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The poem ‘Who are you, little i’ composed by E.E. Cummings depicts a child at the end of the day looking out a window. It’s about nature and how it affects the speaker. The speaker of the poem is the person who “voices” the words, recalling a childhood memory that is closely related to nature. Cummings could be the speaker. The poet addresses himself in the form of his own inner child in the poem “who are you, little I by e. e. cummings. His childlike wonder has surfaced as a result of the inspiration of a brilliant sunrise.
This poem is about nature and how it affects the speaker. This poem is quite short, with only eight lines. The main theme of this poem is the beauty of nature and its positive impact on humans. The speaker of this poem begins by standing near a window and looking out of it at the end of the day.
He enters a nostalgic state, recalling his childhood when he used to sit and contemplate a sunset. He recalls how he used to enjoy such beauty when he was a young boy of about five or six years old. It’s a wonder he still has a youthful aura about him. This presence is capable of appreciating the beauty of the evening. Perhaps he wants to come out of his shell, but his maturity and adulthood prevent him from doing so.
The poet has described a little boy, his surroundings, and his acts of peering (looking) and feeling about beautiful and wonderful nature in a beautiful and wonderful way. He has shared his childhood memories of being close to nature. He has presented himself as a little I or a boy of five or six years old who keeps peering through some high window at the beautiful golden sunset of November and pondering the wonderful transformation of the day into night in a very beautiful way.

domain: artistic


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Join us at the RPAC for movie and popcorn! Different themed movies every month.
Toonie Tuesdays at 7 pm; $2 Family Saturdays at 10:30 am
September’s theme is a tribute to Robin Williams and a celebration of his lesser known films
Good Will Hunting
Tuesday, September 9 at 7:00PM
The most brilliant mind at America’s top university isn’t a student, he’s the kid who cleans the floors. Will Hunting is a headstrong, working-class genius who is failing the lessons of life. After one too many run-ins with the law, Will’s last chance is a psychology professor, who might be the only man who can reach him. Finally forced to deal with his past, Will discovers that the only one holding him back is himself.
Saturday, September 13 at 10:30am
In the world of the emperor penguin, a simple song can mean the difference between a lifetime of happiness and an eternity of loneliness. When a penguin named Mumble is born without the ability to sing the romantic song that will attract his soul mate, he’ll have to resort to some fancy footwork by tap dancing his way into the heart of the one he loves. (animated)
World According to Garp
Tuesday, September 16 at 7:00PM
Robin Williams plays the title character, the son of unmarried, unorthodox feminist Jenny Fields (Glenn Close, in her film debut). Every effort made by Jenny to broaden Garp’s outlook on life — she even arranges for him to spend the night with a hooker (Swoosie Kurtz) — crams more fears and phobias into his psyche. Aspiring to become a novelist, Garp succeeds in this goal at the same time that his mother publishes her first feminist manifesto. Though successful and happily married to college sweetheart Helen Holm (Mary Beth Hurt), Garp remains envious of his fearless mother.
Night at the Museum
Saturday, September 20 at 10:30am
Larry Daley, a down-and-out dreamer whose imaginative ideas have never paid off, finds himself in desperate need of a job. Larry has always believed he was destined for big things. But, he has no idea just how literally gargantuan and hairy a challenge he will face when he grudgingly accepts the supposedly menial graveyard shift as a security guard at the Natural History Museum. On his very first night on the job, Larry is handed an over-sized flashlight and a dog-eared instruction manual, then left all alone in the eerily quiet, cavernous museum. At least, he thinks he’s alone. To his utter astonishment and disbelief, Larry watches in shock and awe as, one by one, the primeval beasts and storied icons that surround him stir magically to life.
One Hour Photo
Tuesday, September 23 at 7:00PM
Semour “Sy” Parrish (Williams) runs the photo processing department at a large discount store; Sy is dedicated to his job, and takes great pride in his work. Sy’s favorite customers are Nina and Will Yorkin (Connie Nielsen and Michael Vartan), an attractive and cheerful young couple with a nine-year-old boy, Jake (Dylan Smith). Sy dotes on the Yorkins and their son whenever they drop off film to be processed — something they’ve been doing quite often ever since Jake was born — and Nina and Will are indulgent of Sy’s attentions, regarding his as a harmless eccentric. What the Yorkins don’t know is Sy is a desperately lonely man with no real life of his own, and he’s been obsessively making copies of their photos, for years, imagining himself to be “Uncle Sy,” a member of the family. Sy’s tenuous hold on reality begins to collapse when he develops a roll of film brought in by a new customer that suggests Will has been unfaithful to Nina; the notion that his ideal family may be falling apart is troubling enough for Sy, and when he loses his job, Sy reaches the breaking point.
Saturday, September 27 at 10:30am
Daniel Hillard (Robin Williams) is an eccentric actor who specializes in dubbing voices for cartoon characters. Daniel is a kind man and a loving father, but he’s a poor disciplinarian and a shaky role model. After throwing an elaborate and disastrous birthday party for his son, Daniel’s wife Miranda (Sally Field) reaches the end of her patience and files for divorce. Daniel is heartbroken when Miranda is given custody of the children, and he’s only allowed to visit them once a week. Determined to stay in contact with his kids, Daniel learns that Miranda is looking for a housekeeper, and with help from his brother Frank (Harvey Fierstein), a makeup artist, Daniel gets the job disguised as Mrs. Iphegenia Doubtfire.
Death to Smoochy
Tuesday, September 30 at 7:00PM
Randolph Smiley has it all–as star of the highest rated kid’s show on TV, “Rainbow Randolph” has a penthouse, a Times Square billboard, cars, boats and all the indulgence that celebrity brings. Randolph also has an alcohol problem and a penchant for taking bribes from stage parents. These under-the-table transactions are quite lucrative until the Feds get wise. Busted and instantly reviled, Randolph is a star no more. Scandals don’t play well to the under-8 demographic. Enter Sheldon Mopes and his alter ego “Smoochy,” a puffy, fuscia rhinoceros. Eager to expand his audience beyond a Brooklyn methadone clinic, Smoochy–with his innocence and unrelenting ethics–is the perfect remedy for what ails the networks. Also, the kids love him! Sheldon soon learns, however, that children’s television is a dangerous world, steeped in corruption, back stabbing and violence.
(The Rotary Performing Arts Centre is located at 3 Elma Street East)


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Synopses & Reviews
Leroy Brown is back in the next six books in the Encyclopedia Brown series. As Idaville's ten-year-old star detective, Encyclopedia has an uncanny knack for trivia. With his unconventional knowledge, he solves mysteries for the neighborhood kids through his own detective agency. But his dad also happens to be the chief of the Idaville police department, and every night around the dinner table, Encyclopedia helps him solve some of the most baffling crimes. With ten confounding mysteries in each book, not only does Encyclopedia have a chance to solve them, but readers are given all the clues as well and can chime in with their own solutions. Interactive and funit's classic Encyclopedia Brown!
Leroy Brown, better known to all as Encyclopedia, is the ten-year-old son of the police chief of Idaville. Whenever Mr. Brown has a particularly difficult case, he brings it home for his son to deal with. A missing ambassador to Latin America, the case of Smelly Nellie and the ambergris, a client who wants to pay with an 1861 Confederate silver half-dollar (worth about $5000) - these and seven more puzzling cases keep Encyclopedia Brown busy.
About the Author
Donald J. Sobol was the author of the highly acclaimed Encyclopedia Brown series and many other books. His awards include a special Edgar Award from the Mystery Writers of America for his contribution to mystery writing in the United States, and the Pacific Northwest Library Association Young Readers' Choice Award for Encyclopedia Brown Keeps the Peace.

