Carotenoid Extraction from Plant Tissues.

Carotenoids are the natural pigments available in nature and exhibit different colors such as yellow, red, and orange. These are a class of phytonutrients that have anti-cancer, anti-inflammatory, anti-oxidant, immune-modulatory, and anti-aging properties. These were used in food, pharmaceutical, nutraceutical, and cosmetic industries. They are divided into two classes: carotenes and xanthophylls. The carotenes are non-oxygenated derivatives and xanthophylls are oxygenated derivatives. The major source of carotenoids are vegetables, fruits, and tissues. Carotenoids also perform the roles of photoprotection and photosynthesis. In addition to the roles mentioned above, they are also involved and act as precursor molecules for the biosynthesis of phytohormones such as strigolactone and abscisic acid. This chapter briefly introduces carotenoids and their extraction method from plant tissue. Proposed protocol describes the extraction of carotenoid using solvents chloroform and dichloromethane. Reverse-phase HPLC can be performed with C30 columns using gradient elution. The column C30 is preferred to the C18 column because the C30 column has salient features, which include selective nature in the separation of structural isomers and hydrophobic, long-chain compounds, and shows the best compatibility with highly aqueous mobile phases. A complete pipeline for the extraction of carotenoids from plant tissue is given in the present protocol.

Appearance of de Gennes length in force-induced transitions.

Using Langevin dynamic simulations, a simple coarse-grained model of a DNA protein construct is used to study the DNA rupture and the protein unfolding. We identify three distinct states: (i) zipped DNA and collapsed protein, (ii) unzipped DNA and stretched protein, and (iii) unzipped DNA and collapsed protein. Here, we find a phase diagram that shows these states depending on the size of the DNA handle and the protein. For a less stable protein, unfolding is solely governed by the size of the linker DNA, whereas if the protein's stability increases, complete unfolding becomes impossible because the rupture force for DNA has reached a saturation regime influenced by the de Gennes length. We show that unfolding occurs via a few intermediate states by monitoring the force-extension curve of the entire protein. We extend our study to a heterogeneous protein system, where similar intermediate states in two systems can lead to different protein unfolding paths.

Septum site placement in Mycobacteria - identification and characterisation of mycobacterial homologues of Escherichia coli MinD.

A major virulence trait of Mycobacterium tuberculosis (M. tb) is its ability to enter a dormant state within its human host. Since cell division is intimately linked to metabolic shut down, understanding the mechanism of septum formation and its integration with other events in the division pathway is likely to offer clues to the molecular basis of dormancy. The M. tb genome lacks obvious homologues of several conserved cell division proteins, and this study was aimed at identifying and functionally characterising mycobacterial homologues of the E. coli septum site specification protein MinD (Ec MinD). Sequence homology based analyses suggested that the genomes of both M. tb and the saprophyte Mycobacterium smegmatis (M. smegmatis) encode two putative Ec MinD homologues - Rv1708/MSMEG_3743 and Rv3660c/ MSMEG_6171. Of these, Rv1708/MSMEG_3743 were found to be the true homologues, through complementation of the E. coli ∆minDE mutant HL1, overexpression studies, and structural comparisons. Rv1708 and MSMEG_3743 fully complemented the mini-cell phenotype of HL1, and over-expression of MSMEG_3743 in M. smegmatis led to cell elongation and a drastic decrease in c.f.u. counts, indicating its essentiality in cell-division. MSMEG_3743 displayed ATPase activity, consistent with its containing a conserved Walker A motif. Interaction of Rv1708 with the chromosome associated proteins ScpA and ParB, implied a link between its septum formation role, and chromosome segregation. Comparative structural analyses showed Rv1708 to be closer in similarity to Ec MinD than Rv3660c. In summary we identify Rv1708 and MSMEG_3743 to be homologues of Ec MinD, adding a critical missing piece to the mycobacterial cell division puzzle.

Entropy by Neighbor Distance as a New Measure for Characterizing Spatiotemporal Orders in Microscopic Collective Systems.

