Toxic effects of organophosphate pesticide monocrotophos in aquatic organisms: A review of challenges, regulations and future perspectives.

In recent times, usage of pesticide, herbicides and synthetic fertilizers in farming lands has made the environment worse. The pesticide residues and toxic byproducts from agricultural lands were found to contaminate the aquatic ecosystem. The misuse of synthetic pesticide not only affects the environment, but also affects the health status of aquatic organisms. The organophosphate pesticide pollutants are emerging contaminants, which threatens the terrestrial and aquatic ecosystem. Monocrotophos (MCP) is an organophosphate insecticide, utilized on crops including rice, maize, sugarcane, cotton, soybeans, groundnuts and vegetables. MCP is hydrophilic in nature and their solubilizing properties reduce the soil sorption which leads to groundwater contamination. The half-life period of MCP is 17-96 and the half-life period of technical grade MCP is 2500 days if held stable at 38 °C in a container. MCP causes mild to severe confusion, anxiety, hyper-salivation, convulsion and respiratory distress in mammals as well as aquatic animals. The MCP induced toxicity including survival rate, behavioural changes, reproductive toxicity and genotoxicity in different aquatic species have been discussed in this review. Furthermore, the ultimate aim of this review is to highlight the international regulations, future perspectives and challenges involved in using the MCP.

Natural quinones induce ROS-mediated apoptosis and inhibit cell migration in PANC-1 human pancreatic cancer cell line.

Pancreatic cancer is one of the most devastating of all malignancies with poor prognosis and high mortality rates worldwide. Thymoquinone, plumbagin and juglone, which are naturally occurring quinones, have been reported for their promising anticancer effect on different cancer cells. However, their mechanism of action and antimetastatic effects are largely unknown against the human pancreatic cancer cell line (PANC-1). In this study, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay revealed a dose-dependent decrease of viability in quinone-treated PANC-1 cells. In addition, the assessment of changes in cells has demonstrated an occurrence of typical apoptotic morphology in treated PANC-1 cells compared with control. Besides this, the apoptosis induction was further quantitatively confirmed through flow cytometry analysis. Furthermore, thymoquinone, plumbagin and juglone were evaluated for their influence on reactive oxygen species (ROS) generation through 2,7-dichlorofluorescein diacetate (DCFDA) staining and they dramatically increased the intracellular ROS level in treated PANC-1 cells, suggesting the critical role of ROS in their apoptosis induction. This study also demonstrated the wound healing potential of these compounds and inhibited PANC-1 cell migration in a time-dependent manner compared with control. This inhibition was correlated with reduced expression of matrix metalloproteinase-9 (MMP-9) in juglone-treated cells detected through gelatin zymography. In conclusion, thymoquinone, plumbagin and juglone significantly inhibited cell growth and induced ROS-mediated apoptosis in PANC-1 cells. In addition, they could be potent antimetastatic agents due to their anti-migratory effect against PANC-1 human pancreatic cancer cells.

Concurrent loss of co-stimulatory molecules and functional cytokine secretion attributes leads to proliferative senescence of CD8(+) T cells in HIV/TB co-infection.

The role of T-cell immunosenescence and functional CD8(+) T-cell responses in HIV/TB co-infection is unclear. We examined and correlated surrogate markers of HIV disease progression with immune activation, immunosenescence and differentiation using T-cell pools of HIV/TB co-infected, HIV-infected and healthy controls. Our investigations showed increased plasma viremia and reduced CD4/CD8 T-cell ratio in HIV/TB co-infected subjects relative to HIV-infected, and also a closer association with changes in the expression of CD38, a cyclic ADP ribose hydrolase and CD57, which were consistently expressed on late-senescent CD8(+) T cells. Up-regulation of CD57 and CD38 were directly proportional to lack of co-stimulatory markers on CD8(+) T cells, besides diminished expression of CD127 (IL-7Rα) on CD57(+)CD4(+) T cells. Notably, intracellular IFN-γ, perforin and granzyme B levels in HIV-specific CD8(+) T cells of HIV/TB co-infected subjects were diminished. Intracellular CD57 levels in HIV gag p24-specific CD8(+) T cells were significantly increased in HIV/TB co-infection. We suggest that HIV-TB co-infection contributes to senescence associated with chronic immune activation, which could be due to functional insufficiency of CD8(+) T cells.

