text:
Man shot outside North Memphis home
MEMPHIS, Tenn. — A North Memphis man is in critical condition at the Regional Medical Center following an overnight shooting.
Police say the man was shot around 11:15 p.m. Thursday while sitting outside a house in the 800 block of Birch St. near Jackson Ave.
Police are still working to identify the gunman and determine what led to the shooting.
They're asking anyone with information on this shooting to call Crime Stoppers at (901) 528-CASH.
According to WREG records, MPD has responded to at least 10 shootings so far in 2017.
Four of those shootings have involved fatalities.
So far, there have been five homicides this year — which is down year to date versus 2016.
By January 13, 2016, MPD had already responded to 12 homicides.


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Tuesday, May 29, 2012
Color Me Happy On a Budget!
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subset: commoncrawl


text:
Dr. Sally Grissom
A brilliant, charismatic scientist from the future. After an accident with her Higgs Inhibitor Array in the year 20█, Sally finds herself in a world that is not her own: America, 1943. Highly opinionated and always ready to make a pop culture reference none of her pre-millennial friends would understand, Sally sticks out like a sore thumb in any group she's in. Determined to solve time travel and save the world, Sally is the physicist trapped in the past that any of us would want to be.
[photo collage by Mo Medved]


text:
Chelsea Clinton and investment banker boyfriend Marc Mezvinsky married on July 31 in a highly secretive affair that cost approximately $3 million. To avoid the media, the event was held on a posh private estate 90 miles north of Manhattan. Questions: 1. What was one of the least expensive items at the wedding? 2. What… Read more »
Posts Categorized: All Articles
HP CEO Resigns
Hewlett-Packard Co.'s Mark Hurd resigned as chief executive officer after an investigation found he had a personal relationship with a contractor who received numerous inappropriate payments from the company. Hurd submitted receipts for expenses ranging from $1,000 to $20,000 over two years, including meals and travel, that should have been labeled as personal and not… Read more »
Who are the most trustworthy companies and why?
Who can you trust? In a recent examination by Audit Integrity, an independent financial analytics company in Los Angeles, the company assessed the true quality of corporate accounting and management practices by looking at more than 100 factors beyond the balance sheet and income statement. Their aim was to identify the measures most highly associated… Read more »
The Costs of Liposuction
Lots of people are curious about the price of liposuction (which is also referred to as lipoplasty and suction lipectomy). Although the procedure sports a somewhat hefty price tag, it is among the cheaper cosmetic surgeries available. Some the of the cheapest areas for liposuction are the arms, buttocks, female back, hips and outer thighs…. Read more »
Multi-Million Dollar Swindle of Four Universities
In one of the most recently uncovered Ponzi cases, a former hedge-fund manager has pleaded guilty to criminal charges in an investment scam in which he bilked as much as $900-million from investors, including four university endowments. According to investigators, the Paul R. Greenwood and his partner Stephen Walsh spent at least $160-million on mansions, horses,… Read more »
Accounting Firms and Social Media
If accounting has the Big 4, then social media has the Big 3: LinkedIn, Facebook and Twitter. Still, many accounting firms and businesses consider LinkedIn as their sole source of social media because the site helps form relationships that often result in more business and turn into referral sources. But what are the benefits and… Read more »
How Low Can the Costs Go?
India has developed the world's cheapest laptop – a touchscreen device which resembles Apple's wildly popular iPad but will cost just £23. The prototype was unveiled this week by Kapil Sibal, the country's human resource development minister, who said 110 million Indian schoolchildren would be the first recipients. Questions: 1. Find the current exchange rates and calculate… Read more »
New Revenue Recognition Standards on the Way for Contractors
Contractors should be educating themselves on the impact of the new proposed revenue recognition standards and the recently published (June 24, 2010) exposure draft pertaining to revenue from contracts with customers. Public comments are due October 22, 2010, and it is expected the standards will be finalized in 2011. Questions: 1. What are some of… Read more »
Mortgage Fairs
Given the last year's trend, is there an end in sight to new record-low interest rates? For example, on July 22, Freddie Mac's Primary Mortgage Market Survey, which provides a snapshot of national average mortgage rates, reported a national average rate of 4.56% with 0.7 points on a 30-year fixed-rate mortgage. At the same time… Read more »
Bank of America: It Depends On How You Define Materiality
Bank of America incorrectly classified as much as $10.7 billion in short-term lending and repurchase deals for mortgage securities as sales. This claim surfaced in a May 13 letter to the SEC where the banking corporation alleges that the transactions were immaterial and that it would be beefing up its internal accounting controls. This letter… Read more »


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A wakeup call from Wyden
Oregon's fearless senior senator is acting like a Democrat this week. He's actually calling for a scale-back of some of the crazy domestic spy powers that were given to the government in the aftermath of 9/11:
While there are several controversial provisions in the USA Patriot Act, the coming debate is likely to center around the "business records" provision. Prior to 9/11, if the FBI or another government agency was conducting an intelligence investigation and wanted to obtain an individual's personal records from a bank, hospital, library, retail store or whatever institution was holding them, the government had to have evidence indicating that the person whose records were sought was a terrorist or a spy. The Patriot Act changed the law to authorize the government to collect any records deemed "relevant to an investigation."...Of course, with Congress blowing every available minute on the health care reform juggernaut, it's not at all clear that any such thing will happen. A rushed examination of the key issues seems much more likely.
I have served on the Senate Intelligence Committee for eight years, and I have yet to see evidence -- classified or otherwise -- that has convinced me that revising the business records provision to include a less intrusive standard would be harmful to U.S. national security. Yet as Congress considers whether to reauthorize this standard -- written in a rush to judgment eight years ago -- some will undoubtedly argue that Congress should just trust that the provision is essential and blindly sign-off on reauthorization. I disagree. While "just trust us" has passed as informed national security debate in this country for eight years, it hasn't resulted in good national security policy....
While the Obama Administration has taken laudable steps to release some information about how certain provisions of the Patriot Act have been used, more needs to be done for Congress to have an informed debate on this issue. Specifically, there is classified information that the public and the majority of my colleagues have not seen that, in my judgment, is essential to understanding the full scope of this issue. I and other senators have raised this problem with the Executive Branch in classified correspondence, and I hope that this information will be declassified soon. If it is not, however, I believe that the Senate should debate it in closed session to ensure that all senators are fully informed before casting their vote.
And then there's the question whether the current occupant of the White House will act like a Democrat on this. At least he can pronounce "terror."


text:
USC's climb up the polls on Sunday appears to again put the Trojans in prime position for a run at the Bowl Championship Series title game.
But it did not exactly move Trojans Coach Pete Carroll. "I'm certainly not going to try and figure it out," he said.
The Trojans' 28-0 victory over Arizona State, combined with losses by several other ranked teams, moved USC from ninth to fourth in the coaches' poll and from eighth to fifth in the Harris Interactive Poll. The two polls and computer rankings are used to determine the Bowl Championship Series standings, which come out for the first time next week.
The Trojans jumped from eighth to sixth in the Associated Press media poll after an uneven performance on offense Saturday.
Carroll maintained that he was not concerned with margin of victory or other issues to sway pollsters.
"There are teams, when they get into that kind of mode where there's conversation about it, those are usually teams that aren't there very often and are making a pitch for their cause," he said. "I don't think we're in that position."


text:
Position Medal
by Wadkin
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The Centers for Medicare & Medicaid Services approved a 3.3% rise in Medicare payments for skilled nursing facilities for fiscal year 2008. The increase will yield approximately $690 million more in payments next year, CMS said.
The rule, which will be published Friday in the Federal Register, bases the new payment on marketbasket data from 2004. Daily rates currently are based on data from fiscal year 1997, according to CMS. The agency uses the marketbasket as a cost-of-living measurement to determine payment increases.
The new SNF pay rates will include a special adjustment to cover the additional services required by nursing home residents with HIV/AIDS, Medicare said.


text:
Mordric is a rune-keeper. And a dwarf. That should already be pretty self-explanatory - he uses his magical powers to accumulate gold and makes us, the pity hobbits-or-humans-or-whatever, envy his wealth whenever his shabby belly catches our sight. Thanks the Valar he sometimes pays a round in the Thorin's Hall Inn and invites poorer fellas for a good Bombur's Beard Lager!
Head: Baingarab, purple dye (Quest: Vol. I, Book 2, Chapter 5: Retake Weathertop; also available as Linen Hat, T2 crafted)
Back: Cloak of the Shining Star, purple dye (Anniversary event reward)


