News Article | March 1, 2017
No statistical method was used to predetermine the sample size. The experiments were not randomized, and the investigators were not blinded to allocation during experiments and outcome assessment. Highly active PSII was isolated from Thermosynechococcus vulcanus and crystallized as described previously with slight modifications4, 27, 28. The final PSII core dimers were suspended in 20 mM Mes (pH 6.0), 10 mM NaCl, 3 mM CaCl , and the final crystallization buffer contained 20 mM Mes, 20 mM NaCl, 40 mM MgSO , 10 mM CaCl , 5–7% polyethylene glycol 1,450, 0.85% n-heptyl-β-d-thioglucopyranoside (Dojindo). No re-crystallization procedure was applied. We screened various sizes of PSII crystals and post-crystallization treatment conditions (see below) to prepare suitable samples for TR-SFX by XFEL, and found that too small crystals did not diffract to a high resolution, whereas larger crystals gave rise to a lower efficiency of the S -state transition induced by the laser excitations. The optimal crystal size was determined to have a maximum length of 100 μm, which diffracted up to a resolution of 2.1 Å by a SACLA–XFEL pulse (Extended Data Fig. 1) and a final population of 46% S state upon 2F illumination (see below). All of the procedures for the preparation, crystallization, pre-flash illumination and diffraction experiments were conducted in the dark or in very dim green light. To prepare a large amount of micro-sized crystals, crystallization was performed in 1.5-ml micro-centrifuge tubes with a sample volume of 50 μl at a concentration of 2.3 mg chlorophyll per ml at 20 °C. The crystals appeared in a few hours; when the crystal size reached a maximum length of 100 μm, 50 μl of the crystallization buffer in which the concentration of PEG1,450 was increased by 1–2% from the crystallization condition was added to stop further growth of the crystals (see Extended Data Fig. 1a for a picture of typical crystals). The crystals were washed several times with this buffer to remove PSII samples that were not crystallized, and stored overnight. Prior to the XFEL experiments, the crystals were transferred to fresh mother liquid containing 10 mM potassium ferricyanide as an electron acceptor for the pre-flashing samples, and 2 mM potassium ferricyanide for the samples without pre-flash (non-pre-flash). The concentration of ‘cryo-protectant’ was then increased to 22% glycerol, 9% PEG1,450 and 9% PEG5,000 MME by stepwise replacement of the mother solution over 1.5 h. Finally, the crystals were carefully mixed with a grease matrix, Super Lube nuclear grade grease (Synco Chemical Co.), and loaded into a high viscosity micro-extrusion injector as described previously13. Although SFX can be performed at room temperature and there is no need to freeze the crystals, we found that this post-crystallization procedure to increase the cryo-protectant concentration was important for obtaining good diffracting PSII crystals. When the post-crystallization procedure was not adequate, some crystals gave rise to larger unit cell dimensions of a = 129.1 Å, b = 228.8 Å and c = 305.4 Å (Extended Data Fig. 6a–c). Diffraction spots from the crystals with this larger unit cell were found to be lower than 3.0 Å resolution, probably owing to the loose crystal packing. Indeed, we found that the PSII dimer in this crystal packing harboured two PsbY subunits (one PsbY per monomer PSII; Extended Data Fig. 6e). On the other hand, only one PsbY was found in the PSII dimer (one monomer contained PsbY whereas the other monomer did not) in the structure with unit cell dimensions of a = 126.5 Å, b = 231.2 Å and c = 287.5 Å analysed in this study (Extended Data Fig. 6f). When the two structures were superimposed, one of the two PsbY subunits in the dimer structure with the larger crystal packing interfered with the adjacent monomer in the structure with the smaller crystal packing (Extended Data Fig. 6d–f). To decrease the possible contamination of the S state in the dark-adapted crystals, PSII micro-crystals were illuminated with one pre-flash before the post-crystallization treatment. An aliquot of 100 μl solution containing micro-crystals of PSII was transferred into a dialysis button, and the pre-flash was provided by a Nd:YAG laser (Minilite-I, Continuum) at 532 nm with a diameter of 7 mm (large enough to cover the entire sample area) at an energy of ~52 mJ cm−2 at the sample position. After the pre-flash illumination, the sample was stored for 1–3 h in the dark while being transferred into the cryo-protectant conditions. This dark-incubation time was long enough to allow the higher S -states (S and S ) to decay into the S state, and therefore to minimize the possible contamination of the S state in the dark-adapted crystals, if there are any. The population of the S state after 2 flashes in the PSII micro-crystals was estimated using light-induced FTIR difference spectroscopy combined with the attenuated total reflection (ATR) method. PSII crystals in the same buffer as the SACLA experiments (including 10 mM potassium ferricyanide) were loaded onto a three-reflection silicon prism and then sealed with a transparent plate and a silicone rubber spacer. For the measurement of standard spectra, the PSII core solution was loaded onto the silicon prism, and the sample temperature was maintained at 20 °C. FTIR spectra were recorded at 4 cm−1 resolution using a Bruker IFS-66/S spectrophotometer equipped with an MCT detector. Flash illumination was provided by a Q-switched Nd:YAG laser (Quanta-Ray INDI-40-10; 532 nm, ~7 ns FWHM, ~21 mJ cm−2). Saturation of the laser flash was confirmed by checking the power dependence of the FTIR signal. After two pre-flashes with subsequent dark adaptation for 30 min, two flashes with an interval of 10 s were applied to the sample and FTIR difference spectra were measured upon each flash. We confirmed that the higher S-states decayed very little during this 10-s interval. This measurement was repeated 10 times to increase the signal-to-noise ratio of the spectra, with each measurement separated by 30 min dark incubation. The FTIR difference spectra obtained upon the 1st (a) and 2nd (b) flashes of PSII complexes in solution (black lines) and crystals (red lines) are shown in Extended Data Fig. 2. The spectra were normalized to the intensity of the amide II band, which reflects the protein amount. The efficiencies of the S-state transitions in the PSII crystals were estimated following the method previously described29, 30. The 1st- and 2nd-flash spectra in the PSII crystal, f (ν) and f (ν), respectively, were fitted with linear combinations of the 1st- and 2nd-flash spectra in solution, F (ν) and F (ν), respectively, as standard spectra: f (ν) = c F (ν); f (ν) = c F (ν) + c F (ν), where c , c , and c are the coefficients of linear combination. The least-squares fitting was performed in the symmetric COO− region (Extended Data Fig. 2b). The coefficients were estimated to be c = 0.73 ± 0.02, c = 0.27 ± 0.01, and c = 0.64 ± 0.02. The efficiencies of the S →S and S →S transitions are expressed as α c and α (c /c ), respectively, and the population of the S state after the second flash is calculated as α 2c . Here, α is the average efficiency of S-state transitions in solution and it was determined to be 0.85 ± 0.01 from the oscillation pattern of the intensity at 1,400 cm−1 (refs 31, 32) obtained by 12 consecutive flashes. Based on this, the population of the S state in the PSII micro-crystals after two flashes is estimated to be 0.46 ± 0.03. Single-shot XFEL data collection was performed using femtosecond X-ray pulses from the SACLA at BL3. The pulse parameters of SACLA were as follows: pulse duration, 2–10 fs; X-ray energy, 7 keV; energy bandwidth, 0.5% (FWHM); pulse flux, ~7 × 1010 photons per pulse; beam size 3.0 μm (H) × 3.0 μm (W); repetition rate, 30 Hz. The PSII crystals mixed with grease were loaded into an injector with a nozzle diameter of 150 μm, and set in a diffraction chamber filled with helium gas in a setup called Diverse Application Platform for Hard X-ray Diffraction in SACLA (DAPHNIS)33. The flow rate was set to 5.6 μl min−1 (5.28 mm s−1) for the 2F data and 2.8 μl m−1 (2.64 mm s−1) for the dark data. The diffraction patterns were recorded using a multiport CCD detector34. Because the excitation laser pulses were provided at 10 Hz and the XFEL pulses had a repetition rate of 30 Hz, each ‘pump-on’ image was followed by two ‘pump-off’ images, which were recorded separately. The pump-on images were used to analyse the 2F-state structure, whereas the diffraction data for the dark-state was collected by a separate run. To advance