Toxicity assessment and screening of tetrodotoxin in the oblong blowfish(Takifugu oblongus)from the Tamil Nadu Coast of Bay of Bengal,India

Objective:To assess the toxicity of the pufferfish Takifugu oblongus,from Chennai coast,Tamil Nadu,India and to detect the presence of Tetrodotoxin(TTX).Methods:The toxicity was evaluated by mouse bioassay using Swiss Albino mice which were expressed in mouse units(MU).Gross anatomical features were observed which is followed by histopathology of the dead mice tissues to establish the toxicity.Instrumental analysis for the presence of tetrodotoxin was also performed through GC-MS and HPLC.Results:The toxicity of ovary was the maximum with 163 MU/g and lowest toxicity was observed in skin with 75.88 MU/g.Histopathological analyses of the dead mice showed various cellular degenerations and inflammations.The amount of Tetrodotoxin detected through GC-MS and HPLC was more reliable and sensitive than the customary mouse bioassay as instrumental analyses were able to detect even nanograms of the toxin.Conclusions:The present study evidently proved that Takifugu oblongus is highly toxic and consumption of the same can pose serious threat for health and possible lethality to humans.

Biosorption of Cd (Ⅱ) and Pb (Ⅱ) Ions by Aqueous Solutions of Novel Alkalophillic Streptomyces VITSVK5 spp. Biomass

Discharge of heavy metals from metal processing industries is known to have adverse effects on the environment.Bio-sorption of heavy metals by metabolically inactive biomass of microbial organisms is an innovative and alternative technology for removal of these pollutants from aqueous solution.The search of marine actinobacteria with potential heavy metal biosorption ability resulted in the identification of a novel alkalophilic Streptomyces VITSVK5 species.The biosorption property of Streptomyces VITSVK5 spp.was investigated by absorbing heavy metals Cadmium (Cd) and Lead (Pb).Physiochemical characteristics and trace metal concentration analysis of the backwater showed the concentrations of different metals were lead 13±2.1 μg L-1,cadmium 3.1±0.3μg L-1,zinc 8.4±2.6μg L-1 and copper 0.3±0.1μg L-1,whereas mercury was well below the detection limit.The effect of pH and biomass dosage on removal efficiency of heavy metal ions was also investigated.The optimum pH for maximal biosorption was 4.0 for Cd (II) and 5.0 for Pb (II) with 41% and 84% biosorption respectively.The biosorbent dosage was optimized as 3 g L-1 for both the trace metals.Fourier transform infrared absorption spectrum results indicated the chemical interactions of hydrogen atoms in carboxyl (–COOH),hydroxyl (–CHOH) and amine (–NH2) groups of biomass with the metal ions.This could be mainly involved in the biosorption of Cd (II) and Pb (II) onto Streptomyces VITSVK5 spp.The results of our study revealed Streptomyces metabolites could be used to develop a biosorbent for adsorbing metal ions from aqueous environments.

Antimicrobial Activity of Ulva reticulata and Its Endophytes

Seaweeds are known to exhibit various antimicrobial properties, since it harbours an enormous range of indigenous bioactive compounds. The emergence of drug resistant strains has directed to the identification of prospective metabolites from seaweed and its endophytes, thereby exploiting the properties in resisting bacterial diseases. The current study was aimed to assess the antimicrobial activity of extracts obtained from Ulva reticulate, for which metabolites of Ulva reticulata and its endophytes were extracted and assessed against human pathogens like Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, and Bacillus subtilis. It was observed that the hexane extract of isolate VITDSJ2 was effective against all the tested pathogens but a significant inhibition was observed for Staphylococcus aureus and Escherichia coli. Further, Gas chromatography coupled with Mass spectroscopy （GC-MS） revealed the existence of phenol, 3, 5-bis （1, 1-dimethylethyl） in the crude hexane extract which is well-known to possess antibacterial activity. The effective isolate VITDSJ2 was identified to be the closest neighbour of Pseudomonas smtzeri by phenotypic and genotypic methods. The crude extracts of the seaweed Ulva reticulata was also screened for antibacterial activity and the hexane extract was effective in showing inhibition against all the tested pathogens. The compound in the crude extract of Ulva reticulata was identified as hentriacontane using GC-MS. The extracts obtained from dichloromethane did not show significant activity in comparison with the hexane extracts. Hence the metabolites of Ulva reticulata and the bacterial secondary metabolites of the endophytes could be used in the treatment of bacterial infections.

Role of aberrant Sonic hedgehog signaling pathway in cancers and developmental anomalies

Development is a sophisticated process maintained by various signal transduction pathways,including the Hedgehog(Hh)pathway.Several important functions are executed by the Hh signaling cascade such as organogenesis,tissue regeneration,and tissue homeostasis,among various others.Considering the multiple functions carried out by this pathway,any mutation causing aberrant Hh signaling may lead to myriad developmental abnormalities besides cancers.In the present review article,we explored a wide range of diseases caused by aberrant Hh signaling,including developmental defects and cancers.Finally,we concluded this mini-review with various treatment strategies for Hh-induced diseases.

Biosorption of Cr(Ⅳ),Cr(Ⅲ),Pb(Ⅱ) and Cd(Ⅱ) from Aqueous Solutions by Sargassum wightii and Caulerpa racemosa Algal Biomass

Heavy metal pollution is one of the most important environmental problems today.Biosorption is an innovative tech-nology that employs biological materials to accumulate heavy metals from waste water through metabolic process or physicochemi-cal pathways of uptake.Even though several physical and chemical methods are available for removal of heavy metals,currently many biological materials such as bacteria,algae,yeasts and fungi have been widely used due to their good performance,low cost and large quantity of availability.The aim of the present study is to explore the biosorption of toxic heavy metals,Cr（VI）,Cr（III）,Pb（II） and Cd（II） by algal biomass obtained from algae Sargassum wightii（brown） and Caulerpa racemosa（green）.Biosorption of algal biomass was found to be biomass concentration-and pH-dependent,while the maximal biosorption was found at pH 5.0 and with the metal concentration of 100 mg L-1.S.wightii showed the maximal metal biosorption at the biomass concentration of 25 g L-1,followed by C.racemosa with the maximal biosorption at 30 g L-1.S.wightii showed 78% biosorption of Cr（VI）,Cr（III）,Pb（II） and Cd（II） ions.C.racemosa exhibited 85% biosorption of Cd（II） and Cr（VI）,and 50% biosorption of Cr（III） and Pb（II）.The results of our study suggest that seaweed biomass can be used efficiently for

Possible therapeutic role of short-chain fatty acids from skin commensal bacteria in UVB-induced skin carcinogenesis

Solar ultraviolet B(UVB)radiation is a major skin cancer-causing agent.Initiation,promotion,and progression are the diverse phases of UVB-induced carcinogenesis.Exposure to UVB causes abnormalities in a series of biochemical and molecular pathways:thymine dimer formation,DNA damage,oxidative stress,inflammatory responses,and altered cell signaling,eventually resulting in tumor formation.The increased skin cancer rates urge researchers to develop more efficient drugs,but synthetic chemotherapeutic drugs have more contrary effects and drug resistance issues,which have been reported recently.The current review focuses on the relationship between microbes and cancer.Human skin acts as a barrier against the external environment and serves as a protective shield for its inhabitant microbiota,collectively called skin microbes.The gut microbiome plays a vital role in cancer therapy.Production of short-chain fatty acids(SCFAs)such as butyrate,acetate,and propionate by intestinal microbes has anti-cancer properties against various cancer cell lines.Yet,the knowledge of SCFAs produced by skin microbes remains yet to be elucidated exhaustively.In this review,we strive to summarize the findings of studies performed to date regarding the anti-cancer properties of SCFA against various cancer cell lines and provide insight into future directions in the skin microbiome field.

