Optimized attention-induced multihead convolutional neural network with efficientnetv2-fostered melanoma classification using dermoscopic images.

Melanoma is an uncommon and dangerous type of skin cancer. Dermoscopic imaging aids skilled dermatologists in detection, yet the nuances between melanoma and non-melanoma conditions complicate diagnosis. Early identification of melanoma is vital for successful treatment, but manual diagnosis is time-consuming and requires a dermatologist with training. To overcome this issue, this article proposes an Optimized Attention-Induced Multihead Convolutional Neural Network with EfficientNetV2-fostered melanoma classification using dermoscopic images (AIMCNN-ENetV2-MC). The input pictures are extracted from the dermoscopic images dataset. Adaptive Distorted Gaussian Matched Filter (ADGMF) is used to remove the noise and maximize the superiority of skin dermoscopic images. These pre-processed images are fed to AIMCNN. The AIMCNN-ENetV2 classifies acral melanoma and benign nevus. Boosted Chimp Optimization Algorithm (BCOA) optimizes the AIMCNN-ENetV2 classifier for accurate classification. The proposed AIMCNN-ENetV2-MC is implemented using Python. The proposed approach attains an outstanding overall accuracy of 98.75%, less computation time of 98 s compared with the existing models.

Link Prediction in Complex Networks Using Average Centrality-Based Similarity Score.

Link prediction plays a crucial role in identifying future connections within complex networks, facilitating the analysis of network evolution across various domains such as biological networks, social networks, recommender systems, and more. Researchers have proposed various centrality measures, such as degree, clustering coefficient, betweenness, and closeness centralities, to compute similarity scores for predicting links in these networks. These centrality measures leverage both the local and global information of nodes within the network. In this study, we present a novel approach to link prediction using similarity score by utilizing average centrality measures based on local and global centralities, namely Similarity based on Average Degree (SACD), Similarity based on Average Betweenness (SACB), Similarity based on Average Closeness (SACC), and Similarity based on Average Clustering Coefficient (SACCC). Our approach involved determining centrality scores for each node, calculating the average centrality for the entire graph, and deriving similarity scores through common neighbors. We then applied centrality scores to these common neighbors and identified nodes with above average centrality. To evaluate our approach, we compared proposed measures with existing local similarity-based link prediction measures, including common neighbors, the Jaccard coefficient, Adamic-Adar, resource allocation, preferential attachment, as well as recent measures like common neighbor and the Centrality-based Parameterized Algorithm (CCPA), and keyword network link prediction (KNLP). We conducted experiments on four real-world datasets. The proposed similarity scores based on average centralities demonstrate significant improvements. We observed an average enhancement of 24% in terms of Area Under the Receiver Operating Characteristic (AUROC) compared to existing local similarity measures, and a 31% improvement over recent measures. Furthermore, we witnessed an average improvement of 49% and 51% in the Area Under Precision-Recall (AUPR) compared to existing and recent measures. Our comprehensive experiments highlight the superior performance of the proposed method.

Short stature and dysmorphic features in Asian Indian siblings with DAAM2-associated steroid-resistant nephrotic syndrome: Expansion of the phenotypic spectrum or a blended phenotype?

Variants in more than 60 different genes, most of which code for podocyte-related proteins, have been found to be associated with monogenic forms of nephrotic syndrome (NS). Biallelic variants in DAAM2, a member of the formin family, were recently identified to cause autosomal recessive (AR) NS type 24 in four unrelated families with steroid-resistant nephrotic syndrome (SRNS). This case report represents only the fifth reported family of DAAM2-associated NS and the first from India, with two sibs who presented with a complex phenotype characterized by steroid-resistant nephrotic syndrome, short stature, dysmorphic facial features, deep-set toenails, myopia, increased thickness of the calvarium of the skull, and sloping ribs. Both sibs were found to have a homozygous likely pathogenic nonsense variant c.196C>T (p.Arg66Ter; NM_001201427.2) in exon 3 of the DAAM2 gene through whole exome sequencing. The dysmorphic features could possibly be part of the DAAM2-related phenotype which has hitherto not been reported or could represent a blended phenotype, with the extrarenal manifestations resulting from a yet to be identified coexisting genetic condition.

