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GTA V Related Footage Surfaces - Kind Of

"Okay, well, you’re probably looking at the title and freaking out.

You should be! But it’s not quite what you were expecting.

A few days ago, a mysterious Twitter feed called the Epsilon Program started tweeting things.

Strange things.

And these things are related to GTA V."

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Proeliator1492d ago

Well... that's interesting.

Derekvinyard131492d ago

Technically this is footage of the game, but not the kind we wanna see. Cool tho

Crazyglues1492d ago

I think the idea was all about GTA V having a full day and night cycle...

So maybe this time around their could be a real time of day aspect to the mission like in Mafia 1 where you could be on a mission in the day but get to the place where you needed to go and it's night time... (so mission could be spoiled if you had to be there by 6:00pm)

it was a really cool feature of Mafia because as you play it had a real time clock speed up. which was like an hour in the real world was like 3 hours in the game... something like that..

-So time actually was a big part of the game, looks like it might be a big factor in GTA V (I think that would be really cool) it was done really well in Mafia, and added to the overall fun of the game, because you had to stay on time. Get things done by a certain time, like get to restaurant by 3:00pm before they left.

So having a real-time day and night cycle could be really cool in GTA V

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guitarded771492d ago

I thought it was cool... allows us to see the dynamic environment and lighting. Looks pretty good. We'll get to see shiz get blowed up soon enough. It's better than nothing, which is what we would have if this video wasn't released today.

@ Crazyglues

I don't mean this in a bad way but when was the last time you played a GTA game? most if not all the stuff you talked about GTA has been doing for years now

inveni01491d ago (Edited 1491d ago )

Oops. Wrong thread. Sorry.

Crazyglues1491d ago (Edited 1491d ago )

yes I played them, I think I just explained it wrong, or people took it wrong, I guess they had to play Mafia 1 to understand what I was talking about...

but anyway this had nothing to do with time it was about some religion that Rockstar is making fun of in the game, so I was way off.. my bad, no big deal it happens..

They explain it here -

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That was the most drawn out way to promote timed missions XD.

+ Show (4) more repliesLast reply 1491d ago
TotalHitman1492d ago

"Everyone is related to everyone else, except for people with red hair - Fact!"

Haha, I love that bit.

1492d ago
Pushagree1492d ago

That was better than the entire Saints Row series combined.

1491d ago
Thantalas1491d ago (Edited 1491d ago )

It includes writing and pictures overlaid on top. It features the 12 Goals of Epsilonism. And a picture of the Epsilon Program's founder.

GuyThatPlaysGames1491d ago

Is this really from GTA5? Is this OFFICIAL footage?

Who knows... but it has the tell-tale signs of the RAGE engine. Could be authentic or a modded GTA4 considering the origins of this video were no doubt from a PC.

betty11barnhill1491d ago

Since I freelance I've been bringing in $90/h… I sit at home and I am doing my work from my laptop. The best thing is that I get more time to spent with my family and with my kids and in the same time I can earn enough to support them... You can do it too. Start here>>>>>>&g t;>>>>>>>& gt;

+ Show (5) more repliesLast reply 1487d ago
Donnieboi1492d ago (Edited 1492d ago )

Looks like a demo for the game engine.

play_the_station1492d ago

i've lurked N4G for a while and rarely do subheadings make it in the title lol, weird thing to point out..

footage looks great thoug, really excited for this!

Donnieboi1492d ago (Edited 1492d ago )

Nah, i'm just kidding. I don't think you can put ellipses (...) in a title on n4g. But the video doesn't really show much in terms of gameplay, looks more like a tech demo for the game engine.

Edit: @Play_the_station:
I changed my first comment, since I was only joking around, I don't want a big hoopla.

play_the_station1492d ago

lol I think it looks pretty nice, watching the day and night cycles has always been a weird pasttime of mine in GTA.... am probably in the minority here haha

contradictory1492d ago

i used to fly the planes in GTA SA
and just watch the days pass by
everytime the same radio tunes...

CaptainFaisal1492d ago

GTA V Rating! Agree for it will be awesome and you will buy it and disagree for if it will suck and you wont get it. Well for me its a must buy on Day 1 however ill buy it on PS4 mainly for graphics and no loading and better multiplayer features... You will know what i mean in 1-2 months cant reveal more :)

Proeliator1492d ago (Edited 1492d ago )

Should be interesting. also, it'll be a shit ton of fun to share shenanigans with the 'share' button!

PersonMan1492d ago

Who says it's coming out for the PS4? and also, I'm sure it will still have load times.

gigoran1492d ago

I like how you left out the "kind of" on the title and preview in order to get hits.

-Gespenst-1492d ago

The water and the clouds look amazing...

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Le Tombeau de... (2008)

Program notes:

The Tombeau is a French musical term from the 17th century describing a memorial piece. My title quite obviously references Ravel's "Le Tombeau de Couperin" and that in itself is homage to Ravel (the music doesn't reference him consciously); however, this piece serves as homage to many composers and musical traditions that have influenced me, and this piece.

The piece begins somberly and deliberately, but goes through a wide range of styles in presenting the various musical motifs of the piece.

These motifs are then freely developed, combined, and recapitulated. This piece was written for Tomo Matsuo.

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Photos: Youths Attack Yahaya Bello At APC Headquarter In Abuja

protest apc secretariat audu abubakar

Nov 30, 2015 – Pictures: Youths Attack Yahaya Bello At APC Secretariat In Abuja

Violence broke out at the APC secretariat in Abuja today as angry youths protest over the nomination of Yahaya Bello as late Audu Abubakar’s replacement.

It took the timely intervention of security agents to smuggle Bello from the angry mob.

Bello was nominated as the new candidate of the APC party after Abubakar’s death but the decision met with strong resistance, prompting the leading party to call an emergency meeting.


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November 23, 2015 By Divine Nwachukwu

Choosing a suitable case study for a research is an essential part of the process of carrying out a research or study. A case study simply put is that geographical location or kind of data that would make your research to be judged powerful. A research is said to be powerful if the case study of that research reflects the hypothesis for the topic under study. In choosing a suitable case study for your research or study, one has to consider:

The topic of the study/research.
In choosing a case study, a lot has to depend on the topic under study. A case study that is not in tandem to the topic may lead to an experiment whose conclusions and recommendation aren’t precise enough to be powerful and thus may lead to accepting a false null or alternative hypothesis. The topic of a study gives or shades light on the type of case study to employ in carrying out a research.

A good example is a topic that has to do with school performance or school environment, employing a case study that falls outside a school environment may actually lead to responses from people who aren’t experienced in academic matters, the consequence of this would simply be that generalizations reached  when the data is analysed may lead to rejection of an alternative hypothesis that should be accepted or acceptance of an alternative hypothesis that should be rejected thus affecting the power or precision of the  research.  So for selecting a suitable case study, a lot has to depend on the topic of the study.

The topic of a research also tells us if a primary data or secondary data will be required for precision of the research. Certain topics especially in econometrics or economics discipline would require secondary method of collecting data e.g. published data instead of the primary means of collecting data like the interview or questionnaire method.
In other disciplines especially social sciences, the topic of a study simply put determines the case study that best fits it. For instance:
For a topic that centers on performance of firms or employees, a suitable case study should or must be an organization and the best form of data collection is through the primary means like the interview method or questionnaire distribution.
In conclusion, the topic of the research best tells us the nature of our case study and the best way to generate the data required for a powerful or précised research.

You may also like: Making Awesome Presentations Developing Outstanding Research Topics How to Write an abstarct How to Write a Project Proposal How To Choose the Right Measurement Instrument

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Nanoscience NY

CUNY NY Skyline logo

Event Home

Speaker Biographies

Poster Abstracts

Organizing Committee

CUNY Advanced Science Research Center
85 St Nicholas Terrace,
New York, NY 10031
June 23rd and 24th 2016

Thursday June 24th 2016

Nano-tools and Bioinspired Tissue Engineering Approaches for the Regeneration of Different Tissues

Rui L. Reis University of Minho

This talk will describe several nano-tools and biomimetic approaches for the regeneration of different tissues. The selection of a proper material to be used as a scaffold or as a hydrogel, in many cases in combination with nanoparticles, to support, hold or encapsulate cells, as well as to control their differentiation, is both a critical and difficult choice. It will ultimately determine the success or failure of any tissue engineering and regenerative medicine (TERM) strategy.

We believe that the use of natural origin polymers is the best option for many different approaches that allow for the regeneration of different tissues. In addition to the selection of appropriate material systems it is of outmost importance the development of processing methodologies that allow for the production of adequate nano delivery systems and scaffolds/matrices.

Furthermore an adequate cell source should be selected. In many cases efficient cell isolation, expansion and differentiation methodologies should be developed and optimized. We have been using different human cell sources namely: mesenchymal stem cells from bone marrow and adipose tissue, cells from amniotic fluids and membranes and cells obtained from umbilical cords.

