Multiple-Drug Resistant Shiga Toxin-Producing E. coli in Raw Milk of Dairy Bovine.

INTRODUCTION: Raw milk may contain pathogenic microorganisms harmful to humans, e.g., multidrug-resistant Escherichia coli non-O157:H7, which can cause severe colitis, hemolytic uremia, and meningitis in children. No studies are available on the prevalence of Shiga toxin-producing E. coli (STEC O157:H7) in sick or healthy dairy animals in the Khyber Pakhtunkhwa Province of Pakistan. AIM: This study aimed to isolate, characterize, and detect antibiotic resistance in STEC non-O157:H7 from unpasteurized milk of dairy bovines in this province. MATERIALS AND METHODS: We collected raw milk samples (n = 800) from dairy farms, street vendors, and milk shops from different parts of the Khyber Pakhtunkhwa Province. E. coli was isolated from these samples followed by latex agglutination tests for serotyping. The detection of STEC was conducted phenotypically and confirmed by the detection of virulence genes genotypically. An antibiogram of STEC isolates was performed against 12 antibiotics using the disc diffusion method. RESULTS: A total of 321 (40.12%) samples were found to be positive for E. coli in this study. These samples were processed for the presence of four virulence genes (Stx1, Stx2, ehxA, eae). Forty samples (5.0%) were STEC-positive. Of these, 38%, 25%, 19%, and 18% were positive for Stx1, Stx2, ehxA, and eae, respectively. Genotypically, we found that 1.37% of STEC isolates produced extended-spectrum beta-lactamase (ESBL) and contained the blaCTX M gene. Resistance to various antibiotics ranged from 18% to 77%. CONCLUSION: This study highlights the risk of virulent and multidrug-resistant STEC non-O157:H7 in raw milk and the need for proper quality surveillance and assurance plans to mitigate the potential public health threat.

Interpretation of adsorption isotherm and kinetics behind fluorene degradation.

The gradual release of slow-degrading polycyclic aromatic hydrocarbons into the environment creates a high level of threat to aquatic and terrestrial life worldwide. Remediation of these PAHs should be designed in such a way that it poses as few or no environmental hazards as possible. In our study, we examined the degradation ability of the synthesized MnO2 nanoparticles against fluorene. The MnO2 nanoparticle prepared was found to be spherical from the SEM analysis. XRD analysis confirms the average crystallite size as 31.8652 nm. Further, the characterization of nanoparticles was confirmed by UV-DRS, FT-IR, DLS, and HPLC techniques. The extent of adsorption potential of the synthesized nanoparticles was established from the batch adsorption studies and the kinetic and isotherm model was interpreted. The antimicrobial properties of the synthesized MnO2 nanoparticles were analyzed.

Calotropis procera: A double edged sword against glioblastoma, inhibiting glioblastoma cell line growth by targeting histone deacetylases (HDAC) and angiogenesis.

