Development of tissue culture and virus-induced gene silencing for Calendula officinalis

Calendula officinalis is grown widely as an ornamental plant across Europe. It belongs to the large. Asteraceae family. In this study, the aim was to explore the possibilities to use Calendula officinalis as a new model organism for flower development and secondary mechanism studies in Asteraceae. Tissue culture of Calendula officinalis was established using nine different cultivars. Murashige & Skoog (MS) medium with four different combinations of plant growth regulators were tested. Of all these combinations, the medium containing 1mg/l BAP, 0.1 mg/l IAA, and 1mg/l Zeatin achieved highest frequency of adventitious shoot regeneration from hypocotyl and cotyledon explants. Virus-induced gene silencing is a recent developed genetic tool for charactering the gene functions in plants, and extends the range of host plants that are not accessible for Agrobacterium transformation. Here, tobacco rattle virus (TRV)-based VIGS technique was tested in calendula (cv. Single Orange). We used TRV carrying Gerbera hybrid phytoene desaturase (PDS) gene fragment to induce PDS silencing in calendula. Vacuum infiltration and syringe infiltration methods both resulted in photo-bleaching phenotypes in leaves, bracts and petals. Loss-of-function phenotypes occurred on calendula 13 days post-infiltration. In conclusion, the data indicates that calendula explants can be regenerated through tissue culture which is a prerequisite for development of stable transformation methods. However, further optimization is still needed to improve the frequency. In addition, VIGS was applied to silence PDS marker gene expression indicating that this method has potential for gene functional studies in future.

Banking psychographics: an Indian empirical study

India has been witnessing an economic boom which fuelling a huge growth in the financial sector especially the banks. The spending power and consumerism has been increasing along with the growth in GDP. The numbers of banks are around 3000 (data according to Reserve Bank of India). With a population base of close to 1.1 billion and a diverse culture that has been dictating the mindset and lifestyle of the population, it has been a challenge for the banks to understand the customer better and hence a the need of the hour is a proper psychographic study of retail banking customers.

Monoclonal Antibodies against Hepatitis C Genotype 3a Virus Like Particle Inhibit Virus Entry in Cell Culture System

The envelope protein (E1-E2) of Hepatitis C virus (HCV) is a major component of the viral structure. The glycosylated envelope protein is considered to be important for initiation of infection by binding to cellular receptor(s) and also known as one of the major antigenic targets to host immune response. The present study was aimed at identifying mouse monoclonal antibodies which inhibit binding of virus like particles of HCV to target cells. The first step in this direction was to generate recombinant HCV-like particles (HCV-LPs) specific for genotypes 3a of HCV (prevalent in India) using the genes encoding core, E1 and E2 envelop proteins in a baculovirus expression system. The purified HCV-LPs were characterized by ELISA and electron microscopy and were used to generate monoclonal antibodies (mAbs) in mice. Two monoclonal antibodies (E8G9 and H1H10) specific for the E2 region of envelope protein of HCV genotype 3a, were found to reduce the virus binding to Huh7 cells. However, the mAbs generated against HCV genotype 1b (D2H3, G2C7, E1B11) were not so effective. More importantly, mAb E8G9 showed significant inhibition of the virus entry in HCV JFH1 cell culture system. Finally, the epitopic regions on E2 protein which bind to the mAbs have also been identified. Results suggest a new therapeutic strategy and provide the proof of concept that mAb against HCV-LP could be effective in preventing virus entry into liver cells to block HCV replication.