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Alaska Native (Inupiat) Elders and children living in northern Alaska share their experiences and stories about the northern lights (aurora borealis). The website features a 23 minute video and a series of shorter videos and interactive animations... (View More) on related topics. (View Less)
Materials Cost: Free
This twelve-minute YouTube video incorporates NASA imagery and visualizations as it traces the history of air quality research.
Materials Cost: Free
This is a video to accompany the Stanford Solar Center's Build Your Own Spectroscope activity. It serves as an introduction to several aspects of the Sun, including information about solar activity, effects of solar activity on Earth and other... (View More) systems, and how astronomers look at the Sun using filters and spectrographs. (View Less)


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Land O Lakes Kennel Club NOHS Results – Sunday, January 7, 2018
NOHS Best in Show: GCHB Sketchbook Rodeo Clown [Dog]
Breed: Standard Schnauzer
Handler: Liz Hansen
Owner: Glenn & Jennifer Kimberlin & Liz Hansen
Breeder: Liz Hansen
Short URL: https://caninechronicle.com/?p=137961
Posted by chesliepickett on Jan 8 2018. Filed under NOHS Show Results. You can follow any responses to this entry through the RSS 2.0. Both comments and pings are currently closed.
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text:
RMB Tracker
What does the internationalisation of the RMB mean for your business? Follow the developing story with our monthly market updates
Follow the internationalisation of China's currency
With over 1,900 financial institutions now using the Renminbi (RMB) for payments with China and Hong Kong, the internationalisation of RMB is a strategically important topic for banks and FIs. How is your business affected? Where should you be targeting resources? Are you sufficiently informed?
The RMB Tracker is a free monthly report from SWIFT Business Intelligence to help you track and understand how RMB is being used across geographies and financial sectors. Based on aggregated message data for RMB transactions available only from SWIFT, the tracker provides key statistics and market insights in an easy-to-access report.
Since 2010, SWIFT has actively supported its customers and the financial industry regarding RMB internationalisation through various publications and reports. The SWIFT RMB Tracker, issued on a monthly basis, provides the community with insights into the local conversation about China's Renminbi and its global adoption.
– Astrid Thorsen, Head of Business Intelligence solutions
Access current and previous RMB Trackers online
The RMB Tracker is available as a pdf report in English, simplified Chinese, traditional Chinese and Japanese. There's also a more detailed PowerPoint presentation pack.
RMB Tracker uses data available only from SWIFT
The RMB Tracker uses aggregated message data (volumes and values) from RMB transactions over SWIFT. This neutral data provides valuable market insights.
Free and easy to access
Current and previous editions of the free monthly RMB Tracker are accessible via our website, in pdf and more detailed PowerPoint versions.
In English, Chinese and Japanese
The RMB Tracker is available in English, simplified Chinese, traditional Chinese and Japanese.
RMB Tracker
Follow the story of RMB internationalisation with our monthly reports.