Benzydamine hydrochloride ameliorates ethanol-induced inflammation in RAW 264.7 macrophages by stabilizing redox homeostasis

Objective:To evaluate the protective effect of benzydamine hydrochloride against ethanol-induced oxidative stress and inflammation in RAW 264.7 macrophages.Methods:RAW 264.7 macrophages were treated with ethanol(100 mM)and benzydamine hydrochloride(7.5μM).The imflammatory status was confirmed by measuring pro-(TNF-αand IL-6)and anti-inflammatory(IL-10)cytokines through ELISA and RT-PCR assays.Reactive oxygen species generation and mitochondrial membrane potential were investigated to study the protective role of benzydamine hydrochloride against ethanol-induced oxidative stress.Apoptosis detection was also investigated using flow cytometry and acridine orange/ethidium bromide staining.Results:Benzydamine hydrochloride significantly decreased the secretion of TNF-αand IL-6,as well as the generation of reactive oxygen species inside the cells,thereby stabilizing the mitochondrial membrane potential and reducing DNA fragmentation.The ethanol-induced cellular necrosis was also reversed by the administration of benzydamine hydrochloride.Conclusions:Benzydamine hydrochloride ameliorates ethanol-induced cell apoptosis and inflammation in RAW macrophages.

Catharanthus roseus:a natural source for the synthesis of silver nanoparticles

Objective:To develop a simple rapid procedure for bioreduction of silver nanoparticles(AgNPs)using aqueous leaves extracts of Catharanthus roseus(C.roseus).Methods:Characterization were determined by using UV-Vis spectrophotometry,scanning electron microscopy(SEM),energy dispersive X-ray and X-ray diffraction.Results:SEM showed the formation of silver nanoparticles with an average size of 67 nm to 48 nm.X-ray diffraction analysis showed that the particles were crystalline in nature with face centered cubic geometry.Conclusions:C.roseus demonstrates strong potential for synthesis of silver nanoparticles by rapid reduction of silver ions(Ag^+ to Ag^0).This study provides evidence for developing large scale commercial production of value-added products for biomedical/nanotechnology-based industries.

Development of a novel nano-biosorbent for the removal of fluoride from water

The study was designed to investigate the use of two sorbents namely(i) Fe3O4 nanoparticles immobilized in sodium alginate matrix(FNPSA) and(ii) Fe3O4 nanoparticles and saponified orange peel residue immobilized in sodium alginate matrix(FNPSOPR) as sorbents for fluoride removal from contaminated water. The synthesized nanoparticles were analyzed and characterized by dynamic light scattering, X-ray diffraction, vibrating sample magnetometry, and scanning electron microscopy with energy dispersive X-ray spectroscopy and Fourier transform-infrared spectrometry. The sorbent matrices were prepared in the form of beads and surface functionalized to enable enhanced sorption of fluoride ions. Batch sorption studies were carried out and the sorption isotherm and reaction kinetics were analyzed. Both the sorbents followed Langmuir model of isotherm and fitted well with Pseudo first order reaction. The maximum sorption capacity exhibited by FNPSA and FNPSOPR was58.24 mg·g-1and 80.33 mg·g-1respectively. Five sorption–desorption cycles exhibited 100%, 97.56%, 94.53%,83.21%, and 76.53% of regeneration of FNPSOPR. Accordingly, it is demonstrated that FNSOPR could be used as a promising sorbent for easy and efficient removal of fluoride from contaminated water with good reusability.The current work suggests a simple and effective method to remove fluoride from contaminated water.

Inhibition of Fish Nodavirus by Gymnemagenol Extracted from Gymnema sylvestre

Viral nervous necrosis(VNN)has emerged to become a major problem in the culture of larval and juvenile marine fish worldwide.Bioactive phytochemicals isolated from commonly available medicinal plants are often screened for their efficacy in controlling fish viral diseases.Occurrence of newer viral strains and resistance to existing antiviral drugs are problems currently as-sociated with treatment of VNN,which necessitates looking for alternate sources for effective antiviral drugs.The aim of the present study was to screen antiviral potential of gymnemagenol(C30H50O4)previously extracted from leaves of Gymnema sylvestre.The fish nodavirus,grouper nervous necrosis virus(GNNV)in infected Sahul Indian Grouper Eye(SIGE)cell lines were used to study the antiviral activity of gymnemagenol under in vitro conditions.The susceptibility of the virus to gymnemagenol was confirmed by measuring the viral titre(TCID50 mL?1)in virus-infected SIGE cells every 24 h.Gymnemagenol at 20 μg mL?1 inhibited the prolifera-tion of GNNV to 53% at the end of the 6th d by inhibiting the proliferation of GNNV-infected SIGE cells.The viable SIGE cells were reduced to 47% as determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.The viral titre(TCID50 mL?1)was also reduced to log 2.8 at the end of the 7th d in gymnemagenol-treated SIGE cells after inoculated with GNNV when compared to untreated control SIGE cell viral titre(log 4.1).Based on our results it can be concluded that gymnemagenol could be used as an antiviral agent against GNNV infection.

