Abu Hasanat Md. Zulfiker

Toad Glandular Secretions and Skin Extractions as Anti-Inflammatory and Anticancer Agents

Toad glandular secretions and skin extractions contain many natural agents which may provide a unique resource for novel drug development. The dried secretion from the auricular and skin glands of Chinese toad (Bufo bufo gargarizans) is named Chansu, which has been used in Traditional Chinese Medicine (TCM) for treating infection and inflammation for hundreds of years. The sterilized hot water extraction of dried toad skin is named Huachansu (Cinobufacini) which was developed for treating hepatitis B virus (HBV) and several types of cancers. However, the mechanisms of action of Chansu, Huachansu, and their constituents within are not well reported. Existing studies have suggested that their anti-inflammation and anticancer potential were via targeting Nuclear Factor (NF)-κB and its signalling pathways which are crucial hallmarks of inflammation and cancer in various experimental models. Here, we review some current studies of Chansu, Huachansu, and their compounds in terms of their use as both anti-inflammatory and anticancer agents. We also explored the potential use of toad glandular secretions and skin extractions as alternate resources for treating human cancers in combinational therapies.

Investigation of Antioxidant and Antimicrobial Potential of Chloroform and Petroleum Ether Extracts of Selected Medicinal Plants of Bangladesh

Aim: To investigate the antioxidant and antimicrobial potential of Chloroform and Pet ether extracts of Manilkara zapota ( MZCE, MZPE ), Polyalthia longifolia ( PLCE, PLPE ), Abroma augusta ( AACE, AAPE ) Ficus hispida ( FHCE, FHPE ), Vitex negundo ( VNCE,

The yield and quality of cellular and bacterial DNA extracts from human oral rinse samples are variably affected by the cell lysis methodology

Recent culture-independent studies have enabled detailed mapping of human microbiome that has not been hitherto achievable by culture-based methods. DNA extraction is a key element of bacterial culture-independent studies that critically impacts on the outcome of the detected microbial profile. Despite the variations in DNA extraction methods described in the literature, no standardized technique is available for the purpose of microbiome profiling. Hence, standardization of DNA extraction methods is urgently needed to yield comparable data from different studies. We examined the effect of eight different cell lysis protocols on the yield and quality of the extracted DNA from oral rinse samples. These samples were exposed to cell lysis techniques based on enzymatic, mechanical, and a combination of enzymatic-mechanical methods. The outcome measures evaluated were total bacterial population, Firmicutes levels and human DNA contamination (in terms of surrogate GAPDH levels). We noted that all three parameters were significantly affected by the method of cell lysis employed. Although the highest yield of gDNA was obtained using lysozyme-achromopeptidase method, the lysozyme-zirconium beads method yielded the peak quantity of total bacterial DNA and Firmicutes with a lower degree of GAPDH contamination compared with the other methods. Taken together our data clearly points to an urgent need for a consensus, standardized DNA extraction technique to evaluate the oral microbiome using oral rinse samples. Further, if Firmicutes levels are the focus of investigation in oral rinse microbiome analyses then the lysozyme-zirconium bead method would be the method of choice in preference to others.

Aqueous and Ethanol Extracts of Australian Cane Toad Skins Suppress Pro-Inflammatory Cytokine Secretion in U937 Cells via NF-κB Signaling Pathway

Toad skin extracts, such as aqueous extracts (AE) of Chinese toad skins, have demonstrated therapeutic benefits for a range of diseases including pain, inflammation, swelling, heart failure, and various types of cancers. In this study, we investigated the anti‐inflammatory potential of an AE (0.1–10 μg/mL) and a 60% ethanol extract (EE; 0.1–10 μg/mL) from Australian cane toad (Bufo marinus) skins and the known bioactive compound, bufotenine (BT; 0.1–10 nM). The assay employed a model of the human monocyte cell line U937 stimulated with lipopolysaccharide (LPS) and phorbol 12‐myristate 13‐acetate (PMA) for the release of tumor necrosis factor (TNF)‐α and interleukin (IL)‐6. We demonstrated that AE, EE, and BT significantly inhibited the release and expression of TNF‐α and IL‐6 in a dose‐dependent manner when the cells were pre‐treated at non‐cytotoxic concentrations. Further investigation revealed that the inhibition of TNF‐α and IL‐6 release and expression was associated with the suppression of nuclear factor (NF)‐kappa (κ)B activation. These results indicate that AE, EE, and BT are strong inflammation inhibitors, thus have the potential for further development as anti‐inflammatory therapeutic agents from a natural source regarded as a feral pest in Australia. J. Cell. Biochem. 117: 2769–2780, 2016.

