Green Synthesis of Chitosan/Silver Nanoparticles Using Citrus paradisi Extract and Its Potential Anti-Cryptosporidiosis Effect.

Cryptosporidium parvum (C. parvum) is one of the most prevalent species infecting humans and animals. Currently, the only FDA-licensed drug to treat cryptosporidiosis is nitazoxanide (NTZ), with no efficacy in immunocompromised hosts. Citrus paradisi (C. paradisi) has demonstrated anti-protozoal activities. This study aimed to investigate the anti-cryptosporidiosis effect of C. paradisi peel extract, either alone or in mediating the green synthesis of chitosan silver nanoparticles (Cs/Ag NPs), compared to NTZ. Mice were sorted into nine different groups. The effectiveness of the treatments was evaluated using parasitology, histopathology, immunohistochemistry, and immunology. C. paradisi outperformed nitazoxanide regarding oocyst shedding (79% vs. 61%). The effectiveness of NTZ Cs/Ag NPs and Citrus Cs/Ag NPs was enhanced to 78% and 91%, respectively. The highest oocyst inhibition was obtained by combining NTZ and Citrus Cs/Ag NPs (96%). NF-κB, TNF-α, and Il-10 levels increased in response to infection and decreased in response to various treatments, with the highest reduction in the group treated with combined NTZ citrus Cs/Ag NPs. Combining C. paradisi with NTZ could have a synergistic effect, making it a potentially effective anti-cryptosporidiosis agent. Utilizing C. paradisi in the green synthesis of Cs/Ag NPs improves the therapeutic response and can be used to produce novel therapeutic antiparasitic drugs.

Extraction of pectins from renewable grapefruit (Citrus paradisi) peels using deep eutectic solvents and analysis of their structural and physicochemical properties.

This study presents an innovative attempt to extract high-quality pectins from grapefruit (Citrus paradisi) peels by using deep eutectic solvents (DESs) as extraction agents. The maximum yield of betaine-citric acid (BC)-extracted pectin (BC-P) reached 36.47 % under the optimum process conditions: an L/S ratio of 25 mL/g, a pH of 2.0, and a temperature of 85 °C for 120 min. The yield of BC-P was significantly higher than HCl-extracted pectin (HCl-P, 8.76 %) under a pH of 2.0. In addition, the structural, physicochemical, and emulsifying properties of the purified pectins (BC-P and HCl-P) and commercial pectin (CP) were comparatively analyzed. Results showed that BC-P exhibited higher RG-I value, more arabinan side-chains, bigger Mw and Mn value than HCl-P. Moreover, the viscosity, G' and G'' of BC-P were significantly higher than those of HCl-P and CP. More importantly, BC-P demonstrated better emulsifying activity and stability compared to HCl-P and CP. When the concentration of BC-P was increased to 1.50 %, a stable emulsion containing a 50 % soybean oil fraction could be obtained. Our results confirmed that DESs can be considered as high-effective agents for pectin extraction. Pectins extracted from grapefruit peels can be as a promising natural emulsifiers that can be used in the food industry.

Grapefruit Root and Rhizosphere Responses to Varying Planting Densities, Fertilizer Concentrations and Application Methods.

Huanglongbing (HLB) disease has caused a severe decline in citrus production globally over the past decade. There is a need for improved nutrient regimens to better manage the productivity of HLB-affected trees, as current guidelines are based on healthy trees. The aim of this study was to evaluate the effects of different fertilizer application methods and rates with different planting densities on HLB-affected citrus root and soil health. Plant material consisted of 'Ray Ruby' (Citrus × paradisi) grapefruit trees grafted on 'Kuharske' citrange (Citrus × sinensis × Citrus trifoliata). The study consisted of 4 foliar fertilizer treatments, which included 0×, 1.5×, 3× and 6× the University of Florida Institute of Food and Agriculture (UF/IFAS) recommended guidelines for B, Mn and Zn. Additionally, 2 ground-applied fertilizer treatments were used, specifically controlled-release fertilizer (CRF1): 12-3-14 + B, Fe, Mn and Zn micronutrients at 1× UF/IFAS recommendation, and (CRF2): 12-3-14 + 2× Mg + 3× B, Fe, Mn and Zn micronutrients, with micronutrients applied as sulfur-coated products. The planting densities implemented were low (300 trees ha-1), medium (440 trees ha-1) and high (975 trees ha-1). The CRF fertilizer resulted in greater soil nutrient concentrations through all of the time sampling points, with significant differences in soil Zn and Mn. Grapefruit treated with ground-applied CRF2 and 3× foliar fertilizers resulted in the greatest bacterial alpha and beta diversity in the rhizosphere. Significantly greater abundances of Rhizobiales and Vicinamibacterales were found in the grapefruit rhizosphere of trees treated with 0× UF/IFAS foliar fertilizer compared to higher doses of foliar fertilizers.

