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Introduction: Bacterial resistance to multiple drugs is increasing at an alarming rate in current era and nanotechnology is one of the effective and novel approaches to overcome drug resistance. Methods: Zinc Oxide Nanoparticles (ZnO NPs) has stronger antibacterial activity and is regarded as bio-safe nanomaterial. The aim of present study is to synthesize the ZnO NPs using Aloe vera leaves extract and to investigate the synergistic effects and antioxidant actions of bio-synthesized ZnO NPs against gram negative bacteria E.coli and K. pneumoniae. The synergistic effect of β-lactam antibiotics (meropenem and ciprofloxacin) was tested along with ZnO NPs by using Kirby’s disc diffusion assay. The antioxidant activity was investigated by α, α-diphenyl-β-picrylhydrazyl (DPPH) method. Results: Results revealed that the antibacterial activity of the selected antibiotics was much enhanced by ZnO NPs than the antibiotics alone. The resistant antibiotic (ciprofloxacin) became sensitive when combined with ZnO NPs. The antioxidant activity reveals that biosynthesized ZnO NPs possess significantly higher (p Conclusion: The findings reveal that biosynthesized ZnO NPs have much more eco-friendly approach. It can act as a strong potentiator of β-lactam antibiotics and put forward the possibility to use them effectively in targeted drug delivery, pharmaceuticals and biomedical fields.

Breast cancer is a major cause of cancer related deaths in women worldwide. Available treatments pose serious limitations such as systemic toxicity, metastasis, tumor recurrence, off-target effects, and drug resistance. In recent years, phytochemicals such as secondary metabolites due to their effective anticancer potential at very low concentration have gained attention. Aim of the study was to evaluate anticancer potential of Citrullus colocynthis and its possible molecular targets on MCF-7, a human breast cancer cell line. Methanolic extract of leaves was prepared and fractionated by solvents (n-hexane, chloroform, ethyl acetate and n-butanol) with increasing polarity. Bioassays and gene expression regulation was conducted to evaluate the anticancer activity, proliferation rate and cell cycle regulation of breast cancer cells treated with extract and its fractions, separately. Results showed a significant anticancer activity of methanolic extract of C. colocynthis and two of its fractions prepared with chloroform and ethyl acetate. Bioassays depicted significant decrease in proliferation and growth potential along with cell cycle arrest of treated cells compared to control untreated cells. Expression regulation of genes further confirmed the cell cycle arrest through significant upregulation of cyclin-CDK inhibitors (p21 and p27) and cell cycle checkpoint regulators (HUS1, RAD1, ATM) followed by downregulation of downstream cell cycle progression genes (Cyclin A, Cyclin E, CDK2). It is concluded that C. colocynthis arrests cell cycle in human breast cancer cells through expression regulation of cyclin-CDK inhibitors and with further research can be proposed for therapeutic interventions.

Nanotechnology-based strategies can be useful tools to combat the alarming challenge of antibacterial resistance. Nanoparticles can be used as alternatives or supplementary in enhancing the effectiveness of the available drugs to control bacterial proliferation. Here, we evaluated the vibrant anti-proliferative role of silver nanoparticle (AgNPs; 31 ± 5 nm) against fish pathogen Pseudomonas aeruginosa (P. aeruginosa) to inhibit the infection in Labeo rohita. Fish was divided into four groups: control untreated (Con), 10 mg/l AgNPs exposed (NPs), P. aeruginosa challenged (Inf) and combinedly treated with AgNPs and P. aeruginosa (NPs + Inf). After 14 days of treatment changes in different bio-indicators including hemoglobin hematological parameters (hemoglobin, total erythrocyte count, hematocrit, total leukocyte count), biochemical parameters (total protein, albumin, globulin), oxidative biomarkers [malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT)] and histological parameters of kidney and liver were observed. We found that infected fish (Inf) showed more symptoms of infection as compared to fish given combined treatment of AgNPs and P. aeruginosa (NPs + Inf). Aforementioned bio-indicators in AgNPs-exposed group (NPs) were comparable to control, showing that AgNPs were not toxic at this concentration. Our results thus show that AgNPs hindered the bacterial proliferation in challenged fish. This was further confirmed by CFU assays of muscle tissues collected from Inf and NPs + Inf groups. The finding of this study suggested AgNPs as potential candidates to prevent bacterial proliferation which could be beneficial for biomedical applications.