A Study of Early Afterdepolarizations in a Model for Human Ventricular Tissue

Sudden cardiac death is often caused by cardiac arrhythmias. Recently, special attention has been given to a certain arrhythmogenic condition, the long-QT syndrome, which occurs as a result of genetic mutations or drug toxicity. The underlying mechanisms of arrhythmias, caused by the long-QT syndrome, are not fully understood. However, arrhythmias are often connected to special excitations of cardiac cells, called early afterdepolarizations (EADs), which are depolarizations during the repolarizing phase of the action potential. So far, EADs have been studied mainly in isolated cardiac cells. However, the question on how EADs at the single-cell level can result in fibrillation at the tissue level, especially in human cell models, has not been widely studied yet. In this paper, we study wave patterns that result from single-cell EAD dynamics in a mathematical model for human ventricular cardiac tissue. We induce EADs by modeling experimental conditions which have been shown to evoke EADs at a single-cell level: by an increase of L-type Ca currents and a decrease of the delayed rectifier potassium currents. We show that, at the tissue level and depending on these parameters, three types of abnormal wave patterns emerge. We classify them into two types of spiral fibrillation and one type of oscillatory dynamics. Moreover, we find that the emergent wave patterns can be driven by calcium or sodium currents and we find phase waves in the oscillatory excitation regime. From our simulations we predict that arrhythmias caused by EADs can occur during normal wave propagation and do not require tissue heterogeneities. Experimental verification of our results is possible for experiments at the cell-culture level, where EADs can be induced by an increase of the L-type calcium conductance and by the application of I-Kr blockers, and the properties of the emergent patterns can be studied by optical mapping of the voltage and calcium.

Cross-Linked, Biodegradable, Cytocompatible Salicylic Acid Based Polyesters for Localized, Sustained Delivery of Salicylic Acid: An In Vitro Study

In order to suppress chronic inflammation while supporting cell proliferation, there has been a continuous surge toward development of polymers with the intention of delivering anti-inflammatory molecules in a sustained manner. In the above backdrop, we report the synthesis of a novel, stable, cross-linked polyester with salicylic acid (SA) incorporated in the polymeric backbone and propose a simple synthesis route by melt condensation. The as-synthesized polymer was hydrophobic with a glass transition temperature of 1 degrees C, which increases to 17 degrees C upon curing. The combination of NMR and FT-IR spectral techniques established the ester linkages in the as-synthesized SA-based polyester. The pH-dependent degradation rate and the rate of release of salicylic acid from the as-synthesized SA-based polymer were studied at physiological conditions in vitro. The polyester underwent surface erosion and exhibited linear degradation kinetics in which a change in degradation rate is observed after 4-10 days and 24% mass loss was recorded after 4 months at 37 degrees C and pH 7.4. The delivery of salicylic acid also showed a similar change in slopes, with a sustained release rate of 3.5% in 4 months. The cytocompatibility studies of these polyesters were carried out with C2C12 murine myoblast cells using techniques like MTT assay and flow cytometry. Our results strongly suggest that SA-based polyester supports cell proliferation for 3 days in culture and do not cause cell death (<7%), as quantified by propidium iodide (PI) stained cells. Hence, these polyesters can be used as implant materials for localized, sustained delivery of salicylic acid and have applications in adjuvant cancer therapy, chronic wound healing, and as an alternative to commercially available polymers like poly(lactic acid) and poly(glycolic acid) or their copolymers.

Development of an ex vivo organ culture model using human gastro-intestinal tissue and Campylobacter jejuni.

Campylobacter jejuni is an important food-borne pathogen. However, relatively little is understood regarding its pathogenesis, and research is hampered by the lack of a suitable model. Recently, a number of groups have developed assays to study the pathogenic mechanisms of C. jejuni using cell culture models. Here, we report the development of an ex vivo organ culture model, allowing for the maintenance of intestinal mucosal tissue, to permit more complex host-bacterium interactions to be studied. Ex vivo organ culture highlights the propensity for C. jejuni to adhere to mucosal tissue via the flagellum, either as discrete colonies or as multicellular units.