High Diagnostic Accuracy of Thyroid-Stimulating Hormone (TSH) Receptor Antibodies in Distinguishing Graves' Disease and Subacute Thyrotoxicosis in the Indian Population.

BACKGROUND AND OBJECTIVE: Thyrotoxicosis is a common clinical condition encountered in endocrine practice. Graves' disease and subacute thyroiditis are the two common causes of thyrotoxicosis and often have overlapping clinical and biochemical features. 99mTc thyroid scintigraphy is the most commonly used confirmatory test to differentiate the two conditions but is not available in the majority of the second-tier cities of India. However, obtaining thyroid stimulating hormone (TSH) receptor antibodies (TSHrAb), another accurate test to differentiate the two conditions, in second-tier cities by outsourcing to labs in major cities is a feasible option nowadays. However, the data on the performance of TSHrAb to differentiate the two conditions in Indian patients is limited. Hence, we have evaluated the diagnostic accuracy of TSHrAb in the Indian population to differentiate Graves' disease and subacute thyroiditis. MATERIALS AND METHODS: This prospective study was conducted on 115 consecutive newly diagnosed thyrotoxicosis patients presenting to the Department of Endocrinology at a tertiary health care centre in India. Clinical parameters like throat pain, duration of symptoms, and grade of goitre were noted. Measurement of total tri-iodothyronine (TT3), total thyroxine (TT4), TSH, TSHrAb, and 99mTc thyroid scintigraphy were performed in all participants. All participants were followed up at least for six months after the recruitment. Increased tracer uptake (>4%) and/or increased thyroid to parotid trace uptake ratio (>2.5) were used to diagnose Graves' disease. RESULTS: Eighty-one and 34 patients were diagnosed with Graves' disease and subacute thyroiditis, respectively. TT3/TT4 ratio had low diagnostic accuracy (area under the curve (AUC): 0.6, best cut-off: 15.6, sensitivity: 53.1%, specificity: 79.4%). TSHrAb had the best AUC (0.9) to distinguish Graves' disease from subacute thyroiditis (cut-off: 2.0 IU/L, sensitivity: 97.5%, specificity: 100%). In contrast, the kit manufacturer's reference range (1.75 IU/L) was slightly more sensitive (98.8%), but less specific (94%). CONCLUSION: The TT3/TT4 ratio is not a good test to differentiate Graves' disease and subacute thyroiditis. TSHrAb is accurate in distinguishing Graves' disease from subacute thyroiditis and a level of 2.0 may be a more accurate cut-off to differentiate the two conditions in the Indian population.

Community-Based Matrix Factorization (CBMF) Approach for Enhancing Quality of Recommendations.

Matrix factorization is a long-established method employed for analyzing and extracting valuable insight recommendations from complex networks containing user ratings. The execution time and computational resources demanded by these algorithms pose limitations when confronted with large datasets. Community detection algorithms play a crucial role in identifying groups and communities within intricate networks. To overcome the challenge of extensive computing resources with matrix factorization techniques, we present a novel framework that utilizes the inherent community information of the rating network. Our proposed approach, named Community-Based Matrix Factorization (CBMF), has the following steps: (1) Model the rating network as a complex bipartite network. (2) Divide the network into communities. (3) Extract the rating matrices pertaining only to those communities and apply MF on these matrices in parallel. (4) Merge the predicted rating matrices belonging to communities and evaluate the root mean square error (RMSE). In our experimentation, we use basic MF, SVD++, and FANMF for matrix factorization, and the Louvain algorithm is used for community division. The experimental evaluation on six datasets shows that the proposed CBMF enhances the quality of recommendations in each case. In the MovieLens 100K dataset, RMSE has been reduced to 0.21 from 1.26 using SVD++ by dividing the network into 25 communities. A similar reduction in RMSE is observed for the datasets of FilmTrust, Jester, Wikilens, Good Books, and Cell Phone.