text:
The hair of all mammals consists of terminally differentiated cells that undergo a specialized form of apoptosis called cornification. While DNA is destroyed during cornification, the extent to which RNA is lost is unknown. Here we find that multiple types of RNA are incompletely degraded after hair shaft formation in both mouse and human. Notably, mRNAs and short regulatory microRNAs (miRNAs) are stable in the hair as far as 10 cm from the scalp. To better characterize the post-apoptotic RNAs that escape degradation in the hair, we performed sequencing (RNA-seq) on RNA isolated from hair shafts pooled from several individuals. This hair shaft RNA library, which encompasses different hair types, genders, and populations, revealed 7,193 mRNAs, 449 miRNAs and thousands of unannotated transcripts that remain in the post-apoptotic hair. A comparison of the hair shaft RNA library to that of viable keratinocytes revealed surprisingly similar patterns of gene coverage and indicates that degradation of RNA is highly inefficient during apoptosis of hair lineages. The generation of a hair shaft RNA library could be used as months of accumulated transcriptional history useful for retrospective detection of disease, drug response and environmental exposure.
Citation: Lefkowitz GK, Mukhopadhyay A, Cowing-Zitron C, Yu BD (2011) The Post-Apoptotic Fate of RNAs Identified Through High-Throughput Sequencing of Human Hair. PLoS ONE 6(11): e27603. https://doi.org/10.1371/journal.pone.0027603
Editor: Johanna M. Brandner, University Hospital Hamburg-Eppendorf, Germany
Received: August 10, 2011; Accepted: October 20, 2011; Published: November 16, 2011
Copyright: © 2011 Lefkowitz et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by grants from the National Institutes of Health AR056667 and the California Institute for Regenerative Medicine RN2-00908, and institutional funds for B.D.Y. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Apoptosis is a cellular program utilized by multicellular organisms to eliminate cells during development or in surveillance of foreign or abnormal cells altered by viral infection or neoplastic transformation [1], [2]. Cells undergoing apoptosis undergo nuclear, chromatin, and additional organelle changes induced by a cascade of molecular signals involving several proteases and deoxyribonucleases [3]. Defects in programmed cell death results in the abnormal accumulation of cells, altered morphogenesis during development, and the persistence of transformed cells in disease.
The hair follicle is a model of apoptosis [4], [5]. Programmed cell death in the hair follicle occurs both during its normal growth and differentiation and during an involutional stage called catagen. Ultrastructural studies demonstrate that hair follicle cells undergo a specialized form of apoptosis called cornification during terminal differentiation [6]. During this process, the nuclear membrane is lost, and the chromatin becomes less coarse. In addition, evidence of DNA damage as detected by direct end-labeling of nicked DNA and indirect double-strand break activity have been identified during early and late stages of hair differentiation [4], [7], [8]. Skin-specific endonucleases such as DNase1L2 target genomic DNA during cornification [3], [9], and in the absence of DNase1L2, nuclear DNA persists in the hair and causes hair fragility [10]. The external hair is defined as the hair shaft and consists largely of the proteinaceous remnants of three cell types [11]. The three hair shaft cell types, the outer cuticle, cortex and central medulla, originate from a self-renewing progenitor population called the matrix [12]. The hair matrix also produces supportive, non-hair shaft cell types, which form a rigid sheath around the hair shaft and enable the hair to exit through the skin. Much of the strength of the hair shaft and sheath come from intermediate filament proteins called keratins and keratin-associated proteins (KRTAPs) which become crosslinked by several enzymes during terminal differentiation [13], [14].
While DNA degradation is a common hallmark of apoptosis, the targeting of RNA for degradation during apoptosis is unclear. The removal of RNA following apoptosis may be of importance as released endogenous RNA appears to activate inflammation and the innate immune response [15]. In viable cells, multiple mechanisms exist to regulate the homeostasis of mRNAs, including nonsense-mediated decay, targeting of AU-rich elements, microRNA-mediated destabilization, and others [16], [17]. During some forms of apoptosis, RNases with broad specificity are upregulated such as IFN-gamma induced RNase L [18]. In addition, specific endonucleolytic activities, which target 28S ribosomal RNA, have been observed during apoptosis and appear to trigger independently of DNA degradation [19], [20]. More recently, an endonuclease RNase III, DICER, has been shown to shift specificities between RNA to DNA during apoptosis [21]. These several observations indicate that during apoptosis, RNA and DNA targeting are distinctly regulated but the final fate of RNAs following apoptosis is unknown.
Here we investigate the extent to which RNAs survive apoptosis by studying the external hair shaft of mice and humans. We hypothesized that pools of small RNAs, in particular, regulatory microRNAs (miRNAs), might preferentially survive destruction by nucleases during the process of cornification, because of their inherent stability [22]. Using RNA sequencing and real-time quantitative PCR, we instead find that many types of RNAs survive apoptosis including miRNAs and mRNAs and persist long after hair shaft formation at several centimeters from the scalp. The post-apoptotic human hair contains the thousands of mRNAs, miRNAs, and other small RNAs and accurately reflects tissue of origin. Finally, we find that the RNA of the post-apoptotic hair shares similar patterns of intragenic coverage as RNA from viable keratinocytes. Hence, the removal and destruction of RNA post-cornification appears to be highly inefficient in the keratinized structure of the hair and potentially provides an ideal tissue for studies of genetic and acquired diseases.
Detection of lineage-specific miRNA and mRNA in mouse hair shafts
In mice, the transcriptional profiles of living hair follicle-specific miRNAs and mRNAs have been previously examined [23], [24]. Based on these expression profiles, we examined whether miRNAs and mRNAs specifically expressed in the hair follicle could also be detected in the external hair shaft and tested whether non-hair shaft transcripts were excluded. As comparison, we utilized whole mouse skin, which contains all lineages of the hair follicle, epidermis, dermis and adipose tissue (Fig. 1A). Using quantitative real-time PCR, miRNAs were readily detected in the hair shaft at 3–4 cycles higher than from equivalent amounts of whole skin RNA (Fig. 1B). Relative to control RNAs, e.g. snoRNA-251, we found evidence for lineage specificity using miRNA expression (Fig. 1C). Significantly, the level of a miRNA (mir-203), which is considered to be epidermal-specific [25], was ten fold less abundant in the mouse hair shaft compared to whole skin. These findings indicate that miRNAs are sufficiently intact in the hair shaft for specific detection by quantitative real-time PCR (qRT-PCR).
(A) Schematic diagram detailing source of tissue used in mouse studies. Hair shaft (HS) refers to the external portion of hair used for RNA isolation. Whole skin (WS) encompasses the epidermis, dermis, and the entire hair follicle, which includes cells fated to become the hair shaft (blue), supporting non-hair shaft cell populations (inner root sheath, orange and dermal papilla, purple). Red circles represent the earliest and latest regions of detected apoptosis. (B) Differential expression of snoRNAs and miRNAs in whole skin versus external hair shaft from 2–3 week old mice. Detection levels are displayed in cycle numbers normalized for equal input total RNA. Higher cycle numbers indicate lower levels of detection. (C) Mouse mir-212 and mir-203 levels normalized to snoRNA-251 levels reveal increased mir-212 and greater than two-fold reduction of mir-203 in mouse hair shaft relative to whole skin. (D) Detection and quantification of cuticle, cortex and medulla-specific transcripts in hair shaft vs. whole skin reveal patterns of enrichment. (E) Assay for dermal papilla-specific transcripts in hair shaft vs. whole skin total RNA. (F) Gel electrophoresis of amplification products of hair shaft and non-hair shaft (non-HS) genes in whole skin and hair shaft.
As hair keratins are abundantly expressed during hair development, we next investigated whether their transcripts can be detected in the external hair shaft. In addition, hair keratin genes are also expressed by specific cell types and identify the cell type of origin for RNAs extracted from external hair (Fig. 1D). Several lineage-specific keratin [13] and central medulla genes [26] could be detected in the post-cornification hair shaft (Fig. 1D, F). Relative to whole skin, Keratin 32 (Krt32), Krt34, and Krt35 were detected at 8.6, 16.1 and 1.5-fold higher in the external hair. Sh3d19, a medulla-specific gene, was also relatively more abundant in the hair shaft. The enrichment of these RNAs in hair shaft relative to whole skin occurs as hair follicle transcripts are diluted by other tissues present in whole skin. As a negative control, genes expressed by tissues that do not contribute cells to the hair shaft were examined. One such tissue, called the dermal papilla, is a closely associated mesenchymal population that supports hair growth but does not itself contribute cells to the growing hair shaft. We found that two dermal papilla-specific genes, Sox2 [27] and Bmp6 [28], were significantly reduced in expression in the hair shaft, only detectable at 6.8 and 3.7 fold levels below whole skin, respectively (Fig. 1E, F). In sum, these studies indicate that miRNAs and mRNAs of at least three cell types persist in the external hair of mice contains both miRNAs and mRNAs and that cornification does not sufficiently destroy these RNAs to prevent their detection.
Generation of A Human Hair RNA-Seq Library
We considered the possibility that residual RNAs in the mouse hair might reflect its rapid growth and potential persistence of cells rather than the low ribonuclease activity during and after cornification. In humans, hair does not contain viable cells [11] and undergoes 1–2 months of maturation prior to its emergence from the scalp, compared to 1 week in mice [29]. While human hair may be a better model to study post-terminal differentiation hair, studies of human hair versus mouse hair posed unique challenges. First, expression data of miRNAs in the living human hair follicle was limited, and second, gene expression biases may arise from individual variation in gene expression or detection biases due to nucleotide polymorphisms. We addressed these limitations by utilizing parallel sequencing of small RNAs [30] isolated from human hair (Fig. 2). To generate a comprehensive library of RNAs in human hair and to account for some aspects of human variation, we pooled RNA from the hair shafts of five individuals, who varied in gender, hair shape, and origin (demographics detailed in Supplemental Methods). Total RNA isolated from external human hair shafts by this method was ligated to adaptors designed for small RNA reads, reverse transcribed, amplified, size selected and analyzed by Illumina/Solexa-based small RNA sequencing.
Small RNA library constructed from RNA pooled from hair shafts of five individuals. (A) Uniqueness of RNA sequences. A large portion of reads can be mapped to 1 or more loci in the human genome (hg18). (B) Distribution of trimmed read lengths demonstrates two peaks at 22 and 29 nucleotides. (C) Types of RNA retained in the human hair shaft. Percentages of genomic aligned RNAs map to known mRNAs, miRNAs, rRNAs, unannotated RNAs, and others. (D) Complexity of miRNA in chart reveals high proportion of LET7 family members. Only miRNAs with 200 or more hits are demonstrated in chart. (E) Complexity of mRNAs in human hair shaft demonstrated in pie chart. (F) Read coverage of hair shaft RNA library of a 60 kb genomic window, representing of a portion of the KRTAP5 gene cluster. Alignment of reads demonstrates enrichment of sequences to transcriptional units and strand preservation of known transcript orientation of KRTAP5-7, 5-8, 5-9, 5-10, and 5-11 genes. These findings confirm absence of contaminating genomic DNA sequences.
From this approach, 13.5 million high-quality reads and 1.2 million unique sequences were obtained. Subsequently, the reads were aligned to the human genome (hg18) and to dedicated small RNA libraries (miRBase and snoBase). A large portion of reads aligned to more than 10 loci or no loci, due to low complexity of sequence, highly repetitive targets or the presence of non-human RNA (Fig. 2A). We also found a high frequency of aligned reads measuring 22 nucleotides, the size of mature miRNAs (Fig. 2B). 7,193 mRNAs were detected with read coverage of 10 or more, and 251 mRNAs were found at 200 or more reads. For alignments that had 200 or more reads, we also identified 449 distinct miRNAs and 339 snoRNAs in the hair shaft library. Percentages of reads of different classes of RNA identified from human genome annotation are shown in Figure 2C.
The vast majority of miRNAs detected in the hair shaft belonged to the LET-7 family, which plays a role in regulation of proliferation, differentiation, and stem cell maintenance [31] (Fig. 2D, Table S1). The remaining 123 miRNAs account for 21.4% of all miRNAs in the hair shaft. Like the mouse hair, the human hair shaft contained numerous mRNAs encoding genes of the keratin associated proteins (KRTAP) or keratins (Fig. 2E, Table S1). Members of the KRTAP family comprise 58.3% of all mRNAs in the hair shaft. Like keratins, KRTAP genes function as support proteins, which become highly crosslinked and participate in forming the shape of the hair shaft [32]. Many characteristics of the small RNA library generated from hair allowed us to verify that the sequences originated from RNA rather than contaminating genomic DNA. First, sequence data showed significantly greater coverage of transcriptional units compared to non-transcribed genomic regions (Fig. 2F). Second, the directional nature of the small RNA library preparation revealed specific alignment of sequence reads to the known transcriptional orientation of individual genes.
Many of the mRNAs found in the hair shaft were not specific to the hair follicle. Several gene annotation searches, including a Set-Distiller batch tool [33], identified strong statistical associations to phenotypes and pathways affecting other organ systems (Fig. 3A). 377 pathways involved in environmental and pharmacologic signals and 391 pathways related to specific human disorders were associated with genes detectable in the hair shaft. Given the large 7,193-gene set used to represent the hair shaft, we attempted to address the specificity of non-hair dataset associations by comparing these results to the comparison of an 11,027-gene set representing normal human epidermal keratinocytes (NHEK). The NHEK dataset was chosen, because it still represents epithelial tissue but is also significantly different from the hair shaft in origin and grown under different conditions. Eighteen signatures (p = 3.7×10−6 to 1.0×10−11) belonging to exposure, disease or medication-induced gene expression showed detection in hair shaft RNA but not NHEK RNA (Fig. 3B–D). These findings suggest that many associations between hair shaft RNAs and non-hair related pathways exist irrespective of the large input gene set. Moreover, these annotation studies suggest that hair shaft tissue may be of significant medical value in the screening of transcribed biomarkers shared with epidermal keratinocytes or uniquely as a source for conditions such as autism, hypertension, and thyroid diseases. Lastly, many of the transcripts present in the hair shaft are themselves genetic targets in human disease (Table S2) or contribute to pathways in involved in response to chemical compounds and drugs (Table S3). The gene ontology characterization of hair shaft RNA profiles provides guidance for the use of the hair shaft to study organismal responsiveness to medications or environmental exposures to chemicals or infections.
(A) Phenotypic association of genes expressed in the hair shaft library. Shown are numbers of genes present in the hair shaft that are associated with organ phenotypes identified by mouse mutations. (B) Environmental exposures pathways associated with genes identified in hair shaft library. Numbers of genes associated with selected exposure pathways are shown. (C) Disorder pathways associated with genes present in hair shaft library. (D) Compound pathways associated with hair shaft RNA library. Gray shading of column bars in all panels indicates lower limits of p-values calculated based on input hair shaft library genes vs. random background genes. Red solid circles denote unique pathway gene sets identified in hair shaft but not keratinocytes.
