral, as well as the synergistic effect of other different compounds. These observations could be a starting point for the building of new anti-leishmanial drugs from natural origins, and which combine different EOs containing Citrus cultivars.

Effects of Nitrogen Deficiency and Resupply on the Absorption of Mineral Nutrients by Tangor Cultivar 'Shiranuhi' (Citrus unshiu × C. sinensis) Grown in a Hydroponic System.

Tangor (Citrus unshiu × C. sinensis) cultivars obtained through interspecific hybridization have gained popularity in various countries, including South Korea. However, tangor has a relatively short history of cultivation and thus has been less well researched in terms of physiology and opping technology compared to mandarin. In addition, although tangor grows more vigorously than mandarin and thus has high nutrient demands and longer harvest time, it is more prone to various physiological disorders. Furthermore, the demand for nutrients could vary depending on the cultivars even within the same species. Thus, the current study examined the effects of nitrogen deficiency and resupply on the uptake of mineral nutrients using a specific cultivar 'Shiranuhi' as a case study. In this study, the tangor cultivar 'Shiranuhi' was cultivated in a hydroponics system, which allows the control of nutrient contents, and changes in mineral nutrient contents under nitrogen deficiency and nitrogen resupply were examined. Based on this, the results show the straightforward relationship between nitrogen and other mineral nutrients under a hydroponic system applying the cultivation of tangor. This implies that the hydroponics system can be applied to tangor cultivation and efficiently and widely expanded in Northeast Asia, and the income of growers will increase through the production of high-quality fruits.

Nucleosides and amino acids, isolated from Cordyceps sinensis, protected against cyclophosphamide-induced myelosuppression in mice.

The material basis of Cordyceps sinensis (Berk.) Sacc has not yet been well understood and natural C. sinensis resources are very rare. The present study aimed to clarify the substance basis and compare the protective effect of natural and artificially-cultivated C. sinensis against cyclophosphamide (CTX)-induced myelosuppression. Both natural and artificially-cultivated C. sinensis effectively improved CTX-induced decrease of peripheral blood counts and hemopoietic growth factors, pathological changes, and apoptosis of bone marrow. Importantly, artificially-cultivated C. sinensis showed similar capacity compared with natural C. sinensis. Uridine (1), adenosine (2), L-pyroglutamic acid (3), lysinonorleucine (4), 1,3,5-trimethoxybenzene (5), D-mannitol (6), L-pyroglutamic acid methyl ester (7), tryptophan (8), and phenylalanine (9) were isolated from bioactivity-guided fraction and identified to attenuate CTX-induced myelosuppression in mice. In conclusions, nucleosides and amino acids represented the effective chemical components in C. sinensis. Artificial cultivation can be used as an effective substitute for natural C. sinensis.

Characterization of the Trans-Epithelial Transport of Green Tea (C. sinensis) Catechin Extracts with In Vitro Inhibitory Effect against the SARS-CoV-2 Papain-like Protease Activity.

This work describes an untargeted analytical approach for the screening, identification, and characterization of the trans-epithelial transport of green tea (Camellia sinensis) catechin extracts with in vitro inhibitory effect against the SARS-CoV-2 papain-like protease (PLpro) activity. After specific catechin extraction, a chromatographic separation obtained six fractions were carried out. The fractions were assessed in vitro against the PLpro target. Fraction 5 showed the highest inhibitory activity against the SARS-CoV-2 PLpro (IC50 of 0.125 µg mL-1). The untargeted characterization revealed that (-)-epicatechin-3-gallate (ECG) was the most abundant compound in the fraction and the primary molecule absorbed by differentiated Caco-2 cells. Results indicated that fraction 5 was approximately 10 times more active than ECG (IC50 value equal to 11.62 ± 0.47 µg mL-1) to inhibit the PLpro target. Overall, our findings highlight the synergistic effects of the various components of the crude extract compared to isolated ECG.

MicroRNA-497 induced by Clonorchis sinensis enhances the TGF-ß/Smad signaling pathway to promote hepatic fibrosis by targeting Smad7.

BACKGROUND: Various stimuli, including Clonorchis sinensis infection, can cause liver fibrosis. Liver fibrosis is characterized by the activation of hepatic stellate cells (HSCs) with massive production of extracellular matrix (ECM). Our previous study showed that the TGF-ß1-induced Smad signaling pathway played a critical role in the activation of HSCs during liver fibrosis induced by worm infection; however, the mechanisms that modulate the TGF-ß/Smad signaling pathway are still poorly understood. Accumulating evidence demonstrates that miRNAs act as an important regulator of activation of HSCs during liver fibrosis. METHODS: The target of miR-497 was determined by bioinformatics analysis combined with a dual-luciferase activity assay. LX-2 cells were transfected with miR-497 inhibitor and then stimulated with TGF-ß1 or excretory/secretory products of C. sinensis (CsESPs), and activation of LX-2 was assessed using qPCR or western blot. In vivo, the mice treated with CCl4 were intravenously injected with a single dose of adeno-associated virus serotype 8 (AAV8) that overexpressed anti-miR-497 sequences or their scramble control for 6 weeks. Liver fibrosis and damage were assessed by hematoxylin and eosin (H&E) staining, Masson staining, and qPCR; the activation of the TGF-ß/Smad signaling pathway was detected by qPCR or western blot. RESULTS: In the present study, the expression of miR-497 was increased in HSCs activated by TGF-ß1 or ESPs of C. sinensis. We identified that Smad7 was the target of miR-497 using combined bioinformatics analysis with luciferase activity assays. Transfection of anti-miR-497 into HSCs upregulated the expression of Smad7, leading to a decrease in the level of p-Smad2/3 and subsequent suppression of the activation of HSCs induced by TGF-ß1 or CsESPs. Furthermore, miR-497 inhibitor delivered by highly-hepatotropic (rAAV8) inhibited TGF-ß/smads signaling pathway by targeting at Smad7 to ameliorate CCL4-induced liver fibrosis. CONCLUSIONS: The present study demonstrates that miR-497 promotes liver fibrogenesis by targeting Smad7 to promote TGF-ß/Smad signaling pathway transduction both in vivo and in vitro, which provides a promising therapeutic strategy using anti-miR-497 against liver fibrosis.

Investigation of cell wall proteins of C. sinensis leaves by combining cell wall proteomics and N-glycoproteomics.

BACKGROUND: C. sinensis is an important economic crop with fluoride over-accumulation in its leaves, which poses a serious threat to human health due to its leaf consumption as tea. Recently, our study has indicated that cell wall proteins (CWPs) probably play a vital role in fluoride accumulation/detoxification in C. sinensis. However, there has been a lack in CWP identification and characterization up to now. This study is aimed to characterize cell wall proteome of C. sinensis leaves and to develop more CWPs related to stress response. A strategy of combined cell wall proteomics and N-glycoproteomics was employed to investigate CWPs. CWPs were extracted by sequential salt buffers, while N-glycoproteins were enriched by hydrophilic interaction chromatography method using C. sinensis leaves as a material. Afterwards all the proteins were subjected to UPLC-MS/MS analysis. RESULTS: A total of 501 CWPs and 195 CWPs were identified respectively by cell wall proteomics and N-glycoproteomics profiling with 118 CWPs in common. Notably, N-glycoproteomics is a feasible method for CWP identification, and it can enhance CWP coverage. Among identified CWPs, proteins acting on cell wall polysaccharides constitute the largest functional class, most of which might be involved in cell wall structure remodeling. The second largest functional class mainly encompass various proteases related to CWP turnover and maturation. Oxidoreductases represent the third largest functional class, most of which (especially Class III peroxidases) participate in defense response. As expected, identified CWPs are mainly related to plant cell wall formation and defense response. CONCLUSION: This was the first large-scale investigation of CWPs in C. sinensis through cell wall proteomics and N-glycoproteomics. Our results not only provide a database for further research on CWPs, but also an insight into cell wall formation and defense response in C. sinensis.