Antioxidant and cytotoxic activities of three species of tropical seaweeds.

BACKGROUND: Three species of seaweeds (Padina tetrastromatica, Caulerpa racemosa and Turbinaria ornata) are widely consumed by Asians as nutraceutical food due to their antioxidant properties. Studies have shown that these seaweeds exhibit bioactivities which include antimicrobial, antiviral, anti-hypertensive and anticoagulant activities. However, investigations into the mechanisms of action pertaining to the cytotoxic activity of the seaweeds are limited. The aim of this study was to determine the antioxidant and cytotoxic activities of whole extracts of P. tetrastromatica, C. racemosa and T. ornata, including the cellular events leading to the apoptotic cell death of the extract treated-MCF-7 cells. Bioassay guided fractionation was carried out and the compounds identified. METHODS: Powdered samples were sequentially extracted for 24 h. Their antioxidant activities were assessed by the DPPH radical, superoxide, nitric oxide and hydroxyl radical scavenging assays. The cytotoxic activity of the extract-treated MCF-7cells was assessed using the MTT assay. The most potent fraction was subjected to bioassay guided fractionation with column chromatography. All the fractions were tested for cytotoxic activity, caspase activity and effect on DNA fragmentation. RESULTS: All three seaweeds showed potent radical scavenging activities in the various assays. The activity of the cellular antioxidant enzymes, superoxide dismutase, catalase and glutathione reductase, in MCF-7 cells, decreased in a time-dependent manner. The partially purified fractions exhibited higher cytotoxic activity, as assessed by the MTT assay, than the whole extracts in the breast adenocarcinoma cell line, MCF-7. LC-MS analysis revealed the presence of bioactive alkaloids such as camptothecin, lycodine and pesudopelletierine. CONCLUSION: Based on the results obtained, all three seaweeds are rich sources of enzymatic and non-enzymatic antioxidants which could contribute to their reported medicinal benefits.

Petroselinum crispum has antioxidant properties, protects against DNA damage and inhibits proliferation and migration of cancer cells.

BACKGROUND: Petroselinum crispum (English parsley) is a common herb of the Apiaceae family that is cultivated throughout the world and is widely used as a seasoning condiment. Studies have shown its potential as a medicinal herb. In this study, P. crispum leaf and stem extracts were evaluated for their antioxidant properties, protection against DNA damage in normal 3T3-L1 cells, and the inhibition of proliferation and migration of the MCF-7 cells. RESULTS: The dichloromethane extract of P. crispum exhibited the highest phenolic content (42.31 ± 0.50 mg GAE g(-1) ) and ferric reducing ability (0.360 ± 0.009 mmol g(-1) ) of the various extractions performed. The extract showed DPPH radical scavenging activity with an IC50 value of 3310.0 ± 80.5 µg mL(-1) . Mouse fibroblasts (3T3-L1) pre-treated with 400 µg mL(-1) of the extract showed 50.9% protection against H2 O2 -induced DNA damage, suggesting its potential in cancer prevention. The extract (300 µg mL(-1) ) inhibited H2 O2 -induced MCF-7 cell migration by 41% ± 4%. As cell migration is necessary for metastasis of cancer cells, inhibition of migration is an indication of protection against metastasis. CONCLUSION: Petroselinum crispum has health-promoting properties with the potential to prevent oxidative stress-related diseases and can be developed into functional food.

Antioxidant activity of Coriandrum sativum and protection against DNA damage and cancer cell migration.