Intra-islet endothelial cell and β-cell crosstalk: Implication for islet cell transplantation

The intra-islet microvasculature is a critical interface between the blood and islet endocrine cells governing a number of cellular and pathophysiological processes associated with the pancreatic tissue. A growing body of evidence indicates a strong functional and physical interdependency of β-cells with endothelial cells(ECs), the building blocks of islet microvasculature. Intra-islet ECs, actively regulate vascular permeability and appear to play a role in fine-tuning blood glucose sensing and regulation. These cells also tend to behave as "guardians", controlling the expression and movement of a number of important immune mediators, thereby strongly contributing to the physiology of islets. This review will focus on the molecular signalling and crosstalk between the intra-islet ECs and β-cells and how their relationship can be a potential target for intervention strategies in islet pathology and islet transplantation.

Synergistic effect of flavonoids combined with antivenom on neutralisation of Naja naja venom

Objective:To evaluate the neutralizing effects of flavonoids on snake venom toxicity by stand-alone and combinatorial approaches.Methods:Synthetic flavonoids were assessed,either individually or in combination with antivenom,for their neutralization of phospholipase A_(2)(PLA_(2)),protease,antioxidant(DPPH)assay and hemotoxic activity.Molecular docking studies were performed to understand possible binding of flavonoids with Naja naja venom PLA_(2).In vivo studies were carried out to confirm the neutralisation effects using a mouse model.Moreover,inhibition of PLA_(2) was monitored using combinatorial approaches.Results:Among the flavonoids used,quercetin and naringenin inhibited PLA_(2)(56%and 45%),protease(71%and 64%),DPPH scavenging(69.0%and 77.5%)and hemotoxic(70%)activities.Molecular docking studies indicated that the flavonoids bind to the substrate-binding site of PLA_(2)(Cys44 and Tyr63).In vivo studies showed a reduction in the venom toxicity level in the presence of naringenin.Additionally,combinatorial studies using the mixture of flavonoid and anti-venom revealed the possibility of synergistic effect(up to 32%enhancement)in neutralising the venom enzymes.Conclusions:These flavonoids can be used as additives for the treatment of snake bites,which may exert synergistic effects in combination with antivenom and decrease the post-therapeutic effects caused by excessive use of antivenom.

Clupeid fish hosts a Peniculus sp.(Pennellidae,Siphonostomatoida, Copepoda)—First report on new host and season dependent prevalence

Pennellid copepod Peniculus fistula fistula(Nordmann, 1832)(Synonym: Peniculus fistula Nordmann, 1832,Aphia ID: 745880), a worldwide distributed species, has been recovered from at least 19 teleost families. The present paper reports for the first time from the Malabar coast(South India), not only the existence of a new host family, Clupeidae, hosting this parasitic copepod species(P. fistula fistula) but also their season dependent hosting. A total of 123 marine fish species, belonging to 77 genera and 38 families surveyed along the Malabar coast, only the clupeid, Anadontostoma chacunda(Hamilton, 1822) was shown to be infected by this copepod species;all the recovered(copepod) parasites were invariably found attached at the mid portion of the caudal fin lobes and lying parallel to the host body, indicating the strict site-specific parasitisation. There is a discrete seasonality in the prevalence(P<0.05) as the sign of infection was noticed during the period from September to May with relatively high prevalence during winter months(November–January). During the monsoon months(June–August), the host fish was found completely free from Peniculus infection. Interestingly, all the 229 recovered specimens(P. fistula fistula) were gravid females having paired uniserrate egg sacs with the length more than its own body length.

