Extraction and quantification of antimicrobial peptides from medicinal plants through TrisNaCl and PBS buffer

Nature has provided plants with their own specific defence system that protect the plant from several traumatic conditions. These include environmental conditions like drought, harsh climate changes, wounding, pathogen attack and other biological as well as a biological stresses. In order to deal with all these harmful occurrences, plants synthesize a wide range of defence factors that include both primary as well as secondary metabolites. Out of these the most popular are the defence proteins which are known as antimicrobial peptides (AMP). These AMPs are actually the pathogenesis-related (PR) defence proteins.  These proteins are activated under the control of defence system of plant whenever triggered by the alarming situation. In the current study Crude protein extraction of four medicinally important plants named as Cassia fistula, Albizia lebbeck, Saccharum officinarum & Cymbopogon citratus was performen. Extraction was done in TrisNaCl and PBS buffer. Quantification of the protein content in the extract was done by Bradford assay. Concentration of protein from TrisNaCl buffer extracts was high as compared to the extracts from PBS buffer. As these proteins play their protective role in defence of the plants against pathogen attack so these extracts can better be used to check the antimicrobial activity of these plants in future to treat several infectious diseases in humans

A comprehensive review on preparation of pure immunoglobulins

Immunoglobulins are also known as antibodies. Plasma cells are responsible for the production of immunoglobulins. Beta cells are activated against a pathogenic attack and facilitated the formation of different types of immunoglobulins naturally. These immunoglobulins are also artificially synthesized by non-specific laboratorial techniques include fractionation precipitation, electrophoretic methods, gel filtration chromatography, ion exchange chromatography, hydrophobic chromatography and by the specific immuno- adsorbent methods. Third generation immunoglobulins are effectively used for therapeutic purpose against viral infections intravenously. Second generation immunoglobulins synthesis involved removal of anti-complement contaminants and IgG aggregates, through enzymatic degradation and chemical modification. The harmful effects of intravenous immunoglobulins can be reduced by using ultraviolet light, P-propiolactone, and pepsin at pH 4. Intravenous immunoglobulin drugs lead to anti-inflammatory and immunomodulatory effects in various infections. The mechanism of action of intravenous immunoglobulins is dependent on the binding between the Fc portion of injected IgG and target cell receptors. Radiolabeling is performed by two methods as in vivo, in which radiolabeled antibodies are incorporated into the body to bind with the antibodies and in vitro method, radioactive material is bound with already formed antibodies. This chapter highlighted the artificial methods adopted for production of radiolabeled immunoglobulins holding significant therapeutic and diagnostic applications

Quality Testing and Antimicrobial Activity of Norfloxacin in Pakistan

Background: There are 400 licensed pharmaceutical companies in Pakistan; one third of Pakistan total consumption of pharmaceuticals is imported. Market potential is good for antibiotics, vaccines, hormones etc. Introduction: High performance liquid chromatography is an important tool for routine determination of antimicrobial drugs with specific emphasis on fluoroquinolones. The quality testing of norfloxacin was also done by Liquid chromatography coupled with mass spectrometry. Antibacterial activity of norfloxacin is good for aerobic gram negative bacteria, including Enterobacteriaceae and Pseudomonas aeruginosa. It has moderate antibacterial activity against gram positive microorganisms, such as staphylococci; anaerobic bacteria are generally resistant.Method: Our investigation focuses on the quality testing of brands of Norfloxacin available in market with variety of trade names. Reverse phase chromatographic technique RP-HPLC was used for the quality testing of norfloxacin at temperature of 35°C and a flow rate of 0.8ml/ml with Isocratic plution. Disk diffusion techniquewas used to check the antibacterial activity of norfloxacin against Proteus mirabilis, Klebsiella pnemoniae, Escherichia coli and Pseudomonas aeruginosa. Different brands of Norfloxacin (noroxin, uracin, qunor, urid , uroquin, webnor, bactinor, uritac , floxacin, norocin and ecoflaxin) from pharmaceutical companies A, B C, D,  F , G , H, I, J & and K were collected from markets of Rawalpindi and Islamabad (Pakistan). The claimed concentration of tablet was 400mg. Result: The quality testing of norfloxacin with sample and standard showed that brands from companies C and F Qunor, Webnor did not qualify the quality testing, they were of medium standard for the treatment of infections while H Company with a brand Uritac stands highest for quality test. Disk diffusion technique showed that all brands of norfloxacin were sensitive against the mentioned bacteria.Conclusion:  This research concluded that special effective measures should be taken by all the pharmaceutical companies in Pakistan to check the quality standards of pharmaceuticals especially antibiotics before manufacturing. If they qualify the quality standards they will be effective to treat infections, if they are of low standard it would be difficult for the clinician to advise the correct amount of dosage for patient.