the PSII samples to the S -state, two consecutive excitation laser flashes were provided from two separate Nd:YAG laser sources (Minilite-I, Continuum) to the sample at t = 0 ms and t = 10 ms (that is, the two flashes were separated by 10 ms) for the pre-flashed samples, and t = 0 ms and t = 5 ms (separated by 5 ms) for the non-pre-flashed samples. To ensure sufficient excitation, each of the pump lasers was split into two beams, and each beam from the same pump laser (pump 1 or pump 2) was focused on the same sample point with an angle of 160° with respect to each other (a nearly counter-propagating geometry). The two beams from pump 1 were used for the first flash illumination, whereas the other two beams from pump 2 were used for the second flash illumination. The pump focal diameter of all beams was set to 240 μm (top-hat) and its energy was 42 mJ cm−2 from each direction. The XFEL pulses were provided 93 μm downstream from the pump focal centre at a time of 10 ms for the pre-flashed samples and 15 ms for the non-pre-flashed samples following the second excitation laser (the total time following the first flash is therefore 20 ms in both cases) (Fig. 1d, e). At the flow rate of 5.6 μl min−1, each pump-illuminated crystal for the pump-on XFEL images was separated by 528 μm, which was long enough to avoid influence from the previous excitation lasers, as the illumination point by the last laser extends to only 120 μm from its centre of illumination. The background of the detector was estimated by averaging dark images and subtracted from diffraction patterns. Diffraction images were filtered by the program Cheetah35, 36 adapted to SACLA, and processed by the program CrystFEL37. The parameters ‘min-snr’, ‘thresholds’ and ‘min-gradient’ used for peak detection during spot finding were as follows: 6, 500 and 10,000, respectively, for the pre-flashed dark data and pre-flashed 2F data; 5, 500 and 10,000 for the non-pre-flashed dark data; and 5, 500 and 5,000 for the non-pre-flashed 2F data. Indexing was performed using DirAx38 and Mosflm39 with peak integration parameters int-radius = 3, 5, 7, where the unit cell information was provided to avoid integration of the diffraction images from crystals with a longer c axis. Before Monte-Carlo integration, the indexed images with either a diffraction_resolution_limit lower than 4.2 Å or a num_peaks less than 400 were discarded. The numbers of total images collected, the hit images filtered by Cheetah, the indexed images and the number of images used for refinement for pre-flashed samples were as follows: 408,071, 76,047, 64,985 and 27,497, respectively, for the dark data; and 273,550, 60,885, 51,482 and 21,680 for the 2F data. The corresponding numbers for the non-pre-flashed samples are 462,343, 70,083, 54,956 and 22,341 for the dark data; and 876,874, 165,463, 63,711 and 23,903 for the 2F data (Extended Data Table 1). The Lorentz factor for still snapshots was applied manually to the averaged intensities of the pre-flashed dark and 2F data sets40. The statistics for the data collection are given in Extended Data Tables 1, 2, which show that our data for both dark and 2F states had a resolution of 2.35 Å for the pre-flashed samples and 2.5 Å for the non-pre-flashed samples, based on the cut-off value of around 50% for CC . We should point out that the overall multiplicities of all our data are very high, and even at the highest resolution shell, the multiplicity exceeded 500. Together with the facts that: (i) the values of CC in the highest resolution shells are reasonably high; (ii) CC decreases gradually without any abnormal disrupt from the low resolution to higher resolution shells; and (iii) the value of CC in the highest resolution shell is reasonably consistent with the value of I/σ(I) (theoretically 0.5 of CC corresponds to 2.0 of I/σ(I), and any substantial deviations from these values are indicators of systematic bias and/or problems in the error model41), we consider that the quality of the data in the present study is sufficiently high to allow us to reveal the small structural changes induced by the flash illuminations. The initial phases up to 4 Å resolution were obtained by molecular replacement with the program Phaser in the CCP4 suite42 using the 1.95 Å resolution XFEL structure of PSII (PDB accession code 4UB6; ref. 5) as the search model, in which the OEC and its direct ligands, Q , waters and glycerol molecules were omitted. After a few cycles of rigid body refinement and subsequent real space density modification using solvent flattening, histogram matching and non-crystallographic symmetry averaging with the program DM in the CCP4 suite42, the electron density map obtained showed features clear enough to allow us to build the model with confidence. Structural refinement was performed with Phenix43 and the model was manually modified with COOT44. After a few cycles of restrained refinement, water molecules were placed in positions corresponding to the water molecules in the higher-resolution structure where positive peaks higher than 3.5σ in the mF − DF map were clearly identified. Then, based on the resulting mF − DF map, water molecules and glycerol molecules were additionally located when positive peaks higher than 3σ were found. Q , the OEC and its direct ligand residues were modelled in the final step. In our previous XFEL structural analysis at 1.95 Å resolution5, the exact positions of the oxo-bridges were identified in the mF − DF map by omitting the oxo-bridged oxygen atoms; however, when the restrained refinement was performed in the same way in this study, the temperature factors of the manganese and/or calcium atoms in OEC were not converged, and gave rise to very high values owing to the limited resolution of 2.35–2.5 Å. Thus, we built the OEC structure using the geometric restraints based on the Mn CaO cluster in the 1.95 Å resolution XFEL structure, and applied tight distance restraints of σ = 0.02 Å to the Mn–O bonds, Ca–O bonds and Mn–ligand residue distances during the refinement. No restraints were given to the Mn–Mn distances and Mn–Ca distances. The R and R values obtained were 0.133 and 0.171, respectively, for the pre-flashed dark state, 0.139 and 0.186 for the non-pre-flashed dark state, and 0.139 and 0.187 for the non-pre-flashed 2F state (Extended Data Table 1). Given the population of the S state estimated from the FTIR measurement, the diffraction data obtained from the 2F illuminated sample is expected to contain those partly from the S and S states of PSII, which means that the resulted electron density would be a mixture of these S-states, including the S state. Even with a high-resolution data set, the structural refinement of the mixed states (or mixed structures) would be challenging, especially when the mixed structures are very similar, as in the case of PSII. As we did not have the structure corresponding to the S state, we refined the pre-flashed 2F data set as a mixture of the S state structure and the 2F state structure45. The occupancies for the S structure and the 2F state structure were set to 0.2 and 0.8, respectively, on the basis of the distributions of the temperature factors. The region in which the structure was refined in multiple states was selected on the basis of the large peaks observed in the isomorphous difference Fourier map, and the rest was refined as a single conformation. As a result, thirty-five amino acid residues, five water molecules, one non-haem iron, one Q and one OEC per monomeric PSII were modelled as multiple states. During the restrained refinement, the coordinate for the S state structure was fixed. Relatively loose geometric restraints for distance, angle and plane with relatively high sigma values (3–10 times larger than the default values) were applied to the OEC and Q during the refinement. Large positive peaks of 10.3σ for monomer A and 9.5σ for monomer B, found at a position with distances of 1.5 Å and 2.3 Å from O5 and Mn1D, respectively, were modelled as a new oxo-oxygen O6 with an occupancy of 0.4, which gave rise to a temperature factor similar to its nearby atoms. This is also consistent with the population of the S -state estimated from the FTIR results. The R and R values thus obtained were 0.129 and 0.176, respectively. The structure factors and atomic coordinates have been deposited in the Protein Data Bank (PDB) with accession numbers 5WS5 and 5WS6 for the pre-flashed dark-stable S state and 2F states, and 5GTH and 5GTI for the non-pre-flashed dark-stable S state and 2F state, respectively. All other data associated with this manuscript are available from the authors on reasonable request.
