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WASHINGTON, May 18, 2017 (GLOBE NEWSWIRE) -- Today Dr. David Veal, executive director of the American Shrimp Processors Association (ASPA), testified before officials from the U.S. Department of Commerce (DOC) and the U.S. Trade Representative (USTR) at a public hearing in Washington, DC to address significant trade deficits in the shrimp industry. “For decades, our shrimp industry has faced surging imports of farm-raised shrimp produced overseas,” Dr. Veal said. “As a result, the U.S. trade deficit in shrimp is substantial and growing." Dr. Veal’s testimony will be considered in the drafting of an “Omnibus Report on Significant Trade Deficits,” which will be presented to President Trump by the DOC and USTR.  ASPA also submitted written comments. The U.S. trade deficit in shrimp totaled $4.49 billion in 2016.  India, Indonesia, Thailand, Vietnam, Mexico, China, and Malaysia are seven of thirteen countries with which the United States ran a significant overall shrimp trade deficit in 2016.  These nations are also nations being investigated in the Omnibus Report.  The deficit of $3.44 billion with these nations is 77 percent of the total shrimp trade deficit.  And, the deficit in shrimp from those seven countries has increased 44 percent since 2012. “The U.S. trade deficit in all seafood was a staggering $10.5 billion in 2016, greater than the deficit in either raw iron and steel or aluminum,” Dr. Veal said in his testimony.  The deficit, he added, is the result of unfair and illegal trade practices, including persistent dumping in the U.S. market and subsidization by foreign governments. On May 2, the International Trade Commission unanimously voted to extend the current antidumping orders on shrimp from China, India, Thailand and Vietnam for an additional five years.  Those orders provide important discipline in the market, Dr. Veal testified, but have limited coverage and still allow large volumes of unfairly traded shrimp to enter the United States. “Unfairly traded imports have distorted trade patterns and hindered the growth and future stability of the domestic industry,” Dr. Veal said. ASPA is currently pursuing a policy agenda to help restore domestic job growth and promote balanced trade, and is optimistic those concerns will be addressed in the Omnibus Report presented to the President. The American Shrimp Processors Association, based in Biloxi, Mississippi, was formed in 1964 to represent and promote the interests of the domestic, U.S. wild-caught, warm water shrimp processing industry along the Gulf Coast with members from Texas, Louisiana, Mississippi, Alabama and Florida. A collective voice of the industry, the ASPA's focus is to promote the interests of shrimp processors, other segments of the U.S. domestic wild-caught shrimp industry and the general public.


WASHINGTON, May 18, 2017 (GLOBE NEWSWIRE) -- Today Dr. David Veal, executive director of the American Shrimp Processors Association (ASPA), testified before officials from the U.S. Department of Commerce (DOC) and the U.S. Trade Representative (USTR) at a public hearing in Washington, DC to address significant trade deficits in the shrimp industry. “For decades, our shrimp industry has faced surging imports of farm-raised shrimp produced overseas,” Dr. Veal said. “As a result, the U.S. trade deficit in shrimp is substantial and growing." Dr. Veal’s testimony will be considered in the drafting of an “Omnibus Report on Significant Trade Deficits,” which will be presented to President Trump by the DOC and USTR.  ASPA also submitted written comments. The U.S. trade deficit in shrimp totaled $4.49 billion in 2016.  India, Indonesia, Thailand, Vietnam, Mexico, China, and Malaysia are seven of thirteen countries with which the United States ran a significant overall shrimp trade deficit in 2016.  These nations are also nations being investigated in the Omnibus Report.  The deficit of $3.44 billion with these nations is 77 percent of the total shrimp trade deficit.  And, the deficit in shrimp from those seven countries has increased 44 percent since 2012. “The U.S. trade deficit in all seafood was a staggering $10.5 billion in 2016, greater than the deficit in either raw iron and steel or aluminum,” Dr. Veal said in his testimony.  The deficit, he added, is the result of unfair and illegal trade practices, including persistent dumping in the U.S. market and subsidization by foreign governments. On May 2, the International Trade Commission unanimously voted to extend the current antidumping orders on shrimp from China, India, Thailand and Vietnam for an additional five years.  Those orders provide important discipline in the market, Dr. Veal testified, but have limited coverage and still allow large volumes of unfairly traded shrimp to enter the United States. “Unfairly traded imports have distorted trade patterns and hindered the growth and future stability of the domestic industry,” Dr. Veal said. ASPA is currently pursuing a policy agenda to help restore domestic job growth and promote balanced trade, and is optimistic those concerns will be addressed in the Omnibus Report presented to the President. The American Shrimp Processors Association, based in Biloxi, Mississippi, was formed in 1964 to represent and promote the interests of the domestic, U.S. wild-caught, warm water shrimp processing industry along the Gulf Coast with members from Texas, Louisiana, Mississippi, Alabama and Florida. A collective voice of the industry, the ASPA's focus is to promote the interests of shrimp processors, other segments of the U.S. domestic wild-caught shrimp industry and the general public.