Role of T Cells in Vaccine-Mediated Immunity against Marek's Disease.

Marek's disease virus (MDV), a highly cell-associated oncogenic α-herpesvirus, is the etiological agent of T cell lymphomas and neuropathic disease in chickens known as Marek's disease (MD). Clinical signs of MD include neurological disorders, immunosuppression, and lymphoproliferative lymphomas in viscera, peripheral nerves, and skin. Although vaccination has greatly reduced the economic losses from MD, the molecular mechanism of vaccine-induced protection is largely unknown. To shed light on the possible role of T cells in immunity induced by vaccination, we vaccinated birds after the depletion of circulating T cells through the IP/IV injection of anti-chicken CD4 and CD8 monoclonal antibodies, and challenged them post-vaccination after the recovery of T cell populations post-treatment. There were no clinical signs or tumor development in vaccinated/challenged birds with depleted CD4+ or CD8+ T cells. The vaccinated birds with a combined depletion of CD4+ and CD8+ T cells, however, were severely emaciated, with atrophied spleens and bursas. These birds were also tumor-free at termination, with no virus particles detected in the collected tissues. Our data indicated that CD4+ and CD8+ T lymphocytes did not play a critical role in vaccine-mediated protection against MDV-induced tumor development.

Pleiotropic fitness effects of the lncRNA Uhg4 in Drosophila melanogaster.

BACKGROUND: Long noncoding RNAs (lncRNAs) are a diverse class of RNAs that are critical for gene regulation, DNA repair, and splicing, and have been implicated in development, stress response, and cancer. However, the functions of many lncRNAs remain unknown. In Drosophila melanogaster, U snoRNA host gene 4 (Uhg4) encodes an antisense long noncoding RNA that is host to seven small nucleolar RNAs (snoRNAs). Uhg4 is expressed ubiquitously during development and in all adult tissues, with maximal expression in ovaries; however, it has no annotated function(s). RESULTS: We used CRISPR-Cas9 germline gene editing to generate multiple deletions spanning the promoter region and first exon of Uhg4. Females showed arrested egg development and both males and females were sterile. In addition, Uhg4 deletion mutants showed delayed development and decreased viability, and changes in sleep and responses to stress. Whole-genome RNA sequencing of Uhg4 deletion flies and their controls identified co-regulated genes and genetic interaction networks associated with Uhg4. Gene ontology analyses highlighted a broad spectrum of biological processes, including regulation of transcription and translation, morphogenesis, and stress response. CONCLUSION: Uhg4 is a lncRNA essential for reproduction with pleiotropic effects on multiple fitness traits.

Privacy Protection of Healthcare Data over Social Networks Using Machine Learning Algorithms.

With the rapid development of mobile medical care, medical institutions also have the hidden danger of privacy leakage while sharing personal medical data. Based on the k-anonymity and l-diversity supervised models, it is proposed to use the classified personalized entropy l-diversity privacy protection model to protect user privacy in a fine-grained manner. By distinguishing solid and weak sensitive attribute values, the constraints on sensitive attributes are improved, and the sensitive information is reduced for the leakage probability of vital information to achieve the safety of medical data sharing. This research offers a customized information entropy l-diversity model and performs experiments to tackle the issues that the information entropy l-diversity model does not discriminate between strong and weak sensitive features. Data analysis and experimental results show that this method can minimize execution time while improving data accuracy and service quality, which is more effective than existing solutions. The limits of solid and weak on sensitive qualities are enhanced, sensitive data are reduced, and the chance of crucial data leakage is lowered, all of which contribute to the security of healthcare data exchange. This research offers a customized information entropy l-diversity model and performs experiments to tackle the issues that the information entropy l-diversity model does not discriminate between strong and weak sensitive features. The scope of this research is that this paper enhances data accuracy while minimizing the algorithm's execution time.

Development and evaluation of a multicomponent bioink consisting of alginate, gelatin, diethylaminoethyl cellulose and collagen peptide for 3D bioprinting of tissue construct for drug screening application.

