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Superbright, Fast X-Rays Can Visualize Just One Layer of Proteins

By MedImaging International staff writers
Posted on 23 Feb 2014

Image: X-ray free-electron lasers (XFELs) can create images (left) that accurately reflect the known structure of proteins determined by conventional methods (right), in this case, three bacteriorhodopsin proteins (Photo courtesy of DOE/Pacific Northwest National Laboratory).

Scientists are now using exceptionally bright and fast X-rays that can capture an image of a protein that rivals traditional techniques with a sheet of protein only one protein-molecule-thick.

A protein’s shape is crucial for understanding how it causes disease or toxicity. Researchers who use X-rays to take images of proteins need a billion copies of the same protein stacked and packed into a well-ordered crystal.

Using a type of laser known as the X-ray free electron laser (XFEL), