Molecular Mechanisms Responsible for In Vitro Cytotoxic Attributes of Conyza bonariensis Extract against Lymphoblastic Leukaemia Jurkat Cells.

BACKGROUND: Conyza bonariensis is known to have anti-cancer properties. OBJECTIVE: The current study investigated the in vitro pro-apoptotic properties of Conyza bonariensis (C. bonariensis) towards human lymphoblastic leukemia Jurkat cells. METHODS: Ariel parts of C. bonariensis were macerated in a non-polar (n-Hexane) solvent. MTS cell viability assay was employed to determine the cytotoxic activity of the extract towards human leukemia Jurket cells and normal Peripheral Blood Mononuclear Cells (PBMCs). The phytochemical composition of the extract was screened using HPLC method. Flow cytometric studies (FACS) were conducted to explore the pro-apoptotic potential of the extract. Western blot studies were employed to identify the molecular targets involved in the induction of apoptosis. RESULTS: The n-hexane extract showed selective cytotoxic activity towards Jurkat cells. FACS analysis indicated that the extract induced early and late apoptosis in Jurkat cells. Western blot studies revealed that the extract downregulated the expression of DNMT1, SIRT1, and UHRF1 with a simultaneous up-regulation of p73 and caspases-3 proteins expression. HPLC characterization of the extract revealed the presence of phenolic compounds. CONCLUSION: Overall, these findings demonstrate that the anti-cancer effects of a Conyza bonariensis extract towards human lymphoblastic leukemia Jurkat cells are due to the modulation of the activity of multiple oncogenic and tumor suppressor proteins. Phenolic contents of the extract are proposed to be responsible for these activities.

A Case of Mixed Type II Cryoglobulinemic Vasculitis Associated With Marginal Zone B-cell Lymphoma.

Our patient is a male in his 40s with a past medical history of sickle cell trait, factor V Leiden mutation, marginal zone B-cell lymphoma, gastric mucosa-associated lymphoid tissue (MALT) lymphoma, and sarcoidosis who presented with the complaint of hemoptysis, dyspnea, abdominal pain, arthralgias, peripheral edema of the lower extremities with petechial rash, and oliguria. Investigations revealed acute kidney injury and bilateral transudative pleural effusion. Serology was positive for elevated rheumatoid factor, low complement components, and cryoglobulins. Renal biopsy showed membranoproliferative cryoglobulinemic glomerulonephritis with deposition of monoclonal IgM and IgG3 with kappa light chain and C3 component. The patient was diagnosed with mixed type II cryoglobulinemic vasculitis in the setting of untreated marginal B-cell lymphoma. He had a complex clinical course, requiring multiple intubations, hemodialysis, and treatment with intravenous immunoglobulin, plasmapheresis, steroids, and chemotherapy, to which he initially responded. During treatment, he developed cardiomyopathy associated with congestive heart failure and passed away due to cardiac arrest. We present a rare case of mixed type II cryoglobulinemic vasculitis secondary to untreated marginal zone B-cell lymphoma in a hepatitis C virus (HCV) negative patient, which has not been reported before.

A Review on Natural Sources Derived Protein Nanoparticles as Anticancer Agents.

Cancer notably carcinoma represents a prominent health challenge worldwide. A variety of chemotherapeutic agents are being used to deal with a variety of carcinomas. However, these delivering agents not only enter the targeted site but also affect normal tissues yielding poor therapeutic outcomes. Chemotherapeutic-associated problems are being attributed to drug non-specificity resulting from poor drug delivery systems. These problems are now being solved using nanomedicine, which entails using nanoparticles as drug delivery systems or nanocarriers. This nanoparticle-based drug delivery system enhances clinical outcomes by enabling targeted delivery, improving drug internalization, enhanced permeability, easy biodistribution, prolonged circulation and enhanced permeability rate, thereby improving the therapeutic effectiveness of several anticancer agents. Natural Protein-based Nanoparticles (PNPs) such as ferritin, lipoprotein, and lectins from natural sources have gained extensive importance at a scientific community level as nanovehicle for effective drug delivery and photo acoustic labeling replacing several synthetic nanocarriers that have shown limited therapeutic outcomes. The bioavailability of PNP, the chance of genetic engineering techniques to modify their biological properties made them one of the important raw material sources for drug delivery research. This current review highlighted different chemotherapeutic agents used in the treatment of some carcinomas. It also focused on the wide variety of natural protein sources derived nanoparticles (NPs) as anticancer delivery of agents for cancer therapy.

Tylophora hirsuta (Wall.) Extracts Ameliorate Diabetes Associated with Inflammation in Alloxan-induced Diabetic Rats.