Effect of Huanglongbing on the Volatile Organic Compound Profile of Fruit Juice and Peel Oil in 'Ray Ruby' Grapefruit.

Along with orange and mandarin, grapefruit production in Florida has declined sharply due to Huanglongbing (HLB), or citrus greening disease, caused by Candidatus Liberibacter asiaticus (CLas). HLB affects the volatile profiles of juice and peel oil in oranges, but there is limited information on grapefruit. In this research, 'Ray Ruby' grapefruit were harvested in 2020 and 2021 from healthy (HLB-) and HLB-affected (HLB+) trees. Peel oil was extracted by hydrodistillation, and the volatiles were analyzed by direct injection of the oil samples into gas chromatography-mass spectrometry (GC-MS). Volatiles in the juice were analyzed by headspace (HS)-solid-phase microextraction (SPME) coupled with GC-MS. HLB significantly altered the volatile profiles of peel oil and juice in 'Ray Ruby' grapefruit. Juice samples of HLB+ fruits had lower decanal, nonanal, and octanal, important citrus juice flavor compounds. HLB+ samples also showed reduced content of nonterpene compounds, other aliphatic and terpene aldehydes, and terpene ketones. Ethanol, acetaldehyde, ethyl acetate, and ethyl butanoate were increased in HLB+ juice samples, indicating an HLB-induced stress response. The most abundant compounds D-limonene and ß-caryophyllene, as well as other sesquiterpenes, were increased in HLB+ juice and peel oil samples. On the other hand, the oxidative/dehydrogenated terpenes were increased by HLB in peel oil but decreased in the juice sample. Nootkatone, the key grapefruit volatile was consistently reduced by HLB in both peel oil and juice samples. The impact of HLB on nootkatone deteriorated the quality of both juice and peel oil in grapefruits.

Anti-inflammatory properties of an aldehydes-enriched fraction of grapefruit essential oil.

Chronic inflammation is linked to the development of numerous diseases and is accompanied by increased cytokine secretion. Macrophages provide a first line of defense against pathogens that under inflammatory stimuli release pro-inflammatory cytokines. The essential oil (EO) fractions obtained from Citrus spp. rich in different compounds have gained the attention of both researchers and users during the last decades. In particular, grapefruit (Citrus paradisi) peel is rich in phenolics and flavonoids with several health benefits, including anti-inflammatory actions. Additionally, its EO consists of a large number of compounds such as monoterpenes, sesquiterpenes, alcohols, aldehydes, esters, and oxides. Among the methods for encapsulating EOs, spray-drying is the main one. In the present study, we aimed to determine the in vitro anti-inflammatory activity of EO from C. paradisi (grapefruit essential oil [GEO]) (whole and fractions) in a lipopolysaccharide (LPS)-induced inflammation model. Results indicate that Fr-GEO and Fr-GEO_SD exert protective effects against LPS-induced inflammation by decreasing gene expression and levels of pro-inflammatory cytokines as IL-6 and TNF-α. Monoterpenes as the most common components, as well as aldehydes and sesquiterpenes, might be responsible for such effects, although a synergistic action is not excluded. Furthermore, a higher percent of aldehydes is linked to improved olfactory properties. Our findings support the anti-inflammatory effects of selected Fr-GEO with a great potential for the development of new nutraceuticals and/or functional food for the treatment of inflammatory-associated diseases. PRACTICAL APPLICATION: The findings of this study support the anti-inflammatory effects of selected Fr-GEO with a great potential for the development of new nutraceuticals and/or functional food for the treatment of inflammatory-associated diseases.

Root-Shoot Nutrient Dynamics of Huanglongbing-Affected Grapefruit Trees.

With huanglongbing (HLB) causing a reduction in fine root mass early in disease progression, HLB-affected trees have lower nutrient uptake capability. Questions regarding the uptake efficiency of certain fertilizer application methods have been raised. Therefore, the goals of this study are to determine if nutrient management methods impact nutrient translocation and identify where in the tree nutrients are translocated. Destructive nutrient and biomass analysis were conducted on field grown HLB-affected grapefruit trees (Citrus × paradisi) grafted on 'sour orange' (Citrus × aurantium) rootstock under different fertilizer application methods. Fertilizer was applied in the form of either 100% soluble granular fertilizer, controlled release fertilizer (CRF), or liquid fertilizer. After three years, the entire tree was removed from the grove, dissected into eight different components (feeder roots, lateral roots, structural roots, trunk, primary branches, secondary branches, twigs, and leaves), weighed, and then analyzed for nutrient contents. Overall, application methods showed differences in nutrient allocation in leaf, twig, and feeder root; however, no consistent pattern was observed. Additionally, leaf, twig, and feeder roots had higher amount of nutrients compared to the other tree components. This study showed that fertilization methods do impact nutrient contents in different components of HLB-affected trees. Further research should be conducted on the impact of different fertilizer application methods and rates on HLB-affected trees.

Explore the antiproliferative phytocompounds from ethanolic extracts of Citrus paradisi against liver cancer cell line by chemical analysis using TLC and FT-IR spectroscopy / Explorar os fitocompostos antiproliferativos de extratos etanólicos de Citrus paradisi contra linhagem de células de câncer de fígado por análise química usando espectroscopia de TLC e FT-IR

The aim of the present study was to evaluate the in vitro antiproliferative activity of ethanolic extract of leaves and fruits Citrus paradisi plant on HepG-2 liver cell lines by MTT (3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2Hterazolium bromide) assay and to isolate and characterize the antiproliferative compounds by TLC (Thin layer chromatography) and FT-IR (Fourier transforms Infrared) spectroscopy. Qualitative phytochemical screening tests were performed to detect phytochemicals compounds from the crude extracts. Antioxidant activity of the plant extracts were characterized by using DPPH (2,2-Diphenyl-1-picrylhydrazyl) free radical scavenging method. The results showed that antioxidant activity using DPPH were found to be increased in a concentration dependent manner and decreased cell viability and cell growth inhibition in a dose dependent manner. The findings from this study indicated that fruit extract exhibited good antiproliferation and antioxidant potential. The seven functional groups of phytocompounds such as carboxylic acid, amine salt, aromatic compounds, cyclic alkene, aldehyde, fluoro compounds and alkene were detected by FT-IR which indicated that fruit extracts of Citrus paradisi possessed vast potential as a medicinal drug especially in liver cancer treatment.

O objetivo do presente estudo foi avaliar a atividade antiproliferativa in vitro do extrato etanólico de folhas e frutos da planta Citrus paradisi em linhagens de células hepáticas HepG-2 por MTT (3- (4, 5-dimetil-2-tiazolil) -2, Ensaio de brometo de 5-difenil-2H-terazólio) e isolar e caracterizar os compostos antiproliferativos por espectroscopia de TLC (cromatografia de camada fina) e FT-IR (infravermelho com transformadas de Fourier). Testes qualitativos de triagem fitoquímica foram realizados para detectar compostos fitoquímicos nos extratos brutos. A atividade antioxidante dos extratos vegetais foi caracterizada pelo método de eliminação de radicais livres DPPH (2,2-difenil-1-picrilhidrazil). Os resultados mostraram que a atividade antioxidante usando DPPH aumentou de uma maneira dependente da concentração e diminuiu a viabilidade celular e a inibição do crescimento celular de uma maneira dependente da dose. Os resultados deste estudo indicaram que o extrato de fruta exibiu bom potencial antiproliferação e antioxidante. Os sete grupos funcionais de fitocompostos, como ácido carboxílico, sal de amina, compostos aromáticos, alceno cíclico, aldeído, compostos de flúor e alceno, foram detectados por FT-IR, o que indicou que extratos de frutas de Citrus paradisi possuíam vasto potencial como medicamento, especialmente no tratamento de câncer do fígado.

Explore the antiproliferative phytocompounds from ethanolic extracts of Citrus paradisi against liver cancer cell line by chemical analysis using TLC and FT-IR spectroscopy

Abstract The aim of the present study was to evaluate the in vitro antiproliferative activity of ethanolic extract of leaves and fruits Citrus paradisi plant on HepG-2 liver cell lines by MTT (3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-terazolium bromide) assay and to isolate and characterize the antiproliferative compounds by TLC (Thin layer chromatography) and FT-IR (Fourier transforms Infrared) spectroscopy. Qualitative phytochemical screening tests were performed to detect phytochemicals compounds from the crude extracts. Antioxidant activity of the plant extracts were characterized by using DPPH (2,2-Diphenyl-1-picrylhydrazyl) free radical scavenging method. The results showed that antioxidant activity using DPPH were found to be increased in a concentration dependent manner and decreased cell viability and cell growth inhibition in a dose dependent manner. The findings from this study indicated that fruit extract exhibited good antiproliferation and antioxidant potential. The seven functional groups of phytocompounds such as carboxylic acid, amine salt, aromatic compounds, cyclic alkene, aldehyde, fluoro compounds and alkene were detected by FT-IR which indicated that fruit extracts of Citrus paradisi possessed vast potential as a medicinal drug especially in liver cancer treatment.

Resumo O objetivo do presente estudo foi avaliar a atividade antiproliferativa in vitro do extrato etanólico de folhas e frutos da planta Citrus paradisi em linhagens de células hepáticas HepG-2 por MTT (3- (4, 5-dimetil-2-tiazolil) -2, Ensaio de brometo de 5-difenil-2H-terazólio) e isolar e caracterizar os compostos antiproliferativos por espectroscopia de TLC (cromatografia de camada fina) e FT-IR (infravermelho com transformadas de Fourier). Testes qualitativos de triagem fitoquímica foram realizados para detectar compostos fitoquímicos nos extratos brutos. A atividade antioxidante dos extratos vegetais foi caracterizada pelo método de eliminação de radicais livres DPPH (2,2-difenil-1-picrilhidrazil). Os resultados mostraram que a atividade antioxidante usando DPPH aumentou de uma maneira dependente da concentração e diminuiu a viabilidade celular e a inibição do crescimento celular de uma maneira dependente da dose. Os resultados deste estudo indicaram que o extrato de fruta exibiu bom potencial antiproliferação e antioxidante. Os sete grupos funcionais de fitocompostos, como ácido carboxílico, sal de amina, compostos aromáticos, alceno cíclico, aldeído, compostos de flúor e alceno, foram detectados por FT-IR, o que indicou que extratos de frutas de Citrus paradisi possuíam vasto potencial como medicamento, especialmente no tratamento de câncer do fígado.

Analysis and Potential Value of Compounds Extracted From Star Ruby, Rio Red, and Ruby Red Grapefruit, and Grapefruit Juice Processing Residues via Steam Explosion.

Culled whole grapefruit (WG) and grapefruit juice processing residues (GP) are currently incorporated into low-cost animal feed. If individual chemical components found within these side streams could be recovered as high-value coproducts, this would improve the overall value of the grapefruit crop. In this study, pectic hydrocolloids, sugars, volatiles, phenolics, and flavonoids were extracted from Star Ruby, Rio Red, and Ruby Red GP and WG using a continuous pilot scale steam explosion system. Up to 97% of grapefruit juice oils and peel oils could be volatilized and contained 87-94% d-limonene. The recovery of pectin, as determined by galacturonic acid content, was between 2.06 and 2.72 g 100 g-1. Of the phenolics and flavonoids analyzed in this study, narirutin and naringin were extracted in the amounts of up to 10,000 and 67,000 µg g-1, respectively.

Chemical Profiling, Toxicity and Anti-Inflammatory Activities of Essential Oils from Three Grapefruit Cultivars from KwaZulu-Natal in South Africa.

