Recent Patents on Biotechnology (v.9, #2)

Recent Patents in Oncolytic Virotherapy by Tauqeer Ahmad, Srividhya Venkataraman, Mounir AbouHaidar, Kathleen L. Hefferon (79-85).
Recent innovative and advanced developments in the diagnosis and treatment of human diseases as well as enhanced in-depth understanding of virus molecular biology have opened novel avenues with respect to the patent landscape. Included are viruses utilized in the development of anticancer agents, agents that are employed against the spread of infectious viral diseases, RNA silencing agents and virus-derived expression vectors that can be used for over-expression of therapeutic proteins or as gene therapy vehicles. The current review describes several recent patents pertaining to virus sequences and their medical and biotechnological applications.

Ovarian cancer continues to present a significant health challenge with little progress being made over the past two decades in reducing the incidence and mortality. More recently, novel therapeutics have emerged as a potential way of improving outcomes for women with advanced ovarian cancer who harbor mutations in genes involved in homologous recombination (HR), most notably BRCA1 and BRCA2. In the United States, Olaparib, a PARP inhibitor, has been recently approved for ovarian cancer patients treated with three or more lines of prior chemotherapy who harbor germline mutations in BRCA1 or BRCA2. As a caveat to Olaparib's FDA approval, BRACAnalysis CDx® was approved as a companion diagnostic test for women with ovarian cancer to determine their BRCA1/2 mutation status and eligibility for treatment. This review article will provide essential background information on hereditary breast and ovarian cancer (HBOC), describe the therapeutic mechanism of PARP inhibitors, and will chronicle the current and emerging homologous recombination deficiency (HRD) assays and their associated patents.

Background: Inter - conversion processes of labile molecules obey similar rules to those of reversible chemical reactions. Solving the corresponding linear differential systems is used along this work, as well as in the preceding version.
Objective: The main purpose of the present mini revue paper is to recall, improve and correct some mathematical methods in determining the optimal values at equilibrium, and remarkable particular rate constants. This part was not proved correctly in my previous work.
Method: In my previous work, the proof of the equality of the concentrations of the main species at equilibrium was not correct. In the current manuscript, we use increasing velocity in order to obtain this first important result. To this aim, one applies Schwarz inequality and the case when equality occurs. In order to determine significant rate constants, we characterize these special values in terms of the norm of the linear operator defined by the matrix of the differential system. In my previous work, the normal probability density function was used. The latter method was not realistic.
Results: Increasing the velocity, one obtains equal optimal values of the concentrations at equilibrium. This method represents a patent in the field. Secondly, characterization of remarkable rate constants (which are also equal) is deduced. The optimal solutions are written explicitly.
Conclusion: Under suitable conditions, the values at equilibrium and the rate constants are equal. The common value at equilibrium equals the common value of the rate constants.

A Case Study of Petroleum Degradation in Different Soil Textural Classes by Reginald B. Kogbara, Josiah M. Ayotamuno, Daniel C. Worlu, Isoteim Fubara-Manuel (108-115).
Background: Patents have been granted for a number of techniques for petroleum biodegradation including use of micro-organisms for degradation of hydrocarbon-based substances and for hydrocarbon degradation in oil reservoirs, but there is a dearth of information on hydrocarbon degradation in different soil textures.
Objective: Hence, this work investigated the effects of different soil textures on degradation of petroleum hydrocarbons during a six-week period.
Methods: Five soil textural classes commonly found in Port Harcourt metropolis, Nigeria, namely sand, loamy sand, sandy loam, silty clay and clay, were employed. The soils were contaminated with the same amount of crude oil and then remediated by biostimulation. Selected soil properties were monitored over time.
Results: Bacterial numbers declined significantly in the fine soil textures after petroleum contamination, but were either unaffected or increased significantly in the coarser soil textures. Hydrocarbon losses ranged from 42% - 99%; the sandy loam had the highest, while the clay soil had the least total hydrocarbon content (THC) reduction. The total heterotrophic bacterial (THB) counts generally corroborated the THC results. Fold increase in bacterial numbers due to remediation treatment decreased with increasing clay content.
Conclusion: The results suggest that higher sand than clay content of soil favours faster hydrocarbon degradation. Hydrocarbon degradation efficiency increased with silt content among soil groupings such as fine and coarse soils but not necessarily with increasing silt content of soil. Thus, there seems to be cut-off sand and clay contents in soil at which the effect of the silt content becomes significant.

Background: Hexavalent chromium (Cr6+) contamination is one of the mejor problems of environmental protection for its carcinogenic effect on human health. Remediation of Cr6+ contaminated environment thus becomes highest priority.
Methods: A Gram positive tiny rod shaped chromate (Cr6+) reducing bacterium strain A1 was isolated from uranium ore collected from Jaduguda, East Singhbhum, Jharkhand. The strain was identified and characterized in terms of its potential to reduce more toxic Cr6+ to its less toxic form for its application in bioremediation of Cr6+ contaminated environments.
Results: 16S rRNA gene based phylogentic analysis identified the strain as Microbacterium oleivorans. Along with Cr6+, the strain showed resistance to other heavy metals including Ag, Cu, Co, Hg, Ni and Zn also. Complete reduction of Cr6+ (750 µ;M) was achieved within 84 h with optimum reduction at pH 9 and 30°C. Effect of different parameters including cell mass concentration, pH, induction with Cr6+, SO4 = ion, heavy metals, etc. on Cr6+ reduction were studied thoroughly at resting cell condition to study its potential towards Cr6+ bioremediation. Chromate reductase gene (chrA) was detected within this strain.
Conclusion: Along with presence of appropriate genetic determinant, efficient Cr6+ reduction ability of the strain indicated its potential for developing redox based Cr6+ remediation system for varied concentrations of Cr6+ under a wide range of environmental conditions. Patent data have suggested the efficient application of Cr6+ reducing bacteria in cleaning up of Cr6+ contaminated environments..