NPS31807, a Standardized Extract from <i>Sphaeranthus indicus</i>, Inhibits Inflammatory, Migratory and Proliferative Activity in Keratinocytes and Immune Cells

Chronic inflammation induced hyper-proliferation, and migration of keratinocytes are pathological hallmarks of psoriasis. Extracts from Sphaeranthus spp. demonstrate pharmacological activity in-vitro and in-vivo. However, the activity in modulating disease relevant pathways in psoriasis has not been reported. In the current study, a standardized herbal extract from Sphaeranthus indicus (NPS31807) was used to study the mechanistic activity under conditions of inflammation, keratinocyte proliferation and migration using cell based and gene expression assays. NPS31807 treatment reduced levels of pro-inflammatory cytokines from human macrophages and activated epidermal keratinocytes in a dose dependent manner. Treatment with NPS31807 diminished NFκB and AP-1 transcription activity in human macrophages. Lowered nuclear translocation of p65 sub-unit in macrophages by treatment confirmed reduced activity of NFκB. Gene expression profiling showed attenuated expression of genes involved with inflammation such as TNF signaling, and angiogenesis by NPS31807. Inhibition of angiogenesis and matrix metalloproteinase production in keratinocytes was confirmed using RTq-PCR assays. Pretreatment with NPS31807 led to significant reduction of STAT3 phosphorylation and mitogen induced cellular migration. NPS31807 induced inhibition of proliferative genes and BrdU uptake in epidermal keratinocytes. In summary, our study provides novel molecular insights into the anti-inflammatory, anti-migratory and anti-proliferative properties of NPS31807. In summary, NPS31807, an extract from Sphaeranthus indicus can be used as therapeutic option in inflammatory and auto-immune conditions such as psoriasis.

Biliary stents for active materials and surface modification:Recent advances and future perspectives

Demand for biliary stents has expanded with the increasing incidence of biliary disease.The implantation of plastic or self-expandable metal stents can be an effective treatment for biliary strictures.However,these stents are nondegradable and prone to restenosis.Surgical removal or replacement of the nondegradable stents is necessary in cases of disease resolution or restenosis.To overcome these shortcomings,improvements were made to the materials and surfaces used for the stents.First,this paper reviews the advantages and limitations of nondegradable stents.Second,emphasis is placed on biodegradable polymer and biodegradable metal stents,along with functional coatings.This also encompasses tissue engineering&3D-printed stents were highlighted.Finally,the future perspectives of biliary stents,including pro-epithelialization coatings,multifunctional coated stents,biodegradable shape memory stents,and 4D bioprinting,were discussed.

Optimization of parameters for cerium(Ⅲ) biosorption onto biowaste materials of animal and plant origin using 5-level Box-Behnken design: Equilibrium, kinetic, thermodynamic and regeneration studies

Response surface methodology （RSM） employing 5-level Box-Behnken design was used to optimize the biosorption of cerium（III） onto biowaste materials of animal and plant origin viz. prawn carapace （PC） and corn style （CS）. Various process parameters viz. pH （A：3.0-9.0）, biomass dosage （B：0.05-0.35 g/L）, initial metal concentration （C：50-350 mg/L）, contact time （D：2-6 h） and temperature （E：20-60 ℃ were chosen for optimization. A log transformation was suggested by the Box-Cox plot in the present case. A low p-value of〈0.0001 validated the significance of the model. Maximum Ce（III） uptake of 218.3 mg/g for PC and 180.2 mg/g for CS was noted under optimum conditions. Among the equilibrium isotherms, Freundlich model was found to be the best fitted one suggesting a heterogeneous mode of biosorption on PC whereas Langmuir model showed the best fit suggesting homogeneous mode of cerium biosorption on CS. This was further confirmed by scanning electron microscopy （SEM）. Kinetic studies showed better applicability of pseudo-first order model suggesting physisorption as phenomena underlying the process. Film-diffusion was suggested by the non-linearity of the Boyd plot. Thermodynamic studies showed that the process was endothermic and spontaneous. FTIR analysis confirmed a major involvement of the participation of amide, amines, ketones and primary alcohol groups during Ce（III） biosorption. EDAX analysis confirmed the major participation of carbon group during Ce（III） biosorption. This was the first report on parameter optimization of Ce（III） biosorption onto biowaste materials using 5-level Box-Behnken experimental design which might be helpful for the recovery of Ce（III） from aqueous environment.

In vitro and in silico studies on fibrinolytic activity of nattokinase： A clot buster from Bacillus sp.