domain: library


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One of the large incense cedars at Little Jimmy Spring. The spring is just off the Pacific Crest Trail near Little Jimmy Trail Camp, in the San Gabriel Mountains near Los Angeles.
Click this link to read more about this photograph.
Click on any of the following categories to see additional photography:
ABSTRACT - QUIRKY - LANDSCAPE - WILDLIFE - INSECTS - WILDFLOWERS - ADVENTURES
ON THE RUN
Copyright © 1995-2023
Gary Valle'. All Rights Reserved.

domain: geography


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The hallmark of classical Hodgkin lymphoma (cHL) is the presence of giant, mostly multinucleated Hodgkin-Reed-Sternberg (HRS) cells. Whereas it has recently been shown that giant HRS cells evolve from small Hodgkin cells by incomplete cytokinesis and re-fusion of tethered sister cells, it remains unsolved why this phenomenon particularly takes place in this lymphoma and what the differences between these cell types of variable sizes are. The aim of the present study was to characterize microdissected small and giant HRS cells by gene expression profiling and to assess differences of clonal growth behavior as well as susceptibility toward cytotoxic intervention between these different cell types to provide more insight into their distinct cellular potential. Applying stringent filter criteria, only two differentially expressed genes between small and giant HRS cells, SHFM1 and LDHB, were identified. With looser filter criteria, 13 genes were identified to be differentially overexpressed in small compared to giant HRS cells. These were mainly related to energy metabolism and protein synthesis, further suggesting that small Hodgkin cells resemble the proliferative compartment of cHL. SHFM1, which is known to be involved in the generation of giant cells, was downregulated in giant RS cells at the RNA level. However, reduced mRNA levels of SHFM1, LDHB and HSPA8 did not translate into decreased protein levels in giant HRS cells. In cell culture experiments it was observed that the fraction of small and big HRS cells was adjusted to the basic level several days after enrichment of these populations via cell sorting, indicating that small and big HRS cells can reconstitute the full spectrum of cells usually observed in the culture. However, assessment of clonal growth of HRS cells indicated a significantly reduced potential of big HRS cells to form single cell colonies. Taken together, our findings pinpoint to strong similarities but also some differences between small and big HRS cells.
Citation: Rengstl B, Kim S, Döring C, Weiser C, Bein J, Bankov K, et al. (2017) Small and big Hodgkin-Reed-Sternberg cells of Hodgkin lymphoma cell lines L-428 and L-1236 lack consistent differences in gene expression profiles and are capable to reconstitute each other. PLoS ONE 12(5): e0177378. https://doi.org/10.1371/journal.pone.0177378
Editor: Francesco Bertolini, European Institute of Oncology, ITALY
Received: August 31, 2016; Accepted: April 26, 2017; Published: May 15, 2017
Copyright: © 2017 Rengstl 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.
Data Availability: Data are available through the GEO database (www.ncbi.nlm.nih.gov/geo/) accession number GSE86477.
Funding: This project was supported by the Deutsche Forschungsgemeinschaft (HA6145/2-1).
Competing interests: The authors have declared that no competing interests exist.
The pathogenesis of classical Hodgkin lymphoma (cHL) has been unsolved for many years. Already around 1900, Dorothy Reed and Carl Sternberg were fascinated by the morphological appearance of the tumor cells, particularly by the usually giant bi- or multinucleated so called Reed-Sternberg (RS) cells [1, 2]. In 1994, it could first be demonstrated that these enigmatic Hodgkin and Reed-Sternberg (HRS) cells constitute a clonal B-cell population . Although it was previously speculated that RS cells develop after fusion of cells , as known from histiocytic giant cells, single cell analyses revealed that these giant multinucleated cells never present more than two rearranged immunoglobulin genes , indicating that RS cells have probably developed from endomitosis as observed in the cHL cell line HDML-2 . Recent studies discovered that giant multinucleated RS cells evolve from small mononucleated Hodgkin cells by incomplete cytokines and re-fusion of tethered sister cells . However, there is also a subset of giant cells containing only one enormous nucleus and not resulting from a re-fusion . In primary cHL samples and the cHL cell lines L-428, KM-H2, and HDLM-2 Hoechst dye-negative side populations—considered as tumor stem cells—could be identified [8, 9]. In culture experiments, these side populations were shown to be able to reconstitute the HRS clone, whereas giant binucleated RS cells failed to proliferate [8, 10]. However, these side populations only represent a small subset of the abundant small HRS cells observed in cell culture. Interestingly, particularly the cHL cell lines L-428 and L-1236 show mono- and multinucleated tumor cells of very variable sizes, including giant tumor cells with sometimes sizes above 100 μm in diameter. Consequently, the aim of the present study was to determine the differences in gene expression profiles, growth kinetics, clonal growth potential, and vulnerability towards treatment of small and giant HRS cells and to gain deeper insight into this particular phenomenon of populations of different cell sizes in cHL cell lines.
Materials and methods
Cell culture and laser microdissection
The cHL cell lines L-428 and L-1236 were obtained from the German Collection of Microorganisms and cell cultures (DSMZ, Braunschweig, Germany) and cultured in RPMI with 10% fetal calf serum. In first experiments, isolation of giant HRS cells in high purity by fluorescence associated cell sorting proved to be difficult, since these cells are very fragile to the sorting procedure. Therefore, laser microdissection was applied as method of choice. Two days after passage, the cells were washed and resuspended in 300 μl phosphate buffered saline substituted with 0.6 μl RNAse inhibitor. Smears of the cell lines were made on membrane slides for laser microdissection and air dried for 15 min. Microdissection was performed on a Microdissection Axiovert 200M microscope (PALM, Bernried, Germany). Cells to be considered as small HRS cells showed a maximum cell area of 350 μm2 in the PALM microscope, whereas big HRS cells had an area of more than 600 μm2. In order to obtain comparable amounts of RNA, the total microdissected areas of small and big HRS cells were adjusted to approximately 700 big and 1500 small HRS cells. Cells were catapulted in PALM adhesive caps and lysed with 2 ml NUGEN Direct Lysis Buffer (NUGEN, Bemmel, The Netherlands).
Gene expression analysis
After lysis of the microdissected cells, RNA was amplified with the WT-Ovation-One-direct-Kit (NUGEN) and hybridized onto Affymentrix Gene Arrays 1.0 ST (Affymetrix, Santa Clara, CA, USA). Gene expression analysis was performed and analyzed as previously described [11, 12]. Gene expression data are available through the GEO database (GEO accession number GSE86477).
Immunohistochemistry, fluorescence microscopy, and primary cases
Immunohistochemistry for SHFM1, LDHB and HSPA8 was performed using the FLEX-Envision Kit (DAKO, Glostrup, Denmark) as described previously . Primary antibodies for SHFM1 (LS-B899, Lifespan Biosciences, Seattle WA, USA, dilution 1:50), LDHB (LS-B4366, Lifespan, dilution 1:100) and HSPA8 (1B5, Enzo Life Sciences, Lörrach, Germany, dilution 1:100) were applied over night at 4°C after heat induced epitope retrieval in citrate buffer (pH6) for 10 min. Primary cHL cases (5 mixed cellularity, 5 nodular sclerosing subtype) were selected from the archives of the Dr. Senckenberg Institute of Pathology, Frankfurt, Germany. The study was approved by the local ethics committee of the Frankfurt University Hospital.
For immunofluorescence, the cHL cell lines L-428 and L-1236 were embedded in 0.5% agarose gel. After formalin fixation and paraffin embedding, 2 μm thick sections were cut, allowing an objective assessment of expression intensities in small and giant HRS cells. The same primary antibodies as well as additional antibodies for LDHB (LS-B6870, Lifespan, dilution 1:400) and HSPA8 (GTX111150, GeneTex, Irvine, CA, USA, dilution 1:100) were used with the Vectafluor Excel Amplified Dyelight 594 Anti-rabbit or Anti-mouse kit (Vector laboratories, Burlingame, CA, USA). Respective negative controls showed no specific staining. Immunofluorescence was evaluated on an Axioskop 2 fluorescence microscope with an AxioCam MRm camera (both Zeiss). Expression intensities were then analyzed using ImageJ software (http://imagej.nih.gov).
Fluorescence activated cell sorting and flow cytometry