News Article | February 23, 2017
Like many living things, a cancer cell cannot survive without oxygen. When young and tiny, a malignancy nestles inside a bed of blood vessels that keep it fed. As the mass grows, however, its demand for oxygen outpaces supply. Pockets within the tumor become deprived and send emergency signals for new vessel growth, a process called angiogenesis. In the 1990s, a popular cancer-fighting theory proposed interfering with angiogenesis to starve tumors to death. One magazine writer in 2000 called the strategy “the most important single insight about cancer of the past 50 years.” It made such intuitive sense. Rakesh Jain viewed angiogenesis through a different lens. Trained as an engineer, not a biologist, Jain was studying tumor vasculature during the height of excitement about drugs that could impede vessel growth. He was bothered by the fact that capillaries that arise in the tumor aren’t normal; they’re gnarled and porous, incapable of effective blood flow in the same way a leaky pipe is lousy at delivering water. The expanding tumor squeezes smaller vessels, making them even less able to transport blood. “The mantra was, ‘Let’s starve tumors,’ ” recalls Jain, director of the Edwin L. Steele Laboratories for Tumor Biology at Harvard Medical School. “I said, ‘No, we need to do the opposite.’ ” In 2001, he published a commentary in Nature Medicine predicting that angiogenesis inhibitors would not entirely shrivel the tumor. Instead, he argued, starving tumors might make them harder to treat. “I was sticking my neck out and saying this is not a good thing to do,” he says. “I had tremendous resistance.” Time has proved him right. Once they came on the market, anti-angiogenesis drugs were not the boon doctors had hoped for. Most disturbing, some patients saw their tumors shrink, only to have the disease return with renewed vengeance. Today, more than a decade after the introduction of the first tumor-starving drug, researchers have a far greater understanding of the role of oxygen deprivation in cancer. Instead of slowing tumors, hypoxia appears to trigger a metabolic panic that can drive growth, drug resistance and metastasis. Rescue proteins called hypoxia-inducible factors, or HIFs, open a bag of tricks so tumors can adapt and outrun the body’s defenses. But there’s now reason for hope: Recent insights into the effects of oxygen deprivation in cancer are sparking new ideas and providing the blueprint for treatments that could short-circuit a cancer’s ability to survive and spread, and help make existing drugs more effective. While the idea of starving cancer made sense, the approach may have underestimated the strength and complexity of a tumor’s resilience. Since oxygen is essential for so much of life, nature equips cells with elaborate safeguards that kick in when the oxygen-rich blood supply dwindles — whether the cells are part of a tumor or part of a muscle straining for one last push of strength. When oxygen levels drop, newly minted proteins stampede throughout the cell, turning on a frenzy of chemical reactions that offer protection from the crisis. Cancer cells distort this natural coping mechanism for their own means. Growing new vessels is just one move in an elaborate strategy. Many changes accompany hypoxia, including: The malignant cells loosen from each other and become less adhesive, ready to break free; tendrils of collagen, a natural binding substance, form and start to reach out to nearby vessels; and proteins appear on the cell surface to pump out lactic acid, a product of the tumor’s switch from primarily aerobic to anaerobic respiration. Researchers now think stopping enough of these and other changes could cripple the cancer. Much of the research focuses on the proteins that are among the first to deploy when a cell senses a danger of asphyxiation. “At zero oxygen, the cell can’t survive,” says Daniele Gilkes of Johns Hopkins University School of Medicine. “Inside a tumor you will see these regions of necrosis,” or dead cells. But those cells that are low on oxygen but still alive will produce new proteins: Key among them are HIF-1 and HIF-2. Both are transcription factors — they help transcribe DNA instructions into RNA. Under normal conditions, the genes that make HIF proteins are mostly silent. Once HIF proteins are made, they turn on genes — Gilkes estimates there are hundreds — that enable cells to live when oxygen concentrations are low. Gilkes’ target of choice is HIF-1. It is not only a first responder, but the protein also appears to be key to cancer’s spread. Tumors with high levels of HIF-1, particularly when concentrated at the invasive outer edge of the mass, are more likely to become metastatic, invading other parts of the body. The reverse is also true: Human tumors transplanted into mice that genetically can’t produce HIF-1 are less likely to spread. The reasons are complicated, Gilkes says, but she considers one thing really interesting: HIF-1 is involved with a lot of enzymes in collagen formation. The collagen appears to provide a means of escape. Last year, in a review in the International Journal of Molecular Sciences, Gilkes described genes, found by her lab group and others, that breast tumors activate to degrade the surrounding environment. In turn, the tumor wraps itself in a stringy web of collagen. As the collagen forms, the strands stretch outward from the tumor and latch onto nearby vessels. “We think cancer cells will find this collagen and use it to migrate and glide.” She calls them “collagen highways.” Her laboratory captured video of human tumor cells migrating along a fibrous strand. “To see them move is really scary.” Once they’ve broken from their home tumor, many types of cancer, including prostate and breast cancers, commonly move into bones. This is no coincidence, Gilkes says. Bones lack the dense thickets of blood vessels that run through soft tissues. That means cancer cells migrating from a hypoxic environment, and therefore already trained for low oxygen, would find hospitable surroundings in the bone. Her lab group is now looking for ways to block collagen formation to close the travel lanes and perhaps keep the cancer from spreading. She and others are also working to find a way to inhibit HIF-1 directly, but so far those efforts have proved challenging. HIF-1’s accomplice, HIF-2, may be a more available target. HIF-2 is a molecule made of two parts that clamp onto DNA to trigger production of other proteins that make tumors tougher to kill. In 2009, structural biologists at University of Texas Southwestern Medical Center in Dallas discovered that the HIF-2 protein had a large cavity. “Usually proteins don’t have holes inside them,” says James Brugarolas, leader of UT Southwestern’s kidney cancer program. With the discovery, researchers began working on a way to use the gap as a foothold for drugs. Now in development, the experimental drug PT2399 slips inside HIF-2 and effectively breaks the molecule in two. Brugarolas and colleagues from six other institutions and the biotech company Peloton Therapeutics Inc. in Dallas published results of the first animal tests of the compound in Nature in November. In mice with implanted grafts of human kidney tumors, PT2399 split HIF-2 and slowed growth in 56 percent of tumors — better than a standard treatment. Brugarolas hypothesizes that the drug worked only about half the time because the other half of tumors relied more heavily on HIF-1. A similar HIF-2–busting drug is now in Phase I safety testing in humans, described in June in Chicago at the annual meeting of the American Society of Clinical Oncology. While Phase I studies are not designed to test whether the treatment works, the drug showed few side effects among 51 people with advanced kidney cancer who took the drug at ever-increasing doses. The patients had already been through multiple types of treatments, one as many as seven. After taking the drug, 16 patients experienced a slowing in disease progression, three more had a partial response and one a complete reversal. Given the dearth of treatments for advanced kidney cancer, Brugarolas says, “this is a big deal.” Still more molecules throw a lifeline to hypoxic tumors in ways that scientists are just beginning to understand. In 2008, pathologist David Cheresh and colleagues at the University of California, San Diego announced a curious discovery in Nature: Depriving cells of vascular endothelial growth factor, or VEGF — the key protein responsible for new vessel growth in a tumor and the main target of drugs that block angiogenesis — could actually make tumors more aggressive. His team went on to discover the same was true for another popular class of drugs, which work by depriving a tumor cell of nutrients in the same way anti-angiogenesis drugs limit oxygen. The drugs, called EGFR inhibitors, were capable of doing the opposite of what was expected: They could make tumors stronger. Cheresh believes that hypoxia — and other stresses of low blood supply, like nutrient deprivation — inflict a wound on the tumor. When normal tissues sustain an injury (like a cut), they immediately enter a period of healing and regeneration. The bleeding stops and the skin grows back. Low oxygen delivers a blow to tumor cells, sending them into a similar state of rejuvenation, he says. “They’re now prepared to survive not only the hypoxia, but everything else thrown at it.” In 2014, Cheresh published his take on why this occurs, at least in some cases, in Nature Cell Biology. He and his team described a molecule called avb3 found on the surface of drug-resistant tumors that appears to reprogram tumor cells into a stem cell–like state. As embers of the original tumor that are often impermeable to treatment, these stem cell–like cells can lie quietly for a time and then reignite. The discovery of avb3 has redefined how Cheresh thinks about resistance. He no longer believes that tumors defy chemotherapy in the way bacteria overcome antibiotics, with only the strongest cells surviving and then roaring back to become dominant. “The tumor cells are adapting, changing in real time,” Cheresh says. In short, his data suggest that when EGFR inhibitors deprive a cell of nutrients, some cells survive not because they are naturally