Javed Iqbal Qazi

Targeting Apoptosis and Multiple Signaling Pathways with Icariside II in Cancer Cells

Cancer is the second leading cause of deaths worldwide. Despite concerted efforts to improve the current therapies, the prognosis of cancer remains dismal. Highly selective or specific blocking of only one of the signaling pathways has been associated with limited or sporadic responses. Using targeted agents to inhibit multiple signaling pathways has emerged as a new paradigm for anticancer treatment. Icariside II, a flavonol glycoside, is one of the major components of Traditional Chinese Medicine Herba epimedii and possesses multiple biological and pharmacological properties including anti-inflammatory, anti-osteoporosis, anti-oxidant, anti-aging, and anticancer activities. Recently, the anticancer activity of Icariside II has been extensively investigated. Here, in this review, our aim is to give our perspective on the current status of Icariside II, and discuss its natural sources, anticancer activity, molecular targets and the mechanisms of action with specific emphasis on apoptosis pathways which may help the further design and conduct of preclinical and clinical trials. Icariside II has been found to induce apoptosis in various human cancer cell lines of different origin by targeting multiple signaling pathways including STAT3, PI3K/AKT, MAPK/ERK, COX-2/PGE2 and β-Catenin which are frequently deregulated in cancers, suggesting that this collective activity rather than just a single effect may play an important role in developing Icariside II into a potential lead compound for anticancer therapy. This review suggests that Icariside II provides a novel opportunity for treatment of cancers, but additional investigations and clinical trials are still required to fully understand the mechanism of therapeutic effects to further validate it in anti-tumor therapy.

Pseudolaric Acid B Induces Caspase-Dependent and Caspase-Independent Apoptosis in U87 Glioblastoma Cells

Pseudolaric acid B (PLAB) is one of the major bioactive components of Pseudolarix kaempferi. It has been reported to exhibit inhibitory effect on cell proliferation in several types of cancer cells. However, there is no report elucidating its effect on glioma cells and organ toxicity in vivo. In the present study, we found that PLAB inhibited growth of U87 glioblastoma cells in a dose-dependent manner with IC50 ~10 μM. Flow cytometry analysis showed that apoptotic cell death mediated by PLAB was accompanied with cell cycle arrest at G2/M phase. Using Western blot, we found that PLAB induced G2/M phase arrest by inhibiting tubulin polymerization in U87 cells. Apoptotic cell death was only partially inhibited by pancaspase inhibitor, z-VAD-fmk, which suggested that PLAB-induced apoptosis in U87 cells is partially caspase-independent. Further mechanistic study demonstrated that PLAB induced caspase-dependent apoptosis via upregulation of p53, increased level of proapoptotic protein Bax, decreased level of antiapoptotic protein Bcl-2, release of cytochrome c from mitochondria, activation of caspase-3 and proteolytic cleavage of poly (ADP-ribose) polymerase (PARP) and caspase-independent apoptosis through apoptosis inducing factor (AIF). Furthermore, in vivo toxicity study demonstrated that PLAB did not induce significant structural and biochemical changes in mouse liver and kidneys at a dose of 25 mg/kg. Therefore, PLAB may become a potential lead compound for future development of antiglioma therapy.

Biotechnological potential and conservatory of extremophiles from climatically wide ranged developing countries: Lesson from Pakistan.

Abstract Technological advances, in developing countries, without considering environmental health issues have generated microsites uninhabitable to organisms including usual bacteria. While extremophilic microorganisms thrive in such locations. It is the high time to isolate and conserve the extremophilic biodiversity from such man made habitats before the environmental awareness treats them to the level of “destructionâ. For biotechnology processes to be geared by employing extremophilic microorganisms, locations characterized with high/low temperature for most part of the year, hypersaline brine and abneutral pH environments may render the respective microbes to do their allotted/required jobs with minimal of extraneous investments. Besides, many known and expected potentials of extremophilic bacteria for biotechnological applications, major attraction lies in operating bioprocesses under non-aseptic conditions for making them economically feasible. This review after giving a thumbnail picture of extremophilies’ known features is centered on the triad of biodiversity richness, environmental availabilities/managements of locations suitable for respective biotechnological bioprocesses and their non-aseptic designing for a developing country like Pakistan.