The HL cell line L-428 was sorted according to the cell size observed in the forward scatter or with respect to the CD15/CD30 expression pattern using a fluorescence activated cell sorter (FACS) Aria II (Beckton Dickinson, Heidelberg, Germany). Sorted populations were confirmed and further analyzed by a MACSQuant analyzer (Miltenyi Biotech, Bergisch Gladbach, Germany). Staining was performed by standard procedure using antibodies against CD15 coupled with FITC (Miltenyi Biotech, Bergisch Gladbach, Germany) and CD30 coupled with PE (Miltenyi Biotech, Bergisch Gladbach, Germany). Sorted cells were cultured separately for 14 days and the composition of small and big cells as well as CD15/CD30 expression pattern was assessed every 3–4 days. For sorting of different cell sizes, the main population was divided according to cell size into equal numbers of small and big cells (each 20%) with a clear gap (60% of the live cells) to delineate the different populations.
Colony forming assays
After cell sorting, 600 cells were pipetted into 3.3 ml RPMI 1640 medium containing 1% methylcellulose, 30% FBS, 2% L-glutamine, 1% penicillin/streptomycin and 0.5 μM β-mercaptoethanol. Duplicates of 1.1 ml were platted into 35 mm culture dishes and incubated for 7 days in a standard cell culture incubator to obtain single clones (colonies) of 10–25 cells. Colonies were then counted manually by conventional microscopy.
Apoptosis assay and Brentuximab Vedotin treatment
After cell sorting, 1 x 105 cells were seeded per ml into 24-well plates with 1 ml of total volume. For assessment of Brentuximab Vedotin induced cell death, the drug was added at total concentrations of 10, 25, or 50 μg/ml and cell numbers as well as the apoptosis rates were assessed using the APC-Annexin V Apoptosis Detection Kit (eBioscience, San Diego, Kalifornien, USA) at 48h.
Size exclusion of HRS cells
To dissect giant cells from the bulk culture, a volume of 80–100 ml L-428 cell culture was filtered through a cell strainer with 50 μm pore size (Fisher Scientific, PA, USA). The cell strainer was then washed two times with 20 ml PBS reducing the amount of contaminating small cells. Next, the cell strainer was inverted and flushed with medium. Giant cells were collected in a 10 cm culture dish and transferred into 24- or 6-well plate for further culture or directly analyzed.
Cell size and surface marker expression correlate with HRS cell subpopulations
First, we were interested to determine if small and big HRS cells differ in their immunophenotype. For this purpose, L-428 cells were sorted according to cell size (small, medium and big, Fig 1) and afterwards analyzed for CD15/CD30 expression (S1 Fig). In the subgroup enriched of big L-428 cells the amount of double positive (DP) cells was increased, whereas the number of CD30 single positive (SP) cells was decreased. This finding implicates that larger sized L-428 cells more often express CD15 together with CD30 than their smaller counterpart. In addition, the opposite experiment was performed. L-428 cells were flow sorted into SP and DP cells (S1 Fig) and further analyzed for cell size via FACS (S1 Fig). In line with the previous experiment, SP cells showed the highest amount of small-sized cells and the DP population had an increased frequency of big-sized cells.
(A) L-428 cells were sorted by cell size (small, middle, big) or after staining for CD15 and CD30 into CD15+CD30+ double positive (DP) and CD15-CD30+ single positive (SP) cells. Frequency of (B) small- and (C) big-sized cells within the three different populations was recorded over time via fluorescence activated cell sorting (FACS).
Both approaches led to the same result, namely that DP L-428 cells are larger in size than the CD30 single positive counterpart. However, neither by sorting according to cell size nor via marker profile, a pure giant or small cell population could be obtained. Cell size and CD15/CD30 expression were monitored in the enriched populations over time. Reanalysis showed that the frequencies of small-sized or big-sized cells were adjusted within all enriched populations after 5–10 days (Fig 1).
Big HRS cells show a significantly reduced clonal growth potential
As a next step, a colony formation assay (CFA) was chosen to elucidate clonal growth potential of individual cells of the sorted subpopulations. In fact, a significantly reduced clonal growth potential could be detected for big-sized as well as DP L-428 cells (Fig 2).
L-428 cells were sorted by (A) cell size (small, middle, big) or (B) CD15/CD30 expression (SP, DP), and subsequently used for a colony formation assay (CFA). After one week, colony forming units (CFU) were counted compared to the bulk population. CFA were performed in duplicates and repeated three times.
The fast reconstitution of HRS cell subpopulations sorted via cell size might be due to the higher fragility of giant HRS cells and sorting of these cells could increase cell death within the big-sized population. Hence, another more gentle strategy to separate L-428 cells by cell size was applied. Cells were filtered through a 50 μm-cell strainer retaining giant cells (> 50 μm in diameter). Interestingly, size-exclusion led to a significantly increased frequency of DP cells in the big-sized population, again confirming that big L-428 cells are more frequently DP (S2 Fig). Conventional microscopy of the isolated giant cell population confirmed the usually big size of the isolated cells. However, small cells were frequently attached to the giant cells (data not shown). These heterogeneous clusters may be responsible for the results at reanalysis after 7 days, when the amount of DP cells within the big-sized population was markedly reduced (S2 Fig), especially as the proliferation capacity of small HRS cells remains more potent. Although size exclusion experiments revealed the most pure HRS-cell subpopulation with regard to cell size, purity was not sufficient enough for further analysis, i.e. gene expression profiling. Hence, a more precise isolation method was chosen for further experiments.
Small and big HRS cells lack major differences in gene expression
Small and giant HRS cells were microdissected from smears of the cHL cell lines L-428 and L-1236. Isolated cells were then processed for microarray analysis. Unsupervised hierarchical clustering of gene expression arrays revealed differences between the four samples investigated. In general, among the differentially expressed probesets between the four samples, small HRS cells showed a higher number of over expressed probesets (mean 672) than giant HRS cells (mean 162, Fig 3). In a supervised comparison of small HRS cells to giant HRS cells from both cell lines, only two significantly differentially expressed genes were found when a stringent filtering with a false discovery rate (FDR) of 0.1 was applied. These were split hand/foot malformation (ectrodactyly) type 1 (SHFM1, 34-fold upregulated, p<0.01) and lactate dehydrogenase B (LDHB, 27-fold upregulated, p<0.01). When less stringent filter criteria (p<0.05 and FDR<0.3) were applied, thirteen genes were found to be upregulated in small compared to giant HRS cells, including six ribosomal proteins and one eukaryotic translation initiation factor (Table 1), suggesting that small HRS cells are more active in protein synthesis. There were no genes that were significantly overexpressed in the giant HRS cells when compared to small HRS cells.
Small HRS cells of both cell lines show more strongly expressed probesets (red color) than giant HRS cells. 958 differentially expressed probesets between the four samples representing a standard deviation of > 1 were considered for the analysis.
SHFM1, LDHB, and HSPA8 proteins are expressed in small and big HRS cells in primary cHL cases as well as in cHL cell lines
Among the differentially expressed genes (Table 1), SHFM1, LDHB, and HSPA8 were selected. Primary cHL cases were immunostained with antibodies against these three proteins and a strong expression was observed in the HRS cells in all cases tested (10/10, Fig 4). To be able to better quantify the expression intensities of SHFM1, LDHB, and HSPA8 proteins, the cHL cell lines L-428 and L-1236 were FFPE embedded and immunofluorescence stainings for SHFM1, LDHB, and HSPA8 proteins were performed (Fig 4). The expression intensities were quantified. However, there were no significant differences in the mean signal intensity per area between small and big HRS cells for SHFM1 (S3 Fig), and the expression differences between small Hodgkin and big RS cells observed for LDHB and HSPA8 applying one antibody, could not be confirmed when a different antibody was used (S3 Fig). This indicates that the differences detected by microarray on transcription level did not convincingly translate into changes at the protein level.
a. LDHB protein expression in small and big HRS cells in a representative primary cHL case (LDHB-immunostaining, 400x). b. SHFM1 protein expression in small and big HRS cells in a representative primary cHL case (SHFM1-immunostaining, 400x). c. HSPA8 protein expression in small and big HRS cells in a representative primary cHL case (HSPA8-immunostaining, 400x). d. Representative example of LDHB expression in small and big HRS cells in the cHL cell line L-428 (LDHB-immunofluorescence staining, 400x). e. Representative example of SHFM1 expression in small and big HRS cells in the cHL cell line L-1236 (SHFM1-immunofluorescence staining, 400x). f. Representative example of HSPA8 expression in small and big HRS cells in the cHL cell line L-428 (HSPA8-immunofluorescence staining, 400x).