Despite substantial investments in anti-glioblastoma (GBM) drug discovery over the last decade, progress is limited to preclinical stages, with clinical studies frequently encountering obstacles. Angiogenic and histone deacetylase inhibitors (HDACi) have shown profound results in pre-clinical studies. Investigating a multicomponent anti-cancer remedy that disrupts the tumor angiogenic blood vessels and simultaneously disrupts HDACs, while inducing minimal side effects, is critically needed. The crude extracts derived from medicinal plants serve as a renewable reservoir of anti-tumor drugs, exhibiting reduced toxicity compared to chemically synthesized formulations. Calotropis procera is a traditional medicinal plant, and its anticancer potential against many cancer cell lines has been reported, however its antiangiogenic and HDAC inhibitory action is largely unknown. The anticancer activity of methanol leaf extract of C. procera was tested in three types of human glioblastoma cell lines. Wild-type and transgenic zebrafish embryos were used to evaluate developmental toxicity and angiogenic activity. A human angiogenic antibody array was used to profile angiogenic proteins in the U251 GM cell line. A real-time reverse transcriptase polymerase chain reaction (RT PCR) assay was used to detect the differential expression of eleven HDAC genes in U251 cells treated with C. procera extract. The extract significantly reduced the proliferation of all three types of GBM cell lines and the cytotoxicity was found to be more pronounced in U251 GM cells, with an IC50 value of 2.63 ± 0.23 µg/ml, possibly by arresting the cell cycle at the G2/M transition. The extract did not exhibit toxic effects in zebrafish embryos, even at concentrations as high as 1000 µg/ml. The extract also inhibited angiogenic blood vessel formation in the transgenic zebrafish model in a dose-dependent manner. The results from the angiogenic antibody array have suggested novel angiogenesis targets that can be utilized to treat GBM. Real-time RT PCR analysis has shown that C. procrea extract caused an upregulation of HDAC5, 7, and 10, while the mRNA of HDAC1, 2, 3 and 8 (Class I HDACs), and HDAC4, 6, and 9 (Class II) were downregulated in U251 GM cells. The cytotoxicity of the C. procera extract on GBM cell lines could be due to its dual action by regulation of both tumor angiogenesis and histone deacetylases enzymes. Through this study, the C. procera leaf extract has been suggested as an effective remedy to treat GBM with minimal toxicity. In addition, various novel angiogenic and HDAC targets has been identified which could be helpful in designing better therapeutic strategies to manage glioblastoma multiforme in human patients.

Assessment of antidiabetic, anti-inflammatory, antioxidant and anticancer activity competence of methonolic extracts of Trianthema ortulacastrum and Andrographis paniculata.

An in-vitro investigation was performed to evaluate and compare the phytochemical, antioxidant, antidiabetic, anti-inflammatory, and anti-lung cancer activities of methanol extracts of aerial parts of Andrographis paniculata and Trianthema portulacastrum. Furthermore studied major functional groups of phytochemicals present in the methanol extracts of these plants through Fourier transform infrared (FTIR) analysis. The results showed that the methanol extract of A. paniculata contain more number of pharmaceutically valuable phytochemicals such as alkaloids, flavonoids, terpenoids, saponin, glycoside, phytosterol, and tannin than T. portulacastrum. Similar way the methanol extract of A. paniculata showed considerable dose dependent antioxidant (DPPH: 63%), antidiabetic (α-amylase: 82.31% and α-glucosidase inhibitions: 72.34%), and anti-inflammatory (albumin-denaturation inhibition: 76.3% and anti-lipoxygenase: 61.2%) activities (at 900 µg mL-1 concentration) than T. portulacastrum. However, the anti-lung cancer activities of these test plants against A549 cells were not considerable. According to FTIR analysis, the A. paniculata methanol extract has a larger number of characteristic peaks attributed to the active functional groups of pharmaceutically valuable bioactive components that belong to different types of phytochemicals. These findings imply that A. paniculata methanol extracts can be used for additional research, such as bioactive compound screening and purification, as well as assessing their potential biomedical uses in various in-vitro and in-research settings.

Biological Control of Hyalomma Ticks in Cattle by Fungal Isolates.

Ticks pose a major threat to cattle health and production in Pakistan because they transmit pathogens of diseases like Babesiosis and Theileriosis. Hyalomma spp., found across Africa, Asia, and Europe, are especially problematic. This study explored biocontrol of Hyalomma spp. using spore-free fungal culture filtrates collected from dairy farm soil in Kohat, Pakistan. Three fungal species of the genera Alternaria, Aspergillus, and Penicillium were isolated, and their filtrates were tested against tick adults and larvae. Filtrate concentrations were prepared at different strengths. Data were taken after the exposure of adults and larvae ticks to various concentrations of the fungal filtrates. Results indicated that at 100% concentration, all fungal filtrates induced 100% mortality in adults and larvae. Decreasing filtrate concentration lowered tick mortality. The lowest concentration caused the least mortality. The effect was time- and dose-dependent. In conclusion, spore-free fungal culture filtrates can provide biocontrol of Hyalomma spp. in a time- and concentration-dependent manner. Further research should explore the active compounds causing mortality and optimal application methods. The process outlined here provides a natural biocontrol alternative to chemical pesticides to reduce tick infestations and associated cattle diseases in Pakistan.