Comparison of metabolism substances in Cordyceps sinensis and Cordyceps militaris cultivated with tussah pupa based on LC-MS.

The objective of our study was to compare the chemical composition of Cordyceps sinensis (C. sinensis) and Cordyceps militaris (C. militaris) cultivated with tussah pupa by using metabonomics technology in order to clarify the similarity and difference of the two medicinal materials from the whole metabolite level. The results showed that there were 25 different metabolites among the 69 metabolites that were highly expressed in C. militaris cultivated with tussah pupa compared with C. sinensis in both positive and negative ion modes. Analysis results of partial differential metabolites pathways indicated that 16 differential metabolites were involved in multiple pathways, such as histidine metabolism, arginine biosynthesis, tyrosine metabolism, glyoxylate and dicarboxylate metabolism, phenylpropanoid biosynthesis, pyruvate metabolism, etc. Therefore, the composition of C. militaris cultivated with tussah pupa had significant advantage comparing with C. sinensis, which demonstrated that high-priced C. sinensis could be substituted with C. militaris cultivated with tussah pupa to some extent. PRACTICAL APPLICATIONS: This study comprehensively compared the chemical composition of Cordyceps sinensis (C. sinensis) and Cordyceps militaris (C. militaris) cultivated with tussah pupa by using metabonomics technology in order to clarify the similarity and difference of the two medicinal materials from the whole metabolite level. The experimental results provide a theoretical basis and scientific support for whether C. sinensis can be substituted with C. militaris cultivated with tussah pupa in clinical practice.

An investigation of Human Clonorchiasis prevalence in an Endemic County in Guangxi Zhuang Autonomous Region, China, 2016.

To effectively promote the implementation of interventions, the identification of high-risk groups and the characteristics of Clonorchis sinensis infection in endemic regions are needed. In a clonorchiasis-endemic area, local residents were randomly enrolled for helminth egg examination in June 2016. The prevalence in subpopulations as well as the knowledge, attitudes, and behaviours and the factors influencing clonorchiasis in egg-positive populations were analysed. A total of 2282 local residents participated in the survey; the C. sinensis prevalence was 48.6% (1109 persons). A higher prevalence was found in males (62.6%) than in females (29.7%). People older than 30 years had the highest prevalence (52.7%-57.6%). Among the 888 persons who were infected with C. sinensis and participated the questionnaire investigation, 19.0% (169/888) knew that it could cause cancer. In addition, 60.6% of people reported that they intended to keep eating raw fish despite knowing the risk of infection. The two primary reasons for continuing to eat raw fish were the disease being regarded as not serious (38.3%) and the belief that anti-parasite medications are effective (39.6%). A total of 94.4% (797/844) of responders reported eating raw fish more frequently in the home than outside of the home. Our study revealed a notably high C. sinensis prevalence in the study area. Awareness of clonorchiasis disease severity should be increased among high-risk individuals and families in highly endemic areas.

Blood orange (Citrus sinensis) as a rich source of nutraceuticals: investigation of bioactive compounds in different parts of the fruit by HPLC-PDA/MS.

This study is part of a wider investigation aimed to sustain the economical value of the by-products generated by the Citrus industry. In particular, the protected geographic indication (PGI) red orange of sicily (known as blood orange) has been analysed by HPLC and by the enzymatic AOAC method. All the by-products contain significant amounts of biologically active compounds (limonoids and flavonoids). The decanted pulps were the most abundant, with the highest amount of flavonoids (130 g/kg) and high amount of limonoids (5.5 g/kg). Seeds were the best source of limonoids with about 10 g/kg. Low amount of anthocyanins were found only in coarse pulps and waste water. The total, the insoluble and the soluble dietary fibre (TDF, IDF and SDF respectively) were also determined. The pulps resulted to be the best source of dietary fibre, based on the amount and on the best insoluble/soluble ratio.

Foodborne zoonotic parasites of the family Opisthorchiidae.