Small and big HRS cells do not show significant differences in susceptibility towards Brentuximab Vedotin treatment
While first experiments could demonstrate differences between small and big HRS cells, GEP analyses in combination with immunohistochemistry-based validation failed to clearly discriminate between both cell types, although cells were separated according to size with the best available method with respect to purity. Therefore, we also assessed their differential drug response. For this purpose, cell sorting of the cHL cell line L-428 was performed, as a high amount of viable HRS cells separated into small and big subpopulations was needed. Fourty-eight hours after treatment with Brentuximab Vedotin, a trend towards higher cell numbers in the subset of small HRS cells of the L-428 could be detected. However, this difference was only significant at a concentration of 25 μg/ml when small HRS cells were compared with the bulk population (Fig 5, unpaired t-test, p = 0.0117) and was never significant when small HRS cells were compared with big HRS cells. There were no significant differences in the apoptosis rate (data not shown). Taken together, small HRS cells are not significantly more susceptible to Brentuximab Vedotin treatment when compared with big HRS cells.
Cells from the cHL line L-428 were sorted according to FSC in subpopulations of small and big cells (each 20% of total live cell population). Total live gates were sorted as control (bulk). Thereafter, subpopulations were seeded at a density of 1 x 105 cells per ml in triplicates and the indicated amount of Brentuximab Vedotin was added. After 48 h cell numbers were determined by FACS. * p = 0.0117, unpaired t-test.
Giant HRS cells are of high importance in HL, because it is suggested that they shape the tumor microenvironment to support disease progression [14–17]. However, these cells display a differentiated end-stage showing a highly impaired ability to proliferate [10, 18–20], which could be confirmed by colony formation assays in the present study. As a consequence, precursor cells, that are commonly neglected during diagnostic analysis of biopsy specimens, must be responsible for the maintenance of the tumor cell clone . It would be desirable to identify such a stem cell-like HRS cell population, as eradication of these cells would display a way to cure patients suffering from HL [22, 23].
Different groups using different isolation strategies described the presence of HL stem cells within HL cell lines, but unfortunately further validation of the disease-promoting properties of these cells is still missing [8, 9, 24]. Moreover, only Jones et al. were able to detect a type of HL stem cells, namely CD20+ HRS cells, also circulating in the blood of HL patients . Although these cells were first identified in HL cell lines L-428 and KM-H2, isolation and functional characterization of this subpopulation was not performed to date. We tested different MACS and FACS strategies to isolate CD20+ L-428 cells and an enrichment of CD20+ HRS cells could be obtained (data not shown). Unfortunately and in contrast to the published data, the sorted population had no enhanced clonal growth or proliferation potential (data not shown). Apart from a stem cell-like subpopulation of HRS cells, it would also be desirable to isolate a highly pure subpopulation of giant HRS cells in order to understand which genes drive these cells into incomplete cytokinesis. Although it has been shown that overexpression of the Epstein-Barr virus (EBV)-encoded latent membrane protein 1 induces formation of multinucleated HRS cells [25, 26], EBV-negative cHL cases likewise present bi- and multinucleated giant tumor cells. In the present study we assessed differences in gene expression between the small and giant HRS cells of the two cHL cell lines L-428 and L-1236. However, apart from the heterogeneity between the two cell lines, only very little differences were observed between small and big HRS cells, indicating that there is no unifying feature between the small and the big HRS cells of these two cell lines.
The most interesting differentially expressed gene found was SHFM1, which interacts with BRCA2 . BRCA2 has been published to impair the completion of cell division by cytokinesis and to be involved in the formation of aneuploid giant cells . However, in an according study these results could not be reproduced . In contrast, SHFM1 knock out variants of the yeast S. pombe also showed a defect in completion of cell division . Downregulation of SHFM1 in giant HRS cells could therefore be related with the giant size and the bi- or multinucleated phenotype of these cells. However, we could not demonstrate any consistent differences in SHFM1 protein levels between small and big HRS cells and therefore the variable size of these cells is probably not influenced by SHFM1 protein expression. In a recent study differences of the localization of Cyclin A in mononucleated Hodgkin and multinucleated RS cells could be observed . Differences in protein localization may of course also contribute to giant cell formation and were not assessed in the RNA expression profiling in the present study.
When the distribution of small and giant HRS cell populations in cell culture was monitored over time, it was noted that after several days a status quo in the fraction of small and big HRS cells was achieved, indicating that generation of big HRS cells follows a constant mechanism and is a phenomenon intrinsic to the entire HRS cell population. Therefore, the lack of major differences in the gene expression of these two cell populations is consistent with the observed capacity to reconstitute the respective other cell type under culture conditions. In line with this, there were no significant differences between small and big HRS cells in the susceptibility towards Brentuximab Vedotin treatment.
In summary, we show that small and giant HRS cells have a strong similarity: They lack major differences in gene expression. The generation of big HRS cells follows a constant mechanism as sorting experiments showed a stable reconstitution of the respective subpopulations, explaining the high similarity found in gene expression analysis. Furthermore, susceptibility towards Brentuximab Vedotin treatment is not significantly different when small HRS cells are compared with big HRS cells. The only biological difference observed between small and big HRS cells is a reduced clonal growth potential of big compared to small HRS cells by colony forming assays.
S1 Fig. Big L-428 cells show the highest amount of DP cells.
(A) L-428 cells were sorted by cell size (small, middle, big) or after staining for CD15 and CD30 into CD15+CD30+ DP and CD15-CD30+ SP cells. (B+C) Compared to the bulk population, populations sorted for cell size were analyzed for CD15 and CD30 expression by FACS. (B) Frequency of CD15-CD30+ SP cells. (C) Frequency of CD15+CD30+ DP cells. (D+E) Compared to the bulk population, DP and SP cells were analyzed for cell size by FACS. Frequency of (E) small-, (F) middle- and (G) big-sized cells. Experiments were repeated three times.
S2 Fig. DP L-428 cells separated by size-exclusion.
L-428 cells were placed onto a 50 μm cell strainer. Cells smaller than 50 μm (small) passed the filter, whereas cells bigger than 50 μm (big) were retained. Subsequently, the separated populations were analyzed for CD15/CD30 expression. (A) Exemplary dot blots of small and big L-428 cells. Frequency of DP (B) and SP cells (C) within the two different populations, compared to the bulk population. (D-E) Frequencies were reassessed after 7 days in culture. Experiments were repeated three times.
S3 Fig. Quantification of immunofluorescence stainings for LDHB, SHFM1 and HSPA8 in small and big HRS cells of the cell lines L-428 and L-1236.
Whereas a significant difference in mean fluorescence intensity between small Hodgkin and big RS cells of the L-1236 was observed with one antibody against HSPA8 (B, GeneTex antibody, p<0.05, t-test), this was not confirmed when a different antibody was applied (A). A significant difference in mean fluorescence intensity between small Hodgkin and big RS cells was also found in the L-428 cell line with an antibody against LDHB (D, antibody LS-B6870, p<0.001, t-test). However, it was not confirmed when a different antibody against LDHB was used (C, antibody LS-B4366). No differences in mean fluorescence intensity were observed for SHFM1 (E).
The authors would like to thank Sabine Albrecht for excellent technical assistance. This project was supported by the Deutsche Forschungsgemeinschaft (HA6145/2-1). We thank Ralf Küppers for helpful discussions and critical reading of the manuscript.
- Conceptualization: MLH BR SH.
- Data curation: CD.
- Formal analysis: CD.
- Funding acquisition: SH.
- Investigation: SK BR CW JB KB.
- Methodology: BR SH SN.
- Project administration: SH.
- Resources: CD MH.
- Software: CD.
- Supervision: SH.
- Validation: BR SK CW KB MH.
- Visualization: SH.
- Writing – original draft: BR SH.
- Writing – review & editing: SH BR SN MLH.
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domain: biology


