Optimization and Scale-Up of Thiarubrine a Production From Hairy Root Cultures of Ambrosia Artemisiifolia.

Thiarubrine A (TA), a plant secondary metabolite synthesized by members of the family Asteraceae possesses antifungal and antiviral properties and is a potential pharmaceutical. The objective of the work was the optimization and scale-up of a bioprocess for the production of TA using hairy root cultures of Ambrosia artemisiifolia. At the inception of the work an established hairy root culture of the model system was used as inoculum in reactor experiments for the development of a protocol for the quantification of TA and in subsequent shake flask experiments to determine optimum culture conditions. TA accumulation closely paralleled growth and then decreased indicating TA degradation. Growth and TA accumulation decreased on increasing the temperature from 25\sp\circC to 30\sp\circC. No TA accumulation was observed in cultures exposed to light and this inhibitory effect was reversible. Elicitation, biotic and abiotic was studied as TA yield enhancement strategy using a statistical method. Biotic elicitation was done with Protomyces gravidus, a pathogen and Botrytis cinereae, a non pathogen of the model system and abiotic elicitation using vanadyl sulfate solution. In all the three cases stimulation of TA accumulation was observed. To obtain highly productive clones the root cultures were disorganized by the addition of phytohormones which facilitated somaclonal variation to provide a pool of variability. Hairy root clones were regenerated from the disorganized phase and a clone with thrice the productivity of the parent clone was achieved. Using the optimal culture conditions obtained, the cultures were scaled-up to a nine liter air sparged reactor with the inoculum concentration of root tips as the basis. Volumetric productivity of TA was of the same order of magnitude in the shake flask and the reactor. The results show that the inoculum concentration of root tips is a key parameter describing root growth and secondary metabolite accumulation at low biomass densities. Experiments performed in a shake flask indicated that manipulation of the inoculum concentration of root tips provides a way of manipulating the kinetics of root growth and TA accumulation early in the run

Carbon Nanotubes Integrated Hydroxyapatite Nano-Composite for Orthopaedic and Tissue Engineering Applications

The reassessment of the literature stipulates that an increasing amount of research in exploring the Hydroxyapatite Carbon Nanotubes (HA-CNT) system for orthopedic application. Chemical precipitation, CNT functionalization, and spray drying are the routinely used methods for CNT dispersal in HA matrix for the application such as bone tissue engineering, nanostructured scaffolds, dental regeneration, myocardial regeneration, and skin regeneration. Although mechanical strength and biocompatibility is a substantial concern for the fabrication of structures. Developing composite and bioceramic scaffolding with different natural and synthetic biomaterials are the futuristic approach in the biomedical engineering field. The problems such as biocompatibility, biodegradability, and mechanical resistance can be solved by combining natural, and artificial biomaterials. The natural biomaterials, such as collagen, cellulose, chitosan, have a close resemblance to the natural extracellular matrix (ECM). These materials are biocompatible, biodegradable. The artificial biomaterials, such as Poly Vinyl Pyrrolidone (PVP), Poly Capro Lactone (PCL), Poly Ethylene Glycol (PEG), and Poly Lactic Acid (PLA) are also the material of choice for the fabrication of the composite materials. Additional effort is necessary to fabricate biocompatible composite scaffolding for tissue engineering. Moreover, vascularization, differentiation, cellular proliferation, and cells to scaffold interaction are the foremost challenges in the area of tissue engineering that remains to overcome

Assessing extent of single stranded DNA damage in oral mucosal cells of patients with oral squamous cell carcinoma and its correlation with TNM staging