In the present study effects of a commercial probiotic, Ecotec on growth, thyroid hormone profile, hematology, immuno-biochemical responses, monocyte morphology and survival against pathogenic challenge in Labeo rohita juveniles have been investigated. Two different concentrations of the probiotic (t1: 8 × 107 CFU/g feed and t2: 16 × 107 CFU/g feed) were administered to the fish. Probiotic treated fish showed a high growth increment as compared to the control. Level of thyroid hormones T3 and T4 was found to be lowered while TSH level was high in probiotic treated groups. Total Leukocyte Count was significantly low in the probiotic supplemented groups. The Differential Leukocyte Count showed abundance of lymphocytes and monocytes in the control group while neutrophils and basophils were abundant in the probiotic treated groups. Biochemical parameters (Albumin and Globulin) were high in probiotic fed groups. Immunological paraameters (Super oxide dismutase, Catalase, Myeloperoxidase and Lysozyme) were significantly high in probiotic fed fish. Monocyte morphology in control group showed a pronounced phagocytic activity in comparison to probiotic treated groups. Infection with Aeromonas hydrophila showed that probiotic treated fish had a better survival rate. In conclusion Ecotec supplementation enhanced growth and improved immunity in Labeo rohita juveniles.

The present study aims to assess the induced nanotoxicity of arsenic nanoparticles (AsNPs) on different organs of fresh water fish Labeo rohita. AsNPs were synthesized by chemical reduction method using sodium arsenite as precursor, ice-cold sodium borohydride as reducing agent, and sodium hydroxide to adjust the solution pH. The synthesized AsNPs were characterized by UV-Vis spectroscopy, X-ray diffraction (XRD), and scanning electron microscopy (SEM) for optical, structural, and morphological investigations. The UV-Vis absorption peaks occurring at around 300 nm indicated the presence of AsNPs in colloidal sample. The rhombohedral crystalline nature and metallic purity of AsNPs with crystallite size of 30 ± 1 nm were confirmed by characteristic peaks of XRD pattern. The SEM micrograph revealed the almost spherical shape and 40 ± 10 nm average size prepared AsNPs. For assessment of induced nanotoxicity, juveniles of Labeo rohita (L. rohita) were exposed to three different concentrations of AsNPs (namely, 1, 10, and 20 mg/L) for 30 days (n = 15 per group), and the control fish was kept untreated. It was observed that the routine behavior activities (such as swimming, mutual interactions, and feed intake) were affected by AsNPs. The growth of AsNPs treated fish was found retarded as compared to the control fish. Total erythrocyte count, total leukocyte count, and hemoglobin and hematocrit values were low in the AsNPs treated fish. Immunobiochemical assays revealed that protein level was altered in the AsNPs treated fish. The levels of antioxidant enzymes catalase and superoxide dismutase were low in the treated fish. The histological alteration induced by AsNPs in liver, gills, and kidneys demonstrated the damage in form of glomerulus shrinkage, vacuolation, inflammation, necrosis, lamellar disorganization, and hemorrhage in comparison with untreated fish. The results of the present study indicate that AsNPs exposure causes behavior, growth, hematology, immunobiochemical, and histological shortcomings in L. rohita.