BACKGROUND: Coriandrum sativum is a popular culinary and medicinal herb of the Apiaceae family. Health promoting properties of this herb have been reported in pharmacognostical, phytochemical and pharmacological studies. However, studies on C. sativum have always focused on the aerial parts of the herb and scientific investigation on the root is limited. The aim of this research was to investigate the antioxidant and anticancer activities of C. sativum root, leaf and stem, including its effect on cancer cell migration, and its protection against DNA damage, with special focus on the roots. METHODS: Powdered roots, leaves and stems of C. sativum were extracted through sequential extraction using hexane, dichloromethane, ethyl acetate, methanol and water. Total phenolic content, FRAP and DPPH radical scavenging activities were measured. Anti-proliferative activitiy on the breast cancer cell line, MCF-7, was assayed using the MTT assay. Activities of the antioxidant enzymes, catalase, superoxide dismutase, glutathione peroxidase, and of the caspases-3, -8 and -9 were assayed on treatment with the extract. Cell cycle progression was analysed using flow cytometry. The scratch motility assay was used to assess inhibition of MCF-7 cell migration. DNA damage in 3 T3-L1 fibroblasts was evaluated by the comet assay. The components in the extract were identified by HPLC and GC-MS. RESULTS: The ethyl acetate extract of C. sativum roots showed the highest antiproliferative activity on MCF-7 cells (IC50 = 200.0 ± 2.6 µg/mL) and had the highest phenolic content, FRAP and DPPH scavenging activities among the extracts. C. sativum root inhibited DNA damage and prevented MCF-7 cell migration induced by H2O2, suggesting its potential in cancer prevention and inhibition of metastasis. The extract exhibited anticancer activity in MCF-7 cells by affecting antioxidant enzymes possibly leading to H2O2 accumulation, cell cycle arrest at the G2/M phase and apoptotic cell death by the death receptor and mitochondrial apoptotic pathways. CONCLUSIONS: This study is the first report on the antioxidant and anticancer properties of C. sativum root. The herb shows potential in preventing oxidative stress-related diseases and would be useful as supplements used in combination with conventional drugs to enhance the treatment of diseases such as cancer.

Black soldier fly (Hermetia illucens) larvae as potential feedstock for the biodiesel production: Recent advances and challenges.

Black soldier fly larvae (BSFL) Hermetia illucens is fastest growing and most promising insect species especially recommended to bring high-fat content as 5th generation bioenergy. The fat content can be fully optimized during the life-cycle of the BSFL through various organic dietary supplements and environmental conditions. Enriched fat can be obtained during the larval stages of the BSF. The presence of high saturated and unsaturated fatty acids in their body helps to produce 70 % of extractable oil which can be converted into biodiesel through transesterification. The first-generation biodiesel process mainly depends on catalytic transesterification, however, BSFL had 94 % of biodiesel production through non-catalytic transesterification. This increases the sustainability of producing biodiesel with less energy input in the process line. Other carbon emitting factors involved in the rearing of BSFL are less than the other biodiesel feedstocks including microalgae, cooking oil, and non-edible oil. Therefore, this review is focused on evaluating the optimum dietary source to produce fatty acid rich larvae and larval growth to accumulate C16-18 fatty acids in larger amounts from agro food waste. The process of optimization and biorefining of lipids using novel techniques have been discussed herein. The sustainability impact was evaluated from the cultivation to biodiesel conversion with greenhouse gas emissions scores in the entire life-cycle of process flow. The state-of-the-art in connecting circular bioeconomy loop in the search for bioenergy was meticulously covered.

Green and eco-friendly approaches for the extraction of chitin and chitosan: A review.

Chitin is one of the most diverse and naturally occurring biopolymers, and it is mainly present in crustaceans, insects, and fungi. Chitosan is derived from chitin by deacetylation process. It is important to note that the conventional chemical method of extracting chitin includes disadvantages and it poses various environmental issues. Recently, the green extraction techniques have perceived substantial development in the field of polymer chemistry. A variety of methods have been successfully developed using green extraction techniques for extracting chitin and chitosan from various resources. It includes the use of ionic liquids (ILs), deep eutectic solvents (DES), microbial fermentation, enzyme-assisted extraction (EAE), microwave-assisted extraction (MAE), ultrasonic-assisted extraction (UAE), subcritical water extraction (SWE), and electrochemical extraction (ECE). In this review, the extraction of chitin and chitosan using greener approaches were summarized. In addition, challenges, opportunities and future perspectives of green extraction methods have also been narrated.

In vivo acute toxicity evaluation and in vitro molecular mechanism study of antiproliferative activity of a novel indole Schiff base ß-diiminato manganeseIII complex in hormone-dependent and triple negative breast cancer cells.