Indolealkylamines from Toad Vertebrates and Sea Invertebrates - Their Identification and Potential Activities on the Central Nervous System

Indolealkylamines (IAAs) are biogenic amines and derivatives of 5-hydroxytryptamine, acting primarily on serotonin receptors. IAAs are often considered the most thoroughly investigated group of aromatic amines in the amphibian skin. On the contrary, at present the detailed knowledge of these compounds in lower organisms is still limited and the biogenic amine receptors, mediating hormonal and modulatory functions, are largely unknown in primitive invertebrates. However, some active research is currently underway investigating this class of biogenic amines. Notably, during the last three decades several investigations have demonstrated the biological activity of endogenous biogenic amines in cnidarians, which are known to be the lowest beings equipped with an effective, even though rudimentary, nervous system. Toads, especially those from the Bufonidae family, constitute a significant part of the amphibian family and are an identified source of IAAs. To date fourteen IAAs have been identified in the skins of toad species. All are 5-substituted IAA derivatives acting mainly on the central nervous system (CNS), with most exhibiting some degrees of 5-HT2A receptor selectivity. This selective ability presents potential for their use in the development of treatments for various disorders such as schizophrenia, depression, anxiety, obsessive-compulsive disorders and chronic pain conditions. There are indications that some IAAs may also show subclass selectivity through binding to multiple 5-HT receptor subtypes. Thus, there exists an additional promising platform for the development of therapeutics targeting multiple 5-HT receptors. In this review, IAAs occurring naturally in various species of toad skins, which have been identified and isolated since 1944 are summarized and comparisons are made with similar biogenic amines recognized in cnidarians to date. Such comparisons highlight the potential to utilize existing knowledge gathered from vertebrates, such as toads in order to improve the understanding of the activities of such compounds in lower invertebrates.

Cane Toad Skin Extract-Induced Upregulation and Increased Interaction of Serotonin 2A and D2 Receptors via Gq/11 Signaling Pathway in CLU213 Cells.

Recent evidences show that activation of serotonin 2A receptors (5‐HT2AR) by agonists is significant in improving therapeutic activity of disease conditions, such as obsessive‐compulsive disorder (OCD). Though the exact molecular mechanism is still not well understood, it is thought to involve agonist‐driven, enhanced expression of 5‐HT2AR in certain areas of brain, such as the pre‐frontal cortex (PFC). Several other reports have also demonstrated association of OCD with lower dopamine receptor (D2R) availability, primarily in the striatum of the brain along with dysfunction of 5‐HT2AR‐D2R heteromer regulation. We thus hypothesized that compound(s) interacting with this molecular mechanism could be developed as drugs for long‐term beneficial effects against OCD. In the present study, we have obtained experimental evidence in cultured neuronal cells (CLU213) that aqueous extract (AE, 50 μg/mL, P < 0.05) of the Australian cane toad skin significantly increased the levels of 5‐HT2AR and D2R protein and mRNA expression. AE was also found to enhance the interaction between 5‐HT2AR and D2R and formation of expression of 5‐HT2AR‐D2R heteromer using co‐immunoprecipitation and Western blot. Further investigation showed the involvement of classical signaling pathway (Gq/11‐PLCβ) along with c‐FOS transcription factor preferentially in 5‐HT2A‐mediated agonist activation. These results obtained demonstrated that AE upregulates 5‐HT2AR by a mechanism that appears to involve Gq/11‐PLCβ signaling pathway and c‐FOS transcription factor activation. We indicate this enhanced 5‐HT2AR and D2R expression and their interaction to induce increased 5‐HT2AR‐D2R heteromer formation by exposure to AE might provide a molecular mechanism to develop potential novel drug candidates to ameliorate OCD symptoms. J. Cell. Biochem. 118: 979–993, 2017.

The Development of Toad Toxins as Potential Therapeutic Agents

Toxins from toads have long been known to contain rich chemicals with great pharmaceutical potential. Recent studies have shown more than 100 such chemical components, including peptides, steroids, indole alkaloids, bufogargarizanines, organic acids, and others, in the parotoid and skins gland secretions from different species of toads. In traditional Chinese medicine (TCM), processed toad toxins have been used for treating various diseases for hundreds of years. Modern studies, including both experimental and clinical trials, have also revealed the molecular mechanisms that support the development of these components into medicines for the treatment of inflammatory diseases and cancers. More recently, there have been studies that demonstrated the therapeutic potential of toxins from other species of toads, such as Australian cane toads. Previous reviews mostly focused on the pharmaceutical effects of the whole extracts from parotoid glands or skins of toads. However, to fully understand the molecular basis of toad toxins in their use for therapy, a comprehensive understanding of the individual compound contained in toad toxins is necessary; thus, this paper seeks to review the recent studies of some typical compounds frequently identified in toad secretions.