Detection of Atomic Scale Changes in the Free Volume Void Size of Three-Dimensional Colorectal Cancer Cell Culture Using Positron Annihilation Lifetime Spectroscopy

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A Trabecular Bone Explant Model of Osteocyte–Osteoblast Co-Culture for Bone Mechanobiology

The osteocyte network is recognized as the major mechanical sensor in the bone remodeling process, and osteocyte-osteoblast communication acts as an important mediator in the coordination of bone formation and turnover. In this study, we developed a novel 3D trabecular bone explant co-culture model that allows live osteocytes situated in their native extracellular matrix environment to be interconnected with seeded osteoblasts on the bone surface. Using a low-level medium perfusion system, the viability of in situ osteocytes in bone explants was maintained for up to 4 weeks, and functional gap junction intercellular communication (GJIC) was successfully established between osteocytes and seeded primary osteoblasts. Using this novel co-culture model, the effects of dynamic deformational loading, GJIC, and prostaglandin E-2 (PGE(2)) release on functional bone adaptation were further investigated. The results showed that dynamical deformational loading can significantly increase the PGE(2) release by bone cells, bone formation, and the apparent elastic modulus of bone explants. However, the inhibition of gap junctions or the PGE(2) pathway dramatically attenuated the effects of mechanical loading. This 3D trabecular bone explant co-culture model has great potential to fill in the critical gap in knowledge regarding the role of osteocytes as a mechano-sensor and how osteocytes transmit signals to regulate osteoblasts function and skeletal integrity as reflected in its mechanical properties.

Feasibility of adopting integrated rice-cum-fish culture system to enhance the development of conventional aquaculture participation in Niger State

In a survey conducted to find out the status of integrated rice-cum-fish culture in Niger State, Nigeria, 0.37 ha of Fadama wetlands was utilized for rice-cum-fish culture and at experimental stage. In the case study of this rice-cum-fish model, the Nile Tilapia (Oreochromis niloticus) was involved. The result was that 1,4720 kg/ha/yr could be produced using chick manure application under rice-cum-fish culture model. The available records reveal that 233,079 ha out of 495,000 ha of estimated Fadama in Niger State was used for rice cultivation in 1997. If 233,079 ha were to be used for integrated rice-cum fish culture, it is estimated that 343,092 mt of fish (Oreochromis niloticus) could be produced per year. The fish demand in Niger State in 2002 was 50,000 mt. The NPK application under rice-cum-fish production gave the best rice production estimated at 43,968.0 kg/ha/yr. The percentage increase in rice yield as well as increase in net income due to introduction of fish was 10.1 % and 54.4% respectively. The culture system is therefore recommended for adoption towards greater participation in aquaculture development by the farmers

Culture performance of Oreochromis niloticus and Sarotherodon galilaeus using similar management strategies

Sarotherodon galilaeus and Oreochromis niloticus are 2 readily available fish species in Nigeria, often cultured interchangeably. The findings are presented of a comparative study conducted to provide information on their performance using similar nutrient loadings without supplemental feed. Results showed a better performance of O.niloticus over S.galilaeus, which may be partly explained by the dietary mechanisms of the fish. Further trials on gut content analyses are required

Study on the biomass yield of Duckweed (Lemna minor) (L.) in hydroponic cultures containing different concentrations of aquaeous extract of chicken manure

The biomass yields of duck week (Lemna minor(L) was monitored in hydroponic media prepared by variously extracting 0.50, 1.00 and 2.00g of dried chicken manure per liter of city water (tap water) supply. The culture media consisting of aqueous extract of the various manure treatments were made up to 12 liters in all cases with tap water as control. Plastic baths of 25 liters capacity with 0.71 super(m2) surface area were used as culture facility. Each bath was stocked at a density of 30g super(m-2) with fresh weed samples (i.e 21.30g/bath). Maximum yields were obtained at all treatment levels and control on day 3 and based on the highest yield of 0.37gm super(-2)d super(-1) (dry matter) obtained at 1.00gL manure treatment which was however not significantly higher (P>0.05) than the 0.36gm super(-2)d super(-1) (dry matter) at 0.05gl super(-1) media manure content, an average manure level of 0.75l super(-1) was selected and used to determine the operational plant density. Thus fresh weights of 30 to 300gm super(-2) was grown in triplicate at 30g intervals for a period of 3 days. A regression equation of Y=2.6720+0.0021x with a corresponding maximum density or operational plant density of 266gm super(-2) and yield of 0.98gm super(-2), d super(-1) (dry matter) were obtained. Further growth trials were carried out at the operational density and manure levels of 0.50, 0.75, 1.00, 1.25, 1.50, 1.75 and 2.00gl super(-1) media manure concentration giving a significantly higher yield (P<0.05) of 17gm super(-2), d super(-1) (dry matter). This yield was however doubled to between 2.21 and 2.24gm super(-2) d super(-1) (equivalent to 7.96 to 8.06mt.ha-1, Yr-1 dry matter on extrapolation) if 25% and 75% respectively of the total weed cover were harvested daily within the experimental period. The role of some dissolved plant nutrients (DPN) were also discussed