Breast cancer is the most frequently diagnosed cancer among women worldwide. Recently, increasing attention has been paid to the anticancer effects of transition metal complexes of indole Schiff bases. ß-diiminato ManganeseIII complex has shown promising cell cycle arrest and apoptosis induction against MCF-7 and MDA-MB-231 breast cancer cells. In this study, time- and dose- dependent inhibitory activity were evaluated using MTT assay after 48 h and 72 h exposure time. In addition, median effect analysis was conducted according to Chou-Talalay method to investigate whether MnIII complex has synergistic effect in combination with chemotherapeutic drugs on inhibiting breast cancer cell growth. The molecular mechanisms underlying its potent antiproliferative effect was determined through bioluminescent caspase-3/7, -8 and -9 activity assays and quantitative expression analysis of cell cycle- and apoptosis-related genes. Furthermore, safety evaluation of MnIII complex was assessed through the acute oral toxicity test in in vivo model. The MTT assay results revealed that it potently reduced the viability of MCF-7 (IC50 of 0.63 ± 0.07 µg/mL for 48 h and 0.39 ± 0.08 µg/mL for 72 h) and MDA-MB-231 (1.17 ± 0.06 µg/mL for 48 h, 1.03 ± 0.15 µg/mL for 72 h) cells in dose- and time-dependent manner. Combination treatment also enhanced the cytotoxic effects of doxorubicin but not tamoxifen on inhibiting breast cancer cell growth. The involvement of intrinsic and extrinsic pathway in apoptosis induction was exhibited through the increased activity of caspase-9 and caspase-8, respectively, leading to enhanced downstream executioner caspase-3/7 activity in treated MCF-7 and MDA-MB-231 cells. In addition, gene expression analysis revealed that MnIII complex exerts its antiproliferative effect via up-and down-regulation of p21 and cyclin D1, respectively, along with increased expression of Bax/Bcl-2 ratio, TNF-α, initiator caspase-8 and -10 and effector caspase-3 in MCF-7 and MDA-MB-231 cells. However, the results did not show increased caspase-8 activity in treated MCF-7 cells. Furthermore, in vivo acute oral toxicity test revealed no signs of toxicity and mortality in treated animal models compared to the control group. Collectively, the promising inhibitory effect and molecular and mechanistic evidence of antiproliferative activity of MnIII complex and its safety characterization have demonstrated that it may have therapeutic value in breast cancer treatment worthy of further investigation and development.

Hyper-Expression of PD-1 Is Associated with the Levels of Exhausted and Dysfunctional Phenotypes of Circulating CD161++TCR iVα7.2+ Mucosal-Associated Invariant T Cells in Chronic Hepatitis B Virus Infection.

Mucosal-associated invariant T (MAIT) cells, defined as CD161++TCR iVα7.2+ T cells, play an important role in the innate defense against bacterial infections, and their functionality is impaired in chronic viral infections. Here, we investigated the frequency and functional role of MAIT cells in chronic hepatitis B virus (HBV) infection. The peripheral CD3+CD161++TCR iVα7.2+ MAIT cells in chronic HBV-infected patients and healthy controls were phenotypically characterized based on CD57, PD-1, TIM-3, and CTLA-4, as well as HLA-DR and CD38 expression. The frequency of MAIT cells was significantly decreased among chronic HBV-infected individuals as compared to controls. Expression of CD57, PD-1, CTLA-4, as well as HLA-DR and CD38 on MAIT cells was significantly elevated in chronic HBV-infected individuals relative to controls. The percentage of T cell receptor (TCR) iVα7.2+ CD161+ MAIT cells did not correlate with HBV viral load but inversely with HLA-DR on CD4+ T cells and MAIT cells and with CD57 on CD8+ T cells suggesting that decrease of MAIT cells may not be attributed to direct infection by HBV but driven by HBV-induced chronic immune activation. The percentage and expression levels of PD-1 as well as CTLA-4 on MAIT cells inversely correlated with plasma HBV-DNA levels, which may suggest either a role for MAIT cells in the control of HBV infection or the effect of HBV replication in the liver on MAIT cell phenotype. We report that decrease of TCR iVα7.2+ MAIT cells in the peripheral blood and their functions were seemingly impaired in chronic HBV-infected patients likely because of the increased expression of PD-1.

Decrease of CD69 levels on TCR Vα7.2+CD4+ innate-like lymphocytes is associated with impaired cytotoxic functions in chronic hepatitis B virus-infected patients.