Three dimensional (3D) bioprinting technology has been making a progressive advancement in the field of tissue engineering to produce tissue constructs that mimic the shape, framework, and microenvironment of an organ. The technology has not only paved the way to organ development but has been widely studied for its application in drug and cosmetic testing using 3D bioprinted constructs. However, not much has been explored on the utilization of bioprinting technology for the development of tumor models to test anti-cancer drug efficacy. The conventional methodology involves a two dimensional (2D) monolayer model to test cellular drug response which has multiple limitations owing to its inability to mimic the natural tissue environment. The choice of bioink for 3D bioprinting is critical as cell morphology and proliferation depend greatly on the property of bioink. In this study, we developed a multicomponent bioink composed of alginate, diethylaminoethyl cellulose, gelatin, and collagen peptide to generate a 3D bioprinted construct. The bioink has been characterised and validated for its printability, shape fidelity and biocompatibility to be used for generating tumor models. Further, a bioprinted tumor model was developed using lung cancer cell line and the efficacy of 3D printed construct for drug screening application was established.

Antibacterial activity of cinnamon and clove oil against wound pathogens.

A wound is a complicated health issue, and it severely affects the injured area because of the growth of many pathogenic microorganisms. Cinnamon and clove oil exhibit antibacterial activity against wound pathogens like Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae were identified by the disc diffusion method. Cinnamon and clove oils are effective antibacterial agents because of their importance in reducing virulence and pathogenicity of drug-resistant bacteria in vivo. The increased frequency in clinically observed cases of antibiotic resistance has been attributed to many factors, such as the misuse and overuse of antibiotics. In some countries, antibiotics are sold over the counter without a prescription; hence, this study aimed to investigate the antimicrobial effect of clove and cinnamon on clinically isolated resistant strains of P. aeruginosa, E. coli, and K. pneumoniae.

Centromere Protein F (CENPF): A novel marker for salivary gland pathology.

Background: Salivary gland tumours are relatively uncommon, and there exists considerable diagnostic difficulty. This is due to individual lesions having diverse histopathological features, presence of number of types and variants, and overlapping histological features in different tumour entities. Aim: The current study aimed at assessing the expression of centromere protein F (CENPF) in benign and malignant salivary gland tumours and to evaluate the efficacy of CENPF as a proliferative marker to aid in the diagnosis of malignancy so that it will help in surgical pathology practice. Materials and Methods: The study group involved 20 cases of benign salivary gland tumours, 20 cases of malignant salivary gland tumours, and 10 normal salivary gland tissues. All the cases were subjected to immunohistochemical analysis for CENPF expression and were assessed by two independent observers and further taken up for evaluation. Statistical Analysis: The results were analysed statistically among different groups using analysis of variance (ANOVA) or Kruskal-Wallis test with Chi-squared test using IBM's Statistical Package for the Social Sciences (SPSS) version 17.0. Results: CENPF expression in normal salivary gland was negative with gradual increase in expression from benign salivary gland tumours to malignant salivary gland tumours. CENPF expression was high in malignant salivary gland tumours. Conclusion: Findings of the study suggest that CENPF can be regarded as a new cell proliferation marker for malignant salivary gland tumours.

Synergistic Induction of Chicken Antimicrobial Host Defense Peptide Gene Expression by Butyrate and Sugars.