Nano-composite rGO-Ag-Cu-Ni mediated photocatalytic degradation of anthracene and benzene.

Polycyclic aromatic hydrocarbons (PAHs) are omnipresent, persistent, and carcinogenic pollutants continuously released in the atmosphere due to the rapid increase in population and industrialization worldwide. Hence, there is an ultimate rise in concern about eliminating the toxic PAHs and their related aromatic hydrocarbons from the air, water, and soil environment by employing efficient removal technologies using nanoparticles as a catalyst. Here, the degradation of selective PAHs viz., anthracene and benzene using laboratory synthesized rGO-Ag-Cu-Ni nanocomposite (catalyst) was studied. Characterization studies revealed the nanocomposites exhibited surface plasma resonance at 350 - 450 nm, confirming the presence of Ag, Cu, and Ni metal ions embedded on the reduced graphene substrate. It was found that the nanocomposites synthesized were spherical, amorphous in nature, and aggregated together with measurements ranging from 423 to 477 nm. An SEM-EDX analysis of the nanocomposite demonstrated that it contained 25.13% O, 14.24% Ni, 27.79% Cu, and 32.84% Ag, which confirms the synthesis of the nanocomposite. Crystalline, sharp nanocomposites of average size 17-41 nm with an average diameter of 118.5 nm (X-ray diffraction and DLS) were observed. FTIR spectra showed that the nanocomposites had the functional groups alkanes, alkenes, alkynes, carboxylic acids, and halogen derivatives. Batch adsorption studies revealed that the maximum degradation achieved at optimum nano-composite concentration of 10 µg/mL, pH value of 5, PAHs concentration of 2 µg/mL and effective irradiation source being UV radiations in the case of both benzene and anthracene pollutants. The degradation of benzene and anthracene followed Freundlich & Langmuir isotherm with the highest R2 value of 0.9894 & 0.9885, respectively. Adsorption kinetic studies under optimum conditions revealed that the adsorption of both benzene and anthracene followed Pseudo-second order kinetics. Antimicrobial studies revealed that the synthesized nano-composite exhibited potential antimicrobial activity against Gram positive bacterium (Bacillus subtilis, Staphylococcus aureus), Gram negative bacterium (Klebsiella pneumonia, Escherichia coli) and fungal strain (Aspergillus niger) respectively. Thus, the synthesized rGO-Ag-Cu-Ni nano-composite acts as an effective antimicrobial agent as well as a PAHs degrading agent, helping to overcome antibiotics resistance and to mitigate the overgrowing PAHs pollution in the environment.

Recent Advanced Synthesis Strategies for the Nanomaterial-Modified Proton Exchange Membrane in Fuel Cells.

Hydrogen energy is converted to electricity through fuel cells, aided by nanostructured materials. Fuel cell technology is a promising method for utilizing energy sources, ensuring sustainability, and protecting the environment. However, it still faces drawbacks such as high cost, operability, and durability issues. Nanomaterials can address these drawbacks by enhancing catalysts, electrodes, and fuel cell membranes, which play a crucial role in separating hydrogen into protons and electrons. Proton exchange membrane fuel cells (PEMFCs) have gained significant attention in scientific research. The primary objectives are to reduce greenhouse gas emissions, particularly in the automotive industry, and develop cost-effective methods and materials to enhance PEMFC efficiency. We provide a typical yet inclusive review of various types of proton-conducting membranes. In this review article, special focus is given to the distinctive nature of nanomaterial-filled proton-conducting membranes and their essential characteristics, including their structural, dielectric, proton transport, and thermal properties. We provide an overview of the various reported nanomaterials, such as metal oxide, carbon, and polymeric nanomaterials. Additionally, the synthesis methods in situ polymerization, solution casting, electrospinning, and layer-by-layer assembly for proton-conducting membrane preparation were analyzed. In conclusion, the way to implement the desired energy conversion application, such as a fuel cell, using a nanostructured proton-conducting membrane has been demonstrated.

Streptomyces Bioactive Metabolites Prevent Liver Cancer through Apoptosis, Inhibiting Oxidative Stress and Inflammatory Markers in Diethylnitrosamine-Induced Hepatocellular Carcinoma.

A safe and effective treatment for liver cancer is still elusive despite all attempts. Biomolecules produced from natural products and their derivatives are potential sources of new anticancer medications. This study aimed to investigate the anticancer potential of a Streptomyces sp. bacterial extract against diethylnitrosamine (DEN)-induced liver cancer in Swiss albino mice and explore the underlying cellular and molecular mechanisms. The ethyl acetate extract of a Streptomyces sp. was screened for its potential anticancer activities against HepG-2 using the MTT assay, and the IC50 was also determined. Gas chromatography-mass spectrometric analysis was used to identify the chemical constituents of the Streptomyces extract. Mice were administered DEN at the age of 2 weeks, and from week 32 until week 36 (4 weeks), they received two doses of Streptomyces extract (25 and 50 mg/kg body weight) orally daily. The Streptomyces extract contains 29 different compounds, according to the GC-MS analysis. The rate of HepG-2 growth was dramatically reduced by the Streptomyces extract. In the mice model. Streptomyces extract considerably lessened the negative effects of DEN on liver functions at both doses. Alpha-fetoprotein (AFP) levels were significantly (p < 0.001) decreased, and P53 mRNA expression was increased, both of which were signs that Streptomyces extract was suppressing carcinogenesis. This anticancer effect was also supported by histological analysis. Streptomyces extract therapy additionally stopped DEN-induced alterations in hepatic oxidative stress and enhanced antioxidant activity. Additionally, Streptomyces extract reduced DEN-induced inflammation, as shown by the decline in interleukin-1 beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α) levels. Additionally, the Streptomyces extract administration dramatically boosted Bax and caspase-3 levels while decreasing Bcl-2 expressions in the liver according to the Immunohistochemistry examination. In summary, Streptomyces extract is reported here as a potent chemopreventive agent against hepatocellular carcinoma through multiple mechanisms, including inhibiting oxidative stress, cell apoptosis, and inflammation.

Phytochemical profiling, anti-hyperglycemic, antifungal, and radicals scavenging potential of crude extracts of Athyrium asplenioides- an in-vitro approach.

This research was aimed to evaluate the phytochemical profile, antifungal, anti-hyperglycemic, as well as antioxidant activity competence of different extracts of Athyrium asplenioides through in-vitro approach. The A. asplenioides crude methanol extract contained considerable quantity of pharmaceutically precious phytochemicals (saponins, tannins, quinones, flavonoid, phenols, steroid, and terpenoids) than others (acetone, ethyl acetate, and chloroform). Interestingly, the crude methanol extract showed remarkable antifungal activity against Candida species (C. krusei: 19.3 ± 2 mm > C. tropicalis: 18.4 ± 1 mm > C. albicans: 16.5 ± 1 mm > C. parapsilosis: 15.5 ± 2 mm > C. glabrate: 13.5 ± 2 mm > C. auris: 7.6 ± 1 mm) at a concentration of 20 mg mL-1. The crude methanol extract also showed remarkable anti-hyperglycemic activity on concentration basis. Surprisingly, remarkable free radicals scavenging potential against DPPH (76.38%) and ABTS (76.28%) free radicals at a concentration of 20 mg mL-1. According to the findings, the A. asplenioides crude methanol extract contains pharmaceutically valuable phytochemicals and may be useful for drug discovery.

Phytotoxicity and cytotoxicity attributes of immobilized Bacillus cereus treated and untreated textile effluents on Vigna mungo seeds and Artemia franciscana larvae.