Context: This study was carried out on the assumption that oral mucosal cells might show DNA damage in oral squamous cell carcinoma (OSCC). Aims: To evaluate the extent of DNA damage in oral smears of patients with OSCC and determine correlation if any of the extent of DNA damage to TNM staging of oral cancer. Settings and design: A randomized controlled study at a regional cancer centre was designed for this project. Smears were taken from lesion proper of 30 patients with OSCC and from the buccal mucosa of 30 normal healthy volunteers. Materials and methods: Collected cells were centrifuged and single-cell gel electrophoresis (SCGE) assay was performed. DNA damage was visualized under a fluorescent microscope. Statistical analysis used : Mean DNA damage levels of both the groups were measured and statistically analyzed with students′ test. The extent of DNA damage was correlated with the TNM stages by employing the one way ANOVA ′F′ technique. Results: High statistical significance (P < 0.0001) was found in DNA damage levels between control and study groups. A stepwise increase in DNA damage levels with high statistical significance (P < 0.005) was also found between all the TNM stages. Conclusions: Statistically significant increased DNA damage levels in OSCC patients and their correlation to clinical staging suggest that comet assay may be used effectively to assess the prognosis of OSCC

Characterization of Con C, a lectin from Canavalia cathartica Thouars seeds

Con C, a lectin from Canavalia cathartica, a wild species of Canavalia was isolated and partially characterized. Mannose-agarose resin was used as an affinity matrix for purification. The lectin showed strong agglutination activity (2.34 AU/ng protein) towards rabbit erythrocytes. Con C agglutinated A, B and O groups of human blood with a preference for A and 0 groups. The native molecular weight of lectin was 62 kDa and the subunit molecular weight was 31 kDa. No carbohydrate moiety was found to be associated with the lectin. Con C showed a broad pH optima of pH 4-8. Total inactivation of lectin activity occurred at 70 degrees C when heated for 10 min. The lectin was found to be mitogenic for mouse-spleen cells (total). N-terminal analysis revealed 94% homology with C ensiformis lectin (Con A). C. cathartica is a legume with high nutritional values and less antinutritional factors and the potential of the same is yet to be exploited. (C) 2006 Elsevier Ltd. All rights reserved

Effect of Vanillin on lipid profile in a model of hyperlipidemia, a preliminary study

288-291To evaluate the effect of vanillin on the lipid profile of high fat diet induced hyperlipidemia in rats, the hyperlipidemia was induced by feeding cholesterol-rich high fat diet for 45 days in wistar rats of either sex. The reduction in the triglycerides and VLDL-C was significant at 200 &amp; 400 mg/kg dose of vanillin compared to atorvastatin group. Reduction in total cholesterol was significant at 200 and 400 mg/kg doses compared to hyperlipidemic control. The results demonstrate that vanillin at a dose of 200 and 400 mg/kg body weight lowers the serum triglyceride, VLDL-C and total cholesterol level significantly in high fat diet induced hyperlipidemic rats. However there was no significant effect on the lipid profile at 100 mg/kg dose. There were no statistically significant changes in the HDL-C and LDL-C levels at any of the given doses

Composition and functional properties of raw and electron beam-irradiated Mucuna pruriens seeds

The proximate, nutritional and functional properties of raw and electron beam-irradiated (0-30 kGy) Mucuna seeds were investigated. Irradiation increased crude protein and crude carbohydrates significantly than crude lipid, crude fibre and energy. Raw seeds were rich in minerals and were not affected by irradiation except for magnesium and phosphorus. Amino acids of raw seeds were comparable to soyabean and FAO/WHO reference pattern and except for 30 kGy, no significant changes were seen in amino acid profile. Raw seeds were rich in unsaturated fatty acids and some of them decreased on irradiation, while linoleic acid steadily elevated (0-14.35 mg g(-1) lipid). In vitro protein digestibility was dose dependent and significantly increased up to 15 kGy. Water and oil absorption capacities and foaming capacity significantly increased on irradiation, while protein solubility decreased (15 and 30 kGy) with an improvement of gelation property of seed flour. The cooking time of seeds significantly reduced on irradiation