Enhanced production of alkaline protease by a mutant of Bacillus licheniformis N-2 for dehairing

The purpose of the present investigations was to improve the yield of alkaline protease for leather dehairing by subjecting the indigenous proteolytic strain Bacillus licheniformis N-2 to various mutagenic treatments viz. UV irradiations, NTG (N-methyl-N-nitro-N-nitrosoguinidine) and MMS (methyl methane sulfonate). After screening on skim milk agar plates, a total of nine positive mutants were selected for shake flask experiments. Among these, the best proteolytic mutant designated as UV-9 showed 1.4 fold higher alkaline protease activity in preoptimized growth medium than the parent strain. The fermentation profile and kinetic parameters such u(h-1), Yp/s, Yp/x, Yx/s, qs, Qs, qp and Qp also indicated the superiority of the selected mutant UV-9 for alkaline protease production over the parent strain and rest of the mutants. The dehairing capability of mutant UV-9 alkaline protease was analyzed by soaking goat skin pieces for different time intervals (3-15 h) at 40 o C. A complete dehairing without degradation of collagen was achieved after 12 h, indicating its commercial exploitation in leather industry.

Exploited application of sulfate-reducing bacteria for concomitant treatment of metallic and non-metallic wastes: a mini review

A variety of multidimensional anthropogenic activities, especially of industrial level, are contaminating our aquatic and terrestrial environments with a variety of metallic and non-metallic pollutants. The metallic and non-metallic pollutants addressed specifically in this review are heavy metals and various compound forms of sulfates, respectively. Direct and indirect deleterious effects of the both types of pollutants to all forms of life are well-known. The treatment of such pollutants is therefore much necessary before their final discharge into the environment. This review summarizes the productive utility of sulfate-reducing bacteria (SRB) for economical and concomitant treatment of the above mentioned wastes. Utilization of agro-industrial wastes and some environmental contaminants including hydrocarbons, as economical growth substrates for SRB, is also suggested and proved efficient in this review. Mechanistically, SRB will utilize sulfates as their terminal electron acceptors during respiration while utilizing agro-industrial and/or hydrocarbon wastes as electron donors/carbon sources and generate H2S. The biogenic H2S will then react vigorously with dissolved metals present in the wastewaters thus forming metal sulfide. The metal sulfide being water insoluble and heavier than water will settle down in the water as precipitates. In this way, three types of pollutants i.e., metals, sulfates and agro-industrial and/or hydrocarbon wastes will be treated simultaneously.

Evodiamine sensitizes U87 glioblastoma cells to TRAIL via the death receptor pathway.

The tumor necrosis factor-α-related apoptosis-inducing ligand (TRAIL) has been shown to selectively induce death in cancer cells without affecting healthy cells. Most glioma cells are resistant to TRAIL-induced apoptosis. Resistance to TRAIL limits its potential use as a drug for therapy of glioma. The present study was conducted to identify bioactive compounds that have the potential to sensitize U87 glioblastoma cells to TRAIL. Evodiamine, a major bioactive compound of the Chinese herb Evodiae fructus, has been reported to sensitize U87 glioblastoma cells to TRAIL. TRAIL and evodiamine, in combination or alone, were used to treat U87 glioblastoma cells. We show that evodiamine treatment inhibited cell growth in a dose-dependent manner; however, TRAIL alone failed to exert any cytotoxic effect. Combining TRAIL with evodiamine significantly increased the apoptotic rate of U87 glioblastoma cells, as compared to evodiamine treatment alone. Further investigation of the mechanism underlying these effects revealed that the evodiamine + TRAIL effect is associated with the increased expression of death receptor (DR)4, DR5, caspase-8 and cleaved caspase-3. The present study demonstrated, for the first time to the best of our knowledge, that evodiamine can sensitize U87 glioblastoma cells to TRAIL via the death receptor pathway. Thus, our results suggest that combined treatment with evodiamine and TRAIL may represent a novel chemotherapeutic strategy for the therapy of glioma.