The physicochemical attributes of textile effluents collected from secondary treatment stage was investigated in this study and also assess the biosorption potential of membrane immobilized Bacillus cereus and free form of Bacillus cereus on textile effluent through bioreactor model study to find a sustainable solution to manage the textile effluent as vital need. Furthermore, the phytotoxicity and cytotoxicity nature of treated and untreated textile effluents on Vigna mungo and Artemia franciscana larvae under laboratory conditions as a novel approach. The textile effluent physicochemical parameter analysis results showed that the properties such as colour (Hazen unit), pH, turbidity, As, Biological Oxygen Demand (BOD), Chemical Oxygen Demand (COD), Cd, Cl, Cr, Cu, Hg, Ni, Pb, SO42-, and Zn were beyond the acceptable limits. Bacillus cereus immobilized on a polyethylene membrane eliminated greater amounts of dye (25.0 ± 1.3, 56.5 ± 1.8, 57.18 ± 1.5, and 54.34 ± 1.7 Hazen unit from An1, Ae2, Ve3, and So4 respectively) and pollutants (As: 0.9-2.0, Cd: 6-8, Cr: 300-450, Cu: 5-7, Hg: 0.1-0.7, Ni: 8-14, Pb: 4-5, and Zn: 4-8 mg L-1) from textile effluent in a week of biosorption investigation using a bioreactor model (batch type) compared to a free form of B. cereus on textile effluent. The phytotoxicity and cytotoxicity study results revealed that the membrane immobilized B. cereus treated textile effluent exposure showed reduced phytotoxicity and minimal cytotoxicity (including mortality) percentage compared with free form B. cereus treated and untreated textile effluents. These entire results conclude that the membrane immobilized B. cereus may considerably minimize/detoxify the harmful pollutants from the textile effluents. A large scale level biosorption approach need to be performed to validate the maximum pollutants removing potential of this membrane immobilized bacteria species and optimal conditions for effective remediation.

The molecular interplay of known phytochemicals as Culex pipiens and Rift Valley fever virus inhibitors through molecular docking.

Infectious diseases transmitted by vectors have claimed millions of lives. The mosquito Culex pipiens is a main vector species of Rift Valley Fever virus (RVFV) transmission. RVFV is an arbovirus that infects both people and animals. No effective vaccine or drugs are available for RVFV. Therefore, it is vital to find effective therapies for this viral infection. Because of their critical roles in transmission and infection, acetylcholinesterase 1 (AChE1) of Cx. Pipiens and RVFV glycoproteins, and nucleocapsid proteins are appealing protein targets. To understand intermolecular interactions, computational screening was carried out using molecular docking. More than 50 compounds were tested against different target proteins in the current study. Anabsinthin (-11.1 kcal/mol), zapoterin (-9.4 kcal/mol), porrigenin A (-9.4 kcal/mol), and 3-Acetyl-11-keto-beta-boswellic acid (AKBA) (-9.4 kcal/mol) were the top hit compounds for Cx. Pipiens. Similarly, the top hit compounds for RVFV were zapoterin, porrigenin A, anabsinthin, and yamogenin. The toxicity of Rofficerone is predicted as fatal (Class II), whereas Yamogenin is safe (Class VI). Further investigations are needed to validate the selected promising candidates against Cx. pipiens and RVFV infection using in-vitro and in-vivo methods.

Ecogenotoxicological studies for an early toxicity screening and monitoring in Epinephalus chlorostigma and Scamberomorus commerson.