WASHINGTON, May 18, 2017 (GLOBE NEWSWIRE) -- Today Dr. David Veal, executive director of the American Shrimp Processors Association (ASPA), testified before officials from the U.S. Department of Commerce (DOC) and the U.S. Trade Representative (USTR) at a public hearing in Washington, DC to address significant trade deficits in the shrimp industry. “For decades, our shrimp industry has faced surging imports of farm-raised shrimp produced overseas,” Dr. Veal said. “As a result, the U.S. trade deficit in shrimp is substantial and growing." Dr. Veal’s testimony will be considered in the drafting of an “Omnibus Report on Significant Trade Deficits,” which will be presented to President Trump by the DOC and USTR.  ASPA also submitted written comments. The U.S. trade deficit in shrimp totaled $4.49 billion in 2016.  India, Indonesia, Thailand, Vietnam, Mexico, China, and Malaysia are seven of thirteen countries with which the United States ran a significant overall shrimp trade deficit in 2016.  These nations are also nations being investigated in the Omnibus Report.  The deficit of $3.44 billion with these nations is 77 percent of the total shrimp trade deficit.  And, the deficit in shrimp from those seven countries has increased 44 percent since 2012. “The U.S. trade deficit in all seafood was a staggering $10.5 billion in 2016, greater than the deficit in either raw iron and steel or aluminum,” Dr. Veal said in his testimony.  The deficit, he added, is the result of unfair and illegal trade practices, including persistent dumping in the U.S. market and subsidization by foreign governments. On May 2, the International Trade Commission unanimously voted to extend the current antidumping orders on shrimp from China, India, Thailand and Vietnam for an additional five years.  Those orders provide important discipline in the market, Dr. Veal testified, but have limited coverage and still allow large volumes of unfairly traded shrimp to enter the United States. “Unfairly traded imports have distorted trade patterns and hindered the growth and future stability of the domestic industry,” Dr. Veal said. ASPA is currently pursuing a policy agenda to help restore domestic job growth and promote balanced trade, and is optimistic those concerns will be addressed in the Omnibus Report presented to the President. The American Shrimp Processors Association, based in Biloxi, Mississippi, was formed in 1964 to represent and promote the interests of the domestic, U.S. wild-caught, warm water shrimp processing industry along the Gulf Coast with members from Texas, Louisiana, Mississippi, Alabama and Florida. A collective voice of the industry, the ASPA's focus is to promote the interests of shrimp processors, other segments of the U.S. domestic wild-caught shrimp industry and the general public.


WASHINGTON, May 18, 2017 (GLOBE NEWSWIRE) -- Today Dr. David Veal, executive director of the American Shrimp Processors Association (ASPA), testified before officials from the U.S. Department of Commerce (DOC) and the U.S. Trade Representative (USTR) at a public hearing in Washington, DC to address significant trade deficits in the shrimp industry. “For decades, our shrimp industry has faced surging imports of farm-raised shrimp produced overseas,” Dr. Veal said. “As a result, the U.S. trade deficit in shrimp is substantial and growing." Dr. Veal’s testimony will be considered in the drafting of an “Omnibus Report on Significant Trade Deficits,” which will be presented to President Trump by the DOC and USTR.  ASPA also submitted written comments. The U.S. trade deficit in shrimp totaled $4.49 billion in 2016.  India, Indonesia, Thailand, Vietnam, Mexico, China, and Malaysia are seven of thirteen countries with which the United States ran a significant overall shrimp trade deficit in 2016.  These nations are also nations being investigated in the Omnibus Report.  The deficit of $3.44 billion with these nations is 77 percent of the total shrimp trade deficit.  And, the deficit in shrimp from those seven countries has increased 44 percent since 2012. “The U.S. trade deficit in all seafood was a staggering $10.5 billion in 2016, greater than the deficit in either raw iron and steel or aluminum,” Dr. Veal said in his testimony.  The deficit, he added, is the result of unfair and illegal trade practices, including persistent dumping in the U.S. market and subsidization by foreign governments. On May 2, the International Trade Commission unanimously voted to extend the current antidumping orders on shrimp from China, India, Thailand and Vietnam for an additional five years.  Those orders provide important discipline in the market, Dr. Veal testified, but have limited coverage and still allow large volumes of unfairly traded shrimp to enter the United States. “Unfairly traded imports have distorted trade patterns and hindered the growth and future stability of the domestic industry,” Dr. Veal said. ASPA is currently pursuing a policy agenda to help restore domestic job growth and promote balanced trade, and is optimistic those concerns will be addressed in the Omnibus Report presented to the President. The American Shrimp Processors Association, based in Biloxi, Mississippi, was formed in 1964 to represent and promote the interests of the domestic, U.S. wild-caught, warm water shrimp processing industry along the Gulf Coast with members from Texas, Louisiana, Mississippi, Alabama and Florida. A collective voice of the industry, the ASPA's focus is to promote the interests of shrimp processors, other segments of the U.S. domestic wild-caught shrimp industry and the general public.


