Identification and Characterization of Active Ingredient Eugenol from Syzygium Aromaticum (Clove Oil) through HPLC and its Phytochemical Analysis

Clove, Syzygium aromaticum, is one of the most valuable, ancient and premium essential oil which has been used as source of spice in agro-food industry and therapeutics for centuries. The aim of the present study was to analyze biological activities of clove oil as well as characterization of its active component Eugenol to make it worthwhile for different food and pharmaceutical formulations. Clove oil extraction was performed by Soxhlet method. Its phytochemical analysis showed the presence of carbohydrates, flavonoids, coumarins, steroids, saponnins, and tannins while proteins and leucoanthocyanins were absent in it. DPPH (2, 2-diphenyl-1-picrylhydrazyl) scavenging assay was performed which showed an increase in percentage inhibition with an increase in concentration of clove oil which confirmed its antioxidant property. Antimicrobial activity of clove oil was tested against three gram positive strains Bacillus subtilis, Staphylococcus aureus and Bacillus thuringiensis and one gram negative strain Escherichia coli by well diffusion and disc diffusion methods. E.coli presented largest zone of inhibition. T-test was applied for statistical analysis of antimicrobial activity. P-value obtained was 0.0215 which is ˂0.05. Aqueous clove nanoparticles were made and were found to have antimicrobial activity against E.coli and B.subtilis. Correlation coefficient (R2) through HPLC was found to be 0.973531. Quantitative estimation showed the presence of 740 ppm eugenol. These properties of clove proved it as a valuable spice in pharmaceutical and agro-food sector

Quantitative estimation of protein in sprouts of 'vigna radiate' (mung beans), 'lens culinaris' (lentils), and 'cicer arietinum' (chickpeas) by Kjeldahl and Lowry methods

Protein scarcity is the most vital cause of long-lasting diseases and even untimely deaths in some developing nations. The application of protein in food is advantageous from the point of view of non-toxicity, biocompatibility, and dietary benefits. This study aimed to determine the protein contents of the sprouts of Vigna radiates (mung beans), Lens culinaris (lentils), and Cicer arietinum (chickpeas) using the Kjeldahl and Lowry methods. The results obtained from the Kjeldahl method identified protein concentrations of 2.54, 2.63, and 2.19%, whereas the Lowry method results identified protein concentrations of 2.96%, 4.10%, and 1.6% in mung beans, lentils, and chickpeas, respectively. In both the methods, lentils were found to have the highest amount of protein followed by mung beans and chickpeas. Both the Kjeldahl and Lowry methods demonstrated good protein values and low variation in the protein amount in the analyzed samples. Furthermore, the methods had greater sensitivity and comparable experimental variability. The outcomes revealed that assays can be applied for protein analysis in legumes. In the context of a lack of suitable standard procedures for evaluating legumes’ compositions, the present study is suitable for food control laboratories. In addition, the studied samples represent a significant source of protein and can be used to fulfil the daily requirements for protein intake and other food applications

Novel Stimuli-Responsive Pectin-PVP-Functionalized Clay Based Smart Hydrogels for Drug Delivery and Controlled Release Application

Stimuli-responsive drug delivery systems are urgently required for injectable site-specific delivery and release of drugs in a controlled manner. For this purpose, we developed novel pH-sensitive, biodegradable, and antimicrobial hydrogels from bio-macromolecule pectin, polyvinylpyrrolidone (PVP), 3-aminopropyl (diethoxy)methyl silane (3-APDEMS), and sepiolite clay via blending and solution casting technique. The purified sepiolite (40 um) was functionalized with 3-APDEMS crosslinker (ex-situ modification) followed by hydrogels fabrication. FTIR and SEM confirmed crosslinked structural integrity and rod-like morphology of hydrogels respectively. The swelling properties of hydrogels could be controlled by varying the concentration of modified clay in pectin/PVP blends. Moreover, the decrease in pH increased the swelling of hydrogels indicating the pH-responsiveness of hydrogels. All hydrogels were degraded after 21 days in phosphate buffer saline pH 7.4 (human blood pH). In-vitro cytotoxicity against 3T3 mouse fibroblast cell line analysis confirmed cytocompatibility of all hydrogels. Ceftriaxone sodium (CTX-S) was selected as a model drug. The release profile of the hydrogel showed 91.82% release in PBS for 2 h in a consistent and controlled manner. The chemical structure of the drug remained intact during and after release confirmed through UV-Visible spectroscopy. Overall, these hydrogels could be used as potential scaffolds for future biomedical applications

Functional Identification of Serine Hydroxymethyltransferase as a Key Gene Involved in Lysostaphin Resistance and Virulence Potential of Staphylococcus aureus Strains

Gaining an insight into the mechanism underlying antimicrobial-resistance development in Staphylococcus aureus is crucial for identifying effective antimicrobials. We isolated S. aureus sequence type 72 from a patient in whom the S. aureus infection was highly resistant to various antibiotics and lysostaphin, but no known resistance mechanisms could explain the mechanism of lysostaphin resistance. Genome-sequencing followed by subtractive and functional genomics revealed that serine hydroxymethyltransferase (glyA or shmT gene) plays a key role in lysostaphin resistance. Serine hydroxymethyltransferase (SHMT) is indispensable for the one-carbon metabolism of serine/glycine interconversion and is linked to folate metabolism. Functional studies revealed the involvement of SHMT in lysostaphin resistance, as ΔshmT was susceptible to the lysostaphin, while complementation of the knockout expressing shmT restored resistance against lysostaphin. In addition, the ΔshmT showed reduced virulence under in vitro (mammalian cell lines infection) and in vivo (wax-worm infection) models. The SHMT inhibitor, serine hydroxymethyltransferase inhibitor 1 (SHIN1), protected the 50% of the wax-worm infected with wild type S. aureus. These results suggest SHMT is relevant to the extreme susceptibility to lysostaphin and the host immune system. Thus, the current study established that SHMT plays a key role in lysostaphin resistance development and in determining the virulence potential of multiple drug-resistant S. aureus