The study was planned to investigate DNA fragmentation in fish to screen aquatic toxicity and in Epinephalus chlorostigma and Scamberomorus commerson collected from Red sea near Jizan, Saudi Arabia from three locations "(Corniche North park: "16.92161, 42.54631; Jizan Port: 16.874, 42.54952" N and Jizan Economic City: 17.26589, 42.34738" ")" were used as a case study for the application of comet assay. The study area of the Red Sea is polluted due to anthropogenic activities and the disposal of wastes from multiple sources. Comet and micronucleus assays were used to detect genotoxicity in these fish species harvested from three sites. The concentration of Pb, Cr, Zn, Mn, Cu, Cd, Sn, and Hg was higher in the water samples collected from the polluted site compared to the non-polluted site of the Red sea. Comet assay for S. commerson showed significant (p < 0.05) genetic damage about 44.33 ± 3.03% DNA in comet tail at site S1. It was subsequently reduced to 31.71 ± 3.52% and 22.11 ± 2.52% at sites S2 and S3. E. chlorostigma also showed significant DNA in comet tail as 17.34 ± 2.19%, 11.87 ± 3.01%, and 36.41 ± 3.98% at site S1-S3, respectively. Significant (p < 0.05) DNA damage was observed in the fishes procured from non-polluted locations and upstream locations. The micronucleus induction in E. chlorostigma was recorded as 23.20 ± 4.19 and 2.20 ± 0.58%, respectively, non-polluted and polluted sites. S. commerson exhibited significant differences between polluted and non-polluted sites (44.80 ± 3.73 and 8.20 ± 2.20‰) polluted and upstream (44.80 ± 3.73 and 20.60 ± 4.02‰), respectively. A significant difference was obtained between E. chlorostigma and S. commerson for nuclear abnormalities S. commerson showed higher frequencies for nuclear deformities than E. chlorostigma. S. commerson showed substantial micronucleus induction frequencies collected from an area of low pollution intensity (upstream). This study showed that E. clorostigma and S. commerson could be successfully used as a bioindicator to determine the health of the Red Sea through the most specific assays such as comet and micronucleus tests as an early warning and to devise the monitoring strategies to ensure a safe supply of fish for human consumption.

Investigating the antiangiogenic potential of Rumex vesicarius (humeidh), anticancer activity in cancer cell lines and assessment of developmental toxicity in zebrafish embryos.

Recent trends in anticancer therapy is to use therapeutic agents which not only kill the cancer cell, but are less toxic to surrounding normal cells/tissue. One approach is to cut the nutrient supply to growing tumor cells, by blocking the formation of new blood vessels around the tumor. As the phytochemicals and botanical crude extracts have proven their efficacy as natural antiangiogenic agents with minimum toxicities, there is need to explore varieties of medicinal plants for novel antiangiogenic compounds. Rumex vesicarius L. (Humeidh), is an annual herbal plant with proven medicinal values. The antiangiogenic potential, and developmental toxicity of humeidh in experimental animal models has never been studied before. The crude extracts were prepared from the roots, stems, leaves and flowers of Rumex vesicarius L. in methanol, chloroform, ethyl acetate and n-hexane. The developmental toxicity screening in zebrafish embryos, has revealed that Rumex vesicarius was not toxic to zebrafish embryos. The chloroform stem extract showed significant level of antiangiogenic activity in zebrafish angiogenic assay on a dose dependent manner. Thirty five (35) bioactive compounds were identified by gas chromatography mass spectrophotometry (GC-MS) analysis in the stem extract of Rumex vesicarius. Propanoic acid, 2-[(trimethylsilyl)oxy]-, trimethylsilyl ester, Butane, 1,2,3-tris(trimethylsiloxy), and Butanedioic acid, bis(trimethylsilyl) ester were identified as major compound present in the stem of R. vasicarius. The anticancer activity of roots, stem, leaves and flowers crude extract was evaluated in human breast cancer (MCF7), human colon carcinoma (Lovo, and Caco-2), human hepatocellular carcinoma (HepG2) cell lines. Most of the crude extracts did not show significant level of cytotoxicity in tested cancer cells line, except, chloroform extract of stem which exhibited strong anticancer activity in all tested cancer cells with IC50 values in micro molar range. Based on these results, it is recommended that formulation prepared from R. vesicarius can further be tested in clinical trials in order to explore its therapeutic potential as an effective and safe natural anticancer product.

Anti-proliferative and anti-inflammatory activities of entophytic Penicillium crustosum from Phoenix dactylifer.