Hepatitis B virus (HBV) infection is a major cause of chronic liver disease that may progress to liver cirrhosis and hepatocellular carcinoma. Host immune responses represent the key determinants of HBV clearance or persistence. Here, we investigated the role of the early activation marker, CD69 and effector cytokines, granzyme B (GrB) and IFN-γ in the exhaustion of innate-like TCR Vα7.2+CD4+T cells, in 15 individuals with chronic HBV (CHB) infection where six were HBV DNA+ and nine were HBV DNA-. The percentage of cytokine-producing T cells and MAIT cells were significantly perturbed in HBV patients relative to healthy controls (HCs). The intracellular expression of GrB and IFN-γ was significantly reduced in MAIT cells derived from HBV-infected patients as compared to HCs, and the levels correlated with the percentage and levels [mean fluorescence intensity (MFI)] of CD69 expression. The total expression of CD69 (iMFI) was lower in CHB patients as compared to HCs. The frequency of CD69+ cells correlated with the levels of cytokine expression (MFI), particularly in CHB patients as compared to HCs. In summary, the polyfunctionality of peripheral T cells was significantly reduced among CHB patients, especially in the TCR Vα7.2+CD4+T cells, and the levels of cytokine expression correlated with functional cytokine levels.

Japanese encephalitis virus disrupts blood-brain barrier and modulates apoptosis proteins in THBMEC cells.

Japanese encephalitis (JE) is a neurotropic flavivirus that causes inflammation in central nervous system (CNS), neuronal death and also compromises the structural and functional integrity of the blood-brain barrier (BBB). The aim of this study was to evaluate the BBB disruption and apoptotic process in Japanese encephalitis virus (JEV)-infected transfected human brain microvascular endothelial cells (THBMECs). THBMECs were overlaid by JEV with different MOIs (0.5, 1.0, 5.0 and 10.0) and monitored by electrical cell-substrate impedance sensing (ECIS) in a real-time manner in order to observe the barrier function of THBMECs. Additionally, the level of 43 apoptotic proteins was quantified in the virally infected cells with different MOIs at 24h post infection. Infection of THBMEC with JEV induced an acute reduction in transendothelial electrical resistance (TEER) after viral infection. Also, significant up-regulation of Bax, BID, Fas and Fasl and down-regulation of IGFBP-2, BID, p27 and p53 were observed in JEV infected THBMECs with 0.5 and 10 MOIs compared to uninfected cells. Hence, the permeability of THBMECs is compromised during the JEV infection. In addition high viral load of the virus has the potential to subvert the host cell apoptosis to optimize the course of viral infection through deactivation of pro-apoptotic proteins.

Immune Responses to Influenza Virus and Its Correlation to Age and Inherited Factors.

Influenza viruses belong to the family Orthomyxoviridae of enveloped viruses and are an important cause of respiratory infections worldwide. The influenza virus is able to infect a wide variety species as diverse as poultry, marine, pigs, horses, and humans. Upon infection with influenza virus the innate immunity plays a critical role in efficient and rapid control of viral infections as well as in adaptive immunity initiation. The humoral immune system produces antibodies against different influenza antigens, of which the HA-specific antibody is the most important for neutralization of the virus and thus prevention of illness. Cell mediated immunity including CD4+ helper T cells and CD8+ cytotoxic T cells are the other arms of adaptive immunity induced upon influenza virus infection. The complex inherited factors and age related changes are associated with the host immune responses. Here, we review the different components of immune responses against influenza virus. Additionally, the correlation of the immune response to age and inherited factors has been discussed. These determinations lead to a better understanding of the limitations of immune responses for developing improved vaccines to control influenza virus infection.

White tea (Camellia sinensis) inhibits proliferation of the colon cancer cell line, HT-29, activates caspases and protects DNA of normal cells against oxidative damage.

Tea (Camellia sinensis) is one of the most consumed beverages in the world. White tea is made from the buds and young leaves of the tea plant which are steamed and dried, whilst undergoing minimal oxidation. The MTT assay was used to test the extract on the effect of the proliferation of the colorectal cancer cell line, HT-29. The extract inhibited the proliferation of HT-29 cells with an IC50 of 87µg/ml. The extract increased the levels of caspase-3, -8, and -9 activity in the cells. DNA damage in 3T3-L1 normal cells was detected by using the comet assay. The extract protected 3T3-L1 cells against H2O2-induced DNA damage. The results from this study show that white tea has antioxidant and antiproliferative effects against cancer cells, but protect normal cells against DNA damage. Regular intake of white tea can help to maintain good health and protect the body against disease.