text:
- Research article
- Open Access
- Open Peer Review
Effects of simvastatin on cell viability and proinflammatory pathways in lung adenocarcinoma cells exposed to hydrogen peroxide
© Gallelli et al.; licensee BioMed Central Ltd. 2014
- Received: 19 April 2014
- Accepted: 18 November 2014
- Published: 29 November 2014
Lung cancer is characterized by a high mortality rate probably attributable to early metastasis. Oxidative stress is involved in development and progression of lung cancer, through cellular and molecular mechanisms which at least in part overlap with proinflammatory pathways. Simvastatin is a statin with pleiotropic effects that can also act as an anti-oxidant agent, and these pharmacologic properties may contribute to its potential anti-cancer activity. Therefore, the aim of this study was to evaluate, in the human lung adenocarcinoma cell line GLC-82, the effects of a 24-hour treatment with simvastatin on hydrogen peroxide (H2O2)-induced changes in cell viability, ERK phosphorylation, matrix metalloproteinase (MMP) expression, innate immunity signaling, NF-κB activation and IL-8 secretion. Cell counting was performed after trypan blue staining, cell proliferation was assessed using MTT assay, and apoptosis was evaluated through caspase-3 activation and Tunel assay. Western blotting was used to analyze protein extracts, and IL-8 release into cell culture supernatants was assessed by ELISA. Our results show that simvastatin (30 μM) significantly (P <0.01) inhibited the proliferative effect of H2O2 (0.5 mM) and its stimulatory actions on ERK1/2 phosphorylation, NF-κB activation and IL-8 production. Furthermore, simvastatin decreased H2O2-mediated induction of the cellular expression of MMP-2 and MMP-9, as well as of several components of the signaling complex activated by innate immune responses, including MyD88, TRAF2, TRAF6 and TRADD. In conclusion, these findings suggest that simvastatin could play a role in prevention and treatment of lung cancer via modulation of important proinflammatory and tumorigenic events promoted by oxidative stress.
- Lung cancer
- NF-κB
- Matrix metalloproteinases
- Innate immunity
- Simvastatin
The respiratory system is remarkably susceptible to oxidative stress because of its peculiar anatomical and functional properties, mainly related to the large area exposed to the external environment. Therefore, the cellular/tissue injury triggered by the oxidant burden generated by air pollutants in association with cigarette smoking plays a pivotal role in the pathogenesis of several inflammatory and proliferative lung disorders, including chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, and also lung cancer.
In particular, inhaled oxidants such as ozone and nitrogen dioxide cause sequestration of inflammatory cells into the pulmonary microcirculation, thus leading to their accumulation within air spaces. Cigarette smoke, which contains many oxidants and free radicals in both its gaseous and particulate phases [1], significantly contributes to recruit macrophages into the airways, as well as to increase neutrophil numbers within lung microvessels. Once recruited and activated, macrophages, neutrophils and eosinophils produce and release reactive oxygen species (ROS) such as hydroxyl radicals and superoxide anion (O2.-), the latter being rapidly converted to hydrogen peroxide (H2O2) by superoxide dismutase (SOD) [2]. ROS may interfere with signal transduction pathways regulating the functions of transcription factors such as nuclear factor κB (NF-κB) and activator protein-1 (AP-1) [3]. NF-κB and AP-1 are responsible for the coordinated expression of several genes that control inflammation, cell proliferation and apoptosis. Within this context a key role is played by mitogen-activated protein kinases (MAPK), whose targets are mainly represented by nuclear transcription factors, also including those involved in oxidative stress. In particular, the ERK1/2 subgroup of MAPK is activated by a MAPK kinase kinase named Raf (most commonly Raf-1), whose activation in turn requires the GTP-bound form of Ras family proteins [4]. Once activated, Raf-1 phosphorylates the MAPK kinases MEK1 and MEK2, that finally stimulate ERK1 and ERK2.
Airborne pollutants and cigarette smoke can induce the bronchial epithelium to acquire a proinflammatory phenotype, characterized by an increased production of autacoids, cytokines, and chemokines [5]. Oxidant-induced phenotypic changes may thus significantly contribute to the key pathogenic role played by bronchial epithelial cells in inflammatory airway disorders such as asthma and COPD. Moreover, ROS may also contribute via several different signalling pathways, including MAPK activation, to development and progression of lung cancer [6].
Lung cancer is the leading cause of neoplastic death worldwide. More than 80% of lung cancer cases belong to the non small cell lung cancer (NSCLC) type, which can be further subdivided into adenocarcinoma (approximately 40% of all NSCLCs), squamous cell carcinoma and large cell carcinoma [7]. The metastatic potential of NSCLC strongly correlates with the cellular expression of matrix metalloproteinases (MMPs), which are regulated by NF-κB and by the metastasis suppressor RECK (reversion-inducing-cysteine-rich protein with kazal motif) [8, 9]. Moreover, the complex cellular and molecular mechanisms underlying the development and progression of NSCLC are also significantly affected by the innate immune system [6]. Indeed, the latter represents the first line of defense against noxious agents such as ROS, which can damage the airway epithelium.
The poor survival rate of patients with NSCLC is mostly due to its high metastatic potential and also to a relative drug resistance [7]. Therefore, new and more effective pharmacological treatments for NSCLC are strongly needed. In this regard, increasing attention is currently being paid to statins because of their capability of inhibiting 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, which is the rate limiting enzyme within the mevalonate pathway. Hence, by blocking the synthesis of mevalonate and its isoprenoid derivatives farnesyl pyrophosphate (FPP) and geranyl geranyl pyrophosphate (GGPP), statins also prevent prenylation-dependent activation of oncogenic Ras proteins [10]. Indeed, statins may have a cytostatic effect on cancer cells, and can prolong the survival of cancer patients [11]. They also act as anti-oxidant, anti-inflammatory and anti-angiogenic factors, and could therefore prevent or inhibit cancer cell growth [12]. Furthermore, we have recently shown that simvastatin is able to induce a decrease in cell proliferation and a significant increase of apoptosis in human NSCLC cell cultures [13], as well as to modulate Ras, MMP-2/9 and NF-κB activity in pulmonary neoplastic tissues obtained from patients undergoing therapeutic surgery for lung cancer [14].
On the basis of the above mentioned considerations, the aim of our present study has been to investigate, in the human lung adenocarcinoma cell line GLC-82 exposed to H2O2, the effects of simvastatin on cell viability and apoptosis, ERK phosphorylation, MMP2/9 and RECK protein expression, NF-κB nuclear content and interleukin-8 (IL-8) secretion. Moreover, we also assessed the effects of simvastatin on H2O2-induced changes in the expression of some important proteins of the signaling network underlying innate immune responses, including MyD88, tumor necrosis factor (TNF) receptor-associated factors (TRAF)2/6, and TNF receptor type 1-associated death domain protein (TRADD).
The anti-caspase-3 monoclonal antibody E83-77 was purchased from Abcam (Cambridge, UK). Trypan blue was purchased from Sigma (St. Louis, MO, USA). Anti-phospho-ERK1/2, anti-NF-κB-p65, anti-RECK, anti-MyD88, anti-TRAF2, anti-TRADD, anti-TRAF6, anti-MMP2 and anti-MMP9 monoclonal antibodies were purchased from New England Biolabs (Beverly, MA, USA); an anti-(total)-ERK1/2 polyclonal antibody was commercially provided by Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Purified crystalline powders of simvastatin were commercially obtained from Sigma (St. Louis, MO, USA), and then dissolved into refrigerated and light-protected DMSO stock solution.
Culture and treatment of human lung cancer cells
GLC-82, a human lung adenocarcinoma cell line [13], was cultured at 37°C, 5% CO2, in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FCS, penicillin 100 U/ml, streptomycin 100 μg/ml, and amphotericin B 25 μg/ml in a humidified 5% CO2 atmosphere. For each treatment, cells were plated in a 100-mm polystyrene dish (Falcon, Becton-Dickinson, Lincoln Park, NJ, USA) and ten ml of supplemented DMEM were then added. When GLC-82 cells grew to about 70% confluence, they were exposed for 2 hours to H2O2 (0.5 mM) and then cells were or not exposed for 24 hour to simvastatin (30 μM). The medium was not changed after treatment. The solvent employed to dissolve this drug was used as a control. After this period, the medium was removed for IL-8 assessment (see later), and cells were processed for protein extraction and immunoblotting.
Cell viability and proliferation
Cell viability was assessed by light microscopy and dye exclusion, using Trypan blue. Cell numbers were evaluated by direct counting using a hemocytometer, in agreement with our previous reports [15–17]. Cell proliferation was investigated by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, based on the conversion by mitochondrial dehydrogenases of the substrate containing a tetrazolium ring into blue formazan, detectable spectrophotometrically. The level of blue formazan was then used as an indirect index of cell density. Briefly, after treatment with either H2O2 and/or simvastatin, cells were exposed to MTT (5 μg/mL) for 150 min at 37°C. The medium was then removed and cells were solubilized with acidified isopropanol and 2% sodium dodecyl sulfate (SDS). After complete solubilization, presence of blue formazan was evaluated spectrophotometrically at a reference wavelength of 650 nm. All experiments were carried out in triplicate.
Protein extraction and immunoblot analysis
Following treatment with simvastatin, cells were lysed for Western blotting in radioimmunoprecipitation assay (RIPA) buffer, as previously described [18]. Moreover, nuclear extracts were obtained using the NE-PER cell fractionation kit (Thermo Scientific, Rockford, IL, USA). Briefly, whole cell lysates or nuclear proteins were then separated on a 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto polyvinylidene difluoride (PVDF) membranes (Amersham Pharmacia, Little Chalfont, UK). Immunoblotting was performed using the above mentioned monoclonal antibodies. After being "stripped", membranes were re-probed with a polyclonal antibody against total (phosphorylated and unphosphorylated) ERK1/2 proteins. Antibody binding was visualized by enhanced chemiluminescence (ECL-Plus; Amersham Pharmacia); intensities of experimental bands were analyzed by computer-assisted densitometry and expressed as arbitrary units, as previously described [19]. These experiments were performed in triplicate.
TUNEL assay
TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labeling (TUNEL) assay was performed in agreement with our previous reports [13]. Briefly, cells were plated on 24 9 24 mm cover slips (Carlo Erba Reagenti, Milan, Italy) and placed in six-well microtitre plates (Corning Incorporated, Corning, NY, USA) in DMEM supplemented with 10% FCS, penicillin 100 U/ml, streptomycin 100 mg/ml, and fungizone 25 mg/ml; 24 h after plating, when cells had reached 50–60% confluence, they were treated or not with H2O2 (0.5 mM) for 2 hours, and then were exposed or not to simvastatin (30 μM) for 24 hours; the incubation medium was not changed. Following treatment, TUNEL assay was performed using MEBSTAIN Apoptosis TUNEL Kit Direct (MBL, Woburn, MA, USA), strictly following instructions provided by the manufacturer. Photographs were acquired using a Leica GRDM confocal microscope (Leica, Wetzlar, Germany).
IL-8 secretion
Culture supernatants were collected and assayed for IL-8 by ELISA using a commercially available kit (Peli-Kine kit; Eurogenetics, Hampton, UK; sensitivity limit, 1 pg/ml), according to manufacturer's protocol. These experiments were performed in triplicate.
Statistical analysis
All data are expressed as mean ± standard error (SEM). Statistical evaluation of the results was performed by analysis of variance (ANOVA). Differences identified by ANOVA were pinpointed by unpaired Student's t test. The threshold of statistical significance was set at P <0.05.
Cell viability
Caspase-3 activation
TUNEL assay
ERK phosphorylation
Matrix metalloproteinase expression
RECK expression
NF-κB nuclear content
MyD88 and TRAF6 expression
TRADD and TRAF2 expression
IL-8 secretion
In this study we evaluated the effects of simvastatin on lung adenocarcinoma GLC-82 cells, exposed to hydrogen peroxide (H2O2). Interestingly, oxidants are capable of exerting a tumorigenic action via molecular mechanisms which at least in part are also activated by proinflammatory pathways [20, 21], and several authors have shown that both oxidative stress and inflammation are involved in the development and progression of cancer [22].
Therefore, we focused our attention on several cellular events triggered by oxidative stress. Within such a research context, it is noteworthy to point out that simvastatin was able to induce a comprehensive inhibition of many phenomena related to lung cancer.
In this regard, Manfredini et al. have previously reported that simvastatin is able to modulate oxidative DNA damage [23], and we also showed that simvastatin significantly decreased H2O2-induced cell proliferation rate, probably through inhibition of Ras-dependent phosphorylative activation of the ERK1/2 subgroup of MAPKs [13]. In fact, H2O2 dramatically enhanced ERK phosphorylation, and such a stimulatory action was markedly reduced by simvastatin. This implies that simvastatin affects ERK function by inhibiting mevalonate-dependent post-translational prenylation of Ras, thus impairing the sequential activation of the Ras/Raf/MEK/ERK signalling cascade [24]. Therefore, this mechanism may be also involved in the observed pro-apoptotic action of simvastatin, documented by the increased expression of active caspase-3 [25].
Interestingly, in our present study the anti-proliferative and pro-apoptotic effects of simvastatin were also paralleled by a drug-induced up-regulation of RECK protein, which plays a pivotal role in inhibiting matrix metalloproteinases. MMP-2 and MMP-9 are responsible for extracellular matrix degradation, and for the consequent neo-angiogenesis, vascular invasion and metastatic potential that characterize malignant tumours [26–28]. Indeed, in many cancers RECK down-regulation is associated with high levels of MMP-9 [9]. Otherwise, preservation of RECK expression in some neoplasms correlates with a relatively low microvascular density as well as with a better prognosis, due to a decreased tendency to metastatic invasion [29]. Consistently with these considerations, our results show that H2O2 down-regulated RECK in GLC-82 cells, whereas simvastatin restored its expression. Moreover, H2O2 enhanced MMP-2 and MMP-9 cellular expression, and these effects of oxidative stress were prevented by simvastatin.
Other important observations reported in the present study refer to further aspects of signal transduction. In particular, our data demonstrate that the noxious effects of oxidative stress on human lung adenocarcinoma cell lines, as well as the potential anti-cancer activity of statins, are also attributable to interferences with the signalling pathways linked to innate immunity [30]. In this regard, it is remarkable that H2O2 induced the expression of MyD88, a key molecular component of the intracellular signalling machinery activated by Toll-like receptors (TLRs), and once again simvastatin abrogated such effect. MyD88 associates with TLRs; upon stimulation, MyD88 recruits IL-1 receptor-associated kinase (IRAK) to TLRs via interaction of the death domains of both molecules [31]. IRAK is activated by phosphorylation and then associates with tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6). Similarly to MyD88, the opposite actions of H2O2 and simvastatin translated into significant increases and decreases in TRAF6 expression, respectively. Therefore, on the basis of this experimental work, we canspeculate that oxidative stress triggers a particular type of tissue injury which is sensed by the surveillance network of innate immunity via TLR stimulation. In fact, it has been demonstrated that H2O2 can prime the responsiveness of the innate immune system through a recruitment of TLRs to cell plasma membrane [32]. However, this chain of molecular events, including the essential step of MyD88 activation, ultimately culminates in the downstream involvement of the transcription factor NF-κB, which is a key effector of the carcinogenic and proinflammatory attitudes of oxidative stress. It can thus be argued that, by interrupting such complex molecular cascades, statins can have relevant anti-cancer and anti-inflammatory properties.
Other important components of the macromolecular complex associated with TNF receptor superfamily, whose stimulation leads to NF-κB activation, include TRADD and TRAF2 proteins [33]. Here it is shown that both TRADD and TRAF2 were susceptible to up-regulation induced by H2O2, and this effect was prevented by simvastatin. Because the above mentioned signaling pathways converge towards NF-κB activation, it was not surprising to detect in GCL-82 cells, as a result of their exposure to H2O2, an increased nuclear content of NF-κB. Indeed, nuclear translocation of this transcription factor is the essential step underlying its activation. Consistently with the overall experimental design of this study, it was well expected to find the observed decrease of NF-κB nuclear levels elicited by simvastatin.
Moreover, a further evidence of the opposite actions of H2O2 and simvastatin was found by analyzing, in GLC-82 cell culture supernatants, the changes referring to the concentrations of IL-8, which acts as a powerful chemoattractant for neutrophils [34]. In particular, IL-8 secretion was up-regulated and down-regulated by H2O2 and simvastatin, respectively. Indeed, this proinflammatory chemokine is encoded by a gene targeted by the transcriptional stimulatory activity of NF-κB [35].
Therefore, taken together these results strongly suggest the existence of a tight cross-talk involving the proliferative, metastatic and proinflammatory mechanisms activated by oxidative stress in GLC-82 cells. Moreover, our data clearly demonstrate that in this lung adenocarcinoma cell line simvastatin can effectively interfere with the complex signal transduction networks underlying such mechanisms. These findings confirm and further extend our previous data obtained in the same GLC-82 cell line, showing that simvastatin was able to significantly decrease cell proliferation and increase apoptosis via inhibition of the ERK1/2 subgroup of MAPKs [13].
With regard to MMP-2 and MMP-9, it is well known that these enzymes promote cancer progression through extracellular matrix and basement membrane degradation, resulting in the exposure of cryptic locations linked to invasion, metastasis and angiogenesis. The 5′ flanking regions of the MMP-2 and MMP-9 genes contain several functional regulatory motifs that bind transcription factors such as NF-κB [36, 37]. Through the interactions between NF-κB and its DNA binding sites, various agents including growth factors and cytokines are able to regulate MMP expression. Hence, the coordinated inhibitory effects of simvastatin on MMP2/9 expression and NF-κB activation imply the involvement of HMG-CoA reductase pathway in the process of metastatic invasion related to NF-κB and MMP activity. In fact, cancer cells usually express high levels of HMG-CoA reductase, which appear to be required by NSCLC cells to satisfy their increased need for isoprenoids and lipids.
Thus, by blocking Ras prenylation, simvastatin could interfere with NF-κB-dependent signaling networks responsible not only for MMP expression, but also for IL-8 synthesis. Indeed, it is reasonable to infer that, by inhibiting Ras/Raf-induced phosphorylative activation of MAPKs, simvastatin may be able to prevent IL-8 production elicited in GLC-82 cells by NF-κB activation due to oxidative stress.
In conclusion, our results show that simvastatin is able to inhibit the effects of H2O2 on lung cancer cells, suggesting that simvastatin is able to induce an antiproliferative, pro-apoptotic and antinflammatory effects in presence of a pro-inflammatory mediator. These data could support the role of simvastatin in the prevention of cancer and lung inflammatory disease (i.e. chronic obstructive pulmonary diseases). In fact to date no definitive reports have been published regarding these diseases [38, 39]. However, in order to validate these preliminary observations further studies are needed, especially including controlled clinical trials.
Daniela Falcone and Monica Scaramuzzino" were supported by a fellowship of the PhD Programme in Molecular Oncology, Experimental Immunology and Development of Innovative Therapies. This research was supported in part by a grant from Sicily Region (PO FESR Sicilia 2007/2013, CUP G73F11000050004 to MN, project "MEPRA", N. 133 of Linea d'Intervento 220.127.116.11).