WASHINGTON, May 18, 2017 (GLOBE NEWSWIRE) -- Today Dr. David Veal, executive director of the American Shrimp Processors Association (ASPA), testified before officials from the U.S. Department of Commerce (DOC) and the U.S. Trade Representative (USTR) at a public hearing in Washington, DC to address significant trade deficits in the shrimp industry. “For decades, our shrimp industry has faced surging imports of farm-raised shrimp produced overseas,” Dr. Veal said. “As a result, the U.S. trade deficit in shrimp is substantial and growing." Dr. Veal’s testimony will be considered in the drafting of an “Omnibus Report on Significant Trade Deficits,” which will be presented to President Trump by the DOC and USTR.  ASPA also submitted written comments. The U.S. trade deficit in shrimp totaled $4.49 billion in 2016.  India, Indonesia, Thailand, Vietnam, Mexico, China, and Malaysia are seven of thirteen countries with which the United States ran a significant overall shrimp trade deficit in 2016.  These nations are also nations being investigated in the Omnibus Report.  The deficit of $3.44 billion with these nations is 77 percent of the total shrimp trade deficit.  And, the deficit in shrimp from those seven countries has increased 44 percent since 2012. “The U.S. trade deficit in all seafood was a staggering $10.5 billion in 2016, greater than the deficit in either raw iron and steel or aluminum,” Dr. Veal said in his testimony.  The deficit, he added, is the result of unfair and illegal trade practices, including persistent dumping in the U.S. market and subsidization by foreign governments. On May 2, the International Trade Commission unanimously voted to extend the current antidumping orders on shrimp from China, India, Thailand and Vietnam for an additional five years.  Those orders provide important discipline in the market, Dr. Veal testified, but have limited coverage and still allow large volumes of unfairly traded shrimp to enter the United States. “Unfairly traded imports have distorted trade patterns and hindered the growth and future stability of the domestic industry,” Dr. Veal said. ASPA is currently pursuing a policy agenda to help restore domestic job growth and promote balanced trade, and is optimistic those concerns will be addressed in the Omnibus Report presented to the President. The American Shrimp Processors Association, based in Biloxi, Mississippi, was formed in 1964 to represent and promote the interests of the domestic, U.S. wild-caught, warm water shrimp processing industry along the Gulf Coast with members from Texas, Louisiana, Mississippi, Alabama and Florida. A collective voice of the industry, the ASPA's focus is to promote the interests of shrimp processors, other segments of the U.S. domestic wild-caught shrimp industry and the general public.


WASHINGTON, March 31, 2017 (GLOBE NEWSWIRE) -- “We are very pleased that President Donald Trump has taken major action today to strengthen U.S. trade enforcement,” said American Shrimp Processor Association Executive Director Dr. David Veal.  “Making it harder for foreign companies to evade legally required duties helps to level the playing field for U.S. companies like ASPA members against unfair foreign competitors,” he added. “For imported shrimp and many other imported products covered by orders, there have been problems with importers disappearing or not paying duties found owed.  Under this order, Customs and Border Protection will have 90 days to devise a plan to protect the revenue where importers pose risks by requiring additional bonding or other security,” said ASPA Gulf Trade Counsel, Eddy Hayes.  “Ensuring that high risk importers cannot import dumped or subsidized imports and then escape liability for duties owed is an important step towards improving the effectiveness of U.S. trade remedies,” he commented. “In an industry like shrimp, where small, family-owned American businesses have to compete with large volumes of unfairly traded imports, foreign duty evasion is a perennial and widespread problem,” commented Dr. Veal.  “When this order is implemented, duty collection should immediately improve.  That means the competitiveness of our U.S. shrimp industry improves and our ASPA members can create more jobs.” The American Shrimp Processors Association (ASPA), based in Biloxi, Mississippi, was formed in 1964 to represent and promote the interests of the domestic, U.S. wild-caught, warm water shrimp processing industry along the Gulf Coast with members from Texas, Louisiana, Mississippi, Alabama and Florida. We are the collective voice of the industry, and our focus is to promote the interests of shrimp processors, other segments of the U.S. domestic wild-caught shrimp industry and the general public.


WASHINGTON, May 02, 2017 (GLOBE NEWSWIRE) -- Today the U.S. International Trade Commission (USITC) voted unanimously to support the domestic shrimp industry’s request to extend the antidumping orders on shrimp from China, India, Thailand and Vietnam for an additional five years.  The USITC voted to remove the orders on Brazil.    “We at the American Shrimp Processors Association (ASPA) are grateful that the U.S. International Trade Commission affirmed evidence of the risk that dumped shrimp imports from these five nations poses to the domestic shrimp industry,” said Dr. David Veal, Executive Director of ASPA.  “We in the domestic shrimp industry look forward to five additional years of relief from unfair foreign trade practices,” Dr. Veal added. “We are pleased that the USITC validated the data and evidence we presented, which we believe clearly showed the harm that these imports would cause to the domestic industry if the orders were revoked,”  noted ASPA Gulf Counsel Edward T. Hayes.  “We are grateful for the strong support of U.S. Senators, Representatives, and other public offices from North Carolina to Texas for their shrimp processors, harvesters and docks across the region,” said Hayes. “ASPA members really appreciate that seventeen senior public officials supported our efforts at the USTIC.  They clearly understood that a successful outcome in this case was vital to the survival of the businesses in the American shrimp industry and to the future viability of the communities in which they live,” said Dr. Veal.  “We are especially grateful for the strong support of those who testified in person at the USITC, Representatives Walter Jones (R-NC), Steven Palazzo (R-MS), Garret Graves (R-LA) and Lt. Governor Billy Nungesser (R-LA),” he added. “ASPA also wants to thank Senators Thad Cochran (R-MS), Richard Shelby (R-AL), Roger Wicker (R-MS), and Bill Cassidy (R-LA)  for all their strong support now and over the years on this and a myriad of other issues,” Hayes noted. The American Shrimp Processors Association (ASPA), based in Biloxi, Mississippi, was formed in 1964 to represent and promote the interests of the domestic, U.S. wild-caught, warm water shrimp processing industry along the Gulf Coast with members from Texas, Louisiana, Mississippi, Alabama and Florida. We are the collective voice of the industry, and our focus is to promote the interests of shrimp processors, other segments of the U.S. domestic wild-caught shrimp industry and the general public.


WASHINGTON, March 16, 2017 (GLOBE NEWSWIRE) -- Today, the U.S. International Trade Commission (USITC) held a public hearing to help decide whether to renew anti-dumping trade enforcement orders against Brazil, China, India, Thailand and Vietnam in place for five more years or whether to sunset those orders. The USITC will hold a final vote on May 2, which will decide their fate.     Public official allies of the domestic shrimp industry today stressed that these orders are critical to restore free and fair trade in shrimp. American Shrimp Processor Association (ASPA) leaders appreciate the strong support of these public officials from the Gulf and South Atlantic regions that are critical to this effort to keep the domestic shrimp industry viable for the long term. “We are so pleased that 14 public officials from across the Gulf and South Atlantic Coast have weighed in so far at the USITC in support of domestic shrimp harvesters, processors and distributors,” said Dr. David Veal, Executive Director of ASPA. “We are grateful that the representatives, senators, and statewide officials support the retention of anti-dumping orders on foreign shrimp.” “In particular, we want to thank Representatives Garret Graves (R-LA), Walter Jones (R-NC) and Steven Palazzo (R-MS), as well as Louisiana Lt. Governor Billy Nungesser (R-LA), who all testified in person today at the USITC,” said Eddy Hayes, Gulf Counsel to ASPA. “Their commitment to trade enforcement and leveling the playing field for the domestic shrimp industry is impressive,” he added. “We are optimistic that the result of this Sunset Review will be the retention of these important anti-dumping orders,” Dr. Veal noted. “Leaders from our Gulf and South Atlantic regions understand the economic and cultural impact of the shrimp industry and are committed to its survival,” Hayes added. So far Senators Bill Cassidy (R-LA), Thad Cochran (R-MS), John Kennedy (R-LA), Richard Shelby (R-AL), Luther Strange (R-AL), and Roger Wicker (R-MS) have all committed to submit either letters or testimony in support of the domestic shrimp industry. On the House side, Representatives Clay Higgins (R-LA), Cedric Richmond (R-LA), Steve Scalise (R-LA), and Randy Weber (R-TX) have all committed to support the domestic shrimp industry. Additional input from other public officials from the region is also welcome. “ASPA is so appreciative of all the support that our government officials have provided. They are an impressive group of leaders for their constituents,” Dr. Veal noted. The American Shrimp Processors Association (ASPA), based in Biloxi, Mississippi, was formed in 1964 to represent and promote the interests of the domestic, U.S. wild-caught, warm water shrimp processing industry along the Gulf Coast with members from Texas, Louisiana, Mississippi, Alabama and Florida. We are the collective voice of the industry, and our focus is to promote the interests of shrimp processors, other segments of the U.S. domestic wild-caught shrimp industry and the general public.