The potential of each type of cells, to be used to develop novel useful regeneration therapies will be discussed. Their uses and their interactions with different nano systems and natural origin degradable scaffolds and smart hydrogels will be described. Examples of the engineering of different tissues will be presented.

Supported Biomembrane Microenvironments of Controlled Composition for Gamma-Secretase Substrate Cleavage Assays

Lane Gilchrist Memorial Sloan-Kettering Cancer Center

The intramembrane protease gamma-secretase is a current target of therapeutic intervention, with pivotal pathological functions within Alzheimer's disease and cancer. Our primary objective is to expand methods for in vitro studies of the intramembrane proteases (IMPs) with the development of a microsphere-supported (proteolipobead) and planar biomembrane platforms. IMPs are found throughout all branches of life and their functions are extremely broad. Despite extensive studies, understanding of IMP regulation and catalysis has been hampered by membrane-associated enzymology. Rhomboids and Secretases are polytopic membrane proteases that are widely conserved in all organisms. The precise reaction mechanisms of the intermembrane proteases remain to be elucidated and furthermore, rigorous analysis of the kinetics of interfacial catalysis in these systems has not yet been undertaken.

High throughput of screening of drug candidates has been carried out in bulk assay systems using cell membrane fragments, solubilized enzymes and is underway in proteoliposomes. A critical barrier to further progress in the study and HTS of gamma-secretase is that such bulk systems do not allow for the direct in situ quantification of enzyme, substrates, or inhibitors or their relative distributions within the structures under assay. We have expanded in vitro models of gamma-secretase to include supported biomembranes, enabling: 1) characterization and verification of biomembrane loading of substrates, inhibitors and protein effectors, 2) studies of co-localization, phase-partitioning and lateral mobility of membrane-bound assay constituents, and 3) the development of flow cytometry-based assays of cleavage. In this work we have characterized the structures, phase localization and compositions of gamma-secretase proteolipobead and planar systems by employing correlative optical and surface microscopy from the micro- to the nanoscale. We have probed the formation of Lo domains in situ, monitored using 3D FRET phase detection. We have probed the phase partitioning of enzyme, substrates and cleavage products in PLB systems with superresolution microscopy. A combination of biomembrane mobility methods are being used to probe the diffusivities and mobile fractions of lipid, substrates and gamma-secretase. The results of phase partioning and diffusivity measurements are being integrated into bottom-up spatial diffusion-reaction models to correlate microenvironment with enzyme function. As new drug targets in cancer and infectious disease involved in regulated intramembrane proteolysis (RIP) based cell signaling are uncovered, it is expected that other IMP enzymes can be subsequently studied and probed at high throughput with this platform

Engraftment and Function of Human Pluripotent Stem Cell-Derived Hepatocyte-like Cells in Mice via 3D Co-aggregation and Encapsulation

Robert E. Schwartz Weill Cornell Medical College

Cellular therapies for liver diseases and in vitro models for drug testing both require functional human hepatocytes, which have unfortunately been limited due to the paucity of donor liver tissues. Human pluripotent stem cells represent a promising and potentially unlimited cell source to derive human hepatocytes. However, the hepatic functions of these human pluripotent stem cells-derived cells to date are not fully comparable to adult human hepatocytes and are more similar to fetal ones. In addition, it has been challenging to obtain functional hepatic engraftment of these cells with prior studies having been done inimmunocompromised animals. In this report, we demonstrated successful engraftment of human induced pluripotent stem cell derived hepatocyte-like cells in immunocompetent mice by pre-engineering 3D cell co-aggregates with stromal cells followed by encapsulation in recently developed biocompatible hydrogel capsules. Notably, upon transplantation, human albumin and ?1-antitrypsin (A1AT) in mouse sera secreted by encapsulated induced pluripotent stem cell derived hepatocyte-like cells/stromal cell aggregates reached a level comparable to the primary human hepatocyte/stromal cell control. Further immunohistochemistry of human albumin in retrieved cell aggregates confirmed the survival and function of iPS-H. This proof-of-concept study provides a simple yet robust approach to improve the engraftment of induced pluripotent stem cell derived hepatocyte-like cells, and may be applicable to many stem cell-based therapies.

Nanoparticle Based Analysis of Biomolecules, Cells and Tissue

Duncan Graham University of Strathclyde

Metallic nanoparticles offer many opportunities in terms of detection including light scattering, surface plasmon resonance and surface enhanced Raman scattering (SERS). We are interested in the optical properties of metal nanoparticles and their potential application in a range of different biological studies. We can make use of the optical properties of nanoparticles in two ways.

1. The nanoparticle can act as an extrinsic label for a specific biomolecular target in the same way as a fluorescent label is used. The advantage of using the nanoparticle is its optical brightness (typically several orders of magnitude more than fluorophores) and the lack of background vibrational signals. Functionalisation of the nanoparticle with a specific targeting species such as an antibody or peptide aptamer allows this approach to be used in a wide range of studies including cell, tissue and in vivo analysis.

2. Nanoparticles can be designed to contain a specific recognition probe designed to cause a change in the aggregation status of the nanoparticles resulting in a discernible optical change when it interacts with its biomolecular target. This allows separation free analysis of specific biomolecular interactions and can be applied to a range of different probe/target interactions such as DNA-DNA, peptide-protein and sugar-protein.

We have been making use of nanoparticles in both of these approaches in conjunction with SERS which is an advanced vibrational spectroscopy. To demonstrate the applicability of the two different approaches examples will be given on the use of nanoparticles for cell imaging in two and three-dimensions, imaging of nanoparticles in tissue and also their ability to report on biological molecules in vitro and in vivo.

Title TBD

Michelle S. Bradbury Memorial Sloan Kettering Cancer Center

Despite recent advances in imaging probe development for biomedicine, the translation of targeted diagnostic platforms remains challenging. Nanomaterials platforms currently under evaluation in oncology clinical trials are largely non-targeted drug delivery vehicles or devices to thermally treat tissue; these are typically not surface modified for targeted detection by clinical imaging tools. New tumor-selective platforms need to satisfy critical safety benchmarks, in addition to assaying targeted interactions with the microenvironment and their effects on biological systems. Coupled with metabolic imaging and analysis tools, such as PET, complete and quantitatively accurate data sets for whole body distributions, targeting kinetics, and clearance profiles of new diagnostic platforms undergoing preclinical testing or transitioning into early-phase clinical trials can be acquired.

In the operating theatre, there is also an urgent need for implementing new image-directed visualization tools that can enhance surgical vision, facilitate minimally invasive surgical procedures, and dramatically alter surgical outcomes of oncological patients. The lack of clear surgical vision impacts the ability of the operating surgeon to accurately and specifically identify the extent of malignancy, microscopic tumor burden, or remnant disease. Collectively, these factors affect therapeutic outcome, prognosis, and treatment management. Newer molecular imaging probe designs coupled with state-of-the-art device technologies, may enhance cancer care, provide real-time imaging guidance, and lead to new, more efficient approaches for early-stage detection and treatment.

Advances in nanotechnology have also fueled a paradigm shift in targeting and safely delivering drugs in conjunction with image-directed approaches. The size, architecture, and chemical composition of particle-based drug delivery vehicles can be fine-tuned to achieve properties optimal for loading and controlled release of therapeutic agents, patient safety/compliance, favorable kinetic profiles, and reducing unwanted side effects. By combining therapeutic particle tracer preparations with quantitative bioimaging approaches, drug delivery, lesion localization, and the extraction of key tumor biologic properties can be achieved for individualizing treatment planning. In turn, dosage regimens needed to achieve therapeutic efficacy might be estimated based on knowledge of drug specific activity and dose, uptake kinetics, and IC50 values.

The ability to flexibly adapt the formulation of clinically-promising drugs to improve their physicochemical and/or biological properties, in combination with metabolic imaging tools, will be important to quantify and establish suitable clinical trial endpoints. Issues relating to solubility, transport, barrier penetration, time-dependent changes in drug uptake, and intratumoral distribution are additional considerations. These properties are often not generally evaluated in the context of drug delivery due to the complexity of the biological systems involved and the inability to serially monitor this process non-invasively in the absence of drug labeling. The future success of molecular medicine will, in part, rest upon our ability to offer improved clinical trial designs addressing the foregoing issues. In conclusion, the adoption of such an approach for image-directed drug delivery in clinical settings will have far-reaching implications for personalizing cancer care in terms of treatment planning, stratification to appropriate trial arms, and response monitoring.