The medicinal potential and volatile composition of different parts of three cultivars of grapefruit (Citrus paradisi) were evaluated for their toxicity and anti-inflammatory activities. Fresh leaf and fruit peel were separately isolated by hydrodistillation for 4 h. The essential oils were subjected to GC/GC-MS analysis for chemical profile. Toxicity of the essential oils in mice were evaluated using Lorke's method, while an anti-inflammatory assay was performed in a rat model using egg albumin-induced oedema. The oils obtained were light yellow in colour, and odour varied from strong citrus smell to mild. Percentage yield of fresh peel oil (0.34-0.57%) was greater than the fresh leaf oil yield (0.21-0.34%). D-limonene (86.70-89.90%) was the major compound identified in the leaf oil, while ß-phellandrene (90.00-91.01%) dominated the peel oil. At a dosage level of 5000 mg/kg, none of the oils showed mortality in mice. An anti-inflammatory bioassay revealed that all the oils caused a significant (p < 0.05-0.01) reduction in oedema size when compared to the negative control group throughout the 5 h post induction assessment period. The study reveals that the oils are non-toxic and demonstrate significant anti-inflammatory activity. Our findings suggest that the leaf and peel oils obtained from waste parts of grapefruit plants can be useful as flavouring agents, as well as anti-inflammatory agents.

Stress-inducible Arabidopsis thaliana RD29A promoter constitutively drives Citrus sinensis APETALA1 and LEAFY expression and precocious flowering in transgenic Citrus spp.

Transgenic 'Duncan' grapefruit (Citrus paradisi Macf.) and 'Valencia' sweet orange (Citrus sinensis [L.] Osbeck) plants ectopically expressing C. sinensis (cv. Washington navel orange) APETALA1 (CsAP1) or LEAFY (CsLFY) genes under control of the Arabidopsis thaliana stress-inducible promoter AtRD29A flowered under non-inductive (warm temperature, well-watered) greenhouse conditions, whereas their wild-type (WT) counterparts did not. The transgenic plants that flowered exhibited no altered morphological features, except the lack of thorns characteristic of juvenile WT plants. The most precocious T0 line, 'Duncan' grapefruit (Dun134-3) expressing the CsAP1 gene, flowered and fruited when it was 4.5 years old and the T1 siblings from this line flowered and fruited when they were just over 18 months old. In contrast, T1 seedlings from three lines of 'Duncan' grapefruit expressing the CsLFY gene flowered within 3 months after germination, but were unable to support fruit development. Transcript levels of corresponding transgenes in leaves were not correlated with earliness of flowering. To further study the activity of AtRD29A, leaves from three 'Carrizo' citrange (C. sinensis × Poncirus trifoliata) rootstock seedlings transformed with the green fluorescent protein (GFP) gene under regulation of the AtRD29A promoter were subjected to drought stress or well-watered conditions. Expression of GFP was not stress-dependent, consistent with the observation of flowering of CsAP1 and CsLFY transgenic plants under non-inductive conditions. Taken together, the results suggest that AtRD29A is constitutively expressed in a citrus background. Despite the loss of control over flowering time, transgenic citrus lines ectopically expressing C. sinensis AP1 or LFY genes under control of the A. thaliana RD29A promoter exhibit precocious flowering, fruit development and viable transgenic seed formation. These transformed lines can be useful tools to reduce the time between generations to accelerate breeding.

Photosensitizing Furocoumarins: Content in Plant Matrices and Kinetics of Supercritical Carbon Dioxide Extraction.

Furocoumarins are a group of plant phytoalexins exhibiting various bioactive properties; the most important of which are photosensitization and alteration of P450 cytochrome activity. Supercritical fluid extraction with carbon dioxide has been proposed as a green alternative for an organic solvent extraction of the furocoumarins. Four plant matrices rich in furocoumarins were extracted with CO2 at a temperature of 80 °C and pressure of 40 MPa, as these conditions were characterized by the highest solubility of furocoumarins. The extracts collected were analyzed using the HPLC method and the results obtained were used for the mathematical modeling of the observed phenomena. The total content of the furocoumarins in the matrices was 4.03-26.45 mg g-1 of dry weight. The impact of the process parameters on the solubility was consistent with the Chrastil equation. The broken plus intact cell model proved to be suitable to describe extraction curves obtained. The research proved the possibility of supercritical carbon dioxide utilization for the extraction of the furocoumarins from plant material and provided valuable data for prospective industrial-scale experiments.

Optimization and comparison of non-conventional extraction technologies for Citrus paradisi L. peels: a valorization approach.