tougher, but because the appearance of avb3 transforms them into drug-resistant stem cells. The good news is that laboratory tests suggest an experimental drug might block this reprogramming, and it may even prevent chemotherapy resistance. A clinical trial will soon begin that combines usual cancer treatment with this avb3-disabling drug, in a one-two punch aimed at reversing or delaying resistance so the treatment can do its job. There are still more ways tumors withstand low oxygen. They start eating leftovers. HIF-1 triggers a switch from oxygen-based aerobic respiration to anaerobic respiration using pyruvate, a product of glucose breaking down. The strategy works in the short term; it’s the reason your muscles keep pumping for a time, even when you’re gasping for air on the last few yards of a sprint. Problem is, anaerobic respiration leaves a trail of lactic acid. A lot of it. “Lactic acid buildup leads to a precipitous drop in pH inside of the tumor,” says Shoukat Dedhar of the BC Cancer Research Centre in Vancouver. To compensate, HIF-1 deploys a fleet of proteins that remove the acid so it won’t accumulate and burn up the cell. Dedhar’s laboratory didn’t start out studying hypoxia. “We had tumors that were readily metastatic and genetically related tumors that couldn’t metastasize,” he says. Those tumors that easily spread were producing HIF-1, along with products from other genes. Searching for the functions of those genes, his group and others found two proteins important in pH balance. The first, MCT-4, acts like a molecular sump pump, bailing out lactic acid. But it’s not enough to normalize the pH, Dedhar says. That job goes to the second protein, carbonic anhydrase 9, or CAIX. “Its job is simply to convert carbon dioxide to bicarbonate, which then neutralizes the acid,” he says. In March 2016, in a review in Frontiers in Cell and Developmental Biology, Dedhar and colleagues described how to improve cancer treatment by taking away some of the tools for hypoxia survival — that is, keeping the cell from neutralizing acid — while simultaneously giving drugs that boost the immune system. His team has developed new compounds that specifically block CAIX. Since CAIX is almost exclusively produced in tumor cells, CAIX inhibitors should theoretically have few side effects. A Phase I safety trial is testing possible drugs now. Harvard’s Jain is still making the case for bathing the tumors in oxygen, giving them more blood, not less. This could keep the tumor from becoming hypoxic and throwing up a new series of defenses, including a flood of angiogenesis-promoting proteins, which produce tormented circulation. When he proposed that concept in 2001, “I thought abnormal vessels were bad,” he says. “I now think they are worse.” His idea is to make tumor vasculature more normal, using the very drugs that he was concerned about almost two decades ago. His research suggests that giving anti-angiogenesis drugs in modest doses will keep the vessels from becoming abnormal, making them less tortured and more capable of normal blood flow (SN: 10/5/13, p. 20). He believes the restored oxygen not only shuts down the hypoxic response that gives the cancer a survival advantage, but also serves as a conduit for chemotherapy drugs and immune cells to penetrate deeper into the tumor. Oxygen is also necessary for radiation to work. His latest experiments take the concept of more oxygen, not less, even further. He combined two chemotherapy drugs with losartan, a generic medicine used to control blood pressure. The result, reported in Nature Communications in 2013, was a delay in pancreatic and breast tumor growth in mice. Another experiment from Jain and colleagues, published in 2016 in Translational Oncology, had similar results. “We are finding every therapy works better when we do this,” he says. A clinical trial is now under way at Massachusetts General Hospital testing whether giving losartan during radiation and chemotherapy will improve results for pancreatic cancer patients. The concept still remains unproven, but Jain has reason for optimism. And he is no longer in the scientific minority. Last May, he received the National Medal of Science from President Barack Obama, who commended Jain for “groundbreaking discoveries of principles leading to the development and novel use of drugs for treatment of cancer.” Jain hopes to see the day, not long in the future, when hypoxic tumors are defeated by giving them the very thing they need the most. This article appears in the March 4, 2017, issue of Science News with the headline, "Deflating cancer: New approaches to low oxygen may thwart tumors."
News Article | February 22, 2017
MALVERN, Pa., Feb. 22, 2017 (GLOBE NEWSWIRE) -- BioTelemetry, Inc. (NASDAQ:BEAT), the leading wireless medical technology company focused on the delivery of health information to improve quality of life and reduce cost of care, today reported results for the fourth quarter and full year ended December 31, 2016. Joseph H. Capper, President and Chief Executive Officer of BioTelemetry, Inc., commented: “2016 was an outstanding year for BioTelemetry. As a result of the effective execution of our strategic initiatives, we recorded record revenue, earnings and adjusted EBITDA, achieving the high-end of our fourth quarter guidance. Additionally, we continued our tradition of innovation and completed three acquisitions, gaining entree into an exciting, new and large market opportunity. Moreover, we serviced approximately 600,000 patients in 2016 alone, representing an almost 8% increase versus the prior year, with our mobile cardiac telemetry (“MCT”) volume experiencing double-digit growth. We also received a positive coverage decision from Anthem, the largest health insurer in the nation, for use of MCT for certain patients. Finally, we obtained FDA clearance on our next generation MCT device, setting the stage for sustained volume growth well into the future. “Looking forward, we are excited about our expansion into digital population health management (“PHM”) through our acquisition of Telcare. We believe PHM represents a large market opportunity, and we are uniquely positioned to assume a leadership position. On the cost side, we made great progress in gaining efficiencies in 2016 and, we will continue to look for additional opportunities. We enter 2017 confident it will be another year of strong financial results and operational successes.” Revenue for the fourth quarter 2016 was $54.0 million compared to $46.8 million for the fourth quarter 2015, reflecting an increase of $7.2 million, or 15.4%. Healthcare revenue increased $3.0 million due to increased patient volumes and higher patient pricing due to a favorable product mix as well as higher MCT Medicare pricing. Research revenue increased $3.4 million due to the acquisition of VirtualScopics during the second quarter. Technology revenue increased $0.9 million due to Telcare as well as sales of our ePatch Holter. Gross profit for the fourth quarter 2016 increased to $33.0 million, or 61.2% of revenue, compared to $28.3 million, or 60.4% of revenue, for the fourth quarter 2015. The increase in gross margin percentage was due to the impact of higher Healthcare pricing, volume efficiencies and cost reductions partially offset by the impact of the acquisitions, which carry lower margins than our existing business. On a GAAP basis, operating expense for the fourth quarter 2016 was $29.6 million, compared to $24.8 million for the fourth quarter 2015. On an adjusted basis1, operating expense for the fourth quarter 2016 was $25.5 million compared to $23.2 million for the fourth quarter 2015, an increase of $2.3 million, or 10.1%. The adjusted operating expense for the fourth quarter 2016 excludes $2.8 million of other charges primarily related to patent litigation and the integration of the current year acquisitions and $1.3 million for a one-time performance bonus paid to a third party in the form of stock-based compensation. The adjusted operating expense for the fourth quarter 2015 excludes $1.6 million primarily related to patent litigation. The increase in adjusted expense was driven by the addition of $2.3 million related to our acquired companies, a $0.8 million increase in bad debt expense and $0.1 million of additional consulting expense related to ongoing product development. These increases were partially offset by a $0.9 million reduction in headcount related expenses. Interest and other loss, net was $0.6 million for the fourth quarter 2016 compared to $0.4 million for the fourth quarter 2015. The increase was due to losses related to the Company’s equity method investment in Wellbridge Health and increased borrowings under the revolving credit facility. During the fourth quarter 2016, the Company released a tax valuation allowance on its net deferred tax assets. Over time, the Company had recorded deferred tax assets, but, due to the Company’s history of losses, management established a valuation allowance to offset these assets. Management has now determined that there is sufficient evidence to conclude that it is more-likely-than-not that the Company will realize these benefits, and, as a result, the Company reduced its valuation allowance accordingly. This reduction resulted in a one-time income tax benefit in the fourth quarter 2016 Consolidated Statement of Operations in the amount of $37.6 million. Without a valuation allowance in place, for GAAP financial reporting purposes, the Company expects its 2017 tax rate to be in the range of 38% to 39%. However, due to the utilization of net operating loss carryforwards, the Company expects 2017 actual cash tax payments to remain in the 3% to 5% range. On a GAAP basis, net income for the fourth quarter 2016 was $40.4 million, or $1.30 per diluted share, compared to net income of $2.8 million, or $0.10 per diluted share, for the fourth quarter 2015. On an adjusted basis1, net income for the fourth quarter 2016 was $7.0 million, or $0.23 per diluted share. This compares to adjusted net income of $4.4 million, or $0.15 per diluted share, for the fourth quarter 2015. Adjusted net income for the fourth quarter 2016 excludes the $37.6 million income tax benefit related to the valuation allowance release, $2.8 million of other charges primarily related to patent litigation and the integration of the current year acquisitions and $1.3 million for a one-time performance bonus paid to a third party in the form of stock-based compensation. Adjusted net income for the fourth quarter 2015 excludes $1.6 million primarily related to patent litigation. As of December 31, 2016, total cash was $23.1 million compared to $19.0 million as of December 31, 2015, a $4.1 million increase. During 2016, the Company used $25.0 million for acquisitions and $10.9 million for capital expenditures, primarily medical devices while generating $38.9 million of cash from operations. In addition, the Company borrowed $14.5 million from our revolving credit facility during the second quarter 2016, repaying $11.5 million of this balance during the fourth quarter 2016. Consolidated days sales outstanding decreased to 45 days as of December 31, 2016, down from 47 days as of December 31, 2015. As of December 31, 2016, the Company had net indebtedness of $2.3 million, comprised of indebtedness of $25.4 million and cash of $23.1 million. 