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Pre-publication history
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The first step in becoming part of the CrossFit Athletic community is being assessed by one of our Coaches. This is an one-hour consultation in which we focus on three key areas – lifestyle, movement patterns and mobility.
When assessing your lifestyle, we like to know more about your exercise history and experience, lifestyle including sleep, nutrition and stress and whether you may have any injuries that may affect your movement patterns.
When assessing your movement patterns, we want to see how your body moves both unloaded and with load. We want to compare your left to your right sides to see how balanced you are; for example, is one leg stronger than the other and what implications will that have on your progress. How is your upper body push and pull – is it balanced? And what about your lower back? We take 10 minutes to assess the strength and endurance of your back muscles to ensure you safely enter our program and not be at risk of injury.
When assessing your mobility, we start with the feet and work our way up. It's important that we assess the critical junctions – the ankle, hips and thoracic – to see how they move and what type of mobility you have in these key areas. Many of our clients sit at a desk in front of a computer all day, so there is a very good chance that the thoracic and hip mobility is limited.
Once we have all of this information, we can then make an educated recommendation on what path you should take before you become a member at CrossFit Athletic. This could be Personal Training to get you up to speed to join our Classes, our On-Ramp Program which builds you up over 3 weeks or straight into our CrossFit Classes.
Our aim is to set you up for success and give you the best possible foundation. And we would not have it any other way.


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Home > Renewable Energy Systems Boston MA
When you turn to us for renewable energy systems and sustainability services for your project or facility in Boston, MA, you'll benefit from our Energy Management & Renewable Generation team's many services, which are proven to provide you with measurable savings. These services include:
For even more energy savings solutions, we can provide further assistance through our Power Distribution & Automation Solutions (PD&AS) department. Our PD&AS team is made up of highly specialized engineers who will listen closely to your needs to provide value-added engineering services and solutions for your power systems, including solar and fuel cell applications.
If you're interested in learning more about the renewable energy systems and services we offer for projects and facilities in Boston, MA, contact Turtle & Hughes today.
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Copyright 2019 Turtle & Hughes, Inc.


text:
Samuel Bozzette
Visiting Scholar
- Profile
Bozzette is an visiting scholar at the School, a senior natural scientist at the RAND and currently serves as vice president of Medical Affairs-Americas at bioMerieuxs.
He is board-certified in internal medicine and infectious diseases, and is a fellow of the American College of Physicians and the Infectious Diseases Society of America. Bozzette is an elected member of the American Society for Clinical Investigation, the Association of American Physicians and the Pacific Council on International Policy (the western affiliate of the Council on Foreign Relations).
He has a strong record of clinical, translational, outcomes and health economics research, resulting in over 150 publications cited over 4,000 times, including over a dozen in the New England Journal of Medicine. Several of these studies significantly changed clinical practice or policy, including related guidelines, rules and laws.
For more information, please visit Samuel Bozzette's personal site.
- firstname.lastname@example.com
- (858) 532-4324
9500 Gilman Dr. #0519
La Jolla , California 92093-0519


text:
I hate most interviews, and I think everyone else does too. They're rarely actionable or insightful.
This interview is uncommonly different. In the last interview Peldi Guilizzoni (Balsamiq Studios) gave detailed advice about how he earned almost a million dollars in revenue in his first year of operation.
In this installment we hear from startup expert Bob Walsh, whose many works for startup founders include:
- Blog: 47 Hats
- Podcast: Startup Success
- New book: Web Startup Success Guide
- Classic book: Micro-ISV: From Vision to Reality
- Online guides and support network for founders: StartupToDo.com
Below Bob digs into his years of experience living with, mentoring, and thinking about tiny startups to tell us what's important for a founder starting a company today.
Q: You've stated that the goal of both your new book (Web Startup Success Guide) and your new tool for entrepreneurs (StartupToDo) is to double an entrepreneur's chance at success: Increase from 10% to 20%. That's a terrific goal — and it certainly seems plausible — but to what do you attribute the other 80% of the risk? Luck? Timing? Ideas? Tenacity? What are the big killers lurking for the new startup founder?
I think the big killers of newborn startups lurk mostly inside the heads of founders. First and foremost, succeeding as a Digital Entrepreneur (my new catch-all keyword for traditional funded startups, unfunded startups, microISVs, indie game developers and content creators like bands and authors. Like it?) means you need to be the most stubborn S.O.B. you know. Perseverance is the single most important attribute you need. Pick any 5 successful software companies you'd like from one-person microISVs up to the largest and trace how their market focus, product, marketing etc. has zigged and zagged from inception. That's staying in the game until you find what works.
Editor's note: Most people think "perseverance" means "being so confident in your ideas that you don't change your mind even when there's reason to doubt." But Bob points out that companies are fluid, and those who survive are stubborn about building something good, not about any one idea.