Traceable and Thermoresponsive Multifunctional Engineered Protein Drug Delivery Agents for Metastatic Breast Cancer

Joseph A. Frezzo NYU Tandon School of Engineering

Theranostics is the field whereby both drug delivery and medical imaging are merged to promote more effective therapies. This is especially important in the delivery of small molecule cancer therapeutics such as doxorubicin (DOX) where clinicians must strike a balance between administering an effective dose while limiting off-target cardiotoxicity risks. In this work, a fluorinated protein is investigated for theranostic use as a chemotherapeutic carrier and 19F MRI agent. The fluorinated protein, CE2-RGD-TFL, is comprised of two functional domains: 1) a coiled-coil domain (C), flanked by two integrin targeting domains, capable of encapsulating small hydrophobic drugs and; 2) two elastin-like peptide domains (E) that impart concentration-dependent thermoresponsiveness. Fluorination imparts interesting thermoresponsive properties upon CE2-RGD. While the circular dichroism analysis reveals that CE2-RGD-TFL is less structured than the wild-type variant, CE2-RGD-TFL coacervates in the physiological range for hyperthermic treatment (39-42oC). Furthermore, fluorination permits greater doxorubicin binding as CE2-RGD-TFL exhibits 49.1% loading of drug while CE2-RGD possesses 17.8% loading. Inversion recovery experiments show little change in R1 values, regardless of temperature or concentration. Car-Purcell-Meiboom-Gill echo train experiments reveal a remarkable dependence in R2 values as a function of concentrations and temperatures. Based on r2/r1 as a function of concentration, there is support for imaging the protein using ultra-short echo time (UTE) pulse sequences. Interestingly, the protein provides an excellent image using UTE at relatively low fluorine concentrations and a short time.

Influence of Surface Chemistry on the Response of Stem Cells via Distinct Fibronectin Adsorption

Ricardo A. Pires University of Minho

Glycosaminoglycans (GAGs) are main building elements of the extracellular matrix where they act in synergism with proteins and are equally critical for the development, growth, function or survival of an organism [1]. GAGs are anionic linear polysaccharides made of repeating disaccharide units, whose negative charge is due to the presence of sulfate groups. These charged units have a crucial role in GAGs interactions with proteins and therefore in key biochemical/signalling processes related to cell functionality and survival. In a previous work we have developed a GAG mimicking platform that is based on self-assembled monolayers (SAM) and allows precise control of the surface exposed hydroxyl (-OH) and sulfonic groups (-SO3H) [2]. We proved that those surfaces are reliable tools to study GAGs interactions with proteins (FGF-2) or cells [2, 3]. Herein, we used the same platform to study the influence of -OH or -SO3H functionalities on fibronectin (FN) adsorption/conformation and the impact of these events on the adhesion and morphology of human adipose derived stem cells (ASCs).

QCM-D data showed that similar amounts of FN were adsorbed on the -OH and -SO3H functionalized surfaces. However, the adsorbed protein underwent different organization on these surfaces and induced distinct ASCs response: it accelerates the cell adhesion on the -OH surfaces and slows down this process on the -SO3H ones. Interestingly, single cell force spectroscopy (SCFS) experiments revealed similar maximum cell adhesion forces for -OH surfaces with or without adsorbed FN, while significantly stronger adhesion was determined for FN-coated -SO3H surfaces, when compared with the non-coated ones. Finally, we have also evaluated the influence of the FN adsorption and organization on the integrin expression of the ASCs cultured on these surfaces. In the case of -SO3H surfaces, the FN coating provoked an overexpression of the ?V?3 integrin and no significant variation of the α5β1 that interacts with the RGD epitotes of FN but also with its synergy domain [4]. On the other hand, FN adsorbed on the -OH surfaces caused underexpression of αVβ3 integrin that recognizes and interact specifically with the RGD domains of FN [4].

Based on these results, we propose a model according to which the RGD epitotes of FN interact with the -OH surfaces reducing their availability to participate in integrin-mediated cell binding. In the case of -SO3H surfaces, the FN interaction with the surface is non-RGD mediated, leaving the RGD epitopes available to interact with the ASCs and to promote the αVβ3-mediated cell binding.

[1] Pashkuleva I and Reis RL, J Mat Chem 2010, 20: 8803-8818; [2] Soares da Costa D et al., J Mat Chem 2012, 22: 7172-7178; [3] Amorim S et al., Langmuir 2013, 29: 7983-7992.; [4] Danen EHJ et al., J Cell Biol 2002, 159, 6: 1071-1086.

We acknowledge the EC H2020 (grant number H2020-TWINN-2015-692333 Chem2Nature) and the European Research Council (grant number ERC-2012-ADG 20120216-321266 ComplexiTE).

Myocardial Delivery of Lipidoid Nanoparticle Carrying modRNA Induces Rapid and Transient Expression

Kevin D Costa Icahn School of Medicine at Mount Sinai

Nanoparticle-based delivery of nucleotides offers an alternative to viral vectors for gene therapy. We report highly ef?cient in vivo delivery of modi?ed mRNA (modRNA) to rat and pig myocardium using formulated lipidoid nanoparticles (FLNP). Direct myocardial injec-tion of FLNP containing 1-10 μg eGFPmodRNA in the rat (n = 3 per group) showed dose-dependent enhanced green ?uorescent protein (eGFP) mRNA levels in heart tissue 20 hours after injection, over 60-fold higher than for naked modRNA. Off-target expression, including lung, liver, and spleen, was <10% of that in heart. Expres-sion kinetics after injecting 5 μg FLNP/eGFPmodRNA showed robust expression at 6 hours that reduced by half at 48 hours and was barely detectable at 2 weeks. Intra-coronary administration of 10 μg FLNP/eGFPmodRNA also proved successful, although cardiac expression of eGFP mRNA at 20 hours was lower than direct injec-tion, and off-target expression was correspondingly higher. Findings were cofirmed in a pilot study in pigs using direct myocardial injection as well as percutane-ous intracoronary delivery, in healthy and myocardial infarction models, achieving expression throughout the ventricular wall. Fluorescence microscopy revealed GFP-positive cardiomyocytes in treated hearts. This nanopar-ticle-enabled approach for highly effcient, rapid and short-term mRNA expression in the heart offers new opportunities to optimize gene therapies for enhancing cardiac function and regeneration.

A New Microscope Objective for Imaging Large Biomedical Specimens with Sub-cellular 3D Resolution

Gail McConnell University of Strathclyde

Optical lenses reached the limit of resolution set by the wavelength of light more than a century ago. However, no attempt was made to achieve the maximum resolution in the case of low-magnification lenses, probably because the visual image would then have contained detail too fine to be perceived by the human eye. Currently available lenses of less than 10x magnification are of simple construction and their numerical apertures (which determine their resolving power) are 0.2 or less, as compared with 1.3 or more in high-power lenses. They are perfectly adequate for the eye or a standard camera, but in the 1980s, confocal microscopy and improvements in camera resolution revealed a need for better low-power lenses: many researchers found that thin confocal optical sections could not be obtained at 4x magnification.

We have developed a novel lens system called the Mesolens, which achieves an N.A. of nearly 0.5 at a magnification of only 4x. When compared with a standard 4x objective, its lateral resolution is 2.5 to 5 times better and its depth resolution (which is vital for confocal or multi-photon microscopy) is 10x better. This unusually large objective lens provides, for the first time, good optical sectioning of specimens as large as entire 12.5-day mouse embryos (>5 mm long) with sub-cellular detail in every developing organ. We are creating at the University of Strathclyde a facility for the development and application of Mesolenses in biomedical science. Progress in the creation of the facility will be presented, together with recent super-wide-field and confocal mesoscopy datasets.

Insights into Molecular Mechanisms of Heart Diseases: Cryo-electron Microscopy Uncovers the Atomic Structure of the Ryanodine Receptor and its Gating Mechanism

Amedee des Georges CUNY Advanced Science Research Center

Cryo-electron microscopy has revolutionized structural biology with its ability to obtain 3D structures of macromolecular complexes without crystals. In particular, it has opened an avenue to the structure of membrane proteins, which are notoriously difficult to crystallize and study by X-ray diffraction. This work sheds light on how RyR1 may be regulated and how known mutations affect its function, a first step towards designing drugs to cure their associated heart and muscular diseases.

Imaging Molecular Structure in Living Animals

Hyungsik Lim Hunter College, The City University of New York.

I will present two stories in this talk. First, I will describe second-harmonic generation for capturing the dynamics of protein conformation in vivo. The significance of our work in neurodegenerative disorders such as glaucoma will be discussed. Second, I will briefly explain our endeavor to interrogate transcription at the single molecule level in living brain.

Fibrous Protein Biomaterials - Designs for Function

David L. Kaplan Tufts University

The study of biomaterials underpins drug delivery systems, medical devices and tissue engineering-regenerative medicine scaffolding. Fibrous proteins, including collagens, elastins and silks provide a useful suite of structural templates upon which to build insight into design-function relationships for biomaterials, as well as useful material systems for the above applications. Traditional trial and error strategies, while useful, extend time frames for discovery and utility. Thus improved experimental strategies to shorten the path from design to utility are needed. Strategies being implemented towards this goal, attained by marrying modeling with experimental platforms, will be discussed. The goal is to develop predictive tools for biomaterial designs, with a current focus on hierarchical features and mechanics.