Variations in Total Phenolic, Total Flavonoid Contents, and Free Radicals’ Scavenging Potential of Onion Varieties Planted under Diverse Environmental Conditions

Genetic diversity and Agro-climatic conditions contribute significantly to the agronomic and morphological features of the food plant species, and their nutraceutical potential. The present study was intended to evaluate the impact of growing conditions on total phenolic and total flavonoid contents, and in vitro antioxidant potential in the bulbs and leaves of onion varieties planted under diverse environmental conditions. Standard analytical methods were used to quantify total phenolic content (TPC), total flavonoid content (TFC), and free radicals’ scavenging/antioxidant capacity. The impact of climatic and soil conditions was assessed using statistical tools. In general, onion varieties cultivated at three different locations viz. Kalar Kahar, Lahore and Swabi exhibited significant variations in TPC and TFC, and antioxidant activities. The bulbs and leaves of Mustang (V1) variety planted at Lahore and Swabi had significantly (p < 0.05), high levels of TPC (659.5 ± 6.59, and 631.1 ± 8.58 mg GAE/100 g, respectively). However, leaves of Red Orb (V2) and bulbs of Mustang (V1), and Golden Orb (V6), harvested from Kalar Kahar depicted the highest concentration of TFC (432.5 ± 10.3, 303.0 ± 6.67, and 303.0 ± 2.52 mg QE/100 g DW, respectively). Likewise, bulbs of V1 planted at Kalar Kahar, Lahore and Swabi exhibited maximum inhibition of DPPH, ABTS, and H2O2 radicals (79.01 ± 1.49, 65.38 ± 0.99, and 59.76 ± 0.90%, respectively). Golden Orb (V6) harvested from Lahore had the highest scavenging of OH radical (67.40 ± 0.09%). Likewise, bulbs of V1 variety planted at KalarKahar and Swabi had significant capacity to scavenge ferric ions (415.1 ± 10.6 mg GAE/100 g DW), and molybdate ions (213.7 ± 0.00 mg AAE/100 g DW). Conversely, leaves of Amazon (V8), planted at Lahore and Swabi depicted significant levels of DPPH, ABTS, H2O2 radical scavenging (90.69 ± 0.26, 63.55 ± 1.06, 51.86 ± 0.43%, respectively), and reduction of ferric ions (184.2 ± 6.75 mg GAE/100 g DW). V6 leaves harvested from Lahore and that of Super Sarhad (V3) from Swabi showed the highest inhibition of OH radical (61.21 ± 0.79%), and molybdate ions (623.6 ± 0.12 mg AAE/100 g DW), respectively. Pearson correlation and principal component analysis revealed strong relationships of climatic conditions, soil properties and elevation with TPC, TFC and free radicals’ scavenging potential in the bulbs and leaves of onion varieties. The variations in the total phenolic and flavonoid contents, and antioxidant potential of different varieties, and their associations with climatic and soil factors revealed the complexity of the growing conditions and genetic makeup that imposed significant impacts on the synthesis of secondary metabolites and nutraceutical potential of food and medicinal plant species

Chemo-thermal Treatment of Tribulus tresstris to Enhance Its Sequestering Potential for Adsorption of Some Heavy Metals from Aqueous Media: Kinetic and Thermodynamic Study

The plant based renewable biosorbents have extensively been investigated for removing water pollutants. The present study describes the sequestering of metal ions by exploiting a low cost biomaterial derived from Tribulus tresstris as sorbent. The batch equilibrium studies have been carried out both with raw and chemically/thermally treated biomaterial as a function of pH, contact time, shaking speed and shaking time to decide the effectiveness of biosorbent. The sorbent was activated chemically by utilizing 0.1M HCl and 0.1M K2CO3. A close muffle furnace was used for thermal treatment of the sorbent. The adsorption capacity was enhanced to 25% by thermal treatment and 54% by chemical treatment because of increase in pore volume and surface area. The greatest sorption was found for particle size of 200 µm with a 0.5 g dosage at pH 6 for 20 min at shaking speed 100 rpm. The FT-IR and SEM study was performed to discover the adsorption capacity of various functional groups and their binding mechanism. The adsorption data demonstrates that Langmuir, Freundlich and Dubinin-Radushkevich isotherm models were very much fitted to describe the adsorption behavio

Variations in Total Phenolic, Total Flavonoid Contents, and Free Radicals’ Scavenging Potential of Onion Varieties Planted under Diverse Environmental Conditions

