Evaluation of mosquito larvicidal activity of green synthesized crystalline silver nanoparticles using leave and fruit extracts of Phyllanthus acidus L. (Phyllanthaceae)

The present study evaluates the potentiality of green silver nanoparticles from the leaves and fruit extracts of Phyllanthus acidus L. against third instar larvae of two vector mosquitoes namely Culex quinquefasciatus and Culex vishnui. Various spectroscopic techniques were used to characterize the synthesized silver nanoparticles (AgNPs). Synthesized silver nanoparticles from both the leaf and fruits of P. acidus were spherical to quasi-spherical in shape and showed Surface Plasmon Resonance (SPR) bands at 420 and 409 nm respectively. In larvicidal bioassay with synthesized AgNPs from the leave of P. acidus, 100% mortality was observed at 20 ppm against third instar larvae of both the mosquito species with LC50 values of 1.64 and 0.87 ppm respectively. 100% mortality was observed in 5 ppm concentration against both Cx. quinquefasciatus and Cx. vishnui in synthesized AgNPs from fruits of P. acidus with LC50 values of 1.62 and 1.24 ppm respectively. The above findings suggest that the AgNPs synthesized from P. acidus leaves and fruit extracts have the potential to be employed in vector mosquito population control

Adipocyte fetuin-A contributes to macrophage migration into adipose tissue and polarization of macrophages

Macrophage infiltration into adipose tissue during obesity and their phenotypic conversion from anti-inflammatory M2 to proinflammatory M1 subtype significantly contributes to develop a link between inflammation and insulin resistance; signaling molecule(s) for these events, however, remains poorly understood. We demonstrate here that excess lipid in the adipose tissue environment may trigger one such signal. Adipose tissue from obese diabetic db/db mice, high fat diet-fed mice, and obese diabetic patients showed significantly elevated fetuin-A (FetA) levels in respect to their controls; partially hepatectomized high fat diet mice did not show noticeable alteration, indicating adipose tissue to be the source of this alteration. In adipocytes, fatty acid induces FetA gene and protein expressions, resulting in its copious release. We found that FetA could act as a chemoattractant for macrophages. To simulate lipid-induced inflammatory conditions when proinflammatory adipose tissue and macrophages create a niche of an altered microenvironment, we set up a transculture system of macrophages and adipocytes; the addition of fatty acid to adipocytes released FetA into the medium, which polarized M2 macrophages to M1. This was further confirmed by direct FetA addition to macrophages. Taken together, lipid-induced FetA from adipocytes is an efficient chemokine for macrophage migration and polarization. These findings open a new dimension for understanding obesity-induced inflammation

Ancestral Variations of the PCDHG Gene Cluster Predispose to Dyslexia in a Multiplex Family

Dyslexia is a heritable neurodevelopmental disorder characterized by difficulties in reading and writing. In this study, we describe the identification of a set of 17 polymorphisms located across 1.9 Mb region on chromosome 5q31.3, encompassing genes of the PCDHG cluster, TAF7, PCDH1 and ARHGAP26, dominantly inherited with dyslexia in a multi-incident family. Strikingly, the non-risk form of seven variations of the PCDHG cluster, are preponderant in the human lineage, while risk alleles are ancestral and conserved across Neanderthals to non-human primates. Four of these seven ancestral variations (c.460A > C [p.Ile154Leu], c.541G > A [p.Ala181Thr], c.2036G > C [p.Arg679Pro] and c.2059A > G [p.Lys687Glu]) result in amino acid alterations. p.Ile154Leu and p.Ala181Thr are present at EC2: EC3 interacting interface of γA3-PCDH and γA4-PCDH respectively might affect trans-homophilic interaction and hence neuronal connectivity. p.Arg679Pro and p.Lys687Glu are present within the linker region connecting trans-membrane to extracellular domain. Sequence analysis indicated the importance of p.Ile154, p.Arg679 and p.Lys687 in maintaining class specificity. Thus the observed association of PCDHG genes encoding neural adhesion proteins reinforces the hypothesis of aberrant neuronal connectivity in the pathophysiology of dyslexia. Additionally, the striking conservation of the identified variants indicates a role of PCDHG in the evolution of highly specialized cognitive skills critical to reading

Vapor of Volatile Oils from <em>Litsea cubeba</em> Seed Induces Apoptosis and Causes Cell Cycle Arrest in Lung Cancer Cells