Relative Similarities in RNA content of Viable Keratinocytes and External Hair
The surprising stability of RNA in the hair shaft led us to examine and identify how RNA read coverages differ between viable and non-viable tissue. We thus compared RNA libraries of the hair shaft to viable NHEK using the same sequencing platform. First, we found that lineage-restricted hair keratin genes were highly enriched in the hair shaft library relative to NHEK [34], [35] (Fig. 4A). Many of the neighboring non-hair shaft keratin genes were absent from the hair shaft library. Comparing the expression of six different cell types in the hair follicle, we concluded that RNA of the human hair shaft library predominantly originate from transcripts of the three hair committed cell types (Fig. 4B, C). We also considered whether RNA from a non-viable source might demonstrate different patterns of intragenic region coverage compared to viable keratinocytes (Fig. 4D). Here, we found a similar pattern of coverage of intragenic regions, e.g. 5′ untranslated regions (UTR), exonic, intronic, and 3′ UTR (Fig. 4E). In both RNA libraries, intronic transcripts, which include 5S rRNA, were the most abundant. Exonic sequences of both libraries revealed similar patterns of coverage, with the exception that the 5′ regions of viable keratinocytes (32.5%) were relatively more enriched than in the hair shaft RNA library (24.2%). Thus, the abundance of detected transcripts and the high degree of similarity in the persistent RNA of the non-viable hair shaft and viable keratinocytes indicate that RNAs are inefficiently targeted by apoptosis during hair.
(A) Cell-type specific detection of human hair keratins (KRT) in hair shaft versus normal human epidermal keratinocytes (NHEK). Pattern of KRT expression in six different hair shaft (HS) and non-hair shaft inner root sheath (IRS) are shown in boxes, where filled boxes indicate ≥50 reads detected. (B) Percent of cell-type specific keratin genes detected reveals degree of enrichment of each hair shaft and non-hair shaft compartment in hair shaft vs. NHEK RNA libraries. (C) Illustration demonstrating the spatial distribution of cell types represented by lineage-specific KRT, including three hair shaft cell types (blue) and three IRS cell types (orange). (D) Genome-wide comparison of intragenic region coverage by hair shaft vs. NHEK RNA libraries reveals similar representation of most intragenic regions. Exonic coverage was greater in viable NHEK compared to hair, while intronic coverage was more extensive in hair. (E) Distribution of sequence coverage within regions of each domain, e.g. 5′ intergenic, 5′-UTR, introns, exons, 3′-UTR and 3′ intergenic regions, are shown as 5% windows from 5′ to 3′.
miRNAs and mRNAs Are Stable In Distal Hair Segments
During hair growth, cells are continuously incorporated into the hair shaft such that as the hair elongates, the progressively older segments of hair are pushed further distally from the scalp. Given the remarkable stability of RNA following cornification in the hair shaft inferred from the large number of detected transcripts and the many similarities to viable keratinocytes, we investigated the stability of RNA at varying distances, using the the scalp as the point of origin. Strands of hair from two individuals were collected, aligned and divided into 2.5 cm segments. We found that RNA could be consistently isolated even at distant regions of the hair shaft (Fig. 5A). At increasing distances, higher cycle numbers were required to detect miRNA and mRNA target genes by real-time PCR, indicating decreased abundance of specific RNA transcripts (Fig. 5B). Nevertheless, even at these higher amplification cycles, specific products were obtained and could be confirmed by the melting curve (Fig. 5C) and gel electrophoresis (not shown). Decay of mRNA and miRNA detection was estimated by examining the increasing cycle numbers over a relative measure of time, in this case, hair length (Fig. 5D). We find that a two-fold reduction of mRNAs and miRNAs occurs over the length of 0.92±0.11 cm and 0.81±0.16 cm, respectively. These studies suggest that the conditions that allow RNAs to be stable during hair formation persist even months after cornification and provide insights into the natural decay of RNAs in the hair.
(A) Total RNA yields from hair shaft at varying distances from the scalp as detected by A260 absorbance. (B) Detection of specific mRNAs and miRNAs identified from hair shaft library at varying distances from scalp, indicated in cycle numbers. Higher cycle numbers reflect reduced levels of detection, particularly at 10 cm. All experiments shown demonstrate at least three replicates and at least two individuals. (C) Melting peaks of KRTAP5-3, MIR-24 and SNO251 as an example of amplified mRNA, miRNA, and snoRNA products from 2.5 to 10-cm segments, respectively. (D) Calculated average half-lifes of mRNAs vs. miRNAs based on cycle numbers using non-linear fit one-phase decay modeling. Because growth rates of hair were not directly assessed in these studies, the distance of hair sampled from scalp was used as a marker of relative time.
In this study, we demonstrate that as a direct remnant of previously living cells, hair retains a vast amount of transcriptional data reflective of incomplete degradation of RNA following apoptosis. In addition to hundreds of miRNAs and snoRNAs, sequencing of hair RNA identified 7,193 unique mRNAs, or an equivalent to over a quarter of all genes in the human genome. Hair RNAs potentially reflect months of expression data temporally deposited along the length of the hair shaft and may be useful for investigations of gene expression and biomarker discovery.
As a model for studying differentiation and cell death, the hair provides insights into the extent to which RNAs are targeted for degradation during cornification. The numerous RNAs identified in the hair shaft suggest that RNA degradation is highly inefficient during cornification. In support of this hypothesis, read coverage between hair shaft versus viable keratinocyte RNA libraries exhibited remarkable similarities in coverage of intragenic regions with small biases toward reduced exonic coverage in hair shaft. Several RNases were detected in the hair shaft RNA library, including RNASE4, 7, 12, 13, RNASEN, and RNASET2, indicating that lack of RNases do not account for the persistence of RNAs in hair. RNA stability in hair might involve other mechanisms such as sequestration of RNases, presence of endogenous RNase inhibitors, protection of RNA by ribonucleoprotein particles or low water content in hair [36]. Understanding the contribution of these potential mechanisms may be important in improving RNA detection and stability in future hair RNA expression studies. The removal of DNA versus RNA during apoptosis may play distinct roles in different tissues [3]. In erythrocytes, corneocytes, and lens fiber cells, where nuclei and DNA are removed during terminal differentiation, persistence of RNA may be important in maintaining cell function. Whether incompletely removed RNA in hair shaft serves a functional purpose is unknown. It is unclear if mRNAs identified in the hair shaft are available for translation as the degree of fragmentation and survival of translational machinery were not examined in this study. In addition, studies aimed at determining whether RNA persists in other cornified tissues such as the nail could be of great importance for studying of diseases that affect localized body sites, such as cancer, infection, injury and exposures.
The discovery of stable mRNAs and miRNAs distant in the hair shaft from the scalp overcomes a formidable obstacle in the application of RNA diagnostics. Currently, RNA studies based on tissue biopsy and phlebotomy are vulnerable to RNA degradation [37]. In the current study, we found that older, more distal regions of hair still contain detectable mRNAs and miRNAs. These findings have several implications. First, the stability of RNA in older portions of hair suggests that RNA may be stable in hair over many months. This attribute differs greatly from other tissues and blood and potentially resolves a problem in storage and transportation of RNA, necessary for RNA-based diagnostic or biomarker studies. Second, temporal changes in gene expression either physiologic or induced by chemical, drug or disease might be stored co-linearly along the length of the hair shaft. Use of this spatiotemporal pattern of RNA deposition might provide a novel approach to studying the natural course or inciting events of disease. In addition to these characteristics, the continuous replacement of hair and its ease of access are advantages to developing diagnostic approaches based on hair RNA.
While possibly ideal for many types of biomedical studies, the use of hair RNA for molecular studies and diagnosis currently has several limitations. At this time, the amount of variation in RNA expression between different individuals, ages, and genetic backgrounds are not known. In addition, differential growth rates of hair in individuals due to differences in genetic background, age, and other factors obscure accurate measurements of time. Recent studies indicate that growth of human hair may vary from 1.3 to 2.2 months per centimeter in different individuals [3]. More accurate measurements might be made possible with the identification of cyclically expressed genes, which could used to normalize differential growth rates [38]. An additional limitation is that since new cells are added to the hair shaft only during active hair growth, it is not yet known what the effects different stages of the hair cycle might have on the pattern and stability of RNA in the hair. In this case, identification of transcripts representative of the final stages of the hair cycle might be required to determine whether retained transcripts reflect specific portions of the hair cycle. Lastly, because the characteristics that contribute to RNA stability in the hair are unknown, it is possible that RNA stability varies in individuals. These unknown aspects may bias the results of RNA detection in the hair.
The use of parallel sequencing of small RNAs provides an atlas of residual transcripts in humans of different genders, populations, and hair type. Sequencing technology provides several informative features valuable to molecular diagnostic studies including transcriptional orientation, multiple means of validation of expression and quantity of transcripts, and sequence data [39], [40]. Sequence data also provide a significant source for the identification of genetic polymorphisms, detection of allele-specific expression differences, and somatic mutations. The extent to which variant alleles are present in hair shaft RNA was not explored in this study. Current obstacles to use of this data for allele-specific studies include the possibility of chemical changes in nucleotides, e.g. cytosine deamination [41], and incorporation errors inherent to RNA polymerases [42]. Further studies are needed to assess the impact of these factors on the fidelity of hair RNA sequences.
In the past, hair analysis has aided in the diagnosis of inherited and acquired diseases [14], [43], [44] and as a phenotypic marker of medication and chemotherapeutic response [45]. Thus while the discovery of post-apoptotic RNA stability in hair provides insights into programmed cell death, these findings also greatly expand the possible applications of hair in medical discoveries in the clinic and the lab. The unique properties of the linear growth of the hair serve as a record of gene activity during organ growth and of the individual's own history. This record may be of great value for studies in organ development, evolution, genetic variation, and population genomics.
Materials and Methods
Animal and Human Specimens
Mouse lines originated from a CF-1 mixed genetic background (Charles River). Human samples were unidentified and pooled. The demographic profiles are shown in Supplemental Methods. Normal human epidermal keratinocytes were grown in EpiLife culture media (Cascade Biologics, Portland, OR) containing 0.06 mM calcium and defined growth supplement. All experiments and informed consents were performed and approved according to the institutional guidelines established by the University of California, San Diego, Institutional Animal Care and Use Committee and the University of California, San Diego, Human Research Protection programs, Protocol ID #091646. Written informed consents were obtained from each participant of the study.
Hair collection, storage, and RNA extraction
For mouse studies, hair from 3-week old animals was trimmed and stored in RNAlater (QIAGEN) solution at −80°C. Hair was visually inspected to verify absence of hair bulbs, which contain viable cells. For human studies, hair was trimmed at a distance of 5 mm from the scalp. Hair was washed in 70% ethanol and water. Two methods were used for RNA extraction. During initial studies in mouse hair and human hair shaft RNA-sequencing, Trizol reagent (Invitrogen) was utilized in combination with mechanical disruption with 1.0 mm zirconia beads (BioSpec Products, Inc.). Subsequently, improved extraction was obtained with the addition of 0.1 M dithiothreitol (DTT) to Trizol reagent or a urea-based RNA extraction buffer [46]. The improved yield of RNA presumably results from the effects of a reducing agent on the highly disulphide crosslinked hair tissue. A more detailed protocol is described in Supplemental methods.
Real-time PCR analysis for mRNAs, snoRNAs, and microRNAs
Primer sequences and miRNA probes are detailed in Supplemental Methods. Primers for mRNAs were designed using Primer3 [47]. Reverse transcription and real time PCR were performed with Maxima First Strand cDNA Synthesis Kit (Fermentas) and Maxima SYBR Green (Fermentas), respectively. For microRNA assays, cDNA was prepared using a RT2 miRNA First Strand Kit (QIAGEN) and amplified with stem-loop primers from SABiosciences/QIAGEN (Table S4) using RT2 SYBR® Green qPCR mastermix (QIAGEN). Real-time PCRs were performed on an ABI 7300 Real Time PCR system and a Roche Lightcycler 480. A comparative CT method was used for quantitative analysis using snoRNA-251 in mouse microRNAs samples, Gapdh for mouse mRNAs. Half-life of mRNAs and miRNAs determined using 2∧deltaCT values normalized to levels at site of origin and Prism (Prism 5, Version 5.0d) non-linear fit first-order decay modeling.
RNA Sequencing and Analysis
Fifty hair shafts, equivalent to 30 mg, were utilized to prepare total RNA using the above methods. One microgram of total RNA isolated from hair was used to generate a small RNA library. Adapter ligation, reverse transcription, amplification, gel extraction and sequencing were performed, according to Illumina protocols for miRNA-Seq (v1.5.0) (Illumina). Sequencing was carried out on an Illumina Genome Analyzer IIx. Genome Analyzer Pipeline was used to generate a FASTQ file from raw RNA-seq data, and quality scores were offset by 64 following Solexa-1.3+ standards. FASTX-Toolkit package, hg18 genome alignment with BOWTIE, was used for pre-processing and alignment. Parameters for alignment are detailed in Supplemental methods, and a list of human hair shaft transcripts are provided in Table S5. Computational processes were carried on Triton Resource at the San Diego Supercomputer Center at UCSD, using a single node of 32 processors and 512GB of RAM. For NHEK to hair shaft RNA library comparisons, representative coverages of intragenic regions in REFSEQ were compared. Each gene in REFSEQ was segmented into annotated 5′-UTR, introns, exons, and 3′-UTR regions; these regions were further broken up into windows of 5% of the length of the region. The hair shaft library was aligned to these regions, and the total coverage of all 5% windows for each region type in each gene was plotted, generating a summary of overall coverage patterns in intragenic regions. Functional and compound associations were performed using Set-Distiller batch tool [33] and Mouse Genome Informatics [48]. Further methods are described in Methods S1 and accompanying References S1.
Supporting Information
Table S1.
Annotation and reads counts of mRNA, miRNA, snoRNAs (>200 reads).
Table S2.
Representation of Genetic Association Database transcripts in hair.
Table S3.
Representation of Novoseek compound-genetic pathways in hair.
Table S4.
Primers and reagents used in PCR analysis.
Table S5.
Annotated reads with >10 reads.
Methods S1.
Supplemental methods.
References S1.
Supplemental references.
We are grateful to Steven Head and the Scripps DNA Core for library preparation and sequencing, Terry Gaasterland for assisting with use of San Diego Supercomputer Center.
Author Contributions
Conceived and designed the experiments: BDY GKL CCZ AM. Performed the experiments: GKL CCZ AM. Contributed reagents/materials/analysis tools: BDY GKL CCZ AM. Wrote the paper: BDY GKL.
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- 48. Blake JA, Bult CJ, Kadin JA, Richardson JE, Eppig JT (2011) The Mouse Genome Database (MGD): premier model organism resource for mammalian genomics and genetics. Nucleic Acids Res 39: D842–848.