Genetic diversity and Agro-climatic conditions contribute significantly to the agronomic and morphological features of the food plant species, and their nutraceutical potential. The present study was intended to evaluate the impact of growing conditions on total phenolic and total flavonoid contents, and in vitro antioxidant potential in the bulbs and leaves of onion varieties planted under diverse environmental conditions. Standard analytical methods were used to quantify total phenolic content (TPC), total flavonoid content (TFC), and free radicals’ scavenging/antioxidant capacity. The impact of climatic and soil conditions was assessed using statistical tools. In general, onion varieties cultivated at three different locations viz. Kalar Kahar, Lahore and Swabi exhibited significant variations in TPC and TFC, and antioxidant activities. The bulbs and leaves of Mustang (V1) variety planted at Lahore and Swabi had significantly (p 2O2 radicals (79.01 ± 1.49, 65.38 ± 0.99, and 59.76 ± 0.90%, respectively). Golden Orb (V6) harvested from Lahore had the highest scavenging of OH radical (67.40 ± 0.09%). Likewise, bulbs of V1 variety planted at KalarKahar and Swabi had significant capacity to scavenge ferric ions (415.1 ± 10.6 mg GAE/100 g DW), and molybdate ions (213.7 ± 0.00 mg AAE/100 g DW). Conversely, leaves of Amazon (V8), planted at Lahore and Swabi depicted significant levels of DPPH, ABTS, H2O2 radical scavenging (90.69 ± 0.26, 63.55 ± 1.06, 51.86 ± 0.43%, respectively), and reduction of ferric ions (184.2 ± 6.75 mg GAE/100 g DW). V6 leaves harvested from Lahore and that of Super Sarhad (V3) from Swabi showed the highest inhibition of OH radical (61.21 ± 0.79%), and molybdate ions (623.6 ± 0.12 mg AAE/100 g DW), respectively. Pearson correlation and principal component analysis revealed strong relationships of climatic conditions, soil properties and elevation with TPC, TFC and free radicals’ scavenging potential in the bulbs and leaves of onion varieties. The variations in the total phenolic and flavonoid contents, and antioxidant potential of different varieties, and their associations with climatic and soil factors revealed the complexity of the growing conditions and genetic makeup that imposed significant impacts on the synthesis of secondary metabolites and nutraceutical potential of food and medicinal plant species

A Comprehensive Analysis and Anti-Cancer Activities of Quercetin in ROS-Mediated Cancer and Cancer Stem Cells

Reactive oxygen species (ROS) induce carcinogenesis by causing genetic mutations, activating oncogenes, and increasing oxidative stress, all of which affect cell proliferation, survival, and apoptosis. When compared to normal cells, cancer cells have higher levels of ROS, and they are responsible for the maintenance of the cancer phenotype; this unique feature in cancer cells may, therefore, be exploited for targeted therapy. Quercetin (QC), a plant-derived bioflavonoid, is known for its ROS scavenging properties and was recently discovered to have various antitumor properties in a variety of solid tumors. Adaptive stress responses may be induced by persistent ROS stress, allowing cancer cells to survive with high levels of ROS while maintaining cellular viability. However, large amounts of ROS make cancer cells extremely susceptible to quercetin, one of the most available dietary flavonoids. Because of the molecular and metabolic distinctions between malignant and normal cells, targeting ROS metabolism might help overcome medication resistance and achieve therapeutic selectivity while having little or no effect on normal cells. The powerful bioactivity and modulatory role of quercetin has prompted extensive research into the chemical, which has identified a number of pathways that potentially work together to prevent cancer, alongside, QC has a great number of evidences to use as a therapeutic agent in cancer stem cells. This current study has broadly demonstrated the function-mechanistic relationship of quercetin and how it regulates ROS generation to kill cancer and cancer stem cells. Here, we have revealed the regulation and production of ROS in normal cells and cancer cells with a certain signaling mechanism. We demonstrated the specific molecular mechanisms of quercetin including MAPK/ERK1/2, p53, JAK/STAT and TRAIL, AMPKα1/ASK1/p38, RAGE/PI3K/AKT/mTOR axis, HMGB1 and NF-κB, Nrf2-induced signaling pathways and certain cell cycle arrest in cancer cell death, and how they regulate the specific cancer signaling pathways as long-searched cancer therapeutics

Performance evaluation of high-performance self-compacting concrete with waste glass aggregate and metakaolin

High-Performance Self-Compacting Concrete (HPSCC) has attracted much attention in recent decades due to its remarkable ability to fill formworks with densely packed reinforcing bars while requiring minimal or no external compaction. Because of the negative environmental impacts of cement and natural aggregates in concrete production, a much more sustainable alternative to manufacturing HPSCC is required. Recycled glass waste is one of the most attractive waste materials that can be used to create sustainable concrete compounds, which is currently a major area of study among researchers. This study aims to develop information not only about the fresh, mechanical, and durability characteristics of HPSCC, evaluate the environmental impact and correlate the crushing strength using a non-destructive approach by utilizing waste glass aggregates at replacement percentages of 0%, 10%, 20%, 30%, and 40%. To improve the performance of the produced HPSCC, Metakaolin was also added. The results of the fresh concrete tests revealed that the substitution of an optimal level of waste glass with Metakaolin provides adequate implementation in flowability, passing ability, and viscosity behaviors. Even though there is a reduction in the mechanical performance with glass aggregates, Metakaolin significantly improved strength and ductility by up to 16.12% and 15.91%, respectively. Furthermore, in most cases, the use of glass aggregates with Metakaolin significantly alters the durability properties of concrete while minimizing the environmental impact as well as the overall project cost. Finally, the NDT assessment demonstrates that the analytical equation can efficiently predict the compressive strength and promising to use for field application