BACKGROUND: Tylophora hirsuta (Wall) has long been used as traditional medicine for the treatment of diabetes. The current study is designed to evaluate the anti-diabetic and anti-inflammatory activity of different extracts of aerial parts of Tylophora hirsuta. METHODS: Sequential maceration was conducted to obtain extracts. Total phenolic contents were determined by the Folin-Ciocalteau method. The anti-oxidant activity was assessed by DPPH free radical scavenging assay. The extracts were tested for its inhibitory activity against α-amylase in-vitro. In-vivo anti-diabetic assay was conducted using alloxan-induced diabetic model and OGTT was conducted on normal rats. ELISA was used to determine the pro-inflammatory cytokines (TNF-α and IL-6). The polyphenolic composition of the extract was analyzed using an HPLC system. RESULTS: Aqueous extract exhibited highest total phenolic contents (985.24± 3.82 mg GAE/100 g DW), antioxidant activity (IC50 = 786.70 ± 5.23 µg/mL), and alpha-amylase inhibition (IC50 =352.8 µg/mL). The aqueous extract of Tylophora hirsuta showed remarkable in-vivo anti-diabetic activity. Results were compared with standard drug glibenclamide. Alloxan induced diabetic mediated alterations in liver function enzymes, renal function determinants, and lipid parameters were significantly restored in aqueous extract treated diabetic rats. A significant reduction in pro-inflammatory cytokines (p<0.001) was observed when compared to the control group. HPLC analysis confirms the presence of quercetin, gallic acid, cinnamic acid, and p-coumaric acid. CONCLUSION: These results showed that Tylophora hirsuta possesses strong anti-diabetic and anti-inflammatory potentials and justify its folklore use for the management of diabetes.

Antioxidant and Wound Healing Potential of Essential Oil from Citrus reticulata Peel and Its Chemical Characterization.

BACKGROUND: Fruit peels are considered as waste and contribute to a major proportion of the biomass. They can be a good source of various therapeutic benefits. Peels biomass of citrus fruits is usually considered as garbage. Such peels may have many important and valuable medicinal components with pharmacological activities. Citrus reticulata, (Rutaceae family, local name tangerine) is a local seasonal fruit in Pakistan, a very good example of wastage of its peels. OBJECTIVE: The study is based on the exploration of a citrus fruit peel derived essential oil, its chemical characterization, identification of various bioactive components and the exploration of pharmacological potentials (antibacterial and wound healing activity). METHODS: Essential oil was recovered by hydro-distillation of freshly collected peels. Chemical constituents of oil were determined by Gas Chromatography-Mass Spectroscopy (GC-MS) analysis. Antioxidant activities were evaluated by total phenolic contents, total flavonoid content, DPPH scavenging activity and reducing power assay. Antibacterial activity was determined using disc diffusion assay. In vivo wound healing potential was determined in rabbits after topical administration of oil. Wound scoring was calculated followed by histological study. RESULTS: GC-MS analysis showed the presence of various components with the greatest proportion of D-Limonene (89.31%). Total flavonoid and phenolic contents were found to be 14.63 ± 0.95 mg CE/g and 17.03 ± 3.24 mg GAE/g respectively, while DPPH activity was found to be 73.32%. A better antibacterial activity was shown against E. coli. In vivo studies showed significant reduction in wound diameter in essential oil treatment groups. Further, the essential oil was found non-irritant in draize scoring. CONCLUSION: The study concluded that essential oil of this fruit peel might be used for antibacterial and wound healing purposes.

Chemical characterization, antioxidant evaluation, and antidiabetic potential of Pinus gerardiana (Pine nuts) extracts.

Methanolic and ethyl acetate extracts of pine nuts were evaluated for in vitro antioxidant and inhibition of alpha-amylase activities. Pine nut extracts were orally administered to alloxan monohydrate-induced diabetes in mice at 250, 500, and 750 mg/kg. Both extracts showed a significant antioxidant and inhibition of alpha-amylase activities. Animal studies showed a decline in fasting blood glucose, hyperlipidemia, and weight loss in diabetic mice. The administration of plant extracts decreased the blood glucose level during the oral glucose tolerance test. Histopathological examination showed a decrease in alloxan-induced lesions in the pancreas, liver, and kidney of animals treated with pine nut extracts. Furthermore, pine nut extract ameliorated oxidative stress-induced hepatotoxicity in diabetic mice. The presence of quercetin, gallic acid, vanillic acid, benzoic acid, syringic acid, m-coumaric acid, and other phenolic compounds might be related to hypoglycemic, alpha-amylase inhibitory, antioxidant, and antihyperlipidemic potential of pine nut extracts. PRACTICAL APPLICATIONS: Dry fruits are rich in dietary nutrients, minerals, and phytochemicals that can be used to treat and prevent lifestyle disease. Pine nuts are edible and have economic importance in addition to several traditional uses such as diuretic, antiseptic, expectorant, antibacterial, antiviral, antifungal, antihypertensive, and antineuralgic properties. This study was conducted to investigate the pine nut extracts for their antioxidant and antidiabetic potentials using in vitro methods and animal disease model. The findings of the present study suggest that the extracts of Pine nuts may be helpful in treating hyperglycemia during diabetes and prevent its complications such as hepatic damage, nephrotoxicity, weight loss, and hyperlipidemia.