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I'm Black. I haven't shared my photo, but in case you had any doubts, I am a Black American woman. Now I am very comfortable in my skin and I celebrate my identity. I love being Black. But my feelings about my identity have evolved over time.
I grew up in Washington, D.C., which is also known as Chocolate City. But in the Maryland suburbs situated just outside of the city, which is where I grew up, there wasn't much diversity in the 80s. Often, I was the only black kid in my class. It was never really an issue for me. I always loved school and I thrived. But I was also somewhat of a "mascot" at school. I remember my mom commenting about how popular I was. "When I walk into the school, everyone knows who I'm there to see. They all know who you are," she bragged. Even as a child, I knew that I wasn't popular because of my glowing personality ('cause it DID glow, even back then!). When you're the only black kid in the class and one of only a few in the entire school, you're difficult to miss. Of course everyone knew my name – I stuck out like a sore thumb.
It's hard being the token black kid. You wear the weight of the world on your shoulders. You field ignorant questions, asked "innocently" and you're not supposed to show anger or aggression in response; you're just supposed to shrug it off and forgive the ignorance, pretending to be grateful that people want to learn more about what makes you different. I'm sure some of the dumb sht*t people said was well intended, but as the old folk say: The Road to Hell is Paved with Good Intentions, so… there's that.
I was in the second grade when I was called a nigger for the first time. I was involved in a fierce game of kickball during recess. I wanted to pitch, and this little boy with white blonde hair who had designated himself the "captain" of our team looked me in the face and said, "We don't let niggers pitch."
I was floored.
The boy, Vernon, was popular. My best friend, also white, was his "girlfriend". When Vernon spit the slur at me, I ran to tell his grandmother, who volunteered at our school, and she let him have it. I felt somewhat vindicated, but then I told my best friend how embarrassed I was that he'd said it. I expected her to side with me, but she didn't. My world was rocked. In addition to my embarrassment, I felt betrayed. It was a sad day.
My parents were fitful when I reported the incident to them. My father was unsurprised, but nevertheless wanted to kill the kid; my mother was shocked and devastated that I was dealing with such a heavy incident at such a young age. She had hoped I would be protected from that ugly word for a little while longer. It came before she was ready. My feelings were hurt. I cried. But, surprisingly, I didn't hate Vernon. I was just confused as to why someone would dislike me because I looked different from them.
It's been almost 30 years since that incident happened, but I remember it like it was yesterday. I vividly see Vernon's round face, his pale skin, colorless hair, and ice blue eyes staring at me with contempt when he called me the n-word. I'm not hurt by it now, but then… then, I was devastated. I recall quite clearly the feeling I had when he said the word. It was as though I'd been punched in the gut. I couldn't breathe, then I felt hot, and finally I, literally, saw red. I've been called a nigger a few times since then, unfortunately, but none of the other incidents hurt quite like the first time. Vernon popped my n-word cherry, so to speak. Thanks, kid. Hooray.
So, that's why I immediately felt like a knife had been driven deep into my chest, opening an old wound, when I saw the story about soccer player Mario Balotelli yesterday. Balotelli, a striker for AC Milan whose parents are Ghanaian, was taunted with racial slurs by the opposing team's fans. Apparently, the slurs were so hurtful, this man broke down and sobbed on the sidelines.
Now, who knows whether this incident was Balotelli's first experience with racial slurs (although I doubt that he's managed to escape other incidents during 20+ years on this Earth), but what we do know is that he was hurt enough to be reduced to tears on the field, which is MAJOR for an athlete since sportsmen are typically known for their bravado and masculinity, making such an emotional spectacle extremely rare. Sure, he's European and maybe they're more in touch with their feelings than American athletes (I'm aware that this is a stereotype and major generalization, but just go with it), but this is obviously a big deal since it's reached us across the pond.
Also this week, Marcus Smart, who plays basketball for the Oklahoma State Cowboys, was criticized for pushing a fan who called him – surprise, surprise – a nigger. I've got to say, I'm never one to encourage physical altercations, but "nigger" is a fighting word, plain and simple. If you are Black, if and when you hear it, you WILL be taken to another level and all bets will be off. So, I freely admit that I do not fault Marcus Smart for reacting the way that he did. He will now have to face consequences and criticism for his actions, but at the time, he did what he felt needed to be done. *tsk tsk* me if you want, but I can respect that and I can relate to it, too. Had I been strong enough to fight Vernon back in the day, we'd have tussled. Trust and believe.
Hours after the Balotelli incident occurred, there is now some question about whether the taunts were really the driving force behind his emotional outburst. But regardless of whether the whole situation really brought a grown man to tears, why would the Napoli fans think it was okay to hurl racial epithets at someone anyway? People deriding your differences instead of celebrating them are hurtful, especially when your differences center on your identity. One cannot change being black, nor hide it (in most cases), and it's particularly frustrating when people hate you for things beyond your control. As such, racial epithets can cut like a knife. They can break you down, and frustrate you enough to make you cry… or fight. I get it, and 7-year-old me gets it, and I need for you to get it, too.
Dear White People (and others):
If you don't want to face any potential repercussions, physical or otherwise, that you may experience as a result of using then n-word, then simply remove it from your vocabulary. It's never okay to use it. Not even as a "joke", not even in the heat of the moment, not even amongst friends, not even ever. The "why" is irrelevant, please just don't use it because we said so.
Black Folk
People: Black folk, White folk, and other folk, please remember that words hurt. Use your mouth to speak LIFE to each other, not to kill, steal, or destroy the self-worth of another human being. Know that a word – ONE WORD – can reduce a grown man to tears. And also know that if you use a (certain) word with the wrong person, it can also get your ass beat. Take heed and govern yourselves accordingly. Please. Life is just too short. Let's be nice to each other while we're here.