Representation of Colorism in Pakistani Dramas: A Critical Discourse Analysis

This research gives an insight into discrimination based on color in Pakistani drama serials. The current study is limited to the analysis of the drama serial Parizaad aired in 2021. It employs Norman Fairclough’s three-dimensional model to explore the connection that lies between different social practices and language. The analysis is delimited to the third dimension which deals with discourse as social practice as theoretical framework for this research. It meticulously unfolds associated prejudices and biases with light and dark skin tones depicted in the dialogues. The study explores particular language, words and strange attitudes of society aimed at the dark-skinned person. This study decodes the main thematic concerns of Parizaad drama highlights the social fabrication of colorism. It is revealed that colorism is deeply rooted in cultural and traditional values. The subject matter deals with the experience of a dark-skinned person through multiple angles, encapsulating multiple points of view. This study also puts an effort to display the social psyche of the central dark-toned character Parizaad. The main findings yielded by this research refer to the utilization of language as a means of exploring the underlying ideology behind Colorism. Within the context of the drama series entitled Parizaad, media discourse thoughtfully addresses the issue of color-based discrimination. The success of Parizaad at the end of the drama challenges the social fabrication. Parizaad gives a powerful message of self-empowerment and self-worth which possesses a positive impact on the society

Children Story Books as a Means of Social and Intellectual Development

The proposed study has explored the interpersonal meta-function in two children story books entitled “Four Feet Two Sandals” and “The Spirit of Cheetah”.  It was aimed to explore what type of social and intellectual messages were being conveyed in these stories. The study was based on qualitative method and Halliday & Mathiessen (2004) modal of interpersonal meta-function was applied as a theoretical framework. Meanwhile, a sample of 10 passages was drawn from the selected story books. Data was analyzed through the lens of content analysis technique. In both aspects of mood and modality, the findings revealed the author’s intention to convey several useful values like sacrifice, friendship, patience, determination to boost the social and intellectual level of the children

Thyroid Dysfunction in Non-Interferon Treated Hepatitis C Patients Residing in Hepatitis Endemic Area

Background. Association of thyroid dysfunction (TD) with interferon treatment of HCV is well known to clinicians. However, a few studies have highlighted the role of hepatitis C virus per se in the development of TD. The aim of this study was to know the prevalence of TD in non-interferon treated HCV infected patients referred for thyroid function testing. Patients and Methods. Among 557 ELISA-positive HCV patients 446 (341 females, 105 males) were selected for this study. Serums FT4, FT3, and TSH were determined by radioimmunoassay method. Results. TD was detected in 15.2% of patients: 9.0% hypothyroidism and 6.3% hyperthyroidism. In increasing order subclinical hypothyroidism, overt hypothyroidism, overt hyperthyroidism, and subclinical hyperthyroidism were found in 4.7%, 4.3%, 3.6%, and 2.7% patients, respectively. Overall TD was more common in female than in male HCV patients but the difference was not significant (16.1% versus 12.4%; p=0.648). Hyperthyroidism and subclinical hypothyroidism were slightly more common in female and overall hypothyroidism and overt hypothyroidism in male patients but the difference was not statistically significant (p>0.05). The incidence of TD was relatively high in patients above 36 years (median age) but the difference was not statistically significant either collectively or in gender base groups (p>0.05). Conclusion. Prior to interferon treatment, HCV infection itself causes biochemical thyroid dysfunction in 15.2% of local HCV patients

Identification of 2′,4′-Dihydroxychalcone as an Antivirulence Agent Targeting HlyU, a Master Virulence Regulator in Vibrio vulnificus

The emergence of antimicrobial resistance and rapid acclimation allows Vibrio vulnificus to rapidly propagate in the host. This problematic pathological scenario can be circumvented by employing an antivirulence strategy, treating Vibrio infections without hindering the bacterial growth. We developed a genome-integrated orthogonal inhibitor screening platform in E. coli to identify antivirulence agents targeting a master virulence regulator of V. vulnificus. We identified 2′,4′-dihydroxychalcone (DHC) from the natural compound library and verified that it decreases the expression of the major toxin network which is equivalent to the ∆hlyU deletion mutant. 2′,4′-DHC also reduced the hemolytic activity of V. vulnificus which was tested as an example of virulence phenotype. The electrophoretic mobility shift assay confirmed that 2′,4′-DHC specifically targeted HlyU and inhibited its binding to PrtxA1 promoter. Under in vivo conditions, a single dose of 2′,4′-DHC protected ~50% wax-worm larvae from V. vulnificus infection at a non-toxic concentration to both V. vulnificus and wax-worm larvae. In the current study, we demonstrated that an orthogonal reporter system is suitable for the identification of antivirulence compounds with accuracy, and identified 2′,4′-DHC as a potent antivirulence agent that specifically targets the HlyU virulence transcriptional regulator and significantly reduces the virulence and infection potential of V. vulnificus