Antimicrobial resistance is a major concern to public health demanding effective alternative strategies to disease control and prevention. Modulation of endogenous host defense peptide (HDP) synthesis has emerged as a promising antibiotic alternative approach. This study investigated a potential synergy between sugars and butyrate in inducing HDP gene expression in chickens. Our results revealed that sugars differentially regulated HDP expression in both gene- and sugar-specific manners in chicken HD11 macrophage cells. Among eight mono- and disaccharides tested, all were potent inducers of avian ß-defensin 9 (AvBD9) gene (p<0.05), but only galactose, trehalose, and lactose obviously upregulated cathelicidin-B1 (CATHB1) gene expression. The expression of AvBD14 gene, on the other hand, was minimally influenced by sugars. Moreover, all sugars exhibited a strong synergy with butyrate in enhancing AvBD9 expression, while only galactose, trehalose, and lactose were synergistic with butyrate in CATHB1 induction. No synergy in AvBD14 induction was observed between sugars and butyrate. Although lactose augmented the expression of nearly all HDP genes, its synergy with butyrate was only seen with several, but not all, HDP genes. Mucin-2 gene was also synergistically induced by a combination of lactose and butyrate. Furthermore, lactose synergized with butyrate to induce AvBD9 expression in chicken jejunal explants (p<0.05). Mechanistically, hyper-acetylation of histones was observed in response to both butyrate and lactose, relative to individual compounds. Mitogen-activated protein kinase, NF-κB, and cyclic adenosine monophosphate signaling pathways were also found to be involved in butyrate- and lactose-mediated synergy in AvBD9 induction. Collectively, a combination of butyrate and a sugar with both HDP-inducing and barrier protective activities holds the promise to be developed as an alternative to antibiotics for disease control and prevention.

Diagnosis and Assessment of Severity of Pediatric Pneumonia Using the Respiratory Index of Severity (RISC) Scoring System.

OBJECTIVE: To evaluate the correlation between the Respiratory Index of Severity (RISC) scoring system and the World Health Organization (WHO) interpretation of chest radiographs, and to evaluate the RISC scoring system in the diagnosis and assessment of severity of pneumonia in children against chest X-ray. METHODS: 241 children presenting to a tertiary care center with a clinical diagnosis of pneumonia that necessitated a chest radiograph to be taken, were enrolled. The RISC scoring was done for all participants by a single pediatrician and chest X-ray graded by a single radiologist as per the WHO interpretation of chest radiographs. RESULTS: There was a statistically significant positive correlation (P=0.02) between the two scores. RISC score of >1 had a sensitivity of 80.3%, and score ≥3 had a specificity of 88.3%, positive predictive value of 61.3% and negative predictive value of 76.8% for diagnosis and to predict severity of pneumonia. CONCLUSION: In a resource-limited setting, RISC scoring can be used to diagnose and predict the severity of childhood pneumonia.

Biofabrication of skin tissue constructs using alginate, gelatin and diethylaminoethyl cellulose bioink.

INTRODUCTION: Biofabrication of skin tissue equivalents using 3D bioprinting technology has gained much attention in recent times due to the simplicity, the versatility of the technology and its ability in bioengineering biomimetic tissue histology. The key component being the bioink, several groups are actively working on the development of various bioink formulations for optimal skin tissue construction. METHODS: Here, we present alginate (ALG), gelatin (GEL) and diethylaminoethyl cellulose (DCEL) based bioink formulation and its application in bioprinting and biofabrication of skin tissue equivalents. Briefly, DEAE cellulose powder was dispersed in alginate solution with constant stirring at 60 °C to obtain a uniform distribution of cellulose fibers; this was then mixed with GEL solution to prepare the bioink. The formulation was systematically characterized for its morphological, physical, chemical, rheological, biodegradation and biocompatibility properties. The printability, shape fidelity and cell-laden printing were assessed using the CellInk bioprinter. RESULTS: The bioink proved to be a good printable, non-cytotoxic and stable hydrogel formulation. The primary human fibroblast and keratinocyte-loaded 3D bioprinted constructs showed excellent cell viability, collagen synthesis, skin-specific marker and biomimetic tissue histology. CONCLUSION: The results demonstrated the successful formulation of ALG-GEL-DCEL bioink and its application in the development of human skin tissue equivalents with distinct epidermal-dermal histological features.

Functional Diversification, Redundancy, and Epistasis among Paralogs of the Drosophila melanogaster Obp50a-d Gene Cluster.