text:
Ahead of the Summer Olympics in Rio and alongside our hatred of personal physical exercise, this designs a winner. Artwork and design copyright Push Merchandising Ltd.

Mug design reads: Im not racist, I love races. Except running. Fuck running.

Our ceramic mugs are all white barring the printed design and are a standard mug size.
A floral printed and slogan Home is Wheverever Im With You cushion, with crotchet and button details; a pretty addition for your bedroom or living room.Fibre Composition: 100% CottonComposition of Pad/Inner Lining: 100% PolyesterWash / Care Instructions: Spot clean onlyDimensions: 30x30cmsColour: Multi
Are you in love with the Coco? This fun little bodysuit is one for the summer. In soft stretch jersey fabric it features a bold white slogan across the front and popper fasten at the gusset. Team with high waist denim shorts and sandals for fun ibiza styling. Approx length 70cm/27.5 (Based on a UK size 8 sample) Mixed fibres Lisa Marie wears a UK size 8/ EU size 36/ US size 4 and her
An Evening With Diana Ross was recorded in L.A. in 1976 and it captures her award-winning one-woman show. Chaptered into sections like The Motown Story and The Supremes it includes hits like "Love Hangover," "Reflections" and "Theme From Mahogany."
Celebrate the joys of true romance with sweet solo-piano renditions of popular love songs. 1. When Im With You 2. Weve Got Tonight 3. Dont Know Much 4. Hello 5. I Know Ill Never Love This Way Again 6. St Elmos Fire (Love Theme) 7. Heaven 8. Babe 9. Tonight I Celebrate My Love 10. All Out Of Love 11. Cant We Try 12. After All 13. When I See You Smile 14. Three Times A Lady
Women’s ‘I’m Not Here’ drop crotch blue jeans from The Fifth. The relaxed fit five pocket jeans feature distressed detail to the knees coin pocket and rear pocket. Fastened with a button and zip fly the jeans are finished with metal hardware zip detail to the hems and a woven brand patch to the waistband. – A.D. Cotton
Step one step two in these fashionable boots by BC Footwear.Made of leather and fabric.Pull on boot.Adjustable ankle and quarter straps with buckle closures.Man-made lining.Lightly padded insole.Man-made outsole.Imported. Measurements:Heel Height: 1 inWeight: 1 lbShaft: 9 1 4 inProduct measurements were taken using size 6, width M. Please note that measurements may vary by size.
\26in Tall, 4 legs covered in sisal, Sisal provides alternate scratching texture your cat will love, Hole in bottom so your cat can climb inside for a cozy home, Poly whiskers provide hours of entertainment for your cat, High quality materials, Excellent craftsmanship. Molly is a unique cat toy that is a favorite among cats and their owners. High quality materials and craftsmanship alone make
This medium (2.5), easy to clean rubber ball whistles in flight; adding another fun element to an what would otherwise be an ordinary game of fetch! Whistler ball is compatible will all Chuckit! Medium ball launchers.
Our best seller is back! If youre looking for a midi length dress cut to sit below the knee then look no further. In our signature stripes or statement floral and crafted from soft jersey this dress with a wide higher neck line its sure to be a go-to favourite.