Indigenous rhizobia associated with native shrubby legumes in Taiwan

Root-nodulating bacteria were isolated and characterized from seven native shrubby legumes growing in Taiwan. Phenotypic characteristics measured included growth rates in various media, colony morphology, and tolerances to extremes of temperature, salt and pH. The isolates were very diverse phenotypically. Among the 83 isolates that were screened, the majority were fast-growing rhizobia. Twenty eight strains tolerated high concentration of salt (4.5% NaCl) and grew well between temperatures of 37 and 45 degrees C. The majority of the strains also tolerated extreme pH in their medium from 3.5 to 12. All strains formed nitrogen fixing nodules, and the highest activity was detected in the legume Hedysarum crinita L. PCR restriction fragment length polymorphism (PCR-RFLP) and sequencing of the small subunit ribosomal RNAs revealed that the majority of the isolates belonged to the genera Rhizobium, Bradyrhizobium and Agrobacterium. Only a single strain represented the genus Sinorhizobium. In addition, a strain related to Burkholderia from the beta-class of the Proteobacteria (CC-CC-5) was found within nodules of the legume Catenaria caudatum. The study contributes to the understanding of symbiotic nitrogen fixation in selected wild legumes that are native to Taiwan and provides insights into the distribution of nodulating and nitrogen-fixing bacteria from other distinct lineages. (c) 2005 Elsevier GmbH. All rights reserved

Uncultured Actinobacteria and Reverse Engineering and Artificial Intelligence Role in Future

12 páginas.- 2 figuras.- referenciasActinobacteria have been known to the world since the mid-eighteenth century, when Ferdinand Cohn portrayed them as ropy critters. In the nineteenth century, these unidentified morphons became famous for their broad array of applications and for causing maladies. Over the last five decades, actinomycetes have been classified and their names have been validly published as representative taxa. Actinobacteria have preserved their overall genetic integrity and kept newer genes that code for bioactive substances, as evidenced by their capacity to produce a myriad of bioactive compounds. As of today, we have hardly cultured 1% of existing microorganisms including Actinobacteria. Some Streptomyces strains were exploited for enhanced production of new bioactive compounds. In times of the bioinformatics era, insufficient and developing laboratory methods for enumeration of potential species are acting as a barrier to obtaining the maximum product in downstream processing. This issue may be addressed by using the reserve-engineering approach. Published reports indicate that new or high production can be achieved through either strain improvement or the remaining microbial dark matter consists of growing hidden gems for an awaited potential application. Reverse engineering of microorganisms, on the other hand, improves the efficacy of a specific target gene to obtain high product value, which can be seen in the expression of the tylosin biosynthetic gene cluster. Hidden gems among the phylum Actinobacteria need to be discovered and reverse engineered for antibiotics and novel drugs. Therefore, advancement in research machine learning and artificial intelligence seems to be incredible tools. Hence, uncultivated Actinobacteria can be used for the benefit of mankind by combining reverse engineering with machine learning (ML) and artificial intelligence (AI).Peer reviewe

Morphological and micro-tomographic study on evolution of struvite in synthetic urine infected with bacteria and investigation of its pathological biomineralization.

Pathological biomineralization in the urinary system leads to urolithiasis. Formation of kidney stones involves a series of events during which they undergo morphological and mineralogical changes. We investigated the mineralization of biogenic struvite (in vitro) and examined the transformation of distinct interior and exterior structure of struvite. In vitro crystallization of struvite was performed in the presence of two bacteria that were originally isolated from the kidney stone patients. Morphological evaluation was carried out using SR-μCT as well as FESEM, XRD and FT-IR. Characteristic internal 3-D morphology and porosity of the stones were studied. For comparison, patient derived struvite stones were used. From the results obtained, we report that the presence of bacteria enhances the crystallization process of struvite in vitro. A series of time-resolved experiments revealed that struvite crystals experienced a significant morphologic evolution from pin pointed structure to X-shaped and tabular morphologies. These X-shaped and unusual tabular habits of struvite resembled biogenic morphologies of struvite. SR-μCT showed similarities between the patient derived and the in vitro derived struvite crystals. In conclusion, these experiments revealed that the bacteria play a major role in the specific morphogenesis of struvite and can able to control the nucleation, modulate crystalline phases, and shape of the growing crystal