All animal experimentation in this study was approved and performed according to the standards of the animal ethics committee at Imperial College London and to UK Home Office regulations (ASPA 1986). C57Bl/6 mice were purchased from Harlan UK Ltd; Col2.3–GFP, Col2.3–CFP18, nestin–GFP and mTmG25 mice were bred and housed at Imperial College London. For imaging experiments, female Col2.3–GFP and nestin–GFP mice >8 weeks old were used. osterix–CreGFP mice were provided by A. McMahon and backcrossed over eight generations into NSG mice and maintained at the Francis Crick Institute, Cancer Research UK26. Equal proportions of male and female osterix–CreGFP mice aged 11–14 weeks were used. T-ALL was generated as previously described27. Briefly, timed matings were established between C57Bl/6 mice and embryos harvested at E14.5. Single-cell suspensions were prepared from whole fetal livers isolated from the embryos. Suspensions were cultured in IL-3, IL-6, and stem cell factor conditioned media with 20% FCS for 3 days. Lin-xE cells were transfected by calcium phosphate with MigR1 plasmids containing either DsRed only or DsRed with NotchICNΔRamΔP as described previously28. We also used GFP-tagged plasmids when required. Supernatants containing recombinant retrovirus were removed and spun by centrifugation onto non-tissue-culture-treated plates coated with 15 μg/ml retronectin (Takara Clontech, CA). Fetal liver cells were cultured in the presence of virus for 3 days and transduction was assessed by flow cytometry. Primary lethally irradiated recipient mice (two doses of 5.5 Gy administered greater than three hours apart) were transplanted with 1 × 106 DsRed+ fetal liver cells by intravenous injection into the tail vein. Recipient mice were maintained on baytril-treated water to prevent infection for >6 weeks post-transplantation. Cohorts of reconstituted mice were the result of three independent fetal liver isolations and three independent transfections. Transformation of Notch-transduced non-malignant cells is highly heterogeneous in vivo, with onset of primary disease ranging from 6–25 weeks28 (Extended Data Fig. 1). More than 4 weeks post-reconstitution, peripheral blood was isolated from mice, red blood cells were lysed, and successful reconstitution determined by presence of DsRed+ cells. Mice reconstituted with NotchICNΔRamΔP-transduced fetal livers were monitored daily for signs of leukaemia onset or other signs of ill health. Mice were euthanized when any one or a combination of the following signs were observed: hunched posture, laboured breathing, weight loss, enlarged lymph nodes and/or spleen, peripheral white blood cell cellularity of 13 × 109 per litre or greater. No experiment exceeded the tumour burden approved by the Home Office and Imperial College ethics committee. Peripheral lymphoid organs were analysed by flow cytometry for DsRed or GFP, CXCR4, CD3, CD4 and CD8 expression. All FACS data was collected on a Fortessa flow cytometer (BD Biosciences, CA). Secondary recipients were sub-lethally irradiated (two doses of 3 Gy administered greater than three hours apart) and injected with 10,000 thawed, Ficoll purified T-ALL blasts and monitored as described earlier. In selected cases 10,000 secondary T-ALL cells were transplanted into tertiary recipients. For therapy experiments, mice were injected i.v. daily with 15 mg/kg dexamethasone sodium phosphate29, 30 (Sellekchem, MA) alone, 0.15 mg/kg vincristine sulfate salt (Sigma) alone or with a combination of 15 mg/kg dexamethasone, 0.15 mg/kg vincristine and 1,000 IU/kg l-asparaginase (medac; obtained from the Imperial College Healthcare NHS Trust Pharmacy). Reconstitution with MigR1 DsRed-transduced cells yields <50% chimaerism. For this reason, analysis of healthy BM cells by microscopy is inaccurate. Therefore, to obtain >95% chimaerism of healthy, red fluorescent BM to be used for imaging control experiments, whole BM mononuclear cells were isolated from femurs, hips and tibia of mTmG donor mice, suspended in phosphate balanced salt solution and administered intravenously to