Engineering with Biomolecular Motors

Jacek Wychowaniec University of Manchester

In the last two decades, significant efforts have been made to develop soft materials exploiting the self-assembly of short peptides for biomedical applications[1]. So-called β-sheet forming peptides are very attractive for the design of biomaterials, in particular hydrogels. Due to the 'simplicity' of the structure formed at the molecular level, the relative robustness of the β-sheet assembly and the ease of functionalization, very stable functional hydrogels with tailored properties, in particular mechanical, can be designed which mimic the cell niche with potential application in a range of fields from tissue engineering and cell culture [2,3] to drug delivery [4,5].

One particular aspect which has attracted significant interest is the ability to use these systems in-vivo. Their low immunogenicity, cell-retention ability and biodegradability make them ideal 3D injectable delivery systems for cell-based therapies. In this context, understanding the shear-thinning and recovery mechanisms of peptide hydrogels is key to their design. The actual structural shear-thinning mechanism is still a matter of debate [6] as two mechanisms are possible: 1. breaking of self-assembled fibres under shear resulting in the breaking of the hydrogels into small isotropic disconnected aggregates (junction strength > fibre cohesive strength) or 2. deformation of the fibrillar network under shear resulting in sliding, disentanglement and alignment of the fibres (junction strength < fibre cohesive strength).

In this project we have explored the shear thinning and recovery properties of series of β-sheet forming peptides with varying hydrophilicities which design is based on the alternation of hydrophobic and hydrophilic residues originally developed by Zhang and co-workers [7,8]. The phase behaviour and shear thinning properties of these peptides was investigated using a range of techniques including oscillatory shear rheology and shear polarised light imaging microscopy. Our results show that the shear thinning mechanism depends on the aggregation propensity of the peptide fibres and the strength of the fibre-fibre interactions which is controlled by the peptide design and overall peptide concentration of the hydrogel. These results give an insight into the deformation mechanism which is important to understand the level of shear cells are subjected to when injected with these materials.

[1] Chem. Soc. Rev., 39, (2010) [2] L. A. Castillo Diaz et al, J. Tissue Eng., 5, 2041731414539344, (2014). [3] L. A. Castillo Diaz et al, in press (2016). [4] D. Roberts et al., Langmuir, 28, 16196-16206, (2012) . [5] C. Tang et al., Int. J. Pharmaceutics, 465, 427-435, (2014). [6] C. Yan et al., Soft Matter, 6, (2010). [7] Zhang et al., J of Biomat. Sci.: Polym. Ed., 9, (1998). [8] Zhang et al, Reactive and Funct. Polym., 41, (1999).

Racemic Hydrogels from Enantiomeric Peptides: Predictions from Linus Pauling

Joel Schneider National Cancer Institute, National Institutes of Health

We have reported that hydrogel materials can be prepared from self-assembling beta-hairpin peptides. For example, the 20-residue peptide MAX1 rapidly self-assembles into a hydrogel network of monomorphic fibrils whose molecular structure was recently determined by solid state NMR. The enantiomer of MAX1, namely DMAX1 assembles affording a hydrogel of identical crosslink density, mesh size, and mechanical rigidity to the MAX1 gel. Surprisingly, the gelation of a 1 wt % equimolar solution of peptide enantiomers occurs more rapidly resulting in a racemic hydrogel network whose mechanical rigidity is over four-fold greater than gels prepared from either pure enantiomer. Keeping in mind that the total amount of peptide in the racemic gel is equal to that of either pure enantiomeric gel, this observation is truly unexpected and suggests that biomolecular chirality, at the level of the monomer, is directly influencing the mechanical properties of the self-assembled hydrogel. We interrogated the self-assembly process and resulting fibrillar and network morphologies of the racemic gel employing CD spectroscopy, isotope-edited FTIR, transmission electron microscopy labeling experiments, small angle neutron scattering, diffusing wave spectroscopy, solid state NMR and molecular modeling to uncover the molecular basis for this behavior. In light of the NMR structure of pure MAX1 fibrils, the mechanism of enantiomeric assembly and their molecular arrangement in the solid state will be presented. The mode of molecular assembly uncovered in our studies was predicted by Linus Pauling in 1953 in the course of deriving models of the pleated β-sheet, a fold ubiquitous in protein structure.

Friday June 24th 2016

Towards Dynamic Hybrid Architectures: Or Can We Make Materials Adaptive?

Joanna Aizenberg Harvard University

Dynamic structures that respond reversibly to changes in their environment are central to self-regulating thermal and lighting systems, targeted drug delivery, sensors, and self-propelled locomotion. Since an adaptive change requires energy input, an ideal strategy would be to design materials that harvest energy directly from the environment and use it to drive an appropriate response. New synthetic approaches that would lead to such adaptive materials present a real challenge for materials chemistry in the 21st century. To address this challenge, I will describe the design of a novel class of reconfigurable materials that, similar to skeletomuscular systems, use a hybrid architecture to interconvert energy between different forms and scales. To specify the materials' functions, we use surfaces bearing arrays of nanostructures put in motion by environment-responsive gels. Their unique topography can be designed to confer a wide range of adaptive optical, wetting, adhesive, anti-bacterial, motion-generating, and other behaviors, similar to their natural counterparts used by lotus leaves to shed water, geckos to stick to surfaces, cephalopods to change color, echinoderms to keep their skin clean, and fish to sense flow. Using both experimental and modeling approaches as well as new fabrication methods, we are developing our ability to take full advantage of the immense potential for energy coupling within these hybrids to create a generation of sustainable, self-reporting, self-adapting materials.

Designing Biomimetics and Smart-materials for Recognition and Capture of Biologics

Cecilia Roque Universidade NOVA de Lisboa

Over the past 40 years monoclonal antibodies and derived structures became the standard binding proteins representing powerful tools in biotechnology and biosensing. Other synthetic and protein scaffolds, with the robustness and versatility required, are being explored by our group to enable the selective capture and oriented immobilization of biomolecules onto surfaces. We employed biological and chemical combinatorial libraries supported by computational design tools to develop robust peptidomimetics against several targets, namely tagged recombinant proteins (1), GFP fusion proteins (2), and virus-like particles (3). Our research has also focused on the development of strategies for the surface coating of magnetic nanoparticles with natural (4) and synthetic polymers and subsequent decoration with the novel affinity reagents (5). These modified magnetically-driven materials are applied to biotechnological and biomedical areas which will be discussed in the seminar.

(1) Pina et al., 2014, ChemBioChem, 15 (10), 1423-1435. (2) Pina et al., 2015, Journal of Chromatography A, 1418, 83-93. (3) Fernandes et al., 2016, Journal of Chromatography A, 1438, 160-170. (4) Palma et al., 2015, Nanoscale (7), 14272-14283. (5) Barroso et al., 2014, Advanced Functional Materials, 24(28), 4528-4541.

Cancer Carbohydrate Nanotechnology: Understanding and Targeting Cell Surface Glycosylation in Disease Therapy and Diagnosis

Adam B. Braunschweig University of Miami

Every eukaryotic cell is coated with a layer of carbohydrates 'termed the glycocalyx' whose glycosylation pattern is cell specific. Because of their rapid division, cancer cells promote immature glycans to the cell surface, and as such display distinct glycosylation patterns that could be exploited for diagnosis or drug delivery. Currently, however, cell-surface carbohydrates are considered 'undruggable targets'. attractive and validated cancer targets that remain outside of the reach of pharmacological regulation, because of (1) the difficulty associated with selectively binding in water saccharides with subtle structural differences, and (2) the primitive understanding of how surface effects contribute to molecular recognition at biological interfaces. This talk will present recent efforts from the Braunschweig group to develop selective molecular receptors for binding non-glucosidic saccharides in water,[1] and developing new printing methods including a platform for 4D organic nanolithography for creating complex cell-surface models.[2-4] These examples demonstrate how emerging tools from supramolecular chemistry and nanoscience can be leveraged to develop new strategies for addressing cancer and understanding how information is communicated within molecular systems and biological networks.[5]

[1] Rieth, et al. Chemical Science, 2013, 4, 357-367. [2] Bian, et al. Journal of the American Chemical Society, 2013, 135, 9240-9243. [3] Bian, et al. Chemical Science, 2014, 5, 2023-2030. [4] Liu, et al. Polymer Chemistry, 2016, DOI: 10.1039/C6PY00283Ht. [5] Xu, et al. Current Opinion in Biotechnology, 2015, 34, 41-47.

3D Cell Culture in Anchored Hydrogel Droplets for High Throughput Drug Screening

Alan M. Lyons ARL Designs LLC

Current drug-discovery approaches are highly inefficient, as <10% of new drugs entering the pipeline receive FDA approval. The tools used today may explain this low success rate as researchers typically study cells cultured in microplates; a non-natural 2D environment. Studying the response of cells cultured in a more physiologically relevant 3D environment has recently shown promise for increasing the drug-discovery success rate. Similarly, isolated lymphocytes, cultured in 3D, are an important source of new biological drugs. Available tools for culturing cells in 3D are challenging to use and difficult to automate, especially for long-term culture studies. The lack of appropriate platforms for handling cells in 3D severely limits the development of 3D cell assays for new drug discovery.

We have developed a new technology, composed of arrays of specially designed surface structures, on which droplets of hydrogels containing cells can be anchored. These structures are built into the AIM lid which is compatible with standard 96 and 384 well microplates. Hydrogel droplets containing cells are dispensed on the AIM lid by standard pipetting or by immersing the surface features into a solution of hydrogel. To form tissues, different types of cells can be juxtaposed by dispensing successive layers of solutions containing different cell types or hydrogel chemistries. After cure, the AIM lid is placed on a standard microplate, immersing the hydrogel droplets into the culture medium or any other solution contained in the microwells. The cells remain within the gel, but nutrients and gases can rapidly diffuse through the nano-porous gels. Medium changes, washing steps, staining and drug introduction are achieved by moving the lid to a new microplate containing fresh solution.

The long term culturing of cells (> 10 days) as well as the diffusion of peptides and proteins in anchored 3D gel droplets was studied as a function of hydrogel composition and cross-link density using optical and high resolution (63x) florescence microscopy. Techniques for dispensing nanoliter droplets of cells in high density arrays (>500 droplets/cm2) will also be described.

P-selectin is a Nanotherapeutic Delivery Target to the Tumor Microenvironment

Yosi Shamay Memorial Sloan Kettering Cancer Center

Disseminated tumors are poorly accessible to nanoscale drug delivery systems due to the vascular barrier which attenuates extravasation at the tumor site. We investigated P-selectin, a molecule expressed on activated vasculature that facilitates metastasis by arresting tumor cells at the endothelium, for its potential to target metastases using this property. We found unexpectedly that P-selectin is expressed on cancer cells in many human tumors. To develop a targeted drug delivery platform, we employed a fucosylated polysaccharide with nanomolar affinity to P-selectin. The nanoparticles targeted tumor vasculature to localize chemotherapeutics and a targeted MEK inhibitor at tumor sites in both primary and metastatic models, resulting in superior anti-tumor efficacy. On non-expressing tumors, we found that ionizing radiation guided the nanoparticles to the disease site by inducing P-selectin expression. Radiation concomitantly produced an abscopal-like phenomenon in unirradiated tumors, suggesting a potential strategy to target disparate drug classes to almost any tumor.

Microfluidics for 3D Tissue Engineering and Personal Health Diagnostics

Samuel K. Sia Columbia University

I will discuss the use of microfluidic techniques for two different applications: controlling 3D microenvironments of cells and tissues, and for developing low-cost point-of-care diagnostics for use in U.S. and in developing countries.

1) A number of microfluidic techniques have been developed in our group for controlling the 3D microenvironments of cells and tissues to high resolution. These techniques are useful for studying microvascularization in a number of organ systems, and for engineering implantable devices.

2) In the second half of the talk, I will discuss the development of lab-on-a-chip devices for personal health in the U.S., and for diagnosing diseases for global health. I will discuss our lab's current efforts, in conjunction with partners in industry, public health, and local governments, to develop new rapid diagnostic tests for use in sub-Saharan Africa.

Sponsor: Biogelx Sponsor: Hunter College Sponsor: Fisher Scientific Sponsor: ACS Biomaterials Sponsor: Gilson Sponsor: Eppendorf Sponsor: ThermoFisher Sponsor: Chem by Cell Press Sponsor: Practichem


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Konoha Comrades!!

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"Konoha Comrades!!"
Chapter 218
(木ノ葉の仲間!!, Konoha no Nakama!!)
Chapter Info
Volume Itachi and Sasuke, Brothers (#25)
Previous "For the Sake of Those Precious"
Chapter Naruto #218
Next "Future and Past"
Arc Sasuke Recovery Mission
Anime Naruto #128
"Konoha Comrades!!" (木ノ葉の仲間!!, Konoha no Nakama!!) is chapter 218 of the original Naruto manga.

Sasuke tries to ignore Naruto and continue on to Orochimaru. Remembering Sakura's earlier failure to persuade Sasuke, Naruto asks why he would turn away from everything and everyone in Konoha. Sasuke responds that he owes them nothing and has finished his business with the village. When Naruto attacks him in anger, Sasuke adds that he didn't become any stronger by staying in Konoha. Orochimaru can give him the power he desires. Naruto reminds him of Orochimaru's actions and his intents to take Sasuke's body. Sasuke does not care, so Naruto promises to bring Sasuke home by force if necessary.


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Coming of the Dark Sage - One Part Down

Shuuren (as Tadashi), Naru

Date: July 11, 2013

Tadashi has moved on to the next part of his training with the Hitokage keeping an eye on him to be sure he doesn't turn to stone, along with Naru being sure the Crawler isn't trying to fully possess his body.

"Coming of the Dark Sage - One Part Down"

Crawler's Cave

Within this cave, it's really hard for Tadashi to really know how long he's been at this training anymore or even what time of day it is. His meditation has carried him pretty far off at times. The only thing keeping him on track with eating, sleeping, and such is when Naru interrupts him. Her help is making this move along a lot faster, though. At this point, he's moved on from simply working on sensing the natural energy to absorbing it into himself. Every so often, he'll start to transform to a rather horrific-looking creature, which leads to a rather hard smack of a rod across his back with a rod by a large Hitokage standing behind him to force it out and put him back to normal. It's not exactly a pleasant process, but it will be beneficial in the end. He has stopped using the ooze, which is a good sign of his progress. Now the practice is keeping the amount of nature energy absorbed at a precise level so it doesn't overwhelm him nor fizzle out.

Having been at this for some time now, Tadashi has quite a few healing bruises on his back. Still, he doesn't seem bothered by that. It's better than dying a stone Hitokage. That same look of focus shows on his face, eyes closed as he goes through this tedious practice. Great power comes with a great price, and this is one that is right up his alley, one that requires great control over that power.

IT was time for Tadashi to get a better focus over this energy, and of course Naru was going to be there to assist him every step up the way. While he meditated Naru was there to pester him. Try to drive his focus away from focusing the chakra, after all for now he was working without distractions, he needed be able to focus this energy and keep it with movement so he could prepare it for combat. "Are you awake?" Narusegawa asked Tadashi, she too sat right infront of him closely enough to keep her voice like a faint whisper. She didn't want to pester to much but she was definitely going to do what she could to slowly gear him out of focus.

At that whisper, the Hitokage would look down at Naru, shaking its head. Tadashi would then turn his head up to look at her, eyelids opening to reveal orbs of pure black. He then blinks a few times, after which his eyes are back to their normal state. "Yes… How long has it been?" he asks, looking at her for a moment before he'd push himself up off the ground to stand and start stretching a bit. This kind of training can make one's body rather stiff. Of course, he stretches out a bit more than most men are able to, his limbs actually coiling around a bit. "It seems to be getting a lot more stable from what I can sense. What do you think?" he asks as he looks at the woman, seeming a bit pleased at his progress. Still, there's a way to go, and they've got things to do once this is done.

"You definitely have made quite a bit of progress… I suppose the question is if you are able to maintain that chakra inside of you. It is more delicate than any kind of chakra I have witnessed, more difficult to grasp than genjutsu," Naru spoke softly, uncrossing her arms while presenting her hands out to him. " It's not time for you to eat and drink yet, I think it would be better if I helped you out with your training by making you a little less stiff… I want you to teach out and grasp my hands," Narusegawa suggested, keeping her eyes peeled on him so he didn't leave her sight. " Come on, it shouldn't be too hard. We need to get you moving,"

"That it is," Tadashi says with a nod. At her next suggestion, he tilts his head slightly then shrugs and says, "Okay." He puts his arms forward, going to grasp Naru's hands then use his rather insane strength to pull her up off the ground, unless she resists or asks him not to. "Now what?" he asks, lifting an eyebrow faintly.

"That it is," Tadashi says with a nod. At her next suggestion, he tilts his head slightly then shrugs and says, "Okay." He puts his arms forward, going to grasp Naru's hands then use his rather insane strength to pull her up off the ground, unless she resists or asks him not to. "Now what?" he asks, lifting an eyebrow faintly, a light smirk gracing his lips. He'd turn his head to crack his neck a bit, letting out a sigh of relief. "I think this is about the most out of touch I've been with the outside world my entire life. You'd almost think I'd be spending some time relaxing while I was away," he says, snickering a bit at that irony.

