A Proposed In-Vivo/In-Vitro Approach to the Toxicological Assessment of Hazardous Waste

Hemocyanins (Hcs) are respiratory, oxygen-carrying metalloproteins that are freely dissolved in the hemolymph of many molluscs and arthropods. The interest in hemocyanins has grown significantly since it was found that they can be successfully used in immunotherapy of neoplastic diseases as non-specific or active stimulators of the immune system. The present study aims to assess the cytotoxicity, in vivo toxicity and antiproliferative activity of hemocyanins isolated from marine snail Rapana venosa (RvH), garden snails Helix lucorum (HlH) and Helix aspersa (HaH). For in vitro safety testing, 3T3 Neutral Red Uptake (NRU) test was used. The experiments for antiproliferative activity of the hemocyanins were performed by MTT assay on a panel of cell lines - a model of breast cancer. The in vivo toxicological assessment was performed by regular clinical examinations of hemocyanin-treated laboratory mice and histopathological analysis of hematoxylin/eosin stained preparations of parenchymal organs. The evaluation of the in vitro cytotoxicity showed that the tested hemocyanins does not induce toxic effects in nontumorigenic epithelial cell lines. In contrast, significant reduction of the viability of human breast carcinoma cell lines was found after treatment with high concentrations of hemocyanins. The in vivo experiments showed no signs of organ and systemic toxicity in the hemocyanin-treated animals. The presented data indicate that Hcs show a potential for development of novel anticancer therapeutics due to their beneficial properties, biosafety and lack of toxicity or side effects. Key words: hemocyanins (Hcs); cytotoxicity; antitumor activity; in vivo biosafety testing.

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In Vitro and In Vivo Antimalarial Activities and Toxicological Assessment of Pogostemon Cablin (Blanco) Benth

Journal of Evidence-Based Integrative Medicine ◽

10.1177/2515690x20978387 ◽

2020 ◽

Vol 25 ◽

pp. 2515690X2097838

Author(s):

Arisara Phuwajaroanpong ◽

Prapaporn Chaniad ◽

Natharinee Horata ◽

Saowanee Muangchanburee ◽

Kantarakorn Kaewdana ◽

...

Keyword(s):

Body Weight ◽

Antimalarial Activity ◽

Ethanolic Extract ◽

Vero Cells ◽

Toxicological Assessment ◽

Pogostemon Cablin ◽

Dehydrogenase Enzyme ◽

Preliminary Study

The aim of this study was to investigate the antimalarial activities and toxicity of Pogostemon cablin extracts. In vitro activities against the chloroquine-resistant Plasmodium falciparum K1 strain were assessed by using the Plasmodium lactate dehydrogenase enzyme (pLDH) assay, while in vivo activity against the Plasmodium berghei ANKA strain in mice was investigated using a 4-day suppressive test. The in vitro and in vivo toxicity were determined in Vero cells and mice, respectively. The ethanolic extract possessed antimalarial activity with an IC50 of 24.49 ± 0.01 µg/ml, whereas the aqueous extract showed an IC50 of 549.30 ± 0.07 µg/ml. Cytotoxic analyses of the ethanolic and aqueous extracts revealed a nontoxic effect on Vero cells at a concentration of 80 µg/ml. Based on a preliminary study of in vitro antimalarial activity, the ethanolic extract was chosen as a potential agent for further in vivo antimalarial activity analysis in mice. The ethanolic extract, which showed no toxic effect on mice at a dose of 2000 mg/kg body weight, significantly suppressed parasitemia in mice by 38.41%, 45.12% and 89.00% at doses of 200, 400 and 600 mg/kg body weight, respectively. In conclusion, this study shows that the ethanolic P. cablin extract possesses in vitro and in vivo antimalarial activity without toxic effects.

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A Review on Recent Researches of Morinda Citrifolia, L. (Noni) as Functional Foods and Medical Herbs

Jurnal Riset Teknologi Industri ◽

10.26578/jrti.v10i2.2569 ◽

2016 ◽

Vol 10 (2) ◽

pp. 172-186

Author(s):

Agus Sudibyo ◽

Tiurlan F. Hutajulu

Keyword(s):