Collective systems self-organize to form globally ordered spatiotemporal patterns. Finding appropriate measures to characterize the order in these patterns will contribute to our understanding of the principles of self-organization in all collective systems. Here we examine a new measure based on the entropy of the neighbor distance distributions in the characterization of collective patterns. We study three types of systems: a simulated self-propelled boid system, two active colloidal systems, and one centimeter-scale robotic swarm system. In all these systems, the new measure proves sensitive in revealing active phase transitions and in distinguishing steady states. We envision that the entropy by neighbor distance could be useful for characterizing biological swarms such as bird flocks and for designing robotic swarms.

Polymer Translocation and Nanopore Sequencing: A Review of Advances and Challenges.

Various biological processes involve the translocation of macromolecules across nanopores; these pores are basically protein channels embedded in membranes. Understanding the mechanism of translocation is crucial to a range of technological applications, including DNA sequencing, single molecule detection, and controlled drug delivery. In this spirit, numerous efforts have been made to develop polymer translocation-based sequencing devices, these efforts include findings and insights from theoretical modeling, simulations, and experimental studies. As much as the past and ongoing studies have added to the knowledge, the practical realization of low-cost, high-throughput sequencing devices, however, has still not been realized. There are challenges, the foremost of which is controlling the speed of translocation at the single monomer level, which remain to be addressed in order to use polymer translocation-based methods for sensing applications. In this article, we review the recent studies aimed at developing control over the dynamics of polymer translocation through nanopores.

Self-Assembly of DNA-Grafted Colloids: A Review of Challenges.

DNA-mediated self-assembly of colloids has emerged as a powerful tool to assemble the materials of prescribed structure and properties. The uniqueness of the approach lies in the sequence-specific, thermo-reversible hybridization of the DNA-strands based on Watson-Crick base pairing. Grafting particles with DNA strands, thus, results into building blocks that are fully programmable, and can, in principle, be assembled into any desired structure. There are, however, impediments that hinder the DNA-grafted particles from realizing their full potential, as building blocks, for programmable self-assembly. In this short review, we focus on these challenges and highlight the research around tackling these challenges.

Dosimetric Influence of Acuros XB Dose-to-Medium and Dose-to-Water Reporting Modes on Carcinoma Cervix Using Intensity-Modulated Radiation Therapy and Volumetric RapidArc Technique.

Aim: We aimed to evaluate the dosimetric influence of Acuros XB (AXB) dose-to-medium (Dm) and dose-to-water (Dw) reporting mode on carcinoma cervix using intensity-modulated radiation therapy (IMRT) and RapidArc (RA) technique. Materials and Methods: A cohort of thirty patients cared for carcinoma cervix was retrospectively selected for the study. Plans were computed using analytical anisotropic algorithm (AAA), AXB-Dm, and AXB-Dw algorithms for dosimetric comparison. A paired t-test and Pitman-Morgan dispersion test were executed to appraise the difference in mean values and the inter-patient variability of the differences. Results: The dose-volume parameters were higher for AXB-Dw in contrast to AAA for IMRT and RA plans, excluding D98%, minimum dose to planning target volume (PTV) and rectum mean dose (RA). There was no systematic trend observed in dose-volume parameters for PTV and organs at risk (OARs) between AXB-Dm and AXB-Dw for IMRT and RA plans. The dose-volume parameters for target were higher for AXB-Dm in comparison to AAA in IMRT and RA plans, except D98% and minimum dose to PTV. Analysis envisaged less inter-patient variability while switching from AAA to AXB-Dm in comparison to those switching from AAA to AXB-Dw. Conclusions: The present study reveals the important difference between AAA, AXB-Dm, and AXB-Dw computations for cervix carcinoma using IMRT and RA techniques. The inter-patient variability and systematic difference in dose-volume parameters computed using AAA, AXB-Dm, and AXB-Dw algorithms present the possible impact on the dose prescription to PTV and their relative constraints to OARs for IMRT and RA techniques. This may help in the decision-making in clinic while switching from AAA to AXB (Dm or Dw) algorithm for cervix carcinoma using IMRT and RA techniques.