There wasn't much for Naru to resist, not that she had the strength to slip out of his grasp anyways. She cringed from the sudden force he had used on her, and while she was pulled up she attempted to pull her arms back to fight for her release. " That…was good but a little painful… I suppose even an oaf like yourself has their needs now don't they?" She asked him curiously, hopefully to withdraw her arms back. If allowed she too ka few stesp back away from him, perhaps two yards until she beckoned him to follow at her. " Keep focusing that energy and come walk towards me,"

"Oaf, huh? Don't think I've been called that before, at least not to my face," Tadashi sound, sounding a bit amused as he easily lets Naru's hands slip from his grasp. For a slender man, he is deceptively powerful. Of course, he didn't exactly answer the question either. At the next request, he blinks a few times, saying, "I don't think you understand how this works. To focus this energy, you have to sit completely still." With that, a rather creepy voice would come from the Hitokage, echoing that of the Crawler. "That's why it's useless in combat, unless you have a partner who can focus the energy for you by sitting still while you move. Once you get this part down, then we will work on that." Tadashi would look up to it and ask, "Mastering that would be the final step then?"

"Oh…so you need something to focus it for you? How about you try that then? Let's see if you can handle that energy be funneled in while you are moving. That's what you need to do right?" Of course Naru was just guessing. Lightly scratching at her head as she attempted to figure out just what was going on. A soft sigh escapes her lips she stodd herself up firmly while attemptign to figure out just what Tadashi would like for her to help him accomplish.

"Trying to skip on to the next phase without this one being complete? And I thought I was the one being impatient," Tadashi says, smirking slightly as he would move to sit back down. "Give me a few more moments to complete this part, and then we'll move on to that part. I've almost got the hang of this." With that, he'd close his eyes again, beginning his meditation as he brings the natural chakra into himself. After a few more moments, a change would begin happening to him again. His muscles would grow more rigid, glowing marks of dark, purple chakra appearing on his face as his fingernails grow long and talon-like. As he opens his mouth to take a breath, his teeth show to have become like the fangs of a beast, then his eyelids reveal those blackened orbs once more. Rather than continuing to twist into a darker form that would endanger him, however, this form stays solid, the natural energy evening out inside him to now be able to be put to use. "Now… I think we can move on."

Unless otherwise stated, the content of this page is licensed under Creative Commons Attribution-ShareAlike 3.0 License


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The Storm Brigade - Storm at Kyuusen

Yoichi (as Hatsu), Michiko

Date: July 22, 2016

Michiko is traveling her way back from Iwagakure to Kumogakure, and she has a little run-in with the Storm Brigade that happens to be a kidnapping!

"The Storm Brigade - Storm at Kyuusen"

Kyuusen Crater

Michiko is making her way through some of the more neutral areas, traveling by land rather than water for a few reasons. The first is that she can travel … Well, not faster, but she can be more aware of her surroundings when she travels by foot. The second is that she was a bit nervous she'd run into the Storm Brigade since they were known to attack islands. Of course, they also staged an attack on Konoha, so they could really appear from anywhere, but Michiko figured her chances of running into them was less. Right now, Michiko is passing through the deserted ruins of Fort Kyuusen, which had been annihilated by some jutsu, likely. She forgets the details right now, more focused on not stumbling and keeping a general eye out. A glance upwards at the sky, and she sees no sign of clouds or storms. It was a good day to travel, at least.

Much like a normal storm, the Storm Brigade's attacks cannot be predicated by location. A group looking to wipe out the shinobi world does not mind taking the battle wherever it needs to go. And it would seem they have a new tactic in how they are attacking… Taking powerful Jounin from the villages that they need to fight back to dishearten them. Okumo Kiji was the first to be taken, but that was basically her own fault since she put herself in position to be taken. Someone else's poison blade stabbing her while trying to stab Hatsu probably didn't help much either.
However the group is keeping an eye on the villages, it seems they have figured out that Michiko was traveling alone on her way back to Kumogakure from Iwagakure. Along the Kyuusen Crater, the clear sky suddenly begins to darken with black clouds. Rain begins to fall upon the land, lightning striking upon the ground as the day turns from completely clear in this area into the eye of a hurricane. No sign of men in black cloaks just yet, but one can basically guarantee they're coming.

RP: Michiko transforms into SIZZLE-IV.
COMBAT: Michiko focuses 8816 stamina to turn it into 12000 usable chakra!
RPCOMBAT: Michiko defends against with a MASTERED-EARTHEN-SENSE…85

Michiko notices the sky darkening to the point where it was far too sudden and unusual. In that instant, she was on the alert, chakra flaring up a bit like a beacon for any sensors that might be around. "Maybe I should have taken up an escort…" she mutters to herself, a chakra aura seeming to flicker around her body much like flames. Michiko cast her senses out along the ground, mapping things out within the mile to see if she can sense much of anything. She couldn't help but think in the back of her mind that it was pointless if they could fly like Seiko, but that didn't halt her attempts.
As the rain came pouring down, Michiko reached up to the hitai-ate around her neck and tugged at its knot to make sure it was secure. A hand dropped to the pouch at her side, but did not move to grab anything from it. Instead, she simply continued on her way across the Kyuusen crater. She was originally going to travel around it, but across seemed like the better option just about now.

COMBAT: Taki focuses 8395 stamina to turn it into 12000 usable chakra!

Even if some of the people might be able to fly, a great number of them are on the ground and rock formations. Dozens of them are incoming and surrounding the Head Ninja, flashes of lightning revealing their black cloaks around in a rather disturbing fashion. Another flash of lightning would reveal them lifting their hands to point at Michiko, firing off dozens of streams of lightning at her with intent to blast her. Seems they don't think she will come along as easily as Kiji did, so they're going to bring her down first or at least weaken her enough to land a long-term stun so she can be brought to their base.

COMBAT: Michiko defends against LIGHTNING-CRASH(84) attack from Taki with a WIND-STEP…84
COMBAT: Michiko defends against LIGHTNING-CRASH(75) attack from Taki with a WIND-STEP…88

Michiko's hair rises as the Lightning crackles around her. Being a Kumogakure shinobi, she was far too used to lightning being used as an attack because … Well, all of Kumo (pretty much) uses lightning. The girl recalls all too well some of the spars she was in throughout her career as a shinobi where she was electrocuted, and she wasn't fond of receiving that experience from an enemy. Michiko seems to disappear, wind cutting through the Lightning as she moves to avoid the attack. "Che…" she mutters, making a few hand seals. The ground is quick to come alive, hands of earth forming and grabbing as many of the Storm Brigade as she can. There were about ten hands in total, all of them working to grab the various individuals tying to attack her. It was far from enough, though, if she was counting the black cloaks correctly.

COMBAT: Taki defends against DEMON-GRAVES(77) attack from Michiko with a SEAL-ENCAMPMENT-WALL…101
COMBAT: Taki defends against DEMON-GRAVES(89) attack from Michiko with a SEAL-ENCAMPMENT-WALL…80
COMBAT: Taki defends against DEMON-GRAVES(76) attack from Michiko with a TENSE…12
COMBAT: Taki ESCAPES from Michiko's stun!

The brigade has a rather interesting method for defending against Michiko's attack. Rather than getting out of the way, a number of them moves through handseals then slam their hands on the ground to create a seal barrier over the ground itself rather than standing upward. This protects a number of them, though a number of them are still pulled into the ground. That, however, doesn't stop those up top from continuing their attack, this time with another element. Michiko would find at least half a dozen jets of water of force extreme enough to cut through stone intending to carve through her body. They are not attacking her in full number yet either, which means they might just be toying with her for now… or simply intending to wear her down.

COMBAT: Michiko defends against PURE-WATER-SPEAR(88) attack from Taki with a EARTH-STYLE-WALL…59
COMBAT: Michiko loses the roll and sustains 2182 damage.
COMBAT: Michiko defends against PURE-WATER-SPEAR(74) attack from Taki with a EARTH-STYLE-WALL…72
COMBAT: Michiko loses the roll and sustains 1996 damage.
COMBAT: Michiko heals Michiko for 1943 with MYSTICAL PALM TECHNIQUE.

Michiko barely has the time to make a sort of follow up attack, as the water bursts out towards her in a powerful jet. She ends up getting some nasty injuries, the water having cut through the earth barrier she formed between herself and the water. It was painful, making red flash in her vision. It was likely only an automatic reaction that she immediately sent healing chakra through her body to heal the worst of the injuries, though she was down for the count for a few moments while she healed herself.

With Michiko unable to followup on her previous attack thanks to being so damaged, the Storm Brigade members she had captured in the earth begin to rise back out of the ground. As they do, their counterparts on the surface shift their handseals, causing the rain that is beating down to begin to form into a number of giant pillars of water that flow down at her like waterfalls from all directions to try and damage her further while restraining her to try and begin the process of capturing her. Not a word has been spoken, no mercy given by either side. It seems even these guys take some battles as serious as they should, possibly a side-effect of their real leader taking the reins after the death of Kano. Whatever the case, they are quite determined to accomplish their mission here today.

COMBAT: Michiko defends against GREAT-WATERFALL(71) attack from Taki with a METAL-DOME…119
COMBAT: Michiko defends against GREAT-WATERFALL(99) attack from Taki with a METAL-DOME…93
COMBAT: Michiko loses the roll and sustains 1477 damage.

Michiko watches the pillars of water appear, and, anticipating a huge attack, she makes a few hand seals that create a dome of metal to surround her. She was hoping that it would keep her safe from the giant waterfall crashing down on her, and it did for a few moments. The initial wave that struck the dome was kept at bay, smashing against the metal, but not denting it. The defense was very strong, apparently. But it can't withstand everything. Michiko had failed to protect the area below her, which allowed the water to sneak into her dome and trap her within. The water rushed up from beneath her, slamming her back into the dome's wall and making her hit her head so that she would see stars for a little bit. @.@

RP: Hatsu transforms into PERFECT-HYDRIFICATION.
COMBAT: Hatsu focuses 5855 stamina to turn it into 8000 usable chakra!

Perhaps the scariest thing about the Storm Brigade is that their leader is someone many consider to be the perfect shinobi. Yoichi was one of the most feared of the Seven Swordsmen ever as well as the Head Hunter of Kirigakure. This means they have access to tactics and knowledge of the ins and outs of shinobi villages and their combat and operation tactics that even the Silence could not match. This makes them capable to attack the villages in ways that other groups could barely dream of… and gives them an edge to even help them against one of the most revered shinobi in the shinobi world.
As Michiko is trapped by the racing waters, another section of the battalion upon her brings their hands forward and fires forth stream after stream of lightning. They are particularly careful with how much damage they do, however, as their goal is to capture not to kill, and a certain other person in their midst might get quite angry if they screw that up.
As the flashes of ligtning fade, out of the water rushing around Michiko forms a figure that haunts the shinobi world as one of its foremost that has turned her back on them… Kaguya-Hozuki Hatsu. The crimson-eyed woman looks over Michiko sternly to survey the damage that has been done to her then stretches her arms forward to wrap her arms around the Head Ninja and pull her close. It might even be comforting to receive the hug were Michiko not further pulled in Hatsu, literally absorbed by her body to be suspended by the pressure of the water of her body in the darkness of her form. Assuming that goes without issue, the storm would at last cease to leave the wreckage and metal of this battle behind. The water created by the jutsu cast flows into the crater and into the areas around it as Hatsu and the Brigade move out to quickly head back to the ocean to move back to their base.

COMBAT: Michiko defends against LIGHTNING-CRASH(84) attack from Taki with a TENSE…17
COMBAT: Michiko loses the roll and sustains 2831 damage.
COMBAT: Michiko ESCAPES from Taki's stun!
COMBAT: Michiko defends against DROWNING-WATER-BLOB(114) attack from Hatsu with a EFFICIENT-METAL-BARRIER…92
COMBAT: Michiko loses the roll and sustains 675 damage.
RP: Michiko reverts to her normal state.

Michiko can only barely withstand the damage of the Lightning, hanging onto consciousness by a thread as the pain races through her body and pretty much steals her breath away. The metal casing was zapped away as the chakra hold on it loosened, which is probably a good thing since Michiko would be free of the water. She is barely breathing, partly because of the effort and partly because she's in so much pain that even the smallest movement is painful. The girl's vision is blurry and surrounded by dark edges that seem to grow. That darkness isn't great enough to disguise the red-eyed woman that's surveying the damage. As Hatsu reaches down, Michiko would, weakly, attempt to fend the woman off by putting her arms in front of her. Chakra races through a bit of the armor she has on her arms, but it isn't going to stop the hug, nor does it stop Hatsu from suddenly absorbing Michiko into her body. It was actually a very strange sensation, being trapped in Hatsu's form… Michiko took a few moments to marvel at it.

RP: Hatsu reverts to its normal state.

If she manages to stay conscious long, Michiko might detect that they've moved into water by the feeling of impact of the water's surface and additional pressure that comes as they head under. The pressure grows more and more as it seems they go deep deep under water, deeper than most shinobi could manage or survive, but they obviously have jutsu for just that. It's probably being inside Hatsu that keeps the pressure from becoming so great that it harms Michiko as they move, a series of constant twists and turns and moving up and down making it impossible to tell where they are heading, but, by the speed and distance they are moving, it is far away from any shinobi nation.

Michiko is fading in and out as the group travels to … wherever it is. Probably their base. It's a bit surprising to Michiko that they hadn't actually killed her yet, but really, she probably would have done the same thing… They pretty much did the same thing when they had taken Kano from the Storm Brigade, even. Their plans were just a bit foiled. While she can still think, the idle thought of how ironic the situation itself is passes through her brain. Not being attached to the earth is disorienting Michiko enough as it is, but adding the twisting and turning of the water was just making that worse… She was getting a headache and starting to hope that she would actually fall unconscious for a longer period of time than a few seconds or minutes.

After a long period of fading in and out and travel somewhere far, far away. At some point when she is out, a set of seal cuffs were placed on her hands. Though they are not chained together, they are designed to keep a constant enough drain on one's chakra to prevent use as well as make it incredibly painful to use chakra at all, which, in Michiko's current state, could easily finish her off. This would be explained at a time when she is awake and would perhaps feel that they have at last left the water, except the pressure still feels like they are far, far under sea level.
Soon enough she would find herself deposited onto a bed in a room that appears to be carved out of a rockface. The room is actually pretty nice with some amenities provided and such. Hatsu gently sets Michiko on the bed and nods to her. "… You should sleep. You will not be escaping this place or sending out any kind of beacon that will reach anywhere."

Michiko recognizes the seal cuffs when she manages to focus enough on the design. She has been studying seals a great deal, even if she doesn't use them often in battle. A small curse escapes her as she just focuses on keeping her wits about her, trying to recover from getting smacked around by the water and electrocuted. Really, she was having a hard time focusing and staying awake, though. Hatsu's words ring through her ears, and it takes far too long for Michiko to process them. And even after she does, the most she can do is eye Hatsu … Sort of. She sorta got mega beat up, after all, and recovering from all that wasn't gonna be the swiftest unless she or another healed her.

Once she has the idea that Michiko understands her predicament, Hatsu nods to her and reaches down to put a hand on her shoulder. "I hope you know I have no malice toward you. I am only doing what must be done. Perhaps you'll see by the time you recover and you can be one of the ones to help me build the new world once we've washed this one away." Withdrawing her hand then, she turns and moves toward the door. "I'll give you some time to think on it." That said, she steps out the door and closes it. Sounds of latches and bars being put in place ring out, letting Michiko know that she has been locked in… Might as well get comfortable.

Michiko has too much pain in her system to react to Hatsu's words visibly, though she does close her eyes. In her mind, a lot of what she's thinking involves cursing Hatsu and the Storm Brigade for kidnapping her while her village needed her. The other small part is saying, 'I'm gonna sleep now'. Once the locks are in place, Michiko chooses to do just that, forcing herself to sleep and get some rest. Even if she isn't the most comfortable right now.

Unless otherwise stated, the content of this page is licensed under Creative Commons Attribution-ShareAlike 3.0 License


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April 2017

Principle Photography by John Jackson

Alan Waddell, Ryan Daniel, Anna Pope, and Sheryl Spencer at O’More College of Design

Linda Ashby and Jamie Carroll at David Lusk Gallery

Denise Stewart-Sanabria, Angie Leahy, and Tiger Gagan at O’More College of Design

The Great Toyzini, Sara Lederach and Katie Wolf at East Side Project Space

Tiffany Foss and Alex Higginson Rollins at Gallery 202

Monica Brown, Misti Wren, Luke brown, and Maria Tipton at Bob Parks Realty The Rymer Gallery

Jamie Carroll at David Lusk Gallery

Kris Griggs and Julie Harvey at Gallery 202

Brian Edmonds, Vicki Sher, and Heath De Hart at Zeitgeist

Sarah Maggart and Kat Friedmann at Channel to Channel

Tim Ross at Bob Parks Realty

Diane Carter and Bill Fahey at Bob Parks Realty

Luke and Olivia Landry at The Arts Company

Ali Katt and Dustin Hedrick at Channel to Channel

Paul Allen, Bobby Bare Jr., and Doni Schroader at Julia Martin Gallery

Charles V. Bennett, Naryan Smith, Cecilia Strand, Katie Kowalski and Jennifer Smith at The Rymer Gallery

Julia Martin, Kevin Doyle and Molly Lahym at Julia Martin Gallery

Ben Lankford at East Side Project Space

Jess and Katora at Tinney Contemporary

Merrilee Challiss at Julia Martin Gallery

The Rymer Gallery

Reygan Pitman and Zander Rosenbaum at The Arts Company

Maliyhah Albayan and LaChanda Q. Akers at Watkins Arcade Gallery

At Brikolai

Curt Thorne, Heather Thorne, Nashville Ballet Artistic Director and CEO Paul Vasterling and Jason Facio; Photograph by Susan Adcock

Nashville Ballet company dancers perform to Muddy Magnolias; Photography by Karyn Photography

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Reigning Cy Young winners battle evenly in a rare matchup

R.A. Dickey leaves with the lead, David Price lasts longer in a game that goes to extra innings More

Mixed results for locals in spring training

Here's a look at how five former area standouts performed in spring training in leading up to Opening Day More

Deal done, R.A. Dickey's trade to Toronto is completed

Nashville native will be the seventh Cy Young Award winner to start the next season with a new franchise More

Price sets career and franchise records with 14 strikeouts

Former Vanderbilt star ties two others for American League's top performance this season More
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Wednesday, August 08, 2007

This Summer's Best Books

At this month’s Cohort we each introduced ourselves by sharing what books (if any) had really blown our doors off this summer. Some folks wanted that list on the blog so here you go.

All the Kings Men by Robert Penn Warren
Red Cactus by Alan Pert
Colossians Remixed By Brian Walsh and Sylvia Keesmaat
Appreciative Inquiry for Congregational Change by Mark Lau Branson
Everything Must Change by Brian McClaren
St. Francis: A Revolutionary Life by ?
On Being Christian by Hans Kung
Legends of the Fall
Harry Potter 1 & 2 by. J.K. Rowling
Teaching to Transgress by Bell Hooks

Thursday, August 02, 2007

Justice in the Burbs Excerpt

Baker Books has put an excerpt of Will and Lisa Samson's Justice in the Burbs: Being the Hands of Jesus Wherever You Live. The book is scheduled for release on August 7th, but you can get the introduction and chapter 1 pdf free. This looks like a very good book and hope that we get a chance to discuss in the next couple of months.

Michael Frost Audio

Many of the ideas from Michael Frost‘s book Exiles: Living Missionally in a Post-Christian Culture, are talked about in sessions that he did in Peterborough, Ontario in 2006. Resonate makes this audio available as a podcast or download from ODEO. I have had problems with the podcast taking a long time to download, but the mp3s downloaded fine.


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My theme for the month is: Secret Diary Of A Serial Killer. Each post will be a new chapter in the story – I’m really excited to finally get this novel idea out of my head and into words. I hope you enjoy it! 🙂

Previous Chapter: Xanthium Start from A: Anna

We were almost back to the house when I started dreaming of killing Carly. It would have been so simple: kill Capinski – check – then kill Carly and stage her body like the others. Leave some crumbs that lead back to Capinski and I’d get away with it because I’d be in charge of the investigation.

I smiled to myself, but a part of me felt sad for missing out on such a thrill.

Still, being with Johnny was a good consolation prize. And if he hadn’t turned up I’m not sure I could have got the better of Capinski while I was so drugged up.

At least burying his body had given me time to detox – my vision seemed fine, and my limbs were fully under my control.

How long was I gone?” I said.

From the time you drove out from the bus station with Capinski, to the time I found you on the floor of the cellar, was around 8 hours,” Johnny said, pulling up to the stone house.

He turned off the engine and faced me, waiting to see what I was thinking.

He took Carly somewhere in that time frame, probably at work,” I said. “In fact, he probably spent some time at work to show his face and give him more time with us in the cellar.”

Johnny nodded. “Your car and phone didn’t move from here until 2 hours before I found you. He probably brought her here, checked on you, then went to dump the evidence while you were still out of it.”

Take me to my place,” I said. “Carly should be asleep for a few more hours, and she’s tied up if she happens to wake up sooner.”

Risky,” said Johnny, raising an eyebrow. But he didn’t argue, he started the car and pulled away from the house.

I need to change out of these clothes, get some cleaning supplies for the blood, and we should deal with Capinski’s car too.”

I can do that,” Johnny said. He glanced over at me when I was quiet for too long. “What are you thinking?”

I was just trying to figure out how to get a confession written from Capinski,” I said, deep in thought. “Maybe I could send an email from his phone?”

If you have a sample of his handwriting, I know a guy,” said Johnny, pulling up behind my house.

I didn’t ask how he knew my address without me telling him.

You know a guy?” I said, and laughed.

He grinned. “Go get changed, I’ll call my guy while I wait.”

I stuffed all of my dirty clothes – covered in spiky seeds, soil, and blood – into a garbage bag. Then pulled on new jeans, a black top, and black running shoes. I tied my hair back in a pony tail, and only then did I notice the bruises on my wrists. Nothing some makeup wouldn’t cover, but that would have to wait.

I grabbed some baggy sweatpants and a t-shirt for Johnny to change into, a few cleaning supplies, then he drove us to the bus station so I could get my car.

He got in with me. “I’ll come and deal with Capinski’s car now,” he said, planning to drive it back from the stone house. “My guy said if you can get him a handwriting sample, he can get you a note within the hour.”

I leaned forward and kissed him. It was brief but left us both smiling.

This ‘someday’ chat better happen soon, you’re killing me here!” I said, laughing, and setting off towards the stone house.

His smile faltered a little, but he didn’t comment.

Reaching back I took the case file from under the passenger seat, and handed one of the basic forms to Johnny.

That has his handwriting. And the address at the top is the station, maybe it could be sent there?”

No problem,” he said. He took a picture of the form with his phone and messaged it to ‘his guy’.

I pulled up to the stone house – it’s grandeur had become lost on me. Johnny had Capinski’s car keys and we planned to meet up at the diner the next day. He knew as well as I did that as soon as Carly woke up, she and I would have to ‘escape’, and the police would have questions for us victims all night.

I waved as he drove away, surprising myself at how eager I was to see him the next day.

Walking into the cellar I could hear the faint sound of Carly’s breathing. She looked so peaceful. I ran my hand along her body, mimicking Capinski’s movement from earlier. She shivered. It was slight, but there.

She was awake.

Next Chapter: Zealous


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Best Natural Weight Loss Resource

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Every Mother Counts

Today I’m sharing Marcella’s birth story over at Ergobaby as part of the Every Mother Counts Birth Story Series.

Every Mother Counts is an organization started by Christy Turlington-Burns (yes, the model). Just like my birth with Marcella, Christy experienced postpartum hemorrhage after her first birth. This complication for both us was managed by our birth professionals expertly, but women in other countries aren’t so lucky. In fact, postpartum hemorrhage often means death for thousands of girls and women around the world and is the leading cause of pregnancy-related death in the world, including in the US.

This information drove Christy to find out why, and she learned that 90% of these deaths are completely preventable. It started with the documentary, No Woman, No Cry. People watched and wanted to know how they could help. Thus Every Mother Counts was born with a mission of arming people with information to help end the needless loss of life of mothers around the world.

You may be wondering what Ergobaby has to do with Every Mother Counts. Well, they have partnered with Every Mother Counts for an exclusive designer series which includes 2 carriers, a diaper bag and a hip & shoulder bag. A portion of the proceeds of every purchase made of this collection will go to prevent pregnancy-related complications around the world.

You can learn more about Every Mother Counts by watching the video embedded below.

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June 19, 2009

Bantu Knots

So, I’ve finally figured out how to do Bantu Knots after a few weeks of trying. The problem is, I never get the consistency I want. One day, they’re perfect, the next I’ll have to do some kind of funky updo to conceal the mess that is my hair!!! Today is that day. I was doing 2 strand twists with perm rods every night. But that took up so much time, that Bantu knots would be a better option. But, with this hot mess on my head, I may go back to the 2 strand twists.

I’m 4 months into my transition and I have about 1 inch of new growth. Although, it seems like I have more than that in the crown area. Everyday, I rehydrate my hair to make it more manageable. I have a 12oz spray bottle filled with Distilled Water, 20 drops of Rosemary EO and 20 drops of Lemon EO. That area being the hardest to comb thru! It works out o.k. I get some breakage and my hair is dry in the morning. So I tried something different.

After watching several youtube.com clips, I saw several people talk about Vegetable Glycerin (I always think of Nitroglycerin when I think of it..not the same). Well, Vegetable Glycerin is supposed to make your hair soft and keep it moisturized. Since I need moisture, I picked up a 32 oz bottle from the health foods store for $20. One video on YouTube said that it should be a 75/25 ratio in your spray bottle. That’s 75% water and 25% vegetable glycerin. That’s what I did last night.

I lightly sprayed my hair, combed it thru and did my Bantu knots. I blow dried the knots a lil and tied my hair up. I woke up this morning and took out my knots and some were still damp. But, my hair is definitely softer!!!! Not only is that, but my curls in my natural hair are more open and loose. I just wish I had more than an inch…

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