Another reason DE's turn back is it's lonely out there. Whether it's creating a software app that you're going to build a global one-person business around, or giving away the Science Fiction stories you write as you build your community so you can launch a book, this is lonely stuff few back in the safe (ha!) warrens of working for some large company would dare do.
Q: You spent two years working on StartupToDo before bringing it to market. Clearly it's the culmination of your years of experience with startups — writing about them, mentoring them, promoting them, and watching them grow, shrink, live, and die. How were you able to convert this knowledge into a software product? What aspects of building a company can be encoded in this manner, and what's still outside the realm of "how-tos?"
When it comes to software, you as the developer have got to really understand both the people who have the problem you want to address and the problem itself. Now maybe you get this understanding working for a company, writing software for clients, or because you are as the saying goes scratching your own itch.
Between the various things I've been doing and my own efforts I started noticing just how much time in this business goes into context work — all of the stuff besides the core of what your are building — and how hard it is, how much you have to sacrifice, especially if you are bootstrapping. I realized that startups reinvent hundreds of wheels and that's an awful way to build businesses. Then one day I read a comment on Darren Rowse's blog by a guy who created a point system to keep himself blogging, consistently, and motivated and I was electrified by the idea putting "actionable guides and discussions" together with "measurable progress you could see, share, and compete with in a friendly way." It took about a year and a half to start learning rails, go off on a tangent (the teachingsells.com approach) get back to learning rails, go off on another tangent (Adobe AIR — good stuff, but not a good fit), get back to learning rails/JS etc. sufficiently well to — with a lot of help — code StartupToDo.com.
There's a lot of value being able to talk about current business issues with your peers and hearing what other people recommend (See the Business of Software Forum and Answers.Onstartups.com), and getting a solid overview/education on one of the many aspects of this business (see Microprenuer Academy). But one of the unfortunate aspects of "forumizing" is it's easy to substitute endless online talk for concrete action. I've really tried to build the first two big chunks of value in my startup to create what I call focused, actionable discussion.
To Do's are not enough. Hopefully the Guides at my startup that I and other community members are writing are more than static recipes.
Q: You're a big advocate for startups using social media because it's essentially free (in dollars). But it costs a lot in time and effort — another scarce resource. Is traditional marketing and advertising completely dead, or is there still room for it in a startup?
Jason, you're right: social media marketing takes lots of time. And depending on your startup's market, traditional marketing (paper spam, email spam, sales visits, shows, conferences, display ads, etc.) may still have some residual value.
But there are three things at work here. First, developed country societies are moving from industrialism where standardized advertising worked to post-industrialism where the Internet empowers people, markets and conversations. Second, the core values that you find among people who came of age after the Internet are markedly different – the ugly corporate truth is traditional marketing and advertising barely influence them while what their personal social networks "think" and how they interact person-to-person (e.g. Zappos) matter far more.
Here's the third: Few startups (let alone Digital Entrepreneurs) have more money than time. It happens to be a very happy circumstance that we can now translate time via social marketing (blogs, Twitter, Facebook and all the rest) into sales. Unfortunately, developers in general hate to market (social or conventional), hence why I'm focusing on making marketing, especially what you do online, ultra easy as the next big feature of StartupToDo.com.
Q: You're well-known for promoting David Allen's Getting Things Done (GTD) system for organization and productivity, and Chapter 8 of your Web Startup Success Guide is devoted to it. How important is "personal productivity" to the success of a startup? We all know people who are completely disorganized yet effective, and others who have everything in order but never seem to accomplish anything. Is organization fundamental to startup success because time is so precious? Or can it just be a distraction?
For 25 or so years when I built software for San Fransisco Bay Area corporations as a contractor, personal productivity was my obsession — the more and better ways I could improve my productivity, the more money I'd make. That's probably why my first commercial product was a Windows desktop personal task manager.
So far, David Allen's GTD methodology of getting tasks out of your head, processing them efficiently and tracking your results is the best productivity theory we have, IMO. If you can find an effective, not obsessive, way to apply that, you're going to have a decided advantage in just about anything you do in this world, including creating your startup.
The key thing about personal organization is it's very personal. Have you ever seen the shot of Al Gore working on three monitors in his office surrounded by stacks of books, paper, and whatnot? That's what works for him, with demonstrable results.
Finding/creating the right tools for you to move tasks to results in a consistent manner is what matters, not the ceremony or form you use.
Presently I use Things on my Mac and iPhone to track tasks, Notebook on my Mac to plan in, and SpeakEasy on my iPhone as a way to capture tasks and thoughts throughout the day I can later process. Also, I find Basecamp comfortable when I need to project management with others and checkvist.com an easy way to collaborate on outlines.
Q: You wait until Chapter 7 of the Web Startup Success Guide to talk about the Unique Selling Proposition (USP) that uniquely describes your company's purpose and niche. Do you believe startups need to determine their USP from the beginning (because it's vital to defining yourself) or is it something you back into over time (because you have to seek your market position)? In either case, how exactly do you go about defining it?
I believe the Internet rewards original content and punishes unoriginal content, be it software, ideas, or products. If you can't explain to the market and yourself who your product or service helps how, you're in trouble. I think that the earlier you can define your startup's USP the better and recently I've found some validation of that approach in what Marc Andreessen and Steve Blank have said about product/market Fit (useful stuff, and yes, I'm writing it up as a S2D Guide!).
That said, the "defining part" is no easy exercise. I wish I could boil it down to a checklist, but there's more to it than that: your USP depends on what you are bringing to the party. Your experience, insight, opinions, values and abilities define your solution to the problem. One thing I think is true: is it's an iterative process —the more times you examine your USP, the sharper, more useful, more effective it becomes.
Q: Your book is clearly targeted at web-based software companies, and perhaps even more specifically at software developers. What of this advice is applicable to other businesses, or at least businesses who want a strong web and social media presence? For the non-technical business owners out there, what parts of the book should they still read, and how do they know which of your advice to apply?
Guilty! :-)
I write/podcast/work for developers who want to build their own software and software companies. Why? Because those are the people I like, and want to help succeed.
But for a while I've been thinking that software startups, microISVs, bands, authors, and others have more in common and share more values online than they do offline. Hence the Digital Entrepreneur meme. Neither The Web Startup Success Guide , the Startup Success Podcast or StartupToDo.com are for programmers as programmers — you won't find a single line of code in the lot of them. So, whether you are or are not a programmer, the stuff I've created over the years will have some value for you.
Any questions?
Thanks Bob! Please leave questions (or arguments!) in the comments to continue the discussion.


text:
Official website of the Department of Homeland Security
The EEO Investigator reviews the complaint to determine if it meets the requirements for acceptance. If the claim is accepted, the assigned EEO Investigator will conduct the investigation within 180 calendar days from the filing date of the claim. The EEO Investigator serves as an unbiased fact gatherer identifying and securing information through interviews of witnesses and review of written records. This information is compiled into a Report of Investigation (ROI), which must be thorough enough to enable an appointed decision maker to make a final decision as to whether unlawful employment discrimination occurred with regard to the claim(s) investigated.


text:
Psychedelic Americana - a celebration of the American song catalog, centered around an interpretation of Allen Ginsberg's song 'Western Ballad
204 backers pledged $23,171 to help bring this project to life.
Replay with sound
pledged of $10,000 goal
Our aim here is to fund my most recent recording project called 'Western Ballad'. My dear friend Mark Bingham and I wrote all of the songs with exception of a traditional hobo song called 'Little Stream of Whiskey' and the title track 'Western Ballad' penned by Allen Ginsberg.
All of the money raised here will allow us to create a beautiful package by working with independent graphic artists and pressing services. I am a completely self funded artist. I do not have a record label. With your help I'll bring this music to as many people as possible.
Questions about this project? Check out the FAQ
Funding period
- (40 days)


text:
Boy, it's been a helluva year for Toyota (TM).
First the brakes go. Now the engines on a coupla hundred thousand Lexuses (Lexii?).
Japan daily Asahi said today repairing all 270,000 automobiles could cost Toyota 20 billion yen ($227 million).
But, let's look at this from an engineering point of view.
I mean, I'm no engineer, but I've met a couple before. And from what I can tell, Toyota has now unintentionally engineered its way out of a problem it engineered itself into:
The first recall was due to failing brakes.
The latest recall is due to failing engines.
Problem solved!
Your brakes go out, just sit back and wait for the motor to cut out.
There, I just saved Toyota $227 million.
You're welcome.
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text:
up to the Verdun Museum and Memorial you pass the Souville Bunker where
the German attack on Verdun was stopped.
next stop is at the "Wounded Lion". This Memorial marks
oficially the point of the clostest German Advance on Verdun. Not quite
right because the German troops came as far as Fort Souville, some
houndred feet closer to the town ~ who cares.
Then make a stop to visit the Verdun Museum and Memorial. The impressive museum offers a phantastic collection of Uniforms, Weaponry, Vehicles and displays. The everyday struggle to survive of the French "Poilou" (Fr. word for soldier) and his counterpart the German "Landser"(Ger. word for soldier).
Here you can get a depressing insight look of the mass murder at Verdun.
the museum head for the ossuary and stop at the destroyed village of Fleurie.
de Douaumont
beginning of the construction was August the 22nd 1920. Marshal Petain
the defender of Verdun was present on this day. It
is 37 metres in length, lang and the bell weighs about 2300 kg.
Inside is a memorial chapel and above the crypts are names of the
battlefields inscribed where the remains of the soldiers were found.
Have a look throu the little windows from where you see the remains of the 130.000 unknown soldiers.
In the evening you should go to one of the many restaurants at the river Meuse and enjoy the beautiful old town that was rebuilt after WWI.
Day 2
breakfast check out of the hotel and leave around 07:30 to have
enough time to see more of the battlefields of Verdun.
give you an idea of what happened there you should imagine that the
preperatory fire of the German attacks for June 22nd and 23rd 1916 the
German artillery fired about 100.000 grenades along the front.
Fort was built in 1885 and reinforced until 1913.
In 1915 the guns were taken out of the Fort and on the 25th of February 1916 the Douaumont is taken by the Germans .
was retaken on October the 24th 1916 at a tremendous cost of French
After visiting the Douaumont you
should have a short visit to Fort Vaux


text:
Rangers are poised to make a formal offer to Hibs as they bid to sign prized midfielder Scott Allan.
The 23-year-old has been a stand-out for the Easter Road outfit men since he joined last summer and his impressive performances have caught the eye of both Rangers, Celtic and Dundee United in the past six months.
United had a bid of £250,000 rejected in January - the only concrete approach made for the former Scotland Under-21 midfielder - and despite a summer of speculation, no clubs have bid as yet for last season's Championship Player of the Year.
However, new Rangers manager Mark Warburton is understood to have watched Allan during pre-season and has been impressed by the creative midfielder, who has scored in both Hibs' friendlies against Berwick and Dunfermline.
Allan has one year to run on his Hibs contract and has said that he is happy to see out the final year of his deal in Edinburgh, insisting that he enjoys working under head coach Alan Stubbs and that he can be improved by him further. Hibs, on their part, are extremely reluctant to sell their star man and Stubbs has said that he'd rather see Allan leave for nothing in the summer than sell him ahead of this season's push for promotion.
Hibs and Rangers, who are the front-runners for this season's Championship, play on Saturday at Easter Road in the first round of the Petrofac Training Cup.


text:
YouTube is a website that everyone knows. It is has been one of the most successful websites, but somehow it has not been a site that has made lots of money in the past. This, however, was before the site was restructured by Google.
Many people have been seeing the YouTube site going through the change. Some people love the website. Other people do not care for the changes. It is designed to make a lot of money this way though. Now YouTube is getting geared up to compete against Netflix and HBO. There are already video links that go straight to the Vevo site. This is what YouTube will need for a movie website as well. Right now people can rent movies on YouTube, but the company needs to actually have an external link from YouTube for new movies rentals.


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Recruiters – how are your influencing skills?
Influencing skills are critical for recruiters, so how can you develop yours?
The ability to persuade or influence others is a key skill in recruitment whether this be influencing a company to use you for their recruitment or persuading a candidate that the job you are recruiting for is right for them.
There are many different ways that you can persuade or influence others. Everyone make decisions for their own reasons, so it is important to consider carefully the person you are dealing with and come from their perspective in order to decide the best method to use.
Here are the most common ways of persuading or influencing others:
Reasoning: Using facts, logic and reasons to put your point of view across. This is best used where you can support your views with good facts, logic and reasons.
Inspiring: Appealing to the other person's emotions, using energy, passion and conviction. This is best used when you are looking for emotional commitment and when the reasoned view is weak.
Asking: Asking questions to encourage the other person to make up their own mind. This is best used when you want the other person to buy into the outcome and when dealing with someone more senior than you.
Complimenting: Making the other person feel good. This is best used when the other person values your views and with people who look up to you.
Making a deal: Offering the other person something in return for what you want. This is best used when you don't mind making a deal and have something to offer as your part of the deal.
Calling in a favour: Getting what you want by calling in a favour. This is best used when you have a very good relationship with the other person.
Comparing to others: Persuading the other person by using the views of people they respect to support your argument. This is best used when the other person is easily swayed by the views of others.
Authority: Influencing by using rules or principles or quoting someone in authority. This should really only be used in exceptional circumstances as it will encourage compliance not commitment.
Forcing: Forcing the other person by using assertive behaviour. This is best used in emergency situations only.
To improve your own ability in this area, think about and consider how you persuade and influence others. Observe the methods that other people in recruitment you respect and admire use and how they use them, so that you can identify good practise. Then decide the methods that you want to use and try them out but be prepared to be flexible and afterwards review how it has gone so that you can identify areas for improvement next time.


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In historic downtown Macon across from the City Auditorium. Located at the intersection of Cotton Avenue and Cherry Street.
430 Cotton Avenue • Macon, GA 31201• 478/743-1368
NOTE: Due to fire damage of 3/13/15, restaurant is closed for repairs.
Monday - Friday: 6:00am - 7:00pm
Saturday: 7:00am - 6:00pm
Closed on Sunday


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Tuesday Nights are "Kids Eat Free" Nights
from 5:00-9:00 p.m.
Buy One (1) Full Priced Entrée and
receive (1) Free Kids Meal for your little one.
*Special Conditions: Eat-in dining only , Child must be 10 years of Age or Younger. Free Meal is a choice of any regular meal from the Kids Menu (additions extra).
(Appetizers, Salads, Pizza, or Sandwiches are NOT Considered Entrées). Thank You.
Make your reservations or just show up!!!