Glomus fasciculatum Fungi as a Bio-convertor and Bio-activator of Inorganic and Organic P in Dual Symbiosis by Rafia Azmat, Neelofer Hamid, Sumeira Moin, Ailyan Saleem (130-138).
Background: Dual symbiosis played an effective role in drought condition and temperature. Furthermore, performed services in absorption of water and solubilization of chief nutrients specially phosphorus for growth of plants. Phosphorous is essential for plant growth in any climatic condition because of central constituent of ATP providing chemical energy for all metabolic reactions of plants.
Method: The goal of this work was to monitor the growth of plant under three climatic conditions in comparison to control plant under Glomus fasciculatum inoculation related with adequate supply of phosphorous.
Results: Results demonstrated that Glomus fasciculatum (VAM) activates the solubilization of P into the anionic form H2PO4- which is highly consumable form by the plants. Minerals including P in soil most regularly solubilized for fixing in plants and continuously changed to highly soluble forms by reaction with inorganic or organic constituents of the soil which are activated in the presence of fungi for continuous availability. Experimental facts and nonstop growth of plants recommended that VAM fungi act as a bio-convertor and bio-activator of soil nutrients, especially of P and their hyphal interaction absorbs soil nutrients and activates insoluble P to soluble one for plant development.
Conclusion: Continuous growth of 18 months old Conocarpus erectus L plant in dual symbiosis supports the proposed idea that phosphorus cycle exists during VAM inoculations, where soil reaction altered in presence of spores that help to solubilize the P which strengthens the plant, activates photo-biological activity and demonstrates the new function of VAM as a recycler for continues growth.

Exploration of Recombinant Streptokinase Degradation Products Under Various pH Reduction Conditions in Downstream Processing by Reza Jalalirad, Alireza Kavianpour, Ahmad Beiroti, Mina Sepahi, Parinaz Tavakoli Zaniani, Firooz Arsalani (139-144).
Background: Methods of producing streptokinase, which can be used in the treatment of myocardial infarction, by hemolytic streptococci and recombinant E. coli have been described in patents since 1955. Degradation products in active pharmaceutical ingredients (APIs) and finished pharmaceutical products are considered as impurities and it is required that these degradation impurities are minimized or rather avoided throughout manufacturing process.
Objective: The aim of this study was to explore the occurrence of rSK degradation during acidification step in downstream processing.
Methods: The polyclonal antibody was produced by immunization of New Zealand white (NZW) rabbit with pure rSK (purity>98%). The solubilized inclusion bodies with various pH values (4.2, 5.0 and 6.0) were analyzed by Western blotting using rSK polyclonal antibody.
Results: Western blot analysis demonstrated the generation of rSK degradation products (with the molecular weight of about 27, 20 and 17 kDa) when the pH value of the solubilized inclusion bodies was reduced to 5.0 and 4.2, while no degradation of rSK observed at pH 6.0.
Conclusion: This study demonstrates that the level of pH reduction in the solubilized inclusion bodies during downstream processing plays an important role in generating rSK degradation products, and substantial post-solubilization degradation of rSK occurs at pH lower than 6.0. Development of these degradation impurities, which cannot be eliminated by subsequent chromatographic purifications, can be exclusively avoided during acidification procedure by appropriate pH adjustment approach in downstream processing.

Background: Pain whether acute or chronic, peripheral or central is believed to originate from inflammation and inflammatory response. The potential toxicity of antinociceptive and antiinflammatory agents such as non steroidal anti-inflammatory drugs (NSAIDs) necessitates sourcing for less toxic alternative drugs. Many natural substances have been used for the management of pain and inflammation with success traditionally.
Methods: The present study sought to investigate phytochemistry, antinociceptive and anti-inflammatory potentials of the methanolic leaves extract of Lannea schimperi. Preliminary phytochemical test was conducted on the extract. Antinociceptive and anti-inflammatory potentials of the extract at doses of 12 and 24 mg/kg were evaluated using acetic acid induced writhing model in mice and egg albumin induced acute inflammation model in rats' respectively. Aspirin at dose of 80 mg/kg was used as the standard drug given to the positive control group, while the drug vehicle was given to the negative control group.
Result: Phytochemical analysis revealed the presence of carbohydrate, glycoside, reducing sugars, triterpene, steroids, cardiac glycosides, flavonoids, phenolic glycosides, alkaloids, tannins, condensed tannins and saponins. The data obtained showed that at both doses the extract significantly (P< 0.05) decreased the acetic acid induced writhing reflex in mice when compared to the negative control group. The result also demonstrated significant (P< 0.05) anti-inflammatory activity of the extract at both doses. However, there was no significant difference (P> 0.05) between the extract treated groups and the group treated with the standard drug (positive control).
Conclusion: The patent data revealed the antinociceptive and anti-inflammatory potentials of the methanolic leaves extract of Lannea schimperi which could be beneficial in alleviating painful inflammatory conditions.

Patent Selections: (153-156).