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So, there's a lot to explain in that image, and it's only the second floor. I've moved the old signal processing consoles down into the 'basement', and the upper floor is left for the planetarium/system map.
The earth and moon should be obvious. Both datasets are from NASA. Earth is the 2013 bathyspheric "blue marble" image, and the moon is albedo corrected. The Sun is textured with the 2012 STEREO image (the first simultaneous full-sun image map) And far off to the left is a pair of Plank Microwave Anisotropy datasets: Background radiation in red, and matter density in blue.
Oh yeah, and there's a skybox too, but it's fake. Looks nice though. There'll be an option to turn it off.
The spheres are not actually spheres - in model space they're cubes. I suppose technically they are 'voxels'. Each sphere is raytraced within the cube with correct perspective, but without creating z-buffer geometry, so "hologram" might also be a good word. The spheres look geometrically perfect, but won't intersect correctly.
Why go to the trouble, compared with simply creating a triangular tesselation mesh for the sphere? Well, each "sphere" voxel is eight vertexes, with no normal or surface buffers. I can have thousand of spheres, millions, so long as they are stacked in non-overlapping minecraft rows.
I'm essentially ready to tackle the next major algorithm, the one which should elevate astromech to a whole new level: Ray Bundle Adjustment.
Long story short: Ray Bundle Adjustment is a tomography technique, think of it as the advanced successor to the 'backprojection' algorithm used to reconstruct CAT scans. Here's a good paper: http://luthuli.cs.uiuc.edu/~daf/courses/Optimization/Papers/bundleadjust.pdf
What the application? Given data coming from multiple viewpoints (such as a ring of solar observatory satellites like SOHO, SDO, or STEREO-A and B) one should be able to reconstruct the 3D structure of the volume.
Near-real-time 3D maps of the Sun and it's near volume (flares, mass ejections) sounds like a neat thing. I expect to have something working within the month.
Another load off my mind is that I found an excellent solution for one of my other problems: sensible rendering of cluster data containing thousands or millions of points. This is some beautiful work, based on the brilliant idea of leaving a step out. Expect to see something very similar in Astromech, soon as I get to it.
Lastly, I'm investigating going nearly fully peer-to-peer for the networking layer using WebRTC. In terms of spec and capabilities for moving video and data streams around, I would be an idiot not too. I'm putting together a small signalling server, and thinking about adaptive mesh "gossip" networks.
Google gave an excellent talk on what WebRTC is all about which I recommend you see, given how much impact that particular spec is going to have on the peer-to-peer internet. Without this technology, I would need some heavy central servers to videoconference a dozen telescopes. Now it can go point-to-point.
That changes many things. Assuming it all works. And it's already in your browser, most likely. There are still Chrome/Moz compatibility issues, but only because the spec is still moving.