Microscopic Swarms: From Active Matter Physics to Biomedical and Environmental Applications.

Microscopic swarms consisting of, e.g., active colloidal particles or microorganisms, display emergent behaviors not seen in equilibrium systems. They represent an emerging field of research that generates both fundamental scientific interest and practical technological value. This review seeks to unite the perspective of fundamental active matter physics and the perspective of practical applications of microscopic swarms. We first summarize experimental and theoretical results related to a few key aspects unique to active matter systems: the existence of long-range order, the prediction and observation of giant number fluctuations and motility-induced phase separation, and the exploration of the relations between information and order in the self-organizing patterns. Then we discuss microscopic swarms, particularly microrobotic swarms, from the perspective of applications. We introduce common methods to control and manipulate microrobotic swarms and summarize their potential applications in fields such as targeted delivery, in vivo imaging, biofilm removal, and wastewater treatment. We aim at bridging the gap between the community of active matter physics and the community of micromachines or microrobotics, and in doing so, we seek to inspire fruitful collaborations between the two communities.

Light-driven carbon nitride microswimmers with propulsion in biological and ionic media and responsive on-demand drug delivery.

We propose two-dimensional poly(heptazine imide) (PHI) carbon nitride microparticles as light-driven microswimmers in various ionic and biological media. Their high-speed (15 to 23 micrometer per second; 9.5 ± 5.4 body lengths per second) swimming in multicomponent ionic solutions with concentrations up to 5 M and without dedicated fuels is demonstrated, overcoming one of the bottlenecks of previous light-driven microswimmers. Such high ion tolerance is attributed to a favorable interplay between the particle's textural and structural nanoporosity and optoionic properties, facilitating ionic interactions in solutions with high salinity. Biocompatibility of these microswimmers is validated by cell viability tests with three different cell lines and primary cells. The nanopores of the swimmers are loaded with a model cancer drug, doxorubicin (DOX), resulting in a high (185%) loading efficiency without passive release. Controlled drug release is reported under different pH conditions and can be triggered on-demand by illumination. Light-triggered, boosted release of DOX and its active degradation products are demonstrated under oxygen-poor conditions using the intrinsic, environmentally sensitive and light-induced charge storage properties of PHI, which could enable future theranostic applications in oxygen-deprived tumor regions. These organic PHI microswimmers simultaneously address the current light-driven microswimmer challenges of high ion tolerance, fuel-free high-speed propulsion in biological media, biocompatibility, and controlled on-demand cargo release toward their biomedical, environmental, and other potential applications.

Order and information in the patterns of spinning magnetic micro-disks at the air-water interface.

The application of the Shannon entropy to study the relationship between information and structures has yielded insights into molecular and material systems. However, the difficulty in directly observing and manipulating atoms and molecules hampers the ability of these systems to serve as model systems for further exploring the links between information and structures. Here, we use, as a model experimental system, hundreds of spinning magnetic micro-disks self-organizing at the air-water interface to generate various spatiotemporal patterns with varying degrees of order. Using the neighbor distance as the information-bearing variable, we demonstrate the links among information, structure, and interactions. We establish a direct link between information and structure without using explicit knowledge of interactions. Last, we show that the Shannon entropy by neighbor distances is a powerful observable in characterizing structural changes. Our findings are relevant for analyzing natural self-organizing systems and for designing collective robots.

Dosimetric validation of Acuros® XB algorithm for RapidArc™ treatment technique: A post software upgrade analysis.