1 The Company believes that providing non-GAAP financial measures offers a meaningful representation of the Company’s performance as they exclude expenses that are not necessary to support the Company’s ongoing business. Please refer to the Company’s “Reconciliation of Non-GAAP Financial Measures” and “Use of Non-GAAP Financial Measures” in this release for additional information. Conference Call BioTelemetry, Inc. will host an earnings conference call on Wednesday, February 22, 2017 at 5:00 PM Eastern Time. The call will be simultaneously webcast on the investor information page of our website, www.gobio.com. The call will be archived on our website for two weeks. About BioTelemetry BioTelemetry, Inc., formerly known as CardioNet, Inc., is the leading wireless medical technology company focused on the delivery of health information to improve quality of life and reduce cost of care. The Company currently provides cardiac monitoring services, original equipment manufacturing with a primary focus on cardiac monitoring devices and centralized cardiac core laboratory services. More information can be found at www.gobio.com. Cautionary Statement Regarding Forward-Looking Statements This document includes certain forward-looking statements within the meaning of the “Safe Harbor” provisions of the Private Securities Litigation Reform Act of 1995. These statements may be identified by words such as “expect,” “anticipate,” “estimate,” “intend,” “plan,” “believe,” “promises” and other words and terms of similar meaning. Such forward-looking statements are based on current expectations and involve inherent risks and uncertainties, including important factors that could delay, divert, or change any of these expectations, and could cause actual outcomes and results to differ materially from current expectations. These factors include, among other things, our ability to successfully integrate acquisitions into our business and the effect such acquisitions will have on our results of operation, effectiveness of our cost savings initiatives, relationships with our government and commercial payors, changes to insurance coverage and reimbursement levels for our products, the success of our sales and marketing initiatives, our ability to attract and retain talented executive management and sales personnel, our ability to identify acquisition candidates, acquire them on attractive terms and integrate their operations into our business, the commercialization of new products, market factors, internal research and development initiatives, partnered research and development initiatives, competitive product development, changes in governmental regulations and legislation, the continued consolidation of payors, acceptance of our new products and services, patent protection, adverse regulatory action, and litigation success. For further details and a discussion of these and other risks and uncertainties, please see our public filings with the Securities and Exchange Commission, including our latest periodic reports on Form 10-K and 10-Q. We undertake no obligation to publicly update any forward-looking statement, whether as a result of new information, future events, or otherwise. Reconciliation of Non-GAAP Financial Measures (In Thousands, Except Per Share Amounts) (a) In the fourth quarter 2016, the Company incurred $2.8 million of other charges primarily due to patent litigation and the acquisitions completed in the current year. In the fourth quarter 2015, the Company incurred $1.6 million of other charges primarily related to patent litigation. (b) In the fourth quarter 2016, the Company incurred $1.3 million for a one-time performance bonus paid to a third party in the form of stock-based compensation. This is a nonrecurring expense for the Company and is the only time in the Company’s history when such a bonus was awarded. There are no additional agreements outstanding of this nature. (c) During the fourth quarter 2016, the Company released a tax valuation allowance on its net deferred tax assets. This release resulted in a one-time income tax benefit in the fourth quarter 2016 Consolidated Statement of Operations in the amount of $37.6 million. This benefit is nonrecurring and not indicative of the Company’s ongoing results or future tax position. Reconciliation of Non-GAAP Financial Measures (In Thousands, Except Per Share Amounts) (a) In 2016, the Company incurred $8.6 million other charges primarily due to patent litigation and the acquisitions completed in the current year. In 2015, the Company incurred $6.1 million of other charges primarily due to patent litigation and other legal fees. (b) In 2016, the Company incurred $1.3 million for a one-time performance bonus paid to a third party in the form of stock-based compensation. This is a nonrecurring expense for the Company and is the only time in the Company’s history when such a bonus was awarded. There are no additional agreements outstanding of this nature. (c) During 2016, the Company released a tax valuation allowance on its net deferred tax assets. This reduction resulted in a one-time income tax benefit in the 2016 Consolidated Statement of Operations in the amount of $37.6 million. This benefit is nonrecurring and not indicative of the Company’s ongoing results or future tax position. Use of Non-GAAP Financial Measures In addition to the results prepared in accordance with generally accepted accounting principles in the United States, or GAAP, this press release also includes certain financial measures which have been adjusted and are not in accordance with generally accepted accounting principles (“Non-GAAP financial measures”). These Non-GAAP financial measures include adjusted income from operations, adjusted net income, adjusted net income per diluted share and adjusted EBITDA. In accordance with Regulation G of the Securities and Exchange Commission, the Company has provided a reconciliation of these Non-GAAP financial measures with the most directly comparable financial measure calculated in accordance with GAAP. These Non-GAAP financial measures are not intended to replace GAAP financial measures. They are presented as supplemental measures of our performance in an effort to provide our stakeholders better visibility into the Company’s ongoing operating results and to allow for comparability to prior periods as well as to other companies’ results. Management uses these Non-GAAP financial measures to assess the financial health of the Company’s ongoing operating performance. Management encourages our stakeholders to consider all of our financial measures and to not rely on any single financial measure to evaluate our performance. Adjusted income from operations, adjusted net income and adjusted net income per diluted share exclude Other charges, a one-time performance bonus paid to a third party in the form of stock-based compensation and the release of the Company’s valuation allowance. By excluding expenses that are considered not necessary to support the ongoing business or which are nonrecurring in nature, the Company believes that these Non-GAAP financial measures offer a meaningful representation of the Company’s ongoing operating performance. Patent litigation expense is a component of Other charges. We view patent litigation as an extreme measure not typically required in our industry to protect a company’s intellectual property and which has not been common practice for the Company. The Company commenced patent litigation proceedings after the Company uncovered specific evidence of four distinct cases of misappropriation and infringement. The Company can choose to resolve the outstanding matters and terminate the expense at any time. Also included in Other charges are transaction related expenses which include integration costs, primarily professional fees and severance, which are not part of the ongoing operations, and therefore, not reflective of the Company’s core operations. The Company also excluded a one-time performance bonus paid to a third party in the form of stock-based compensation. This is the first time in the Company’s history that such a bonus was offered and issued and the expense is nonrecurring. There are no additional agreements outstanding of this nature. Finally, the Company excluded the nonrecurring benefit from the release of its tax valuation allowance from its non-GAAP financial measures. In addition to adjusted income from operations, adjusted net income and adjusted net income per diluted share, we also present adjusted EBITDA. This Non-GAAP financial measure excludes income taxes, interest, Other charges, other one-time items such as the tax valuation allowance release, depreciation and amortization and stock compensation expense. EBITDA is a widely accepted financial measure which we believe our stakeholders use to compare our ongoing financial performance to that of other companies. Adjusting our EBITDA for Other charges and other one-time items is a meaningful financial measure as we believe it is an indication of our ongoing operations. In addition, we also add back stock compensation expense because it is non-cash in nature. Other companies in our industry may calculate adjusted EBITDA in a different manner.