Large multigene families, such as the insect odorant-binding proteins (OBPs), are thought to arise through functional diversification after repeated gene duplications. Whereas many OBPs function in chemoreception, members of this family are also expressed in tissues outside chemosensory organs. Paralogs of the Obp50 gene cluster are expressed in metabolic and male reproductive tissues, but their functions and interrelationships remain unknown. Here, we report the genetic dissection of four members of the Obp50 cluster, which are in close physical proximity without intervening genes. We used CRISPR technology to excise the entire cluster while introducing a PhiC31 reintegration site to reinsert constructs in which different combinations of the constituent Obp genes were either intact or rendered inactive. We performed whole transcriptome sequencing and assessed sexually dimorphic changes in transcript abundances (transcriptional niches) associated with each gene-edited genotype. Using this approach, we were able to estimate redundancy, additivity, diversification, and epistasis among Obp50 paralogs. We analyzed the effects of gene editing of this cluster on organismal phenotypes and found a significant skewing of sex ratios attributable to Obp50a, and sex-specific effects on starvation stress resistance attributable to Obp50d. Thus, there is functional diversification within the Obp50 cluster with Obp50a contributing to development and Obp50d to stress resistance. The deletion-reinsertion approach we applied to the Obp50 cluster provides a general paradigm for the genetic dissection of paralogs of multigene families.

Citral Induced Apoptosis through Modulation of Key Genes Involved in Fatty Acid Biosynthesis in Human Prostate Cancer Cells: In Silico and In Vitro Study.

The isomers of citral (cis-citral and trans-citral) were isolated from the Cymbopogon citratus (DC.) Stapf oil demonstrates many therapeutic properties including anticancer properties. However, the effects of citral on suppressing human prostate cancer and its underlying molecular mechanism have yet to be elucidated. The citral was isolated from lemongrass oil using various spectroscopic analyses, such as electron ionized mass spectrometry (EI-MS) and nuclear magnetic resonance (NMR) spectroscopy respectively. We carried out 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to evaluate the cell viability of citral in prostate cancer cells (PC-3 and PC3M). Furthermore, to confirm that PC3 undergoes apoptosis by inhibiting lipogenesis, we used several detection methods including flow cytometry, DNA fragmentation, Hoechst staining, PI staining, oil staining, qPCR, and Western blotting. Citral impaired the clonogenic property of the cancer cells and altered the morphology of cancer cells. Molecular interaction studies and the PASS biological program predicted that citral isomers tend to interact with proteins involved in lipogenesis and the apoptosis pathway. Furthermore, citral suppressed lipogenesis of prostate cancer cells through the activation of AMPK phosphorylation and downregulation of fatty acid synthase (FASN), acetyl coA carboxylase (ACC), 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR), and sterol regulatory element-binding protein (SREBP1) and apoptosis of PC3 cells by upregulating BAX and downregulating Bcl-2 expression. In addition, in silico studies such as ADMET predicted that citral can be used as a safe potent drug for the treatment of prostate cancer. Our results indicate that citral may serve as a potential candidate against human prostate cancer and warrants in vivo studies.

4(3H)-Quinazolinone derivatives: Promising antibacterial drug leads.

Emergence of drug resistance has created unmet medical need for the development of new classes of antibiotics. Discovery of new antibacterial agents with new mode of action remains a high priority universally. 4(3H)-quinazolinone, a fused nitrogen heterocyclic compound has emerged as a biologically privileged structure, possessing a wide range of biological properties viz. anticancer, antibacterial, antitubercular, antifungal, anti-HIV, anticonvulsant, anti-inflammatory and analgesic activities. Promising antibacterial properties of quinazolinones have enthused the medicinal chemists to explore and develop this fused heterocyclic system for new antibacterial agents. Utilization of quinazolinone core for the design and synthesis of new antibacterial agents has recently gained momentum. This review aims to provide an overview of the structures and antibacterial activity of various 4(3H)-quinazolinone derivatives covering various aspects of in vitro and in vivo pharmacological activities and structure activity relationships (SARs).

Phyto-assisted synthesis of zinc oxide nanoparticles using Cassia alata and its antibacterial activity against Escherichia coli.