AIM: To validate the Acuros® XB (AXB) algorithm in Eclipse treatment planning system (TPS) for RapidArc™ (RA) technique following the software upgrades. MATERIALS AND METHODS: A Clinac-iX (2300CD) linear accelerator and Eclipse TPS (Varian Medical System, Inc., Palo Alto, USA) was used for commissioning of AXB algorithm using a 6 megavolts photon beam. Percentage depth dose (PDD) and profiles for field size 2 cm × 2 cm, 4 cm × 4 cm, 6 cm × 6 cm, 10 cm × 10 cm, 20 cm × 20 cm, 30 cm × 30 cm to 40 cm × 40 cm were taken. AXB calculated PDDs and profiles were evaluated against the measured and analytical anisotropic algorithm (AAA)-calculated PDDs and profiles. Test sites recommended by American Association of Physicists in Medicine task group (AAPM TG)-119 recommendation were used for RA planning and delivery verification using AXB algorithm.Dosimetric analysis of AXB calculated data showed that difference between calculated and measured data for PDD curves were maximum <1% beyond the depth of dose maximum and computed profiles in central region matches with maximum <1% for all considered field sizes. Ion-chamber measurements showed that the average confidence limit (CLs) was 0.034 and 0.020 in high-gradient and 0.047 and 0.042 in low-gradient regions, respectively, for AAA and AXB calculated RA plans. Portal measurements show the average CLs were 2.48 and 2.58 for AAA and AXB-calculated RA plans, with gamma passing criteria of 3%/3 mm. CONCLUSIONS: AXB shows excellent agreement with measurements and AAA calculated data. The CLs were consistent with the baseline values published by TG-119. AXB algorithm has the potential to perform photon dose calculation with comparable fast calculation speed without negotiating the accuracy. AAPM TG-119 was successfully implemented to access the proper configuration of AXB algorithm following the TPS upgrade.

Dosimetric impact of Acuros XB on cervix radiotherapy using RapidArc technique: a dosimetric study.

BACKGROUND: Acuros XB (AXB) may predict better rectal toxicities and treatment outcomes in cervix carcinoma. The aim of the study was to quantify the potential impact of AXB computations on the cervix radiotherapy using the RapidArc (RA ) technique as compared to anisotropic analytical algorithm (AA) computations. MATERIALS AND METHODS: A cohort of 30 patients previously cared for cervix carcinoma (stages II-IIIB) was selected for the present analysis. The RA plans were computed using AA and AXB dose computation engines under identical beam setup and MLC pattern. RESULTS: There was no significant (p > 0.05) difference in D95% and D98% to the planning target volume (PTV); moreover, a significant (p < 0.05) rise was noticed for mean dose to the PTV (0.26%), D50% (0.26%), D2% (0.80%) and V110% (44.24%) for AXB computation as compared to AA computations. Further, AXB estimated a significantly (p < 0.05) lower value for maximum and minimum dose to the PTV. Additionally, there was a significant (p < 0.05) reduction observed in mean dose to organs at risk (OARs) for AXB computation as compared to AA, though the reduction in mean dose was non-significant (p > 0.05) for the rectum. The maximum difference observed was 4.78% for the rectum V50Gy, 1.72%, 1.15% in mean dose and 2.22%, 1.48% in D2% of the left femur and right femur, respectively, between AA and AXB dose estimations. CONCLUSION: For similar target coverage, there were significant differences observed between the AAA and AXB computations. AA underestimates the V50Gy of the rectum and overestimates the mean dose and D2% for femoral heads as compared to AXB. Therefore, the use of AXB in the case of cervix carcinoma may predict better rectal toxicities and treatment outcomes in cervix carcinoma using the RA technique.

Impact of acuros XB algorithm in deep-inspiration breath-hold (DIBH) respiratory techniques used for the treatment of left breast cancer.