Vitamin A ◽

Vitamin C ◽

Biological Effects ◽

Morinda Citrifolia ◽

In Vivo Studies ◽

Cancer Disease ◽

Potential Health ◽

Toxicological Assessment

Morinda Citrifolia, L. known as Noni or Mengkudu is planting belonging to the family of Rubiaceae. A number of major components have been identified in leaves, roots, fruits of Noni plant, such as scopoletin, octanoic acid, vitamin C, iridoid, terpenoids, alkaloids, anthraquinones, beta-sitosterol, carotene, vitamin A, flavone glycosides, alizarin, amino acids, acubin, austin, caproic acid, caprylic acid and putative procyonine. Its use as a botanical dietary supplement has grown tremendously in recent years. The results of epidemiological studies suggest that the Noni consumption may help prevent several chronic diseases, including cancer disease, cardiovascular disease, type 2 diabetes mellitus, heart disease, artherosclerosis, blood vessel problem, gastric ulcer, drug addiction, muscle ached and pein. Several studies have also demonstrated anti-inflammatory, antioxidant, antimicrobial, analgesic, and immunologicalactivity. Based on a toxicological assessment, Noni juice was considered as safe. Although, a large number of in vitro and to a certain extent, and in vivo studies demonstrated a range potentially beneficial effects, clinical information data are still lacking completely. Therefore, to what extent the information findings from experimental pharmacological studies is not complete at present, so this article reviews potential health benefits for consumptions, its biological effects and looking for a new informationthat needs to be explored in detail before a recommendation can be madeABSTRAKMorinda citrofolia, L (mengkudu) merupakan jenis tanaman yang termasuk dalam golongan Rubiaceadan buahnya dikenal dengan nama Noni atau mengkudu. Beberapa komponen utama dalam tanaman tersebut telah diidentifikasi, mulai dari bagian akar, daun, dan buah, seperti kandungan scopoletin, asam oktanoad, vitamin C, iridoid, terpenoid, alkaloid, anthraquinon, beta-sitosterol, karotene, vitamin A, flavon glikosida, alizarin, asam amino, acubin, austin, asam kaproat, asam kaprilat dan putativ prokseronin. Buah tersebut akhir-akhir ini telah sukses banyak dimanfaatkan sebagai diet suplemen. Hasil studi secara epidemiologi menyatakan bahwa konsumsi mengkudu dapat membantu mencegah beberapa penyakit kronis, seperti penyakit kanker, kardiovaskular, diabetes tipe 2, penyakit jantung, artherosklerosis, masalah pembuluh darah, pencernaan, dan sakit otot. Beberapa studi juga menunjukkan bahwa mengkudu dapat berfungsi sebagai anti inflamasi, antioksidan, antimikroba, analgesik, dan bersifat immunlogis.Berdasarkan kajian secara toksikologi, buah dan jus mengkudu dinyatakan aman untuk dikonsumsi. Penelitian secara in vitro pada beberapa jenis penyakit tertentu sedang diperluas, dan penelitian secara in vivo menunjukkan bahwa mengkudu mempunyai rentang potensi pengaruh yang baik bagi kesehatan, meskipun data informasinya secara klinis kurang lengkap. Oleh karena itu, untuk mengetahui apa yang sudah ditemukan dari hasil penelitian secara farmalogis yang belum lengkapsaat ini, maka dalam tulisan ini membahas potensi keuntungan kesehatan bila dikonsumsi, pengaruh biologinya dan pencarian informasibaru yang perlu dikaji lebih rinci sebelum rekomendasi ditetapkan.Kata kunci: Mengkudu (Morinda citrifolia, L), pangan fungsional, rempah medis.

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Toxicological Evaluation of Human Adipose-derived Mesenchymal Stem Cells (hADSCs) and hADSCs-derived Exosomes

10.21203/rs.3.rs-626854/v1 ◽

2021 ◽

Author(s):

Yang Zhou ◽

Bo Zhao ◽

Xin-Liao Zhang ◽

Yi-jun Lu ◽

Jian Cheng ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Human Adipose Tissue ◽