News Article | February 23, 2017
David Nolte, PhD accepted Purdue University’s 2016 Outstanding Commercialization Award during the 12th annual Inventors Recognition Reception at Purdue Research Park of West Lafayette, Indiana. The top commercialization award is given annually to a faculty member in recognition of outstanding contributions to, and success with, commercializing discoveries from Purdue research. “This award is truly an honor. Purdue’s outstanding model for commercialization of intellectual property should serve as an inspiration for other universities around the world,” Nolte said. “I’m humbled to accept this as just one member of the greater Purdue community that includes a host of the world’s top minds doing an enormous amount of world-changing research.” Nolte holds a total of 14 patents and co-founded the company Animated Dynamics Inc. (ADI). With Nolte’s expertise, ADI has patented a bio-dynamic imaging technology called Motility Contrast Tomography (MCT) to improve targeting of chemotherapy treatments. MCT technology provides the ability to evaluate response (or non-response) of a cancer patient’s tumor to various chemotherapy agents before prescribing a cancer-treating drug. In preparation for commercialization of ADI’s Onco 4D™ test, study sites are currently being recruited for clinical trials targeting breast and bladder cancers. The trials will enroll certain breast biopsy patients, and bladder cancer patients undergoing a trans-urethral resection of bladder tumor (TURBT) procedure. The information gathered during the study will validate the Onco4D™ predictive algorithms for these clinical settings, and contribute to improved cancer care for millions of breast and bladder cancer patients. “It is my pleasure to be associated with Dr. Nolte and the other ADI founders, Drs. John Turek and Ran An. Their innovation and on-going research will provide a profound benefit in terms of physicians’ ability to achieve improved outcomes for their cancer patients,” said Ted Schenberg, CEO of Animated Dynamics. About Animated Dynamics, Inc. Animated Dynamics invented Motility Contrast Tomography (MCT), an advanced form of digital holography which measures kinetic activity at the nano-scale. MCT provides an unprecedented view into the physical processes at work within living tissue, allowing researchers and clinicians to quantify the phenomic signature of tissue in three dimensions and measure response to stimuli over time (i.e. four dimensions). The company’s Onco 4D™ test for phenomic chemotherapy selection is expected to become clinically available during 2017. More information is available at http://www.anidyn.com. Contact: media(at)anidyn(dot)com.
News Article | February 28, 2017
ALEXANDRIA, Va., Feb. 28, 2017 (GLOBE NEWSWIRE) -- As the global leader in Remote Weapon Station (RWS) development, manufacture, and support, KONGSBERG recently unveiled their latest system developments at the Orbital ATK Bushmaster User Conference. The Bushmaster demonstration took place on 1 February, and showcased the PROTECTOR LW30, with a M230 LF cannon and the PROTECTOR Medium Caliber (remote) Turret (MCT) with both 30mm and Super 40mm cannons. The centerpiece of the event was the demonstration of the PROTECTOR MCT's flexibility to “upgun” from a standard 30mm configuration, to a 40mm cannon with no changes to the turret structure. The demonstration included live fire with both 30mm and 40mm high explosives and programmable airburst munitions, giving the audience proof of the increased effects on a variety of targets. “Our ability to quickly change from the standard 30mm configuration to Super 40, in less than 60 minutes, in an austere environment, says volumes about the flexibility of the system and the effectiveness of its design. The demonstration not only shows how the MCT-30 system is currently being delivered to the Stryker Program, but also how it can support future needs for increased capabilities and lethality across a variety of formations,” said Pam Willgohs, Executive Vice President for Medium Caliber Turrets. In addition, KONGSBERG demonstrated its PROTECTOR LW30, firing in moving and static scenarios. The PROTECTOR LW30 was able to demonstrate full station functionality and mobility, firing without physical restraints on the system and successfully engaging targets out to ~1,000 meters. “This is our second demonstration of the system for an international audience. The fact that we are able operate the system fully, without physical safety restraints, speaks to the maturity of the system and the control software,” said Alf Borstad, Executive Vice President for RWS USA. He went on to add, “In this case, we successfully demonstrated the ability to leverage the U.S. Common Remotely Operated Weapon System (CROWS) family of systems to support new weapons development. Our U.S. Army customer challenged us to design a system that leverages the U.S government (USG) (owned) Technical Data Package (TDP) and the expansive CROWS supply chain (and industrial base). We feel we have done that, but also delivered on the performance and capability expectations of the end customer. This successful demonstration proves the ability for the USG to continue to realize the fruit of its substantial investment in the CROWS infrastructure. This will be specifically advantageous for qualification of the system, but also aid in continued growth for our depot partners already sustaining CROWS.” About KONGSBERG Protech Systems (KPS): KPS is the world's leading supplier of Remote Weapon Stations, providing flexible solutions that meet our customers' specific requirements. Through world class innovation, program execution and customer understanding, we provide high tech systems for enhanced situational awareness and protection of personnel and property in high-risk areas. KPS proudly manufactures and supports all U.S. customer products from its Johnstown, PA facility. KONGSBERG (OSE-ticker: KOG): KOG is an international, knowledge-based group delivering high technology systems and solutions to clients within the oil and gas industry, merchant marine, defense and aerospace. KONGSBERG has 7,700 employees located in more than 25 countries and total revenues of NOK 17.0 billion in 2015.
News Article | February 1, 2016
This could be the big year when Elon Musk will disclose more details of the anticipated SpaceX project Mars Colonial Transporter (MCT). The MCT can take about 100 people to Mars via a system that consists of reusable rockets, launch vehicles and space capsules that can safely return to Earth. The news comes from when Musk answered some questions regarding his and the company's plans at the StartmeupHK Festival in Hong Kong recently. "I'm hoping to describe that architecture later this year at IAC ... and I think that will be quite exciting," Musk said (video). IAC is the International Astronautical Congress, and it will take place on Sept. 26-30, 2016 in Guadalajara, Mexico. The expected topics that are going to be discussed there range from super rockets to spacecraft that can accommodate a considerable number of occupants - the MCT. Musk first announced the MCT in 2015 during a Reddit Ask Me Anything session, saying he will unveil the plans for the project later in that year. "The Mars transport system will be a completely new architecture. [I'm] hoping to present that towards the end of this year. Good thing we didn't do it sooner, as we have learned a huge amount from Falcon and Dragon," he said. However, after some time, he announced that the reveal has been moved to 2016 instead. The MCT makes use of two vital components: one booster nicknamed BFR (for "Big F'n Rocket") that can carry 100 tons worth of cargo and a habitat-slash-landing module that can land on the surface of Mars and make the trip back to Earth in one piece after it's refueled at the red planet. It's worth mentioning that Musk has been vocal about his plans to colonize Mars, and this effort is a big step toward that ambition. In December, engineer and "tinker" John Gardi published a sketch of how the MCT could look like. Just to be clear, Gardi does not work for SpaceX, and he has made some spot-on predictions of certain designs before. Watch the video below to see Musk at the 2016 StartmeupHK Venture Forum.