<div><p>Non-small cell lung carcinoma (NSCLC) is a major killer in cancer related human death. Its therapeutic intervention requires superior efficient molecule(s) as it often becomes resistant to present chemotherapy options. Here we report that vapor of volatile oil compounds obtained from <em>Litsea cubeba</em> seeds killed human NSCLC cells, A549, through the induction of apoptosis and cell cycle arrest. Vapor generated from the combined oils (VCO) deactivated Akt, a key player in cancer cell survival and proliferation. Interestingly VCO dephosphorylated Akt at both Ser⁴⁷³ and Thr³⁰⁸; through the suppression of mTOR and pPDK1 respectively. As a consequence of this, diminished phosphorylation of Bad occurred along with the decreased Bcl-xL expression. This subsequently enhanced Bax levels permitting the release of mitochondrial cytochrome c into the cytosol which concomitantly activated caspase 9 and caspase 3 resulting apoptotic cell death. Impairment of Akt activation by VCO also deactivated Mdm2 that effected overexpression of p53 which in turn upregulated p21 expression. This causes enhanced p21 binding to cyclin D1 that halted G1 to S phase progression. Taken together, VCO produces two prong effects on lung cancer cells, it induces apoptosis and blocked cancer cell proliferation, both occurred due to the deactivation of Akt. In addition, it has another crucial advantage: VCO could be directly delivered to lung cancer tissue through inhalation.</p> </div

Time dependent inhibition of Akt phosphorylation by VCO.

<p>(<b>A</b>, <b>B</b>) Immunoblot analysis of Akt phosphorylation at Thr³⁰⁸ (A) and Ser⁴⁷³ (B) in A549 treated cells with VCO for the indicated time period (upper panel). Fold change represents the protein level of the VCO treated cells relative to the control cells. Bands were quantified by densitometric analysis where pAkt level was then normalized to the total Akt level (lower panel). β-actin served as loading control. (<b>C</b>, <b>D</b>) Immunoblot analysis of pPDK1 Ser ²⁴¹ (C) and mTOR (D) was done at different time hour (0 h, 12 h, 24 h, 36 h) exposure of VCO to A549 cells (upper panel). Bands were quantified by densitometric analysis where pPDK1 or mTOR level was then normalized with β-actin which is represented by folds change (lower panel). Figures are representative of three independent experiments, *p<0.01, **p<0.001 versus control (0 h).</p

VCO induces apoptosis in A549 lung cancer cells.

<p>(<b>A</b>) Annexin-Cy3 (red) and 6-CFDA (green) double staining of apoptotic cells was examined by fluorescence microscopy where VCO treated A549 cells showed both green and red stains and control (untreated) cells stained green only. (<b>B</b>) Percentage of apoptotic A549 cells was measured at different time points (0 h, 12 h, 24 h, 36 h) with VCO treatments. (<b>C</b>) Mitochondrial membrane potential was observed in control and VCO exposed (36 h) A549 lung cancer cells by JC-1 staining assay. (<b>D</b>) Apoptotic DNA fragmentation was observed by VCO treated A-549 cells on 1.5% agarose gel electrophoresis. Data are presented as means ± SEM of three independent experiments. *p<0.05, **p<0.01 versus control (0 h). Bar represents 20 µm.</p

Effect of VCO on the viability of A549 cells by MTT assay.

<p>(<b>A</b>) Cell viability of A549 lung cancer cells were measured when exposed to vapors of different dilutions (10⁶ to 10²) of crude oil for 72 h by using MTT assay and the data was expressed as % of cell survivability relative to control. (<b>B</b>) Chemical structures of four most available compounds (C1- Citronellal; C2- neo-isopulegol; C3- isopulegol; C4- citronellol) isolated from <i>Litsea cubeba</i> seed essential oil. (<b>C</b>) Percentage of cell death was observed when A549 cells were exposed individually with these compounds for 72 h. (<b>D</b>) Effect of VCO (C2∶C3∶C4 as 1∶1∶1) and C4 on cell death at 72 h was observed by MTT assay, which was visualized by microscopic images. (<b>E</b>) Cell survivability was measured at different time intervals (24, 48, 72 h) with VCO exposure on A549 cells. (<b>F</b>) Western blot of Akt phosphorylation at Thr³⁰⁸, Ser⁴⁷³ and total Akt in A549 cells treated without (Con) with C1, C2, C3, C4 and VCO for 36 hours. β-actin served as internal loading control. Values are means ± SEM of 3 individual experiments. *p<0.05, **p<0.01 versus control and #p<0.05 versus C4.</p

VCO induces apoptotic cell death by activating caspase cascade.

<p>(<b>A</b>) A549 cells were exposed with VCO for 36 h followed by staining of mitochondria with Mitotracker (red) and cytochrome c with FITC conjugated anti-cytochrome c antibody (green). (<b>B</b>) Immunoblot analysis was done by using anti-cleaved caspase-9 or caspase-3 antibodies in A-549 cells incubated in the presence of VCO at 0 h, 24 h, 36 h time intervals. β-actin used as internal control. (<b>C</b>) A549 cells were exposed with VCO for indicated time periods and on termination of exposure, cells were lysed and caspase 3 activity was measured in DTX multimode detector by using proluminescent caspase 3 as the substrate. (<b>D</b>) PARP cleavage was observed in VCO exposed cells by immunoblot analysis using anti-PARP antibody. β-actin used as loading control. Values are means ± SEM of three independent experiments, *p<0.01, **p<0.001 versus control (0 h). Bar represents 20 µm.</p