recipient mice at a dose of 2 × 106 cells/mouse. Recipient Col2.3–GFP or C57Bl/6 mice had been lethally irradiated (two doses of 5.5 Gy irradiation greater than 3 h apart) immediately before the transplant, and were maintained on baytril-treated water to prevent infection >6 weeks post-transplantation. Intravital microscopy was performed using a Leica SP5 and a Zeiss LSM 780 upright confocal microscope with a motorized stage. The SP5 was fitted with the following lasers: Argon, 546, 633 and a tunable infrared multiphoton laser (Spectraphysics Mai Tai 690-1020). The Zeiss LSM 780 was fitted with the following lasers: Argon, 561, 633 and a tunable infrared multiphoton laser (Spectraphysics Mai Tai DeepSee 690-1040). Signal was visualized with a Leica HCX IRAPO L ×25 water immersion lens (0.95 N.A) and a W Plan-Apochromat ×20 DIC water immersion lens (1.0 N.A). Collagen bone second harmonic generation signal and GFP and CFP signals were generated through excitation at 840 and 870 nm and detected with external detectors. Internal detectors were used to collect DsRed and Cy5 signal (and on some occasions, GFP). Prior to surgery, mice were administered analgesia with buprenorphine (0.1 mg/kg intraperitoneally (i.p.)). Anaesthesia was induced in mice with 4% isoflurane mixed with pure oxygen. This was gradually reduced to approximately 1% as anaesthesia stabilized. Surgery to attach the headpiece was then performed as described previously16. Large three-dimensional ‘tile scans’ of the entire BM cavity space were acquired by stitching adjacent, high-resolution z-stack images using a surgically implanted imaging window that ensures steady positioning of mice on the microscope. The calvarium has been demonstrated to be equivalent to the long bones such as the femur with regards to haematopoietic stem cell frequency, function and localization14, 22, and is the only BM compartment that allows longitudinal imaging through minimally invasive surgery16, 31. Blood vessels were highlighted by i.v. injection of 50 μl of 8 mg/ml 500 kDa Cy5-Dextran (Nanocs, MA). Cy5-Dextran was re-injected every 1–2 h to maintain blood vessel signal and cross reference for registration of blood vessel data in time-lapse analysis. For repeated imaging, protective intrasite gel (Smith & Nephew) was applied to the imaging window to preserve the bone integrity and prevent scar formation. The window was bandaged, and mice were allowed to recover from anaesthesia. Owing to the lock-and-key mechanism of the imaging window16, mice could then be re-anaesthetized and accurately repositioned on the microscope stage and the same BM areas re-imaged. After each imaging, analgesia was administered via oral buprenorphine in raspberry jelly at a dose of approximately 0.8 mg/kg. Microscopy data was processed using multiple platforms. Tile scans were stitched using Leica Application Systems (LAS; Leica Microsystems, Germany) and ZEN black (Zeiss, Germany) softwares. Raw data were visualized and processed using Fiji/Image J. Simulated data was prepared using FIJI macros to create, and overlay z-stack images on original tile-scan data. Using the internal random number algorithm, spheres matching the size of T-ALL cells (11–15 μm) were placed at random x,y,z coordinates. Simulated data FIJI macro is available on request. Automated cell segmentation, distance and volume measurements were performed in Definiens (Definiens Developer 64, Germany) using local heterogeneity segmentation32 to isolate osteoblast and nestin cells as well as vasculature, and a combination of seed detection algorithm and morphological growing and shrinking operations to detect leukaemia cells. Definiens rulesets for these functions are available upon request. Distance measurements from this segmentation were performed as described previously32. Cell tracking was performed using Imaris (Bitplane, Switzerland) and the FIJI plugin MTrackJ. For accuracy in cell tracking data, videos were registered when required before using four-dimensional data protocols implemented in Fiji33. Three-dimensional data rendering and measurement of cell division distances were performed in Volocity (Perkin Elmer, MA) and Definiens (Definiens Developer 64, Germany). T-ALL samples were harvested from bone marrow and FACS sorted based on fluorescent protein expression (DsRed or GFP) unless infiltration of bone marrow was complete. Control samples for microarray were prepared by FACS sorting for splenic CD4+ T cells, CD8+ T cells and CD4+CD8+ thymocytes from 8–14-week female C57Bl/6 mice. RNA was purified from samples using the Qiagen RNeasy mini kit (Netherlands) as per the manufacturer’s instructions. Purified RNA was prepared for hybridization using the Genechip WT Plus reagent kit (Affymetrix, CA) as per the manufacturer’s instructions and hybridized with genechip Mouse gene 2.0 ST array (Affymetrix, CA) by the MRC Genomics Facility (Imperial College London). Analysis was performed using R version 3.1.1. Data were normalized and summarized to ‘core’ level using the RMA method from the oligo package (version 1.30.0)34. Annotation was downloaded from the Affymetrix NetAffx Query website. Differential expression was determined using limma version 3.22.7 (ref. 35). Genes with Benjamini–Hochberg-adjusted P value < 0.05 and absolute log-fold-change >1 were deemed significant. Heatmaps of gene expression were generated with pheatmap package version 1.0.2. Primary human T-ALL samples were obtained from Barts Hospital (London) after informed consent via a protocol approved by the East London Research Ethics Committee and carried out in accordance with the principles of the Helsinki declaration (see Supplementary Table 2 for details), before treatment being administered to the patients. Primary cells from two distinct patients were immunophenotyped, and CD45+/CD7+/CD4−/low/CD8−/low cells sorted and infused i.v. in non-conditioned osterix–CreGFP/NOD/SCID/γ recipient mice. Primary xenograft transplantation was assessed via peripheral blood sampling and/or BM aspiration. BM and spleen-derived primary xenografts were infused i.v. in non-conditioned NOD/SCID/γ secondary recipient mice for therapy experiments. Intravital imaging was performed as described earlier. Human T-ALL cells were labelled by injecting 10 μg of PE-conjugated human CD45 antibody (clone HI30, Biolegend) 15–30 min before the imaging session. For dexamethasone therapy experiments, mice were treated with daily injections of 15 mg/kg i.v.30. Number of human T-ALL cells in therapy experiments was quantified using reference beads as described previously36. Hips and tibias were harvested and post-fixed overnight in periodate-lysine-paraformaldehyde fixative, at 4 °C. Bones were then washed with 0.1 M phosphate buffer, cryoprotected in sucrose (10–30% gradient), for 48 h, frozen in optimal cutting temperature compound (TissueTek) and stored at −80 °C. Sections were cut in a Leica Cryostat, using the Cryojane tape transfer system (Leica Microsystems) and stored at −80 °C. For staining, slides were re-hydrated in PBS, permeabilized in 0.1% Triton X-100, blocked in 5% goat serum and incubated with primary antibodies overnight, at 4 °C. After washing in PBS, slides were incubated with secondary antibodies, counter-stained with DAPI (Invitrogen), washed in 0.1% Triton X-100 and mounted using Prolong Diamond antifade (Invitrogen). The following antibodies were used: Alexa Fluor 647 mouse anti-Ki-67 (B56, BD Biosciences, 1:50), PE-conjugated human CD45 antibody (HI30, BD Biosciences, 1:100), rabbit anti-cleaved caspase-3 (Asp175, Cell Signaling, 1:100), goat anti-rabbit IgG Alexa Fluor 633 (Life Technologies, 1:400). TUNEL labelling was performed to detect apoptotic cells, according to the manufacturer’s instructions (DeadEnd Colorimetric TUNEL System, Promega). Images were obtained using a Zeiss LSM 780 upright confocal/two-photon combined microscope and analysed using Fiji/ImageJ. Cell counting was performed manually using the FIJI plugin Cell Counter. BM from human T-ALL xenotransplanted, untreated and treated mice was harvested and stained with DAPI (Invitrogen) and FITC mouse anti-Ki-67 set (BD Biosciences), according to the manufacturer’s instructions. Cells were analysed by flow cytometry and absolute numbers were obtained using reference beads as described previously36. T-ALL engraftment and infiltration was confirmed via peripheral blood sampling and/or tibia puncture. Once mice presented with signs of ill health (as described earlier), mice were euthanized and bones were digested with a DNase I/Collagenase (Sigma) solution. The total number of Osx–GFP+ cells was assessed by flow cytometry analysis using counting beads (CountBright, Life Technologies). Samples were obtained from patients after informed consent had been obtained, under full ethical approval by the Peter MacCallum Cancer Centre Human Research Ethics Committee. De-waxed human trephine biopsy sections (3 μM) were stained with osteocalcin antibody (Abcam ab93876, Cambridge), counterstained and mounted for viewing. All areas of each section were monitored for visible osteoblasts. The sample size required for the experiments was estimated based on the results of preliminary data. Blinding or randomization for animal experiments were not necessary due to the nature of the experiments. Statistical differences between the means of two data groups was determined by using two-tailed unpaired Student’s t-test, and P values < 0.05 were considered significant. Multiple group comparisons were performed using ANOVA with a Bonferroni correction, P values < 0.05 were considered significant.