domain: hobby


text:
The Day Paul McCartney Was Knighted
On March 11, 1997, at the age of 54, former Beatles and Wings member Paul McCartney received one of Britain's highest honors when he was knighted by Queen Elizabeth II. The centuries-old ceremony, officially bestows the title of "Sir" upon all fortunate enough to receive the designation.
It was not the first time that McCartney had received high recognition from his homeland. In October 1965, McCartney, along with fellow band members John Lennon, George Harrison and Ringo Starr, were given Member of the British Empire medals, much to the shock of the British establishment at the time. Lennon returned his MBE medal in 1969 as a war protest.
Despite having been a part of one of the biggest bands in the history of music and being one-quarter responsible for having created Beatlemania all over the world, McCartney admitted to some nervousness heading into the ceremony.
"Proud to be British, wonderful day and it's a long way from a little terrace in Liverpool," McCartney told reporters.
The ceremony was attended by three of McCartney's four children with his wife Linda, who was absent from the ceremony as she was in the midst of her battle with breast cancer. Thirteen months later, on April 17, 1998, Linda sadly lost her fight and succumbed to the disease.
Musicians receiving the British knighthood was a still somewhat of a rarity when McCartney received his designation in 1997. Cliff Richard was actually the first musician to have been given a knighthood in 1995 but since then, many of music's royalty have received the title including Bono (U2, 2007), Elton John (1998) and the Rolling Stones' Mick Jagger (2003).
In an interesting twist, David Bowie allegedly declined the knighthood in 2003, reportedly stating that it was not what he had spent his life working for.
Rock Stars Who Have Been Knighted

domain: news


text:
Copyright ©2007-2020 The Biker Lawyers The determination of the need for legal services and the choice of a lawyer are extremely important decisions and should not be based solely upon advertisements or self-proclaimed expertise. Click here for the full disclaimer.
The Nashville Police Department has reported that a motorcycle-sport utility vehicle collision that occurred at around 7:15 p.m. on Wednesday, July 20 on River Road near Indian Creek Road in Nashville resulted in the death of one individual.
The victim was identified as a 17-year-old girl from Iowa. She was riding on the back of her father’s Harley Davidson motorcycle. Her father was driving the vehicle at the time of the accident. He is in critical condition, but has stabilized.
According to preliminary investigators, the motorcycle was navigating a westbound curve on River Road near Indian Creek Road when it collided with a Subaru Forester that was headed east.
Police officers said they found no reason or evidence to believe that the accident was caused by alcohol intoxication or substance impairment.


Queen Seon-deok has SBS running scared
The upcoming Angel’s Temptation
Wow. It’s one thing to admit your competition is tough. But it’s another thing entirely to rearrange your station’s prime-time broadcasting schedule specifically to avoid going up against said competitor, as SBS is doing next month to give its dramas a shot against Queen Seon-deok.
Starting in October, they will actually air their Monday-Tuesday drama in the 9pm hour, rather than the standard prime-time 10pm hour where Seon-deok has been so dominant.
This is a pretty drastic — one might say desperate — move. It’s not unusual for rival stations to fiddle with drama premiere dates, pushing back a new show until a strong competitor wraps (as Ja Myung Go did to avoid going up against East of Eden and Boys Before Flowers). But shifting its prime-time schedule entirely is a big surprise. On one hand, it’s good that their drama(s) will be getting a shot to survive, but on the other, it feels like an admission of weakness.
Ja Myung Go, Dream
Of course, SBS’s official statement says that they have been wanting to make this move for a while based on ratings research, but they’re hardly fooling anyone. It is a highly unusual decision to break from tradition and instead offer cultural and educational programs in the 10pm hour as counterprogramming against KBS and MBC’s prime-time dramas. (The other two stations air the news in the 9pm hour.)
Several of SBS’s highly anticipated dramas have crumbled against Seon-deok, such as its big-budget historical epic series Ja Myung Go, which was pulled early, and Dream, which is limping along with 4% ratings. Seon-deok‘s six-week extension stretches its run through the end of 2009, and SBS must be impatient to breathe some life into their schedule rather than waiting until next year.
Therefore, the new 9pm schedule starts on October 12. It’s too late to save Dream, but the schedule goes into effect with SBS’s next Monday-Tuesday offering, the makjang revenge drama Angel’s Temptation. It’s no surprise SBS is unwilling to sacrifice a probable hit with a name writer whose last drama (Wife’s Temptation) was a 40% ratings sensation earlier this year.
Angel’s Temptation stars Han Sang-jin (of Sons of Sol Pharmacy, pictured at top) and Bae Soo-bin (Brilliant Legacy) playing the same character: the character’s wife (Lee So-yeon) marries him purely out of a revenge scheme, and he comes back from an accident and plastic surgery with a new face and a desire to counter his wife’s revenge with his own.
I know this news doesn’t affect us international viewers, who don’t influence (or likely keep track of) ratings wars. But I’m pretty surprised with SBS. They may gain ratings but they sure lose face…
Via My Daily


Thorough and well-written article. I completely agree with the speed of installation advantages of a wireless system. Many years ago, I had to set up a wired network to our factory floor and the time required to plan and install this older technology was significant (not to mention the time needed to troubleshoot improper hardware node connections). In many instances, wireless technology will prove to be faster and much more flexible than existing wired systems.
I see combatting the noisy industrial environment, wirelessly, is making some strides. I think I feel confident enough in the tech these days to use wireless in the machine shop. The little risk may have a huge payoff in mobility.
Artificially created metamaterials are already appearing in niche applications like electronics, communications, and defense, says a new report from Lux Research. How quickly they become mainstream depends on cost-effective manufacturing methods, which will include additive manufacturing.
Sharon Glotzer and David Pine are hoping to create the first liquid hard drive with liquid nanoparticles that can store 1TB per teaspoon. They aren't the first to find potential data stores, as Harvard researchers have stored 700 TB inside a gram of DNA.
Focus on Fundamentals consists of 45-minute on-line classes that cover a host of technologies. You learn without leaving the comfort of your desk. All classes are taught by subject-matter experts and all are archived. So if you can't attend live, attend at your convenience.