News Article | November 30, 2016
Reductions observed in frequency and total duration of major medical events NOVATO, Calif., Nov. 30, 2016 (GLOBE NEWSWIRE) -- Ultragenyx Pharmaceutical Inc. (NASDAQ:RARE), a biopharmaceutical company focused on the development of novel products for rare and ultra-rare diseases, today announced positive 78-week data from the Phase 2 study of UX007 (triheptanoin) in patients with long-chain fatty acid oxidation disorder (LC-FAOD). The frequency and duration of major medical events were reduced significantly during treatment with UX007, and patients demonstrated improved exercise tolerance and quality of life during the study. “The reduction in frequency and duration of hospitalizations and other major medical events suggests a clinically meaningful improvement for patients and is consistent with the data from the prior retrospective compassionate use study,” said Emil D. Kakkis, M.D., Ph.D., Chief Executive Officer and President of Ultragenyx. “These results are encouraging and we continue to further develop the Phase 3 study design and endpoints before meeting with regulators and initiating the study in 2017.” The major clinical event (MCE) rate aggregates events from the 29-patient open-label Phase 2 study related to hypoglycemia, cardiomyopathy and rhabdomyolysis. For this study, events that qualified included those that led to a hospitalization, emergency room visit, or an emergency intervention at home. There was a 48.1 percent reduction (p=0.0208) in the mean annualized rate of MCEs and a 50.3 percent reduction (p=0.0284) in the mean annualized duration of all MCEs after 78 weeks of treatment, compared to the mean annualized number and duration of events in the 18 to 24 months prior to treatment with UX007. Among the event subtypes, rhabdomyolysis was the predominant MCE and there were fewer hypoglycemia events and only a few cardiomyopathy events. There was a reduction in the mean annualized rates and total duration of all events for rhabdomyolysis, cardiomyopathy, and hypoglycemia events after initiation of treatment with UX007. These findings were generally comparable to those observed in the retrospective compassionate use study previously conducted by Ultragenyx with partial reduction in rhabdomyolysis and near complete reduction of hypoglycemia events. Eight patients performed the twelve minute walk test (12MWT) at baseline and at Week 60, and showed a 29.7 percent improvement (mean increase: +199.8 meters; median: 149.5; min,max: -122, 1000) from a baseline of 673.4 meters. These results demonstrate that the increase in distance walked observed at 18 weeks was maintained through 60 weeks. The cycle ergometry test at 78 weeks was inconclusive and uninterpretable due to missing data as a result of scheduling conflicts, withdrawal of consent, intercurrent illness and other factors. Health Related Quality of life was assessed in the Phase 2 study using age appropriate SF-10™ (5-17 years, n=3) and SF-12v2® (>18 years, n=5) Health Surveys. Significant improvements in the impaired physical summary measures (SF-12v2 Physical Component Summary and SF-10 Physical Health Summary) reported at the start of the study were observed after 78 weeks of treatment with UX007. Five of the 29 enrolled patients discontinued treatment over the course of the study. One patient discontinued due to diarrhea in week 1, which resolved within a few days of discontinuation, and four patients withdrew consent (weeks 1, 8, 8, 78) for reasons not attributed to treatment with UX007. There were no deaths. There was one previously reported treatment-related serious adverse event for moderate gastroenteritis with vomiting. This patient completed the study and maintained dosing throughout the event, which has now resolved. Overall, 19 patients (66%) had treatment-related adverse events, most of which were mild-to-moderate in nature. The most common treatment-related adverse events were diarrhea, abdominal/gastrointestinal pain, and vomiting. Some gastrointestinal events were managed by adjusting dosing or dosing with food. The most common adverse events, including those not deemed treatment-related, were diarrhea, rhabdomyolisis, vomiting, viral infections, gastrointestinal disorders, headache and fever. The single-arm, open-label Phase 2 study evaluated 29 pediatric and adult patients across three main symptom groups (musculoskeletal, liver/hypoglycemia, and cardiac). Patients needed to have moderate to severe FAOD with significant disease in at least one of these domains or a frequent medical events history in order to enroll. The study began with a four-week run-in period to assess baseline data while on the standard of care therapy including medium-chain triglyceride (MCT) oil, if applicable. Twenty-five patients completed 24 weeks of treatment, at which point the acute effects of UX007 treatment on the muscular aspects of the disease were evaluated. Twenty-four patients opted to continue to be treated for a total of 78 weeks with one who withdrew consent late during this period on turning 18 years old. Safety and quality of life assessments were also measured throughout the study. These Phase 2 results are based on open-label uncontrolled treatment referenced to baseline run-in period using careful medical chart reviews for each patient, but the lack of a randomized parallel control group does limit definitive conclusions about efficacy and safety. The majority of patients enrolled presented with musculoskeletal disease compared to a limited number who presented with liver and cardiac symptoms. Patients spanned a wide age range from ten months to 58 years old. Patients with four LC-FAOD genotypes were enrolled: twelve (41%) with VLCAD, ten (35%) with LCHAD, four (14%) with CPT-II, and three (10%) with TFP. Prior to initiating treatment with UX007, 27 of the 29 patients were on the standard of care MCT oil therapy. UX007 was then titrated to a target dose of 25-35% of total daily caloric intake. Patients performed only the assessments that were appropriate and valid for their age when they entered the study; therefore not all patients performed all assessments included in the study design. Due to the small number of patients who completed a number of the assessments in this Phase 2 study, the results would need to be confirmed in a larger controlled study. LC-FAOD are a group of autosomal recessive genetic disorders characterized by metabolic deficiencies in which the body is unable to convert long-chain fatty acids into energy. The inability to produce energy from fat can lead to severe depletion of glucose in the body, and serious liver, muscle, and heart disease, which can lead to hospitalizations or early death. LC-FAOD are included in newborn screening panels across the U.S. and in certain European countries. LC-FAOD patients are currently treated with the avoidance of fasting, low-fat/high carbohydrate diets, carnitine, and medium-chain triglyceride (MCT) oil, a medical food product. Despite current therapy, many patients have significant metabolic events including hospitalizations and mortality due to LC-FAOD. UX007 is a purified, pharmaceutical-grade form of triheptanoin, a specially designed synthetic triglyceride compound, created via a multi-step chemical process. It is an investigational medicine intended to provide patients with medium-length, odd-chain fatty acids that can be metabolized to increase intermediate substrates in the Krebs cycle, a key energy-generating process. Unlike typical even-chain fatty acids, UX007 can be converted to new glucose through the Krebs cycle, potentially providing an important added therapeutic effect, particularly when glucose levels are too low. Ultragenyx is a clinical-stage biopharmaceutical company committed to bringing to market novel products for the treatment of rare and ultra-rare diseases, with a focus on serious, debilitating genetic diseases. Founded in 2010, the company has rapidly built a diverse portfolio of product candidates with the potential to address diseases for which the unmet medical need is high, the biology for treatment is clear, and for which there are no approved therapies. The company is led by a management team experienced in the development and commercialization of rare disease therapeutics. Ultragenyx's strategy is predicated upon time and cost-efficient drug development, with the goal of delivering safe and effective therapies to patients with the utmost urgency. For more information on Ultragenyx, please visit the company's website at www.ultragenyx.com. Except for the historical information contained herein, the matters set forth in this press release, including statements regarding expected timing of release of additional data, and expected timing of discussion of Phase 3 design with regulatory authorities and initiation of a Phase 3 study are forward-looking statements within the meaning of the "safe harbor" provisions of the Private Securities Litigation Reform Act of 1995. Such forward-looking statements involve substantial risks and uncertainties that could cause our clinical development programs, future results, performance or achievements to differ significantly from those expressed or implied by the forward-looking statements. Such risks and uncertainties include, among others, the uncertainties inherent in the clinical drug development process, including the regulatory approval process, the timing of our regulatory filings and other matters that could affect sufficiency of existing cash, cash equivalents and short-term investments to fund operations and the availability or commercial potential of our drug candidates. Ultragenyx undertakes no obligation to update or revise any forward-looking statements. For a further description of the risks and uncertainties that could cause actual results to differ from those expressed in these forward-looking statements, as well as risks relating to the business of the company in general, see Ultragenyx's Quarterly Report on Form 10-Q filed with the Securities and Exchange Commission on November 8, 2016, and its subsequent periodic reports filed with the Securities and Exchange Commission.
News Article | February 15, 2017
VisualSP has invited Microsoft's Bill Baer to explain the benefits of SharePoint Server 2016, the investments Microsoft has made to improve SharePoint, and why a migration might make sense for an organization. The webinar, titled SharePoint Server 2016: What's in it for Me? will be held February 22, 2017, 12:00 – 1:00pm EST. During this extensive introduction to SharePoint Server 2016, attendees will learn about the improvements Microsoft has made in: Bill will discuss how Microsoft brought innovation from SharePoint Online experiences to the 2016 on-premises server environment. He also discusses the introduction of MinRole to server farm topology, which enables a role-based installation. Baer is also expected to talk about patching without downtime. During the presentation, he'll provide a walkthrough of the hybrid SharePoint experience. He is a Senior Technical Product Manager and Microsoft Certified Master for SharePoint in the SharePoint product group at Microsoft. He was previously a Hewlett-Packard Technology Solutions Group MVP, with a background in infrastructure engineering and enterprise deployments of SharePoint Products and Technologies. VisualSP is inviting all interested parties to this informative live event to find out "What's in it for me?". The company develops contextual, on-demand SharePoint training and help system for on-premises versions, starting with SharePoint 2007. Recently, VisualSP released its cloud training solution for Office 365, which provides guidance for all apps in the suite, including SharePoint Online. To learn more about the Training for Office 365 product, visit visualsp.com. Webinar: SharePoint Server 216: What's in it for Me? Date: 2/22/17 Time: 12:00 – 1:00 PM EST Register: Save your seat ***** VisualSP® has been a leader in e-learning/performance support integration with SharePoint since 2004. The company introduced its on-demand, inline Help System for SharePoint to assist end users in developing mastery over common SharePoint tasks. In 2016, VisualSP expanded its technology solutions to online Microsoft products. The company is led by Microsoft MVP and MCT Asif Rehmani. To learn more, visit http://www.visualsp.com.
News Article | February 15, 2017
CTA approval in Spain for Phase I AM101 clinical study evaluating safety of adjuvanted, modified house dust mite allergy vaccine CTA approval in Spain for Phase I AM101 clinical study evaluating safety of adjuvanted, modified house dust mite allergy vaccine Allergy Therapeutics ( : AGY), the fully integrated specialty pharmaceutical company specialising in allergy vaccines, today announces that the Phase I clinical study investigating the safety and tolerability of Acarovac MPL (monophosphoryl lipid A) has received Clinical Trial Application (CTA) approval in Spain. House dust mites are a major cause of perennial allergic rhinitis and allergic asthma1. Acarovac MPL builds on the strong foundation of technologies employed in the successful Pollinex® Quattro range of subcutaneous allergy immunotherapies, and builds on the demonstrated efficacy2 of the existing successful product platform of Acarovac Plus™, the fastest growing "named-patient product" in Allergy Therapeutics' Spanish subsidiary. Acarovac MPL is the only house-dust mite immunotherapy in development utilising MCT® (microcrystalline tyrosine), a natural, biodegradable depot, and the adjuvant MPL for the treatment of perennial allergic rhinitis making this vaccine unique in a $1.5 billion per annum market3. CTA approval has been granted and the Company now expects this formulation to begin Phase I studies immediately, to assess the safety and tolerability of two different dose regimens. Both treatment regimens include a two-to-four week initial up-dosing phase but differ in the duration of the later maintenance phase. The Phase I trial of 32 patients is expected to last one year, and to be delivered on-time in-line with the Company's stated strategic plan when funding the programme in November 2015. If the Phase I trial is successful, the Company expects to launch Acarovac MPL in Spain on a named-patient basis. Commenting on the CTA approval, Manuel Llobet, Chief Executive Officer of Allergy Therapeutics, said: "By using the successful biodegradable adjuvant system of MCT® and MPL, integral in our seasonal marketed vaccines, in a perennial house dust mite vaccine, we aim to improve both safety and efficacy for the thousands of patients who suffer year-round due to house dust mite allergy. Acarovac Quattro will provide convenience to our patients and doctors and more efficient pharmacoeconomics. Effectively, the dosing regime in Acarovac Quattro has the potential to improve the convenience, adherence and compliance that is essential for a successful treatment, and builds on the acceptance of the existing Acarovac Plus platform." About Allergy Therapeutics Allergy Therapeutics is an international specialty pharmaceutical company focussed on the treatment and diagnosis of allergic disorders including immunotherapy vaccines that cure disease. The Company sells proprietary products and third party products from its subsidiaries in nine major European countries and via distribution agreements in an additional ten countries. Formed in 1999 out of Smith Kline Beecham, Allergy Therapeutics is headquartered in Worthing, UK with MHRA-approved manufacturing facilities. The Company employs c.495 employees and is listed on the London Stock Exchange ( : AGY). For more information, please see www.allergytherapeutics.com. References 1. Calderón M et al., Respiratory allergy caused by house dust mites: What do we really know? J Allergy Clin Immunol. 2015 Jul;136(1):38-48 2. Roger, A., Depreux, N., Jurgens Y., Heath M, Garcia G., Skinner M, A novel and well tolerated mite allergoid subcutaneous immunotherapy: Evidence of clinical and immunologic efficacy. Immunity, Inflammation and Disease, 2014; 2 (2); 92-98 3. QYR Pharma report. September 2016 This information is provided by RNS The company news service from the London Stock Exchange
News Article | March 1, 2017
Leader in pure nutrition adds health aiding oils and weight management supplements to its arsenal to help customers reach their fitness potential ORLANDO, FL--(Marketwired - Mar 1, 2017) - NutraKey, creator of research-based, high-quality health and wellness products, today announces new performance-optimizing products to its supplement lineup: CLA 1250, MCT Oil, Turmeric Complex, and Fish Oil. NutraKey's new top-of-the-line products further extend the company's reach into the supplement market, as each product is designed to coincide with daily routines to help fitness enthusiasts reach performance goals and increase their overall health and wellness. "Our vision for the future is to allow customers to define their own path to health, while also providing innovative formulas tailored to specific fitness goals," said Chris Wagner, Chief Executive Officer of NutraKey. "By combining our passion for fitness with dedication to pure nutrition, we aim to make the journey to improved health easier for everyone." NutraKey's four new supplement offerings help to bridge the nutritional gap and focus on a path towards well-being: CLA 1250 is a Conjugated Linoleic Acid supplement well-known for supporting fat loss. An Omega-6 fatty acid naturally found in beef, dairy and vegetable oils, NutraKey's CLA 1250 mg softgels not only contribute to weight loss, but help in building lean muscle, as well as being a general health aid. CLA 1250 is available in 90 and 180-count options for $14.99-$23.99. MCT stands for medium chain triglycerides and is comprised of primarily capric fatty acids, known for its fat burning and energy sustaining powers. NutraKey's MCT Oil is available in two different forms: softgels and oils. When MCT oil is metabolized in the body, it behaves more like a carbohydrate than a fat. NutraKey has a variety of different flavored oils, such as lemon and vanilla, or can be unflavored, available in a 16-oz. option. NutraKey's MCT Oil softgels are available in 90 and 180-count options for $14.99-$23.99. Turmeric is widely known for its powerful anti-inflammatory, natural medicinal properties. Turmeric is used to treat a variety of health conditions, as well as aid in immune and joint support. NutraKey's Turmeric Complex 1500 mg softgels are available in 90 and 180-count options for $12.99-$19.99. Fish Oil contains a natural source of powerful Omega-3s, providing multiple benefits when looking to build muscle, lose fat or maintain a healthy diet. Fish Oil is known for joint, immune and mood support and maintaining a healthy cholesterol level. NutraKey's Fish Oil softgels, available in 90 and 180-count options for $10.99-$18.99, help support workout quality and overall health. All products are available online at https://nutrakeyhealth.com/. To find a store near you, visit our store directory at https://nutrakeyhealth.com/find-a-store/. About NutraKey: NutraKey is a global nutritional supplement company founded in 2010, specializing in performance-enhancing products to help consumers build muscle, boost testosterone, increase fat loss, and aid in overall health and wellness. NutraKey's robust line and flavorful products set the standard in nutritional supplements, providing pure, raw, and nutritional ingredients to help consumers of all fitness levels reach their performance goals.