Several foodborne zoonotic trematodes of the family Opisthorchiidae have been reported to infect people, specifically two genera, Clonorchis and Opisthorchis. Three species Clonorchis sinensis, Opisthorchis felineus and Opisthorchis viverrini are the most extensivley studied of the Opisthorchiidae. At least 680 million people worldwide are at risk of infection of these liver flukes through the consumption of raw or partially cooked freshwater cyprinid fish. An estimated 45 million people in Europe and Asia are currently infected with these liver flukes. Of these, the 35 million are infected with C. sinensis mainly in China, 10 million with O. viverrini in Southeast Asia, and 1.2 million with O. felineus in Eastern Europe and Russia. These liver flukes have been proven to be causative agents of bile duct cancer or cholangiocarcinoma (CCA). A multidisciplinary program should be implemented involving comprehensive research on molecular genetics, diagnosis, treatment, prevention, as well as educational and control programs should progressively be introduced and applied in endemic regions of O. viverrini, O. felineus and C. sinensis throughout their ranges, hence, opisthorchiasis and clonorchiasis free communities can be realised globally.

Identification and Analysis of the Tegument Protein and Excretory-Secretory Products of the Carcinogenic Liver Fluke Clonorchis sinensis.

Liver fluke proteins, including excretory-secretory products (ESPs) and tegument proteins, are critical for the pathogenesis, nutrient metabolism, etiology and immune response of liver cancer. To understand the functions of various proteins in Clonorchis sinensis physiology and human clonorchiasis, the ESPs and tegument proteins of C. sinensis were identified. Supernatants containing ESPs from adult C. sinensis after culture for 6 h were harvested and concentrated. The tegument was detached using a freeze/thaw method and successively extracted using various extraction buffers. The outer surface proteins of C. sinensis were labeled with biotin, and the biotinylated proteins were purified. The ESP, tegument and labeled outer surface proteins were identified and analyzed by high-resolution LC-MS/MS. The identified proteins were compared with those of other flukes, and the protein functions associated with pathogenesis, carcinogenesis and potential vaccine antigens and drug targets were predicted and analyzed. A total of 175 proteins were identified after the 6-h culture of C. sinensis ESPs. A total of 352 tegument proteins were identified through sequential solubilization of the isolated teguments, and a subset of these proteins were localized to the surface membrane of the tegument by labeling with biotin. Thirty identified proteins, including annexins, actin and tetraspanins, were identified as potential immunomodulators and promising vaccine antigens. Interestingly, among the 352 tegument proteins, as many as 155 were enzymes, and most were oxidoreductases, hydrolases or transferases. A comparison of the outer surface proteins of C. sinensis with those of other flukes indicated that flukes have some common outer surface proteins, such as actin, tetraspanin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and annexin. Granulin, thioredoxin peroxiredoxin, carbonyl reductase 1 and cystatin were identified in the C. sinensis proteome and predicted to be related to liver disease and cancer. The analysis of the C. sinensis proteome could contribute to a more in-depth understanding of complex parasite-host relationships, improve the diagnosis of clonorchiasis and benefit research on the pathogenesis and development of novel interventions, drugs and vaccines to control C. sinensis infection.

Sweet and bitter oranges: An updated comparative review of their bioactives, nutrition, food quality, therapeutic merits and biowaste valorization practices.

Sweet and bitter oranges are two of the most commercially-important fruit with a total world production of 75.4 Mt, well-recognized for their unique sensory characters in addition to multiple nutritive and therapeutic attributes due to their highly-valued bioactive ingredients. Hence, their differential qualitative/quantitative phytochemical make-ups are presented for better utilization as therapeutic agents. Sweet orange exhibits therapeutic applications as being effective anti-diabetic, anti-obesity, and hypocholesterolemic agents. Whereas, for anti-osteoporotic products and intestinal dysbiosis treatment, bitter orange is more preferred. Moreover, the review recapitulates on different valorization practices of citrus bio-wastes and utilization of their bioactives as therapeutic agents and in functional food industry. Sweet orange waste functions as a fat replacer and preservative to increase food shelf life with better organoleptic attributes than bitter orange. The detailed action mechanism and safety of Citrus bioactives, as well as processing technologies to further improve its effects are posed as future research perspectives.