Soft Agar ◽

Toxicological Evaluation ◽

Toxicological Assessment ◽

Toxicological Profile ◽

Treatment Agent

Abstract Background: Human adipose tissue-derived stem cells (hADSCs) are considered an ideal source of cells for regenerative medicine. Mesenchymal stem cells derived-exosomes (MSC-Exos) are being opined as new cell-free therapeutics for numerous human diseases. For future clinical applications, the safety of allogenic hADSCs and hADSCs-derived exosomes (hADSCs-Exos) needs to be addressed and verified in pre-clinical animal models. This study sought to evaluate the toxicity of hADSCs and hADSCs-Exos by performing in vivo and in vitro toxicological assessments.Methods: We used IVIS to track the biodistribution of GFP-labeled hADSCs and the PKH26-labeled in a mouse model. The tumorigenicity of hADSCs and hADSCs-Exos was analyzed by soft agar colony formation assay and nude mice tumorigenicity test in vitro and in vivo. The acute animal toxicity and allergenicity test were used to explore the toxicological proﬁle of hADSCs and hADSCs-Exos in mice.Results: We found that hADSCs-Exos accumulated faster in the tissues of mice and were also cleared more rapidly compared to hADSCs. Both hADSCs and hADSCs-Exos have little risk of tumorigenicity, and hADSCs-Exos had lower toxicity and lower immunogenicity than hADSCs.Conclusion: Our study is the first to compare the safety between hADSCs and hADSCs-Exos, and revealed that hADSCs-Exos are safer for application as systemic therapy, without complications in toxicological assessment, and have a better prospective utility as a treatment agent and for drug delivery.

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Toxicological assessment of Euglena gracilis strain Eu029 shows no adverse effects in vivo and in vitro

Toxicology Research and Application ◽

10.1177/2397847318761672 ◽

2018 ◽

Vol 2 ◽

pp. 239784731876167

Author(s):

Jennifer M Symonds ◽

Nicole Beauchamp ◽

Takuto Takeuchi ◽

Koji Yamada ◽

Kohei Atsuji ◽

...

Keyword(s):

Euglena Gracilis ◽

Metabolic Pathways ◽

Micronucleus Assay ◽

Food Ingredient ◽

Toxicological Assessment ◽

Acute Toxicity Study ◽

Adverse Effect Level ◽

Effect Level

Euglena gracilis is a single-celled organism capable of photosynthesis and heterotrophy. Euglena sp. have long been studied in the laboratory for its metabolic pathways, cell motility, and ease of culture. The safety of E. gracilis strain eu029 (EG029) for use as a food ingredient was assessed in a bacterial reverse mutagenesis assay (Ames), rec assay, in vivo micronucleus assay, acute toxicity study in mice, 13-week toxicology in rats, and a teratology study in mice and rats. EG029 was not genotoxic. The No Observed Adverse Effect Level (NOAEL) in the 13-week study was greater than 1000 mg/kg/day, the highest dose tested. Teratogenicity studies did not find any defects in fetal development or effects to maternal health in rats at 1000 mg/kg/day, the highest dose tested.

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Exploiting Cancer Metal Metabolism using Anti-Cancer Metal- Binding Agents

Current Medicinal Chemistry ◽

10.2174/0929867324666170705120809 ◽

2019 ◽

Vol 26 (2) ◽

pp. 302-322 ◽

Cited By ~ 8

Author(s):

Angelica M. Merlot ◽

Danuta S. Kalinowski ◽

Zaklina Kovacevic ◽

Patric J. Jansson ◽

Sumit Sahni ◽

...

Keyword(s):

Metal Ion ◽

Antitumor Agents ◽

Metal Ion Binding ◽

Toxicological Assessment ◽

Metal Metabolism ◽

Metal Levels ◽

Anti Cancer ◽

Binding Agents

Metals are vital cellular elements necessary for multiple indispensable biological processes of living organisms, including energy transduction and cell proliferation. Interestingly, alterations in metal levels and also changes in the expression of proteins involved in metal metabolism have been demonstrated in a variety of cancers. Considering this and the important role of metals for cell growth, the development of drugs that sequester metals has become an attractive target for the development of novel anti-cancer agents. Interest in this field has surged with the design and development of new generations of chelators of the thiosemicarbazone class. These ligands have shown potent anticancer and anti-metastatic activity in vitro and in vivo. Due to their efficacy and safe toxicological assessment, some of these agents have recently entered multi-center clinical trials as therapeutics for advanced and resistant tumors. This review highlights the role and changes in homeostasis of metals in cancer and emphasizes the pre-clinical development and clinical assessment of metal ion-binding agents, namely, thiosemicarbazones, as antitumor agents.