Natural sources have been and will remain an inspiration source for modern chemistry. The current study investigates the antiproliferative and anti-inflammatory action of the ethyl acetate fraction of Penicillium crustosum from Phoenix dactylifera. This paper reports the isolation of P. crustosum from leaves of P. dactylifera and the antiproliferative activities of ethyl acetate fraction on cancer cells. To reach this goal, the anti-proliferation and cytotoxicity effects were evaluated by MTT and LDH assay respectively. The quantitative real time PCR technique was used to investigate IL-6 and IL-8 gene expression. Our results revealed higher anti-proliferative activity against HepG2 (82µg/ml) than MCF7 (126µg/ml) and inhibited the migration of the cell lines. The ethyl acetate fraction significantly altered LDH levels and reduced IL-6 transcript expression on MCF7 cell line but not in HepG2 cell line which could be specific anti-inflammatory drug in breast cancer cell line. These results suggest that Phoenix dactylifera extract has a potent anti-proliferative and anti-inflammatory action. Further investigation to isolate the active compounds and mode of action is required.

Synthesis of Novel Class of N-Alkyl-isatin-3-iminobenzoic Acid Derivatives and Their Biological Activity in Zebrafish Embryos and Human Cancer Cell Lines.

Isatin (1H-indole-2,3-dione) and many of its derivatives are reported to have pharmacological properties. In this study, we report the synthesis and biological activity of a new class of N-alkyl-isatin-3-iminobenzoic acid derivatives prepared via the condensation of N-alkyl isatin with 4-aminobenzoic acid by conventional, microwave, and ultrasonic methods. Microwave irradiation yielded the products in a shorter reaction time with higher yields and purities. The compounds were screened in zebrafish embryos, and also in three human cancer cell lines (MCF7, HepG2, and Jurkat) and one normal human cell line i.e., human foreskin cell line (HFF-1). Two compounds (3c, 3f) were found to be highly effective against hematopoiesis in live zebrafish embryo at 10 µM concentration. The developmental stage-dependent treatment indicated that these compounds interfered with the differentiation of hemangioblasts to hematopoietic cells in zebrafish embryos. The comparative screening of semaxanib (SU5416) (a known isatin derivatives), to compounds synthesized in this study, revealed the contrasting effects of these two classes of isatin derivatives on zebrafish hematopoiesis. Most of the N-alkyl-isatin-3-iminobenzoic acid derivatives were toxic on cancer and non-cancer tested human cells lines, however, the compounds 3c and 3f specifically affected the cell viability of Jurkat cells (human hematological cell line) with least IC50 values of 16.5 and 7.8 µM. The structure-activity relationship (SAR) analysis indicated that the substitution pattern of the isatin at the 5-position was vital for activity. The in vivo and in vitro biological activities of these compounds suggested their potential use as pharmaceutical compounds for human leukemia treatment.

Biological screening of novel derivatives of valproic acid for anticancer and antiangiogenic properties.

BACKGROUND: Valproic acid (VPA) is a potent anticancer and antiangiogenic agent. However, design and synthesis of chemical derivatives with improved antiangiogenic and anticancer activities are still necessary. In this study a library of novel derivatives of VPA was synthesized and tested. METHODS: A human liver cancer cell line (HepG2) and a human normal embryonic kidney cell line (HEK 293) were exposed to various concentrations of VPA derivatives for 24 hours and cell viability was checked by MTT colorimetric assay. Anti-angiogenic properties were evaluated in transgenic zebrafish embryos. RESULTS: N-valproylglycine derivatives suppressed survival almost 70% (p value 0.001) in HepG2 cells but only 10-12% in HEK 293 cells (p value 0.133). They also suppressed angiogenic blood vessel formation by 80% when used between 2-20 µM in zebrafish embryos. Valproic acid hydrazides showed moderate level of anticancer activity by affecting 30-50% (p value 0.001) of cell viability in HepG2 cells and 8-10% in HEK293 cells (p value 0.034). CONCLUSION: The majority of compounds in this study showed potent and stronger antiangiogenic and anticancer activity than VPA. They proved selectively toxic to cancer cells and safer for normal cells. Moreover, these compounds inhibited developmental angiogenesis in zebrafish embryos. Based on the fact that liver is a highly vascularized organ, in case of liver carcinoma these compounds have the potential to target the pathological angiogenesis and could be an effective strategy to treat hepatocellular carcinoma.