The phenolics from grapefruit peel were obtained by ultrasound assisted extraction (UAE) and enzyme-assisted extraction (EAE). Extraction parameters were optimized using response surface methodology to maximize the yield of total phenolic content (TPC) and total flavonoid content (TFC). The optimized extracts from UAE and EAE were then compared with conventional solvent extraction for their TPC, TFC, antioxidant activity and phenolic composition. The best optimized conditions for UAE was obtained at 33.12 min extraction time, 71.11% amplitude and a solvent-solid (SS) ratio of 39.63 mL/g. The optimized parameters for EAE were 4.81 h extraction time, 0.9% enzyme concentration and 40 mL/g SS ratio. Similar values of experimental and predicted TPC and TFC at optimized conditions indicates the suitability of the quadratic model in optimizing the extraction parameters. Further characterization of extracts suggested EAE as most efficient process in extracting bioactive compounds.

Extraction of diverse polyphenols in relation with storage periods of Citrus paradisi CV. Shamber through HPLC-DAD technique using different solvent.

In this article, Citrus paradisi, (Shamber) an exceptional source of Vitamins A and C and full of nutrients, selected for extraction of diverse polyphenols including dietary flavonoids and essential flavonoids by HPLC-DAD technique using various solvents. These essential targeted compounds also analyze after keeping different storage periods and compare with fresh fruits for better efficacy of these compounds. The highest number of phenolic compounds including gallic acid, chlorogenic acid sinapic acid, ferulic acid, myricetin, quercetin, and kaempferol extracted in methanol solvent leading to the new compounds of tetra-O-methylscutellar and heptamethoxy flavone. The essential flavonoids determined by polyethersulfone filter and insoluble precipitation separated by the dimethyl sulfoxide. The results showed that the methanolic extraction exhibited higher essential flavonoids including nobiletin, sinensetin, tangeritin, and tetra-O-methylscutellarein and heptamethoxy flavone. The RP-HPLC analysis exposed the maximum number of nutritional flavonoids like naringin, hesperidin, total flavones, glycosyl. Moreover, it observed that dietary flavonoids and phenolic compounds of stored fruits were unaffected in 30 days of storage periods while minor variations were pragmatic during 60-90 days storage. The investigation revealed that C. paradisi proves to be the valuable resource of different phenolic compounds and flavonoids which are effective against various oxidative stresses in the human body.

Effect of different concentrations of pulverized mesocarp of Citrus paradisi Macf on the morphology and glass transition temperature of spray-dried lemon juice powder.

The aim of this research was to evaluate the effect of different concentrations of pulverized mesocarp of Citrus paradisi Macf as a drying aid, on morphology, particle size and glass transition temperature of spray-dried lemon juice powder. Five concentrations of grapefruit mesocarp (0.4%, 0.8%, 1.2%, 1.6% and 2.0% (w/w)) and maltodextrin DE 10 (1.2%, w/w) were evaluated as encapsulant agents. The obtained data were evaluated by one-way ANOVA using Statistix version 8.0. For the means separation, the Tukey's test was applied with a 95% reliability. The morphology of the particles was described. According to the results, by applying different levels of coating agent in lemon juices, powder particles with different sizes (from 3.07 to 6.20 µm) and shapes (spherical, irregular and shrunken or reduced) are obtained; however, their glass transition temperature is not modified, finding values between 37.43 and 38.64°C.

Effect of different concentrations of pulverized mesocarp of Citrus paradisi Macf. on the bromatological characteristics of spray-dried lemon juice powder.

The aim of this research was to evaluate the effect of different concentrations of pulverized mesocarp of Citrus paradisi Macf. as a drying aid on the bromatological characteristics and yield of spray-dried lemon juice powder. Five concentrations of grapefruit mesocarp encapsulant (0.4, 0.8, 1.2, 1.6, and 2.0% (w / w)) and maltodextrin DE 10 (1.2%, w / w) were evaluated as encapsulant agents. The highest yield (46.76%) was obtained with 1.2% of grapefruit encapsulant. Water activity and ash content were inversely proportional to the added encapsulant concentration. The highest moisture value was obtained with 0.4% and the highest soluble solids with 2.0%. For all treatments, the pH level did not change, except at 0.4% (it was lower). The concentrations of the encapsulants significantly affected the evaluated characteristics, except for the proteins.

Inhibition of X-linked inhibitor of apoptosis protein enhances anti-tumor potency of pure total flavonoids on the growth of leukemic cells.