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Advice About Peregrine Nest Boxes (1)
I think there may be peregrines trying to nest on the building where I work. What should I do?
Have a look at this excellent leaflet and also this Guidance note written by David Morris of London Peregrines. Peregrine Nest-Box Advice to Owners and Managers and Peregrine Falcons – What to do if they're on your building


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Friend's Email:
Subject: I have found a DVD that I think you would enjoy
|Tank Girl
Ws Sub
Actors: Lori Petty, Ice-T, Naomi Watts, Don Harvey, Jeff Kober
Director: Rachel Talalay
Genres: Action & Adventure, Comedy, Science Fiction & Fantasy
R 2001 1hr 44min
If you're into in-your-face visuals, outrageous action sequences and non-stop explosive laughs, this is your "rip-roaring power surge of a movie" (L.A. Weekly). The year's 2033 and since a humongous meteor hit earth, the ... more »
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Movie Reviews
DVD's are supposed to contain MORE....
Kimberly Wells | Shreveport, LA USA | 07/15/2002
(4 out of 5 stars)
"I adore this movie in a completely irrational, fan girl way. I know it's corny, I know that some people will hate it, and think it's stooopid. It fits the comic book style of punchy, fast action. Lori Petty quite effectively becomes the punk-girl she ought to be. But like other reviewers, I am quite, quite disappointed in the DVD version of this film. From the trenches you hear grumblings that there was a lot of edited stuff that COULD HAVE been put on the DVD. Why don't DVD makers do more than reprint what we already have on video for us to (silly consumers we sometimes are) just rush out to buy anyway? The picture quality of the DVD is not significantly higher than that of the video-- the only real bonus is that you can do still pauses and get some cool screen shots for that conference paper presentation you're doing... There is nothing more than the video on this DVD. Not even any nifty "Making of Tank Girl" footage, or sketches of other rejected cartoon scenes, or whatever! Even an oldie/goodie Cherry 2000 has more than this one, and surely the DVD makers could have done better. I'll just hope that Men In Black is correct, and someday, when we get the next alien form of media (I guess I'll have to buy the White album again) the next makers of whatever futuristic thing we'll all have to rush out and buy will heed this advice-- put more on your big media! There's no real reason for me to excercise my insane consumerism if you don't fill up my big old plate!"
Pure entertainment
Lee Gaiteri | Syracuse, NY USA | 05/04/2001
(5 out of 5 stars)
"Tank Girl is, make no mistake, a campy B sci-fi. It lives up to the genre perfectly, keeping a light, humorous tone in a world so dark that it would make most of us hide under the bed. It's probably no surprise that someone like Tank Girl would survive the apocalypse.Lori Petty's character is perfectly witty, a girl who plays it large and in charge even when things are at their worst. She can kill an armed man and then answer the irritated stares of his buddies with "What?" While McDowell's character tries to break her in for a mission, she spends all her time coming up with lines to really get under his skin--and succeeds.The complaints of some accusing McDowell of hamming up the role of the villain are unfounded; he not only plays the role perfectly, but does it so well that I don't think anyone else could have done it. He's playing the stereotypical megalomaniac bad guy, yes, but on top of that he adds a touch of class and intelligence. His repartee with Tank Girl proves he's nearly a match for her wit with flippant remarks. He's a very bright guy with an ingenious solution to an ongoing problem, who's simply flummoxed by the dauntlessly rude spirit of his opponent. McDowell is brilliant enough as an actor to get all these characteristics across. Bravo.The plot gives us a nice, reasonable adventure without being overly simplistic; the campy tone prevents it from getting any more complex. The off-the-wall antics of Tank Girl bleed over at times from the character to the movie itself, which gets a bit goofy (in a good way) in parts where you'd never otherwise expect it to. In the middle of the film they break into a musical number in the silliest of venues--it might not be logical but it's a whole lot of fun. All this is carried along by a soundtrack pretty well suited to the film, arranged appropriately enough by Courtney Love.For sheer entertainment (though not a family film, if that wasn't obvious), Tank Girl is really worthwhile. The DVD is noticeably light on features, but for the price I can live with it. If you enjoy a movie that doesn't take itself seriously, Tank Girl is for you."
Tank Girl DVD! no extras???
Jason karlson | HULL | 02/10/2001
(5 out of 5 stars)
"From the first minute I saw the movie I was hooked, the charcters are fantastic and the film just so rich in design and content.It's also one of the funniest things i've seen in ages with the humour pouring from it!I was however sad to hear that many scenes had been cut from the film and that a DVD would never be released. I started up an internet protest to get MGM to release it on DVD with the footage and here it is! This DVD is out due to the fans who protested for it!I am still angry that have not included ANY extras or the cut footage and after much arguing with MGM i dont think they ever will replace the footage! Anyway check out www.tankgirldvd.8m.com to see what bits they have not included on this disk and sign our online from to try to get them to put more stuff on the disk! theres still time!!"
ULTIMATE chick flick....
Darkcat_ak | Alaska | 03/17/2005
(5 out of 5 stars)
"Okay, I didn't just like this movie, I love, LOVE this movie. Yes, its campy, yes the plot line is bizarre, No I can't get a guy to watch it with me (but they won't watch Earth Girls are Easy either) but I just love this movies humor and girl power without being GI Jane.
Yeah, the Rippers (aka Kangaroo people) are a little scary, but they are genetically engineered kangaroo mutants who happen to be trying to find their creator. (I got over the Rippers scariness when Rebecca and Jet were high on helium and then doing that stupid dance.)
I think about this movie when I am down because most of the movie had me laughing my head off. But I guess this tells you about my personality because this movie isn't for everyone. Don't watch this movie expecting to learn anything or have a plot to follow. Just sit back, and enjoy the cartoons...and the fact that one of the main characters is a tank!
While I didn't know this was a comic first, I'm curious to see them now. But since I generally won't watch a movie if I've read the book, I can understand why the people that read the comic didn't like the film. To them I have to say I'm sorry, because I still adore this cartoon/movie. But I will admit that knowing now that this was supposed to be an Australian flick explains a lot...I always thought this had kind of a Mad Max feeling too it.
BTW, it really would have been cool if they had done outtakes etc on this movie, so I do agree there..."


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Polyform Products, the manufacturer and owner of the esteem Sculpey polymer clay is pleased to announce the appointment of That's Original Ltd as its new and exclusive distributor in the UK. Sculpey has a huge consumer fan base in the UK and its products are well known by the UK craft retailers.
"This is an exciting new chapter in our development of the UK market and partnership with UK craft retailers. We've been impressed by the way That's Original has worked with other well known brands and look forward to seeing Sculpey benefit from a new exciting and passionate approach," comments Wayne Marsh, International Sales Director for Polyform.
That's Original will now carry the full Polyform range of products including Sculpey, Premo, Super Sculpey, Bake Shop, Bake n Bend and Souffle.
For more information visit www.sculpey.com or contact That's Original on email@example.com or +44 (0)2381 159 093.
Posted: 26 May 2016
Look through our regularly updated list of trade suppliers and wholesalers.