AIM: To investigate the impact of Acuros XB (AXB) algorithm in the deep-inspiration breath-hold (DIBH) technique used for treatment of left sided breast cancer. BACKGROUND: AXB may estimate better lung toxicities and treatment outcome in DIBH. MATERIALS AND METHODS: Treatment plans were computed using the field-in-field technique for a 6 MV beam in two respiratory phases - free breathing (FB) and DIBH. The AXB-calculations were performed under identical beam setup and the same numbers of monitor units as used for AAA-calculation. RESULTS: Mean Hounsfield units (HU), mass density (g/cc) and relative electron density were -782.1 ±â€¯24.8 and -883.5 ±â€¯24.9; 0.196 ±â€¯0.025 and 0.083 ±â€¯0.032; 0.218 ±â€¯0.025 and 0.117 ±â€¯0.025 for the lung in the FB and DIBH respiratory phase, respectively. For a similar target coverage (p > 0.05) in the DIBH respiratory phase between the AXB and AAA algorithm, there was a slight increase in organ at risk (OAR) dose for AXB in comparison to AAA, except for mean dose to the ipsilateral lung. AAA predicts higher mean dose to the ipsilateral lung and lesser V20Gy for the ipsilateral and common lung in comparison to AXB. The differences in mean dose to the ipsilateral lung were 0.87 ±â€¯2.66 % (p > 0.05) in FB, and 1.01 ± 1.07% (p < 0.05) in DIBH, in V20Gy the differences were 1.76 ±â€¯0.83% and 1.71 ±â€¯0.82% in FB (p < 0.05), 3.34 ± 1.15 % and 3.24 ± 1.17 % in DIBH (p < 0.05), for the ipsilateral and common lung, respectively. CONCLUSION: For a similar target volume coverage, there were important differences between the AXB and AAA algorithm for low-density inhomogeneity medium present in the DIBH respiratory phase for left sided breast cancer patients. DIBH treatment in conjunction with AXB may result in better estimation of lung toxicities and treatment outcome.

Mechanical Coupling of Puller and Pusher Active Microswimmers Influences Motility.

Active self-propelled colloidal populations induce time-dependent three-dimensional fluid flows, which alter the rheological (viscoelastic) properties of their fluidic media. Researchers have also studied passive colloids mixed with bacterial suspensions to understand the hydrodynamic coupling between active and passive colloids. With recent developments in biological cell-driven biohybrid microswimmers, different type biological microswimmer (e.g., bacteria and algae) populations need to interact fluidically with each other in the same fluidic media, while such interactions have not been studied experimentally yet. Therefore, we report the swimming behavior of two opposite types of biological microswimmer (active colloid) populations: Chlamydomonas reinhardtii (C. reinhardtii) algae (puller-type microswimmers) population in coculture with Escherichia coli (E. coli) bacteria (pusher-type microswimmers) population. We observed noticeable fluidic coupling deviations from the existing understanding of passive colloids mixed with bacterial suspensions previously studied in the literature. The fluidic coupling among puller- and pusher-type microswimmers led to nonequilibrium fluctuations in the fluid flow due to their opposite swimming patterns. Such coupling could be the main reason behind the shift in motility behaviors of these two opposite-type swimmer populations suspended in the same fluidic media.

Multifunctional magnetic hairbot for untethered osteogenesis, ultrasound contrast imaging and drug delivery.

To explore novel materials graded for biological functions is one of the grand challenges and ambitions of robotics. In this study, the design, development, and external guidance of micron-sized hair-derived robots (hairbots) are shown as autologous cargo carriers for guided drug delivery, untethered osteogenesis, and sonographic contrast agents. Having biogenic origin, the hairbots show excellent biocompatibility, as demonstrated with cell adhesion, spreading and proliferation. External magnetic fields are used to enhance differentiation of mesenchymal stem cells (MSCs) into bone like cells, which can be used as magnetic therapy for bone healing. Effect of external magnetic forces was simulated by COMSOL Multiphysics® modelling software. The action of hairbots as osteoconductive material triggering osteogenic differentiations of MSCs is studied via calcium signaling by fluorescence microscopy. Further, by exploiting the hollow medullary region, the proposed hairbots are designed to perform theranostic dual functions (therapy + diagnostic) - as Doxorubicin drug delivery vehicle, and for Ultrasound contrast imaging. Harnessing sensing and actuation due to magnetic capabilities of hairbots, for enhanced biological functionality shown herein, provides a novel material in the search of new multifunctional microrobots.