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Welcome to our mini blog series that will focus on a number of locations around north Wales. Starting with Trearddur Bay on Holy Island then moving over on to Anglesey, Snowdonia and the Llyn Peninsular. We have kept the mini guide concise and short so its an ideal for a reference for your stay. Whether … More Trearddur Bay
We headed out for a quick walk around The dingle nature reserve in Llangefni (Nant Y Pandy), Anglesey. It’s a 25 acre woodland site in the center of Anglesey. The council have created a fantastic board walk area that leads all the way up towards a salmon leap and follows up all the way to … More The Dingle Nature reserve, Llangefni Anglesey.


text:
1). https://lms.latrobe.edu.au/mod/resource/view.php?id=4017633(this link contains the assignment description)
2.)https://lms.latrobe.edu.au/mod/resource/view.php?id=4017634(this is the rubric)
3.)https://lms.latrobe.edu.au/mod/resource/view.php?id=4017635(this is the link to the FAQS i thought u might need it)
4.)https://lms.latrobe.edu.au/mod/resource/view.php?id=4017636(this is an example of one of the assignments from last year)
Below is the general description of the assignments with the links to articles.
Individual written report research critique
Due Dates:0900, 7 September 2020
Word Limit: (1,750 Words)
Critique the two papers listed in the assessment tile below. You will need to locate the full text of the papers. These papers should then be critiqued and an essay written in which you compare and contrast the two research articles by:
Once you have critically appraised each paper, you should consider what these two papers contribute, relative to each other, to the clinical issue being considered.
Your critique needs to be appropriately referenced using APA (7th) guidelines.
Submit your assessment via the Turnitin link in the Assessments area on LMS for grading. It will be graded via Grademark and returned to you via the Turnitin link.
The articles to be critiqued for the assessment are:
Hghaug, G., Skr, R., Tran, T. N., & Schou-Bredal, I. (2019). Nurses experiences with newly acquired knowledge about medication management: A qualitative study.Journal of Nursing Management, 27(8), 1731-1737. doi:10.1111/jonm.12864
Huckels-Baumgart, S., Niederberger, M., Manser, T., Meier, C. R., & Meyer-Massetti, C. (2017). A combined intervention to reduce interruptions during medication preparation and double-checking: a pilot-study evaluating the impact of staff training and safety vests.Journal of Nursing Management, 25(7), 539-548. doi:10.1111/jonm.12491


Prepared by : Wigdan Nadeem
Introduction Theory of concurrency Models Key terms related to concurrency Concurrency areas Concurrency control Advantages and Disadvantages References
Concurrency is the ability of a database to allow multiple users to affect multiple transactions. This is one of the main properties that separate a database from other forms of data storage like spreadsheets.
Concurrency theory has been an active field of research in theoretical computer science. One of the first proposals was Carl Adam Petri's seminal work on Petri Nets in the early 1960s. In the years since, a wide variety of formalisms have been developed for modeling and reasoning about concurrency.
A number of formalisms for modeling and understanding concurrent systems have been developed, including but not limited to: - The Parallel Random Access Machine. - The Actor model - Computational bridging models such as the BSP model.
- Operating systems: The most common topic where concurrency is taught and addressed is in the context of operating systems design and implementation. - Scientific and high-performance computing: For many decades, the HPC and scientific computing areas were the primary source of work on parallelism. - Databases and servers: Server-side applications that needed to handle multiple concurrent users.
Any application that is run on a laptop or desktop system with multiple cores can use concurrency for performance gains. - Games - Desktop applications - Development tools - Image and data processing
To ensures that correct results for concurrent operations are generated, while getting those results as quickly as possible. Simultaneous execution of transactions over a shared database can create several data integrity and consistency problems: - Lost Updates. - Uncommitted Data. - Inconsistent retrievals.
- Pessimistic concurrency control: assumes that...
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Oklahoma House passes bill that would provide alternative ways to carry out death penalty
The Oklahoma House passes House Bill 1679, which would provide alternative ways to carry out death penalty.
The measure provides that the available manners of execution are to be lethal drug, nitrogen hypoxia, firing squad, any method not prohibited by the United States Constitution.
The manner of carrying out the punishment shall be selected by the Director of the Department of Corrections.
Stay with KOCO 5 for updates.


Photo: Gary Newbold, Extreme Networks
In today's highly connected and overly stimulated world, it is exceedingly difficult to measure user engagement, improve the mobile and social user experience and ensure the integrity of mission-critical networks.
However, the latest network-powered application analytics, ASIC-based technology addresses these complex requirements by integrating with network data that carries context of users, devices, locations and applications in use. ASIC—application-specific integrated circuit—is an integrated circuit customised for a particular use rather than intended for general-purpose procedure.
Breakthrough ASIC technology is able to capture network data and then aggregate, analyse, correlate, characterise it then finally report on it. These reports provide insights into how and why systems and users perform. Massive volumes of application usage data can be collected, analysed and reported to help focus organisations on improving user experience, engagement and organisational efficiency.
Such solutions provide visibility into application use across the network, helping organisations in four ways:
- Improving the experience of connected users.
- Enhancing organisations' understanding of user engagement.
- Optimising application performance.
- Protecting against malicious or unapproved system use.
Organisations using the technology are able to deliver significant improvements to their critical technology assets. The new solutions leverage breakthrough technology to bridge the gap between IT network management, security and business intelligence through a single integrated view. They help organisations improve user experience, while meeting the growing needs of network users and improving network performance. The benefits can be realised on networks from any Ethernet switching vendor.
Although the technology is new to Singapore, it is already operating successfully in the USA. Michelle McKenna-Doyle, CIO of the National Football League, said: "We want to deliver the most immersive and connected experience for our fans at the stadium and the new ASIC-based solution can help enhance the fan experience through its advanced analytics capabilities. In delivering insight on user application preference and usage, our teams are able to best use Wi-Fi technology to deliver an exceptional game day experience."
One version of this technology is the official Wi-Fi analytics solution for the National Football League (NFL), available to all 32 teams. The solution is currently deployed across four NFL stadiums, and provided the Wi-Fi analytics for the NFL at Super Bowl XLVIII.
"With the explosion of enterprise mobility, social and collaborative applications, network infrastructure is now viewed as an enterprise IT asset and a key contributor to optimising business applications and strategic intelligence," said Rohit Mehra, vice president, network infrastructure, IDC. "Such solutions are unique in terms of providing rich application analytics integrated within a network management platform, so enterprise IT is enabled with deep visibility and control of data, applications and users, for better business decisions, easily and efficiently."
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The Boys of Winter
The 1980 U.S. men’s hockey team was a ragtag bunch of amateurish blue-collar guys who were led by a charismatic coach. When they went skate-to-skate with the Russian men’s hockey juggernaut at the height of the Cold War, they managed to pull off a victory, which we’ve come to know as “The Miracle on Ice.” Sportswriter Wayne Coffey’s book gives an ice-level view of the action, weaving profiles of the players and coach with a play-by-play of the game for an exhilarating read.
In the 1930s, as tensions mounted toward the Second World War, speed was paramount and bobsledding was the sport du jour at the 1932 Olympics in Lake Placid, New York. Dangerous and thrilling, the sport required great skill and even greater courage. In the midst of the Great Depression, Americans followed the stories of the four-man bobsledding team—Jay O’Brien, Tippy Grey, Eddie Eagan, and Billy Fiske—as they dealt with bothersome officials and competed against the opposing German team.
Fire on the Track
In the 1928 Olympic Games in Amsterdam, Betty Robinson competed in what was only her fourth-ever organized track meet, yet she crossed the finish line first and was named the fastest woman in the world. Her career was nearly dashed, however, when a plane she and her cousin were piloting crashed, and she was pronounced dead. But Robinson wasn’t quite dead, and though she could barely walk, she earned herself a spot on the 1936 Olympic team with grit and determination. An incredible story of the power of persistence and how one woman’s pioneering role paved the way for other female athletes.
Jules Boykoff, a former soccer player for the U.S. Olympic team, dives into the political history that often surrounds the Olympic Games, from their 19th-century origins to today. Modern-day Olympics are plagued by corporate control, and the building of Olympic villages often displace indigenous people from their land. This no-holds barred history of the modern Olympic Games provides insight into the backdrops that athletes have been competing against for decades.
The Boys in the Boat
Daniel James Brown
This New York Times-bestseller tells the improbable story of nine working-class men from the American West who formed an eight-oar crew team and defeated elite rowing teams from the U.S. East Coast, going on to compete in the Olympic Games in Nazi Germany in 1936. The individual stories of the young men are woven with the tale of their rise from obscurity to their triumph over Hitler’s German team in Berlin. An inspiring underdog story that will have readers white-knuckled with suspense.
Age Is Just a Number
Four-time Olympic gold medalist Dara Torres was a 41-year-old new mother when she came out of retirement and staged her Olympic comeback in Beijing. In this memoir, she writes about what it took for her to dive back in to her sport, being older in a younger person’s game, having a new take on competition, self-doubt, and perseverance, and what it felt like to have America cheer her on.
There’s nothing like watching the Opening Ceremony of the Olympic Games when that familiar music plays and athletes from all over the world gather to compete on an international stage. It’s inherently inspiring and goosebump-inducing. As the world prepares for the start of the 2022 Winter Olympics in Beijing, China (and then Milan and Cortina d’Ampezzo, Italy in 2026!), we’re grabbing one of these stirring stories about triumph at the Games and the quest for the gold medal. From historic games set against the backdrop of Nazi Germany to memoirs by modern competitors, each book is sure to inspire awe in its reader.
Featured Image: Heritage Images/Getty Images