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Antimalarial Efficacy and Toxicological Assessment of Extracts of Some Ghanaian Medicinal Plants

Journal of Parasitology Research ◽

10.1155/2019/1630405 ◽

2019 ◽

Vol 2019 ◽

pp. 1-9 ◽

Cited By ~ 7

Author(s):

Michael Konney Laryea ◽

Lawrence Sheringham Borquaye

Keyword(s):

Plasmodium Falciparum ◽

Medicinal Plants ◽

Acute Toxicity ◽

Plant Extracts ◽

Mammalian Cells ◽

Toxicity Tests ◽

Bidens Pilosa ◽

Toxicological Assessment

The economic costs associated with morbidity and mortality due to malaria and malaria associated complications in many sub-Saharan countries and other malaria endemic regions of the world are huge. Reports of emergence of parasite resistance to current malaria drugs have complicated malaria treatment and require the development of new therapeutic agents. The folkloric use of medicinal plants for the management of malaria is well documented. This work evaluated the antiplasmodial activities and toxicity of some medicinal plants used to treat malaria and malaria-like symptoms in Ghana. Plant extracts were obtained by cold maceration in 70% ethanol. Antiplasmodial efficacies were assessed in vitro against 3 strains of Plasmodium falciparum strains (FCM, W2, and CAM06) and in vivo via the 4-day suppressive test in Plasmodium berghei infected mice. Cytotoxicity and acute toxicity were assessed in mammalian cells and mice, respectively. All extracts were active against at least one of the Plasmodium falciparum strains in in vitro evaluations with IC50’s in the range of 4–116 μg/mL, whereas Bidens pilosa extracts, with a chemosuppression rate of 75%, was the most active plant in the in vivo experiments. All plant extracts displayed very weak to no cytotoxicity against the mammalian cell line used and exhibited very good selectivity towards the Plasmodium parasites. Syzygium guineense and Parinari congensis extracts were the most toxic in the acute toxicity tests. Altogether, the results indicate that the medicinal plants do possess impressive antiplasmodial properties and provide scientific basis for their use in traditional herbal medicine.

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Prospecting the insect model, Galleria mellonella, for gut-related pathobiology

10.23889/suthesis.56984 ◽

2020 ◽

Author(s):

◽

Helena Emery

Keyword(s):

Gastrointestinal Tract ◽

Galleria Mellonella ◽

Communicable Disease ◽

Oral Route ◽

Cordyceps Sinensis ◽

Shellfish Poisoning ◽

In Vivo Models ◽

Toxicological Assessment

Animal research has contributed immensely to medical and scientific advances over the last century, and continues to play important roles in enhancing our understanding of infectious and non-communicable disease development, and the search for treatments. The mouse, for example, shares ~95% of human genes and is the most widespread vertebrate model in use. Since the late 1980s, there has been several UK and EU directives (e.g., 2010/63/EU) to improve the welfare of animals considered essential for experimentation, and to link directly with the principle of the 3Rs, to Replace, Reduce and Refine animal use. Additionally, animal maintenance, husbandry, compliance with legislation and licencing, and staff training are costly and time-consuming. Hence, there is much to gain from developing alternative in vivo models and complementary in vitro, in chemico and in silico tools. Larvae of the wax moth Galleria mellonella represent one such surrogate to rodents, and have been used successfully to study microbial isolates for virulence traits, putative antibiotic therapies, and more recently, toxicological assessment. There is an abundance of practical and biological advantages to selecting G. mellonella over rodents and traditional non-mammalian fruit flies and nematodes (which are described in Chapter 1), but one area lacking in knowledge is their applicability for studies of gut pathobiology. Therefore, the aim of this thesis was to evaluate the usefulness and accuracy of G. mellonella larvae as a model for gut specific toxins and pathogens when administered through an oral route (gavage). A series of whole-organism (phenotype), cellular, biochemical, microbiological and microscopy methods were used to interrogate the gastrointestinal tract of G. mellonella in the absence and presence of chemicals and microbes known to cause gastropathy in rodents and humans. First, the transferability of the indomethacin restraint/ulcer assay was established in G. mellonella, with levels of tissue deterioration and enhanced leakiness reminiscent of rodents (Chapter 2). Second, the rearing of insects on nutraceuticals Cordyceps sinensis and bovine colostrum alleviated gut damage caused by indomethacin, and improved survival outcomes when challenged with the enteric pathogen Campylobacter jejuni (Chapter 3). Third, oral administration of shellfish poisoning toxins (okadaic acid and azaspiracids 1-3) to G. mellonella, interfered with tissue integrity and microbial stability of the gastrointestinal tract, and produced comparable LD50 levels to their rodent counterparts (Chapter 4). The results presented here go beyond establishing synonymous damage phenomena between G. mellonella larvae and vertebrates (Chapter 5), but adds new knowledge to the structure and function of the lepidopteran alimentary canal, the cytopathology of emerging marine toxins, and how diet invariably influences a host’s capacity to recover from subacute chemical and microbial disruptors.

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