Validation of the RapidArc Delivery System Using a Volumetric Phantom as Per Task Group Report 119 of the American Association of Physicists in Medicine.

AIM: This study validated the RapidArc (RA) delivery using a volumetric ArcCHECK phantom as per the guidelines proposed in Task Group Report 119 from the American Association of Physicists in Medicine Task group 119 (AAPM TG 119). This study also investigated the impact of the Acuros XB (AXB) algorithm in comparison to analytical anisotropic algorithm (AAA) on the RA dose calculations in the homogeneous medium of the ArcCHECK phantom. MATERIALS AND METHODS: A volumetric ArcCHECK phantom along with AAPM TG 119 tests was used to evaluate the RA plans and verify the dose delivery for photon beam of 6 MV energy. RESULTS: The RA planning results were comparable and satisfied the planning criteria stated in the TG 119 report for all test cases. The average percentage gamma passing rates for the AAA-calculated plans were 98.5 (standard deviation [SD]: 0.6), 98.5 (SD: 1.3), and 98.1 (SD: 2.0) and for the AXB-calculated plans were 95.1 (SD: 1.8), 96.1 (SD: 1.3), and 94.0 (SD: 0.9) for the Clinac-iX (6 MV) and TrueBeam (TB)-STx (6 MV_filtered beam [FB] and 6 MV_flattening filter-free beam [FFFB]), respectively. For ion chamber measurements, the average percentage dose differences for the AAA-calculated plans were 1.5 (SD: 2.5), 2.7 (SD: 1.4), and 1.4(SD: 2.7) and for AXB-calculated plans were 2.3 (SD: 1.6), 3.2 (SD: 1.5), and 2.3 (SD: 2.0) for Clinac-iX (6 MV) and TB-STx (6 MV_FB and 6 MV_FFFB), respectively. CONCLUSION: Thus, the ArcCHECK can successfully be utilized for the validation of the RA delivery. The AXB has potential to perform dose calculations comparable to those of the AAA for RA plans in the homogeneous medium of the ArcCHECK phantom.

Dosimetric Analysis of Unflattened (FFFB) and Flattened (FB) Photon Beam Energy for Gastric Cancers Using IMRT and VMAT-a Comparative Study.

PURPOSE: To evaluate the feasibility of flattening filter free beam (FFFB) for the treatment of gastric tumors and to review their benefits over 6MV flatten beam (6MV_FFB). METHODS: Fifteen patients with histologically proven gastric carcinoma were selected. CT scans with slice thickness of 0.3 cm were acquired and planning target volume (PTV) and organ at risk (OAR) were delineated. Plans were made retrospectively for each patient for the prescription dose of 45 Gy/25 fractions to the PTV. Four isocentric plans were compared in the present study on Varian TrueBeam linear accelerator (Varian Medical Systems, Palo Alto, CA, USA). RESULTS: PTV D98% was 44.41 ± 0.12, 44.38 ± 0.13, 44.59 ± 0.14, and 44.49 ± 0.19 Gy for IMRT 6MV_FFB, IMRT 6MV_FFFB, VMAT 6MV_FFB, and VMAT 6MV_FFFB respectively. 6MV_FFFB beam minimizes the mean heart dose Dmean (P = 0.001). VMAT dominates over IMRT when it came to kidney doses V12Gy (P = 0.02), V23Gy (P = 0.015), V28Gy (P = 0.011), and Dmax (P < 0.01). VMAT has significantly reduced the doses to kidneys. It was analyzed that 6MV_FFFB significantly reduces the dose to normal tissues (P = 0.006 and P = 0.018). VMAT significantly reduces the TMU, which is required to deliver the similar dose by IMRT (P < 0.01). CONCLUSIONS: Unflattened beam spares the organs at risk significantly to avoid the chances of secondary malignancies and reduces the intra-fraction motion during treatment due to provision of higher dose rate. Hence, we conclude that 6MV unflattened beam can be used to treat gastric carcinoma.

Graphene Oxide Synergistically Enhances Antibiotic Efficacy in Vancomycin-Resistant Staphylococcus aureus.