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Extracts from this document introduction comparison of 'once upon a time' by gabriel okara and 'a martian sends a postcard home' by craig raine gabriel okara was born in 1921 in ijaw country in the niger delta, in nigeria. Tamara background information television programs once upon a time portrayed by sonequa martin-green character information full name tamara other names her personality cunning, untruthful, plotting, uncaring, intelligent, aspiring, cunning, ruthless, murderous, elusive, brooding, dark. Lyrics to once upon a dream song by lana del rey: i know you, i walked with you once upon a dream i know you, that look in your eyes is so familiar a. I think that nadine gordimer uses irony in her short story once upon a time” to send her readers the main idea of the story in a more interesting way than just directly explaining it. Once upon a time summary by gabriel okara the poem “once upon a time” written by gabriel okara illustrates the changes a father has seen in him throughout his.
In the problem of the month once upon a time, students use measurement, number properties, and circular geometry to solve problems involving time and angles. Sergio leone goes a long way with once upon a time in the westperhaps too long once upon a time in the west isn't exactly a race down the track and running 2 hours and 45 minutes doesn't help its cause. Home once upon a time: the adaptation of grimm’s fairy tales in the eponymous television series it also bases its analysis on quotations and their importance. Once upon a time fan club 2,287 likes 3 talking about this welcome to once upon a fan club (fan page closing after season 6.
Ennio morricone’s score for once upon a time this music represents the very best of the art and i thank you for your intensive analysis of “once upon a time. We're all about sexy captain hook on once upon a time, and his relationship with emma after going through hell and back together this season, they have proven. The fairy tale drama debuted sunday to abc's strongest ratings in the 18-49 demographic in nearly three years of regular programming abc’s once upon a time opened to strong ratings sunday, lifting abc to its best performance in the time period with regular programming in nearly three years in the 18-49 demographic. Once upon a time in china is a 1991 wuxia martial arts film directed by tsui hark, a long time collaborator with john woo the protagonist is doctor wong. Who here is a once upon a time fan have you been following what's going on in the swan queen community, and trying more .
Find and follow posts tagged once upon a time on tumblr. Themes childhood dreaming black boy forgive my guilt once upon a time forgive my guilt alienation test match sabina park dreaming black boy theme for. By: firas krekshi and sara lizzio once upon a time question #1 photo analysis this is the passage that we chose, it reminded us of certain fairy tales. Once upon a time in the west iconic score that deserves its place in film music folklore a review by james southall films don't come much more highly-regarded than once upon a time in the west, sergio leone's western-to-end-all-westerns.
Once upon a time, kalenjin nation had straight thinking leaders, ignore murkomen july 19, 2018 leave a comment. Once upon a time tv episode reviews, analysis and news. High noon (1952) title sequence often imitated but never surpassed, the influence of once upon a time in the west’s iconic opening is still felt to this daydirector quentin tarantino opened his 2009 film inglourious basterds with a sequence entitled “once upon a time in nazi-occupied france,” introducing the movie’s primary villain in.
Once upon a time is an american fairy tale drama television series created by edward kitsis and adam horowitz, who also serve as executive producers alongside steve. A christmas carol: novel summary, free free study guides and book notes including comprehensive chapter analysis, once upon a time. Once upon a time, there were some people called the rushers of din each night as they slept, sweet dreams were delivered to them from sunny frivoli, while nightmares came to them from the mysterious. Once upon a time in the west is a 1968 spaghetti western film about a mysterious stranger with a harmonica who joins forces with a notorious desperado to protect a beautiful widow from a ruthless assassin working for the railroad.
Ouat fans is a once upon a time a thorough analysis of the costumes we’ve seen on the show ouat fashion finds ouatfashionfindscom once upon a time. 6 analyze the story's final paragraph in detail how does it help to elucidate the theme i found this story to be incredibly satirical the author seemed to be making fun of typical children's stories, and implemented several common phrases like once upon a time or they lived happily ever after. In the context of once upon a time in the west, ennio morricone’s score for once upon a time in the west i would be interested in an analysis of this music.
Coit-essay nadine gordimer nadine gordimer once upon a time someone has written to ask me to contribute to an anthology of stories for children. Topic: - critical analysis of three poem refugee mother and child, new york, and once upon a time.

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I have a similar idea...
I opened my E-Mu XBoard25 keyboard and found out that there is a perfect place with definitely enough space to implant an Axoloti, namely above the battery compartment.
Therefore, it seems very attractive to me to make a real, stand-alone synthesizer of this device.
I'm not yet sure if I make the connection via MIDI or USB-MIDI, and how to unite the power supplies. And some more mechanical considerations are to be made, like a back plate design. But it should be relatively easy.
Maybe I can even retain all the original functionality as well.
I think we could collect and share here our findings about the suitability of all sorts of MIDI devices, couldn't we?
What I think would be very attractive for implanting is the Behringer BCR2000. Does anyone here have one and could check if there is space left inside?
"Axolotify your MIDI device!"