Efficacy of Locally Isolated Lactic Acid Bacteria Against Antibiotic-Resistant Uropathogens

Background: Antibiotic resistance represents a serious global health threat to public health, so infections such as pneumonia and urinary tract infection (UTI) are becoming harder to treat. Therefore, it is necessary to develop an action plan to restrain the problem of antibiotic resistance. One approach in UTI control could be the use of lactobacilli because these indigenous inhabitants in human intestine have been found to play an important role in protecting the host from various infections. Objectives: We sought to check the efficacy of locally isolated Lactobacillus species to eradicate antibiotic-resistant pathogenic bacteria causing UTI. Materials and Methods: Lactic acid bacteria isolated from spoiled fruits and vegetables and grown in MRS medium were screened against multi-drug-resistant Candida albicans, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, and Enterococcus fecalis. Results: Fifty-four lactic acid bacteria were isolated from spoiled fruits and vegetables, of which 11 Gram-positive and catalase-negative Lactobacillus isolates were identified by carbohydrate assimilation profiles as Lactobacillus acidophilus, L. paracasei, L. delbrueckii, L. casei, L. helveticus, L. brevis, L. salivarius, L. fermentum, L. rhamnosus, L. animalis, and L. plantarum. The latter organism had the highest abundance of all the samples, so its isolates were also verified through 16S rRNA gene sequencing. The isolated Lactobacilli were screened against multi-drug-resistant uropathogens, viz. C. albicans, P. aeruginosa, K. pneumoniae, E. fecalis, and E. coli. The growth inhibition zone (GIZ) was over 10 mm against all the uropathogenic test organisms, where L. fermentum and L. plantarum strains demonstrated remarkable inhibitory activities against E. coli and E. faecalis, with a GIZ up to 28 mm. The susceptibility test to 16 antibiotics showed multidrug resistance (3 to 5 antibiotics) among all the tested uropathogens. Conclusions: The obtained results revealed that all the Lactobacillus isolates displayed antimicrobial activity against 6 out of 7 antibiotic-resistant uropathogens, indicating that these bacteria could represent a good ecological plan for the control and prevention of UTI.

Lignocellulose for ethanol production: a review of issues relating to bagasse as a source material.

Fossil fuels reservoirs have been declared to serve mankind’s needs more for a very limited time period. This notion has already initiated scientific search for alternatives. Amongst renewable resources for emerging biotechnological strategies to produce high energy-less volume fuels, cellulose is the most abundantly synthesized but stable carbohydrate of the biosphere. Cellulose has earlier been taken into account for chemical/biological saccahrification and subsequent biological conversion of the monomeric sugars to ethanol. The stable nature of the substrate and some of the monomeric products’ fermentation difficulties have been the major hardles. But because of its ubiquitous nature and being the most abundantly available renewable resource, research on the utilization of cellulose for obtaining the biofuel has continued and has been representing by diverse fields. Following the recognition of different bacteria and yeasts and various kinetics of the process involved in its saccahrification, the substrate is increasingly being worked out by different laboratories. Biotechnological endeavors are in fact reshaping the economics of different countries. Production of high grade sweeteners with low caloric values by the contemporary biotechnological processes is likely to influence the conventional sucrose production negatively. The raw material for sugar industry would be available for other products such as ethanol. At present, cellulosic waste of such industries for example, sugarcane bagasse, may be targeted for sacharification and ethanologenesis. Various aspects regarding the nature of the cellulosic substrate and its potential for obtaining the biofuel are covered in this review.

Microbial production and industrial applications of keratinases: an overview

Massive production of keratinaceous byproducts in the form of agricultural and industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This review summarizes the potential utility of some bacterial and fungal species for the production of keratinase using a variety of keratinaceous wastes as growth substrates. The application of microbial keratinases in waste management; animal feed, detergent, and fertilizer manufacturing; and leather, cosmetic, and pharmaceutical industries is also abridged in this review.

Molecular characterisation of Bacillus chitinase for bioconversion of chitin waste

In this work chitin was extracted chemically from shrimp shells. Seventeen Bacillus isolates were screened for chitinolytic activity. The chitinolytic strains of Bt. were screened at different temperatures and pHs for their hydrolytic potentials. By using a pair of specific primers, endochitinase gene was amplified from SBS Bt-5 strain through PCR, and then cloned into pTZ57 TA cloning vector and transferred in Escherichia coli DH5α strain. The sequenced gene (GenBank Accession No: HE995800) consists of 2031 nucleotides capable of encoding 676 residues. The protein consisted of three functional domains with a calculated molecular mass of 74.53 kDa and a pI value of 5.83. The amino acid sequence of chi gene showed 99% similarity to the genes of Bt MR11 endochitinase, Bt serovar kurstaki chitinase (kchi), Bt strain MR21 endochitinase and Bacillus cereus B4264.