Zinc oxide, an established inorganic metal oxide in nanoparticles form exhibits tremendous anti-bacterial activity. The present study focuses on determining the anti-bacterial activity of green synthesized zinc oxide nanoparticles (ZnO NPs). Results clearly validate the effective synthesis of spherical shaped nanoparticles with average size range of 60-80 nm. SEM and EDAX data buttresses the results obtained by XRD pattern in terms of size and purity. ZnO NPs exhibited dose-dependent anti-bacterial activity against Escherichia coli (E. coli) and the IC50 value was calculated to be around 20 µg/mL. Growth kinetics study was conducted in the presence of nanoparticles which demonstrated the bacteriostatic effect of ZnO NPs. The study recommends the potential use of ZnO NPs in industries like food, pharmaceutical, agriculture, cosmetic industries for its anti-bacterial activity.

Biosynthesis of zinc oxide nanoparticles usingMangifera indica leaves and evaluation of their antioxidant and cytotoxic properties in lung cancer (A549) cells.

Green synthesis is an eco-friendly approach to nanoparticle production, which eliminates the use of toxic chemicals, high temperatures, and costly equipment needed for traditional physical and chemical synthesis methods. This eco-friendly approach was used in the present study to biosynthesize zinc oxide nanoparticles (ZnO NPs) from Mangifera indica (mango) leaves which were then evaluated for their antioxidant activity and cytotoxic effects on lung cancer A549 cells. Synthesized ZnO nanoparticles were characterized using UV-vis spectroscopy, XRD, SEM, and EDX analyses. The XRD and SEM analyses showed 45-60 nm as the size of synthesized nanoparticles, the pure crystal form of ZnO NPs, and the shape of the NPs as nearly spherical and hexagonal quartzite. The antioxidant potential of nanoparticles was estimated using a DPPH free radical scavenging assay. The percent of viable cells was inversely proportional to the concentration of ZnO nanoparticles at 25 µg/mL concentration. The MTT assay used for cytotoxicity evaluation depicted the significant cytotoxic effect of ZnO NPs against the A549 lung cancer cell line. The drop in the proportion of viable A549 cells after exposure to ZnO NPs was comparable to the effects of the standard drug used i.e. cyclophosphamide. Antioxidant activity of NPs was increased by increasing the concentration of NPs. The present biosynthesis approach is rapid, inexpensive and eco-friendly and it yielded highly stable ZnO NPs with significant antioxidant and anticancer potential. This is the first report of M. indicia -mediated synthesis of ZnO NPs as antioxidant and, anticancer agents for the treatment of lung cancer and subsequent therapeutic applications.

High Throughput Screening for Natural Host Defense Peptide-Inducing Compounds as Novel Alternatives to Antibiotics.

A rise in antimicrobial resistance demands novel alternatives to antimicrobials for disease control and prevention. As an important component of innate immunity, host defense peptides (HDPs) are capable of killing a broad spectrum of pathogens and modulating a range of host immune responses. Enhancing the synthesis of endogenous HDPs has emerged as a novel host-directed antimicrobial therapeutic strategy. To facilitate the identification of natural products with a strong capacity to induce HDP synthesis, a stable macrophage cell line expressing a luciferase reporter gene driven by a 2-Kb avian ß-defensin 9 (AvBD9) gene promoter was constructed through lentiviral transduction and puromycin selection. A high throughput screening assay was subsequently developed using the stable reporter cell line to screen a library of 584 natural products. A total of 21 compounds with a minimum Z-score of 2.0 were identified. Secondary screening in chicken HTC macrophages and jejunal explants further validated most compounds with a potent HDP-inducing activity in a dose-dependent manner. A follow-up oral administration of a lead natural compound, wortmannin, confirmed its capacity to enhance the AvBD9 gene expression in the duodenum of chickens. Besides AvBD9, most other chicken HDP genes were also induced by wortmannin. Additionally, butyrate was also found to synergize with wortmannin and several other newly-identified compounds in AvBD9 induction in HTC cells. Furthermore, wortmannin acted synergistically with butyrate in augmenting the antibacterial activity of chicken monocytes. Therefore, these natural HDP-inducing products may have the potential to be developed individually or in combinations as novel antibiotic alternatives for disease control and prevention in poultry and possibly other animal species including humans.