Feeding and waste management strategies in the culture of Clarias gariepinus

Nutrient digestibility and amino acid availability were assessed in sharp-toothed catfish, Clarias gariepinus, fingerlings fed diets containing soyabean flour (SF) - Poultry meat meal (PMM) blends (25:75. 50:50, and 75:25) and 0.5 of 1.0%, Cr sub(2)0 sub(3). There was agreement between the pattern of overall protein digestibility and average amino acid availability despite the variability in individual amino acid availability the best dry matter, lipid and protein digestibility coefficients, and amino acid availability values were obtained with diets containing 0.5% Cr sub(2)0 sub(3). Chromic Oxide inclusion level appeared to affect nutrient availability. Increased marker level resulted into decreased nutrient digestibility coefficients. Similarly, these diets generated lower fecal crude protein than those with 1.0% Cr sub(2)0 sub(3). However, the latter group recorded higher protein retention efficiency. Dry mailer and lipid of diets containing more soyabean flour seemed to be more digestible than those of poultry meat meal. Similar trend was observed for the apparent availability of the amino acids. This investigation has indicated that low level of marker was better in digestibility study. Utilization of more SF than PMM in the diets of this catfish was more beneficial and should be encouraged in the feed industries producing catfish diets towards a better feed and waste management strategies in this aquaculture operation

Effect of fish meal in formulated diet on the growth of Heterotis niloticus in semi-intensive pond culture system

The aim of the investigation is to know the percentage of fish meal required to support the best growth of Heterotis niloticus in a semi intensive pond culture system. To achieve this, feed was formulated with equal percentages of blood meal, and corn meal and varying levels of fish meal. The experiment was in four treatments. Results showed that the mean weight gained was directly proportional to the quantity of fish meal made available to the fish fence 31.58g, 33.79g, 45.15g and 51.24g were recorded for treatments I, II, III and IV respectively. Result from this study when compared with previous works, shows that size of the water body to a greater extent affects the growth. The availability of fish meal in the feed made it more acceptable to the fish and hence a commensurate conversion in to flesh. The analysis of variance showed that there is significant difference in the growth performance in the treatments

A study of women's participation in fish food security and poverty alleviation in Lagos and Ogun States

Women, all over the world have contributed in various ways to the social, political and economic development of the Society. In fact, the World Resource Institute recognizes that "women have profound and preserve effect onn the well-being of their families, communities and local ecosystems" (Gamble and Well 1997:211). Women constitute more than 50 percent of the Agricultural (Fisheries being a sub sector), labour force. A study on Women in Fisheries showed that they participate in all aspects of the sector (capture, culture, processing, marketing research, training and Extension services). This paper reports the result of the study on women's contributions in the development of the Fisheries Industry particularly their roles in Fish Food Security, Poverty Alleviation and high rates of women's adoption of Fisheries technologies. The Case-study research methodology is used to study the "How" and "Why" Women's Contribution in Fish Food Security and Poverty Alleviation is at the index level recorded for the gender. The study made use of "Case Study" Research Instrument; documents, interview, artefacts, direct observation and archival records. The sampling techniques were purposive for research audiences and simple random for fisher-folks in the chosen locations. Analysed data showed among others that in Fisheries Research women occupy very important positions as Heads of Division/Section, Fisheries Liasion/Extension Officers and Fisheries Laboratory Chiefs etc. The paper also gave results of women production, processing, marketing and other services statistics; it also discusses the "whys" of women's low capacity in fisheries development of the nation and finally suggested ways in improving women's optimal capacity utilization in fisheries development