Macrophages are phagocytic cells from the innate immune system, which forms the first line of host defense against invading pathogens. These highly dynamic immune cells can adopt specific functional phenotypes, with the pro-inflammatory M1 and anti-inflammatory M2 polarization states as the two extremes. Recently, the process of macrophage polarization during inflammation has been visualized by real time imaging in larvae of the zebrafish. This model organism has also become widely used to study macrophage responses to microbial pathogens. To support the increasing use of zebrafish in macrophage biology, we set out to determine the complete transcriptome of zebrafish larval macrophages. We studied the specificity of the macrophage signature compared with other larval immune cells and the macrophage-specific expression changes upon infection. We made use of the well-established mpeg1, mpx, and lck fluorescent reporter lines to sort and sequence the transcriptome of larval macrophages, neutrophils, and lymphoid progenitor cells, respectively. Our results provide a complete dataset of genes expressed in these different immune cell types and highlight their similarities and differences. Major differences between the macrophage and neutrophil signatures were found within the families of proteinases. Furthermore, expression of genes involved in antigen presentation and processing was specifically detected in macrophages, while lymphoid progenitors showed expression of genes involved in macrophage activation. Comparison with datasets of in vitro polarized human macrophages revealed that zebrafish macrophages express a strongly homologous gene set, comprising both M1 and M2 markers. Furthermore, transcriptome analysis of low numbers of macrophages infected by the intracellular pathogen Mycobacterium marinum revealed that infected macrophages change their transcriptomic response by downregulation of M2-associated genes and overexpression of specific M1-associated genes. Among the infection-induced genes, a homolog of the human CXCL11 chemokine gene, cxcl11aa, stood out as the most strongly overexpressed M1 marker. Upregulation of cxcl11aa in Mycobacterium-infected macrophages was found to require the function of Myd88, a critical adaptor molecule in the Toll-like and interleukin 1 receptor pathways that are central to pathogen recognition and activation of the innate immune response. Altogether, our data provide a valuable data mining resource to support infection and inflammation research in the zebrafish model.

Macrophages are phagocytic cells from the innate immune system, which forms the first line of host defense against invading pathogens. These highly dynamic immune cells can adopt specific functional phenotypes, with the pro-inflammatory M1 and anti-inflammatory M2 polarization states as the two extremes. Recently, the process of macrophage polarization during inflammation has been visualized by real time imaging in larvae of the zebrafish. This model organism has also become widely used to study macrophage responses to microbial pathogens. To support the increasing use of zebrafish in macrophage biology, we set out to determine the complete transcriptome of zebrafish larval macrophages. We studied the specificity of the macrophage signature compared with other larval immune cells and the macrophage-specific expression changes upon infection. We made use of the well-established mpeg1, mpx, and lck fluorescent reporter lines to sort and sequence the transcriptome of larval macrophages, neutrophils, and lymphoid progenitor cells, respectively. Our results provide a complete dataset of genes expressed in these different immune cell types and highlight their similarities and differences. Major differences between the macrophage and neutrophil signatures were found within the families of proteinases. Furthermore, expression of genes involved in antigen presentation and processing was specifically detected in macrophages, while lymphoid progenitors showed expression of genes involved in macrophage activation. Comparison with datasets of in vitro polarized human macrophages revealed that zebrafish macrophages express a strongly homologous gene set, comprising both M1 and M2 markers. Furthermore, transcriptome analysis of low numbers of macrophages infected by the intracellular pathogen Mycobacterium marinum revealed that infected macrophages change their transcriptomic response by downregulation of M2-associated genes and overexpression of specific M1-associated genes. Among the infection-induced genes, a homolog of the human CXCL11 chemokine gene, cxcl11aa, stood out as the most strongly overexpressed M1 marker. Upregulation of cxcl11aa in Mycobacterium-infected macrophages was found to require the function of Myd88, a critical adaptor molecule in the Toll-like and interleukin 1 receptor pathways that are central to pathogen recognition and activation of the innate immune response. Altogether, our data provide a valuable data mining resource to support infection and inflammation research in the zebrafish model.

The current study aimed to evaluate lipid profile parameters in patients with chronic kidney disease (CKD). This case-control study was conducted at the Zoology Department of Lahore College for Women University, Lahore, Pakistan, from September 2016 to June 2017 and analysed lipid profiles of patients with CKD attending different hospitals of Lahore. Of the randomly collected 100 blood samples, 50 were healthy, while the remaining 50 were patients with CKD. Lipid profile parameters were analysed using Hitachi 704 Analyser. For data analysis Chi-square test was used by means of SSPS 17. Results showed that all the parameters, including high density lipoprotein, triglycerides and low density lipoprotein were considerably altered in patients with CKD as compared to the control group. The values of triglycerides and LDL parameters were higher, while HDL cholesterol was lower in patients with CKD as compared to controls.