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We work in IT service sector. We have School ERP, Pharmacy ERP, Hospital ERP Solutions, CDR Analysis System, Payroll+ HRM Solutions and many more. Relevant Skills and Experience We work in IT service sector. We have S More
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Doctor's Appt on Short Notice
What a great site this is!
This actually happened to me a few years back. I was about 35 or so, and I was getting chest pains in the morning. So of course I immediately thought heart problems. But they were not the classic symptoms, so I let it go for a few days. But the chest pains persisted, and my wife called the doctor. He was a bit alarmed, and had me come in that same day after all his regular appointments. When I got there it was late, and the receptionist and the nurses had already left. It seemed to be only the doctor and me in his offices. He had me take off my shirt, and he examined my chest, listened, asked me lots of questions, took some blood, etc, etc. All of these tests and procedures seemed to me to be quite normal for a person with chest pains. But then he asked me to pull down my pants and my shorts, and to turn around and bend over the examining table. He was a doctor that did not say too much, and I did not ask him what a prostate exam had to do with chest pains, I just did as he asked. But I was wondering why he wanted to do a butt exam. And it was only him and I in the office. Then when he did not put on rubber gloves, I got the picture. And it was exciting! I decided to let him do whatever he wanted, and I was going to encourage him. So I pushed my butt out to him a little bit, and kind of wiggled it a bit to let him know I liked what he was doing. And although I had never gotten a hard-on getting a prostrate exam before, this time just thinking about what could happen I immediately started getting really hard! And then he got some KY lube and started rubbing it on my asshole. But the way he rubbed it was nothing but sexy! In little circles, very softly, round and round. Then he would slip one finger in just a little bit, about an inch, and wiggle it a little, then slip it out again and start with the circles again. Well, by this time I was hard as a rock, and leaking like crazy. Then he reached around, took hold of my cock very gently, turned me around to face him, and gave me the greatest blowjob I have ever gotten. It did not take me long to come long and hard, right in his mouth. It was incredible to look down and see this older doctor that I did not know that well, with my dick in his mouth. Strangely, after it was all over, he got up looking very serious, and just told me to put my clothes back on. He said that what I had was simply a contracting esophagus, and gave me a prescription! Soon after this we moved to another town and I never saw him again, but he got me started in my bi-ways!
Posted on:
2003-05-19 00:00:00 |


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It's no secret that your canine or feline friend hates going into its carrier when it's time for a visit to the vet or on a long car trip, so the Expandable Dog Carrier by Petsfit helps to ease the stress of being in such a confined area by offering an expanding design aesthetic. Looking like a traditional soft carrier, the Expandable Dog Carrier by Petsfit is able to be unzipped on the side to almost double the size of the bag. The expansion is fully vented to ensure your pooch gets optimal ventilation.
When it's time to move, the Expandable Dog Carrier by Petsfit easily collapses back to normal size and features a shoulder strap for easy carrying.
Expanding Accordion Dog Carriers
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The Expandable Dog Carrier by Petsfit Gives Your Dog Extra Space
- By: Michael HemsworthOct 31, 2015
References: amzn.to & thegadgetflow
References: amzn.to & thegadgetflow


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Trailer: Stephen King's 'IT' Comes To Life Again
After being in development from 2009 into 2015, IT is finally ready to see the light of day … and scare the pants off the Losers Club.
Who's ready for a good scare?
For those not already familiar with the novel or the 1990 miniseries, IT follows the seven self-proclaimed members of The Losers Club (Finn Wolfhard, Jaeden Lieberher, Chosen Jacobs, Jack Dylan Grazer, Sophia Lillis, Wyatt Oleff, and Jeremy Ray Taylor) who first face Pennywise (Bill Skarsgård), an evil otherworldly entity abducting children and masquerading as a clown, in the sewers under Derry, Maine as pre-teens before returning to finish Pennywise off as adults 30 years later. This version of IT is updated to the 1980s from the 1950s, and it's split into two films – the first (this one) focusing on the kids, and the second (with a TBA release date) focusing on them as adults.
While IT was originally set to be directed by Cary Fukunaga off of a script co-written with Chase Palmer, Fukunaga stepped out of the director's chair over story disagreements with New Line Cinema in mid-2015 and was replaced shortly after by Mama director Andrés Muschietti, who is directing off of what appears to be the Fukunana-Palmer script with rewrites by Gary Dauberman.
This first trailer is light on Pennywise, instead choosing to hide him in sewers, behind balloons, and in slideshows, and ratchets up the tension and horror by focusing on the kids. Wolfhard and Lieberher are no strangers to horror/fantasy, as the former is on Netflix's Strangers Things and the latter was in last year's Midnight Special. This new adaptation looks to do Stephen King proud – and he's confirmed it to producer Seth Grahame-Smith and Twitter.
Andy Muschietti's remake of IT (actually it's Part 1–The Losers' Club) succeeds beyond my expectations. Relax. Wait. And enjoy.
— Stephen King (@StephenKing) March 7, 2017
IT creeps into theaters on September 8, 2017.


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Join us in our mission to inform the game community about mental health issues, to provide education about mental disorders and mental illness prevention, and to reduce the stigma of mental illness.
Donate to Take This
Want to help us fund the AFK Room program and our educational and awareness efforts? Here's how you can help:
Take This is on Patreon, where we raise funds to support the Psychomancer Project, which trains and provides clinical staff and quiet spaces to gaming conventions around the world. Join a growing movement of people who know that there is no shame in coping with mental health issues, that reaching out for help is a sign of great bravery, and that this industry and this hobby are better when we're all here to support one another.
With your donation, you can bolster programs that raise awareness and educate people about mental health. You'll also support our efforts to make the game world a healthier, safer place, particularly for people coping with mental health issues. You can make a one-time donation or provide long-term support with a recurring donation.
For those interested in donating larger amounts, Take This offers sponsorship opportunities for programs including the AFK Room. Interested? Contact us for details.
Each time we open the doors of an AFK Room, we first make sure the room quiet, welcoming, and staffed with clinicians and other volunteers who can help our visitors in their moments of need -- whether they need a breather from a hectic convention or a safe place to cope with mental health issues.
We also make sure each room is well-stocked. To that end, we keep an up-to-date wishlist of all the items required to make each AFK Room exactly what show attendees and exhibitors need. Purchasing the coloring books, tissues, gaffers tape and other tools on the list helps us put together a calm, secure space everywhere we go.
Amazon Smile and Humble
When you're shopping at Amazon or Humble, you can select Take This as your charity of choice. A small portion of eligible purchases at Humble and smile.amazon.com will be donated to Take This. You can do the same anywhere that works with the PayPal Giving Fund.
Other Opportunities
Have something else in mind? Want to collaborate on a fundraising campaign? We're always open to new ideas for raising funds and awareness for our lifesaving mission, so get in touch.
Stream For Take This
Are you a streamer? Are you interested in supporting Take This? You can easily raise funds for Take This while streaming using Tiltify. Just click "Start Fundraising" to get started.
If you'd like to get more involved with streaming for Take This, contact us for information on our upcoming Streaming Ambassador program.
Volunteer With Take This
Generally, we also have a Booth of Hope where we can accept donations and offer loot and information to Take This supporters. Volunteers are always needed to staff those booths, especially at our less-established shows.
Sign up to be informed when we have volunteer opportunities in your area, and please be sure to let us know if you have clinical psychiatric experience. If you know anyone else who might be interested, please share the signup link.
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Welcome to the JH & Sons Paving's blog. Our goal is to keep you informed about proper asphalt maintenance, signs of wear, when to replace your asphalt, how to prevent damage, and how to prolong the life of your driveway or parking lot.
With decades of experience in the industry, JH & Sons Paving specializes in paving, seal coating, patio design, walkway installation, parking lot services, asphalt millings, and excavating. If you're looking for a true professional to repair, replace, or maintain the asphalt of your driveway or parking lot, call JH & Sons Paving for a free estimate on all of our services.


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It's easy for Liberals like me to become incensed at Trump supporters who show up at rallies primed for a fight, but we do well to remember that each of them has a reason, a reason that makes sense to them, to be angry and hostile. You might say they are each like a bear with a broken tooth. Though the pain of the tooth is totally invisible to me, it is nonetheless provoking behaviour which, for the most part, is uncharacteristic in the bear as a rule.
"Do not avoid contact with suffering or close your eyes before suffering. Do not lose awareness of the existence of suffering in the life of the world. Find ways to be with those who are suffering including personal contact and visits, images, sounds. By such means, ...awaken yourself and others to the reality of suffering in the world. If we get in touch with the suffering of the world, and are moved by that suffering, we may come forward to help the people who are suffering."