Flavonoids, a vast group of polyphenols widely distributed in plants, are known to possess a range of biological activities and potential anti-tumor effects. X-linked inhibitor of apoptosis protein (XIAP) promotes the progression of leukemia by preventing tumor cells undergoing apoptosis. The present study investigated the potential effects and underlying mechanisms of pure total flavonoids from Citrus paradisi Macfad (PTFC) on human U937 cells, and explored the effects of short hairpin (sh)RNA-mediated XIAP knockdown on the anti-cancer effects of PTFC. Western blotting was used to determine level of apoptosis-associated effectors following PTFC treatment. A lentiviral vector of RNA interference of XIAP gene was constructed to downregulate XIAP expression. MTT assay and flow cytometry were used to determine the effects of PTFC separately or combined with XIAP-shRNA on inhibition and apoptosis of U937 cells, respectively. Treatment with PTFC effectively inhibited leukemic cell proliferation in a dose- and time-dependent manner. PTFC induced apoptosis of U937 cells in a dose-dependent manner, at a particular concentration range, by decreasing XIAP expression levels and activating caspases-3, -7 and -9. PTFC treatment combined with XIAP-shRNA additionally demonstrated a marked increase in cell apoptosis, compared with PTFC or XIAP-shRNA alone (P<0.05). Therefore, these findings suggest that PTFC inhibits growth and induces apoptosis in U937 cells in vitro. Furthermore, suppression of XIAP expression enhances these effects.

Bioactivity-guided isolation of potent anxiolytic compounds from leaves of Citrus paradisi.

CONTEXT: Citrus fragrances have been attributed with mood-enhancing properties by aromatherapists. Leaves of this plant have been reported to exert anti-anxiety activity. Till date, no specific phytoconstituent responsible for this has been identified. OBJECTIVE: Isolation of anxiolytic constituent of Citrus paradisi using bioactivity-guided fractionation. MATERIALS AND METHODS: Leaf extracts of four varieties of C. paradisi in petroleum ether, chloroform, methanol and water were evaluated for anti-anxiety activity in mice using elevated plus-maze apparatus. Because of activity in methanol extract, it was used for safety evaluation/acute toxicity studies in animals. Bioactive fraction of methanol extract was subjected to column chromatography and structure of the isolated compound was elucidated by melting point, ultraviolet, infrared, nuclear mass reactor and mass spectroscopy. The isolated constituents were further evaluated for anti-anxiety activity using light/dark model and hole-board model of anxiety. RESULTS: Results showed no mortality at a dose up to 2000 mg/kg body weight that indirectly reflects the safety profile of the leaf extracts. Fractionation of methanol extract led to the isolation of four flavonoids (rutin, quercetin, kaempferol and myricetin). The isolated compounds exhibited significant anxiolytic activity in different animal models. CONCLUSION: The study confirms the presence of four flavonoids responsible for anti-anxiety activity.

Evaluation of Antibacterial Activity of Three Selected Fruit Juices on Clinical Endodontic Bacterial Strains.

INTRODUCTION: The increasing problem of antibiotic drug resistance by pathogenic microorganisms in the past few decades has recently led to the continuous exploration of natural plant products for new antibiotic agents. Many consumable food materials have good as well as their bad effects, good effect includes their antibacterial effects on different microorganisms present in the oral cavity. Recently, natural products have been evaluated as source of antimicrobial agent with efficacies against a variety of microorganisms. METHODOLOGY: The present study describes the antibacterial activity of three selected fruit juices (Apple, Pomegranate and Grape) on endodontic bacterial strains. Antimicrobial activity of fruit juices were tested by wel l diffusion assay by an inhibition zone surrounding the well. The aim of the study was to evaluate the antibacterial activity of three fruit juises on different endodontic strains. RESULT: Agar well diffusion method was adopted for determining antibacterial potency. Antibacterial activity present on the plates was indicated by an inhibition zone surrounding the well containing the fruit juice. The zone of inhibition was measured by measuring scale in millimeter. Comparision between antibacterial efficacy of all three fruit juices against Enterococcus feacalis and Streptococcus mutans was observed with significant value of P ≤ 0.05. CONCLUSION: The results obtained in this study clearly demonstrated a significant antimicrobial effect of apple fruit juice against Enterococcus fecalis and Streptococcus mutans. However, preclinical and clinical trials are needed to evaluate biocompatibility & safety before apple can conclusively be recommended in endodontic therapy, but in vitro observation of apple effectiveness appears promising.