The current study highlights a new polyvalent inhibitor approach based on Vancomycin conjugated with graphene oxide (Van@GO) against a Vancomycin-resistant Staphylococcus aureus (VRSA) strain. Physicochemical characteristics of the prepared Van@GO composites were studied using UV-vis and FTIR spectroscopy techniques. Characterization results confirm the attachment of Vancomycin to the graphene oxide. A significant inhibition of VRSA growth is achieved by Vancomycin when presented as Van@GO. The polyvalent inhibition activity of Van@GO was characterized by performing bacteriological experiments along with scanning electron microscopy. Results clearly exhibit the enhanced inhibition activity of Van@GO compared to Vancomycin alone against VRSA. The high surface area of GO facilitates high loading and multivalent interaction of conjugated Vancomycin leading to polyvalent inhibition. Further, we found that Van@GO significantly reduces the motility of VRSA via inducing oxidative stress compared with untreated samples. Our findings highlight the importance of Van@GO as an effective polyvalent inhibition recipe for VRSA.

Cancer cells biomineralize ionic gold into nanoparticles-microplates via secreting defense proteins with specific gold-binding peptides.

Cancer cells have the capacity to synthesize nanoparticles (NPs). The detailed mechanism of this process is not very well documented. We report the mechanism of biomineralization of aqueous gold chloride into NPs and microplates in the breast-cancer cell line MCF7. Spherical gold NPs are synthesized in these cells in the presence of serum in the culture media by the reduction of HAuCl4. In the absence of serum, the cells exhibit gold microplate formation through seed-mediate growth albeit slower reduction. The structural characteristics of the two types of NPs under different media conditions were confirmed using scanning electron microscopy (SEM); crystallinity and metallic properties were assessed with transmission electron microscopy (TEM) and x-ray photoelectron spectroscopy (XPS). Gold-reducing proteins, related to cell stress initiate the biomineralization of HAuCl4 in cells (under serum free conditions) as confirmed by infrared (IR) spectroscopy. MCF7 cells undergo irreversible replicative senescence when exposed to a high concentration of ionic gold and conversely remain in a dormant reversible quiescent state when exposed to a low gold concentration. The latter cellular state was achievable in the presence of the rho/ROCK inhibitor Y-27632. Proteomic analysis revealed consistent expression of specific proteins under serum and serum-free conditions. A high-throughput proteomic approach to screen gold-reducing proteins and peptide sequences was utilized and validated by quartz crystal microbalance with dissipation (QCM-D). STATEMENT OF SIGNIFICANCE: Cancer cells are known to synthesize gold nanoparticles and microstructures, which are promising for bioimaging and other therapeutic applications. However, the detailed mechanism of such biomineralization process is not well understood yet. Herein, we demonstrate that cancer cells exposed to gold ions (grown in serum/serum-free conditions) secrete shock and stress-related proteins with specific gold-binding/reducing polypeptides. Cells undergo reversible senescence and can recover normal physiology when treated with the senescence inhibitor depending on culture condition. The use of mammalian cells as microincubators for synthesis of such particles could have potential influence on their uptake and biocompatibility. This study has important implications for in-situ reduction of ionic gold to anisotropic micro-nanostructures that could be used in-vivo clinical applications and tumor photothermal therapy.

Identification and functional annotation of mycobacterial septum formation genes using cell division mutants of Escherichia coli.

The major virulence trait of Mycobacterium tuberculosis is its ability to enter a latent state in the face of robust host immunity. Clues to the molecular basis of latency can emerge from understanding the mechanism of cell division, beginning with identification of proteins involved in this process. Using complementation of Escherichia coli mutants, we functionally annotated M. tuberculosis and Mycobacterium smegmatis homologs of divisome proteins FtsW and AmiC. Our results demonstrate that E. coli can be used as a surrogate model to discover mycobacterial cell division genes, and should prove invaluable in delineating the mechanisms of this fundamental process in mycobacteria.