BACKGROUND： Nattokinase （NK） is a serine protease enzyme of the subtilisin family. It exhibits a strong fibrinolytic activity. The fibrinolytic enzymes from Bacillus sp. have attracted interest as thrombolytic agents because of their efficiency in the fibrinolytic process including plasmin activation. METHODS： In the present study, VIT garden soil was collected and subjected to isolation process in order to screen for the NK production. Screening for NK enzyme was performed by radial caseinolytic assay. The production of NK enzyme was done in two different production medium for comparative studies. The NK enzyme was purified by gel permeation chromatography. The activity of the purified NK was checked by clot lysis and casein digestion assay. To investigate the structural basis of NK and fibrinogen interaction and also to identify the best binding mode, molecular dynamics and docking studies were performed. RESULTS： Based on the morphological and biochemical characterization, the isolate was identified as Bacillus sp. The overall purification fold of NK was about 3 with the specific activity of 664U/mg and 9.9% yield. Homogeneity of the purified enzyme was analyzed and confirmed by the single band obtained in SDS-PAGE. Molecular weight of the purified protease was estimated as 25 kDa. Purified NK enzyme exhibited 97% of effective clot lysis activity. The NK was docked in to the knob region of the fibrinogen at its binding site using Dock server. A total of 26 residues of fibrinogen and 29 residues of NK constitute the interface region. However, 9 residues offibrinogen （THR238, MET264, LYS266, ARG275, THR277, ALA279, ASN308, MET310, and LYS321） and 8 residues ofNK （GLY61, SER63, THR99, PHE189, LEU209, TYR217, ASN218, and MET222） are involved in intact binding. CONCLUSIONS： A significant amount of NK enzyme was obtained from Bacillus sp. The docking analysis revealed that the NK and fibrinogen adopt an extended binding pattern and interacts with the crucial residues to exhibit their activity.

Screening and molecular characterization of Serratia marcescens VITSD2： A strain producing optimum serratiopeptidase

The current work was attempted to isolate and characterize the serratiopeptidase producing Serratia sp. Among the 10 bacterial isolates 7 strains were identified as Serratia sp. Out of 7 strains one showed potent proteolytic activity and selected for further studies. Based on the morphological, biochemical and molecular characterization, the potent isolate （RH03） was identified as Serratia marcescens （GenBank accession number： KC961637） and the strain was designated as Serratia marcescens VITSD2. The production of serratiopeptidase was carried out in trypticase soya broth and the enzyme was partially purified using ammonium sulfate precipitation and dialysis. The specific activity was determined by casein hydrolysis assay and was found to be 12.00, 21.33, and 25.40 units/rag for crude, precipitated and dialysed samples. The molecular weight of the protease was determined by SDS-PAGE and it was found to be 50 kDa. The antibacterial activity of the produced serratiopeptidase showed moderate activity against Pseudomonas aeruginosa MTCC No. 4676 （12 mm） and Escherichia coli MTCC No. 1588 （15 mm）.

Synthesis, antibacterial and anti-inflammatory activity of bis(indolyl)methanes

A series of bioactive bis（indolyl）methanes are synthesized by one-pot green reaction of indole with various substituted aldehydes by microwave irradiation under solvent free conditions. The antibacterial activity against Staphylococcus aureus and anti-inflammatory activity of the synthesized bis（indolyl）-methanes are evaluated in vitro and compared to standard drugs tetracycline and diclofenac,respectively. The majority of the compounds showed good antibacterial and anti-inflammatory activity.Interestingly, compounds 3j, 3i, 3k and 3g exhibited much higher anti-inflammatory activity than the standard diclofenac drug and thus qualify for clinical trials to be used as an anti-inflammatory compound.

Melanoma therapeutics: a literature review

Melanoma is a relentless type of skin cancer which involves myriad signaling pathways which regulate many cellular processes.This makes melanoma difficult to treat,especially when identified late.At present,therapeutics include chemotherapy,surgical resection,biochemotherapy,immunotherapy,photodynamic and targeted approaches.These interventions are usually administered as either a single-drug or in combination,based on tumor location,stage,and patients'overall health condition.However,treatment efficacy generally decreases as patients develop treatment resistance.Genetic profiling of melanocytes and the discovery of novel molecular factors involved in the pathogenesis of melanoma have helped to identify new therapeutic targets.In this literature review,we examine several newly approved therapies,and briefly describe several therapies being assessed for melanoma.The goal is to provide a comprehensive overview of recent developments and to consider future directions in the field of melanoma.