Silver nanoparticles (AgNPs) of different shapes and sizes were prepared by solution-based chemical reduction routes. Silver nitrate was used as a precursor, tri-sodium citrate (TSC) and sodium borohydride as reducing agents, while polyvinylpyrrolidone (PVP) was used as a stabilizing agent. The morphology, size, and structural properties of obtained nanoparticles were characterized by scanning electron microscopy (SEM), UV-visible spectroscopy (UV-VIS), and X-ray diffraction (XRD) techniques. Spherical AgNPs, as depicted by SEM, were found to have diameters in the range of 15 to 90 nm while lengths of the edges of the triangular particles were about 150 nm. The characteristic surface plasmon resonance (SPR) peaks of different spherical silver colloids occurring in the wavelength range of 397 to 504 nm, whereas triangular particles showed two peaks, first at 392 nm and second at 789 nm as measured by UV-VIS. The XRD spectra of the prepared samples indicated the face-centered cubic crystalline structure of metallic AgNPs. The in vitro antibacterial properties of all synthesized AgNPs against two types of Gram-negative bacteria, Pseudomonas aeruginosa and Escherichia coli were examined by Kirby–Bauer disk diffusion susceptibility method. It was noticed that the smallest-sized spherical AgNPs demonstrated a better antibacterial activity against both bacterial strains as compared to the triangular and larger spherical shaped AgNPs.

The zebrafish (Danio rerio) is increasingly used as a model for studying infectious diseases. This nonmammalian vertebrate host, which is transparent at the early life stages, is especially attractive for live imaging of interactions between pathogens and host cells. A number of useful fluorescent reporter lines have recently been developed and significant advances in RNA sequencing technology have been made, which now make it possible to apply the zebrafish model for investigating changes in transcriptional activity of specific immune cell types during the course of an infection process.Here we describe how to sequence RNA extracted from fluorescently labeled macrophages obtained by cell-sorting of 5-day-old zebrafish larvae of the transgenic Tg(mpeg1:Gal4-VP16);Tg(UAS-E1b:Kaede) line. This technique showed reproducible results and allowed to detect specific expression of macrophage markers in the mpeg1 positive cell population, whereas no markers specific for neutrophils or lymphoid cells were detected. This protocol has been also successfully extended to other immune cell types as well as cells infected by Mycobacterium marinum.

Deficiency in Src homology region 2 domain-containing phosphatase 1/protein tyrosine phosphatase nonreceptor type 6 (SHP1/PTPN6) is linked with chronic inflammatory diseases and hematological malignancies in humans. In this study, we exploited the embryonic and larval stages of zebrafish (Danio rerio) as an animal model to study ptpn6 function in the sole context of innate immunity. We show that ptpn6 knockdown induces a spontaneous inflammation-associated phenotype at the late larval stage. Surprisingly, glucocorticoid treatment did not suppress inflammation under ptpn6 knockdown conditions but further enhanced leukocyte infiltration and proinflammatory gene expression. Experiments in a germ-free environment showed that the late larval phenotype was microbe independent. When ptpn6 knockdown embryos were challenged with Salmonella typhimurium or Mycobacterium marinum at earlier stages of development, the innate immune system was hyperactivated to a contraproductive level that impaired the control of these pathogenic bacteria. Transcriptome analysis demonstrated that Kyoto Encyclopedia of Genes and Genomes pathways related to pathogen recognition and cytokine signaling were significantly enriched under these conditions, suggesting that ptpn6 functions as a negative regulator that imposes a tight control over the level of innate immune response activation during infection. In contrast to the hyperinduction of proinflammatory cytokine genes under ptpn6 knockdown conditions, anti-inflammatory il10 expression was not hyperinduced. These results support that ptpn6 has a crucial regulatory function in preventing host-detrimental effects of inflammation and is essential for a successful defense mechanism against invading microbes.