Delonix regia a Folklore Remedy for Diabetes; Attenuates Oxidative Stress and Modulates Type II Diabetes Mellitus.

BACKGROUND: The folkloric profile of Delonix regia demonstrates that it can be used in the management of diabetes. OBJECTIVE: The present study was conducted to evaluate the safety profile of the aerial part extracts of Delonix regia and their antidiabetic potential along with improvement in oxidative stress. MATERIALS AND METHODS: Phytochemical screening, total phenolic, and flavonoid contents along with in-vitro antioxidant and alpha-amylase inhibitory activities were determined. HPLC analysis, acute toxicity, glucose tolerance, in-vivo antidiabetic effect along with the influence on biochemical, oxidative stress parameters, and comet assay of the active extract were performed and assessed. RESULTS: Total phenolic (831.6±0.002 mg/g GAE) and flavonoid (361.4±0.002 mg/g QE) contents were found to be higher in the methanolic extract. Inhibitory concentration IC50 indicated better results for the methanolic extract in DPPH (47.6µg/mL) and alpha-amylase inhibitory (14.61µg/mL) assays. HPLC analysis of the methanolic extract confirmed the presence of quercetin, gallic acid, caffeic acid, cinnamic acid, ferulic acid, and p-coumaric acid. Acute oral toxicity exhibited no mortality and morbidity during the 24h period. The methanolic extract showed better tolerance to glucose. Streptozotocin- nicotinamide (55-110 mg/kg) induced hyperglycemia declined along with improvement in hematological, biochemical parameters and oxidative stress markers (SOD, CAT, H202) in a dose-dependent manner. The maximum effect was recorded at 500mg/kg dose. Comet assay was performed for genotoxic studies and it was observed that the methanolic extract of Delonix regia showed the maximum genoprotective effect at 100µg/mL. CONCLUSION: The findings suggest that the methanolic aerial part extract of Delonix regia exhibited hypoglycemic, antioxidant, and hypolipidemic activities. The antidiabetic effect was comparable to glibenclamide suggesting its therapeutic use as a natural anti-diabetic remedy.

Preliminary study on phosphate solubilizing Bacillus subtilis strain Q3 and Paenibacillus sp. strain Q6 for improving cotton growth under alkaline conditions.

BACKGROUND: Low phosphorus availability limits crop production in alkaline calcareous soils in semi-arid regions including Pakistan. Phosphate solubilizing bacteria may improve crop growth on alkaline calcareous soils due to their ability to enhance P availability. METHODS: Twenty rhizobacterial isolates (Q1-Q20) were isolated from rhizosphere of cotton and characterized for their growth promoting attributes in vitro. The selected phosphate solubilizing isolates were further screened for their ability to improve cotton growth under axenic conditions (jar trial). The phosphorus solubilization capacities of selected strains were quantified and these strains were identified through 16S rDNA sequencing. RESULTS: Isolates Q2, Q3, Q6, Q7, Q8, Q13 and Q14 were able to solubilize phosphate from insoluble sources. Most of these isolates also possessed other traits including catalase activity and ammonia production. The growth promotion assay showed that Q3 was significantly better than most of the other isolates followed by Q6. Maximum root colonization (4.34 × 106 cfu g-1) was observed in case of isolate Q6 followed by Q3. The phosphorus solubilization capacities of these strains were quantified, showing a maximum phosphorus solubilization by Q3 (optical density 2.605 ± 0.06) followed by the Q6 strain. The strain Q3 was identified as Bacillus subtilis (accession # KX788864) and Q6 as Paenibacillus sp. (accession # KX788865) through 16S rDNA sequencing. DISCUSSION: The bacterial isolates varied in their abilities for different growth promoting traits. The selected PGPR Bacillus subtilis strain Q3 and Paenibacillus sp. strain Q6 have multifarious growth promoting traits including ability to grow at higher EC and pH levels, and phosphorus solubilizing ability. These strains can efficiently colonize cotton roots under salt affected soils and help plants in phosphorus nutrition. It is concluded that both strains are potential candidates for promoting cotton growth under alkaline conditions, however further investigation is required to determine their potential for field application.