News Article | December 15, 2016
Site: www.prweb.com

Who’s the ideal candidate for liposuction? It really depends on where you are getting your information from and what your expectations are for your results. Trevor Schmidt PA-C, the owner and liposuction expert at MyShape Lipo has a very liberal view on this subject, which he discusses in his live interview on the FOX 5 More Show. The American Society of Plastic Surgeons (ASPS) claims the best lipo candidates are “Adults within 30% of their ideal weight who have firm, elastic skin and good muscle tone.” By this definition, you should be near your ideal weight in order to get your fat removed through liposuction. Unfortunately, this eliminates nearly all the people that actually want liposuction to remove their unwanted fat. Schmidt believes that anyone who has excess fat that they want removed and has realistic expectations is a good candidate for liposuction. “I remove fat for a living, who am I to tell someone that they have too much fat for me to remove,” says Schmidt. “If they understand and have realistic expectations, then I think they are a good candidate for lipo.” According the the Center for Disease Control and Prevention, “2 out of 3 adults are considered overweight and 1 in 3 adults is considered obese.” So by the definition of the ideal candidate by ASPS, nearly 2/3 of the adult population who struggle with fat are eliminated as candidates. “It’s absurd to tell someone who struggles with their accumulation of fat, that they are not a good candidate to get their fat removed,” says Schmidt. “We have had amazing, life changing results with treating larger individuals. The more fat we can remove, the bigger the change the patient will experience.” Tina Gray was not an ideal candidate in that she was overweight and she is approaching her 60’s. Despite this, she was able to achieve very good results from liposuction. In a single procedure she was able to reduce her waist by 4 inches and eliminate the part of her belly that folds over. She also reduced her inner thighs by 2 inches each and eliminated the rubbing she experienced. “I’m ecstatic, my belly is nearly flat and I am able to fit into clothes that were previously too small,” says Tina. “I feel so much more confident, I wish I would have done this sooner.” At MyShape Lipo, most of their patients are not the “ideal candidate” set forth by ASPA. Instead they remove large volumes of fat to offer dramatic results for those that have bigger problems and have struggled with diet and exercise. Their patients are expecting to get significantly smaller. Most of the larger patients are expecting only to look better in their clothes as opposed to getting into a bathing suit or showing off their belly. Many already have irregularities or loose skin and are not expecting these issues to completely go away. These people are looking for improvement, not perfection and they are happy with a significant reduction is size. In the case of Tina, she had a substantial reduction in her belly. In fact the fold of loose skin and fat in the front of her belly has retracted and disappeared completely. She’s been able to fit into her clothes better and experienced improvements in her functionality. Hygiene is easier now that her belly doesn't fold over. “I’ve actually found that our larger patients tend to be much happier with their results,” says Schmidt. “Their improvement goes far beyond cosmetic results including functionality, mobility and confidence.” MyShape Lipo is a Liposuction and Fat Transfer specialty clinic located in Las Vegas, NV. Trevor Schmidt PA-C, the owner and liposuction specialist has performed over 15,000 liposuction procedures on all shapes, sizes and ages of individuals. He has the experience to get the most dramatic, smoothest and most consistent results for his patients. They offer free Body Shape Analysis, Call Now 702-818-5476. View their photo gallery online at http://www.MyShapeLipo.com.

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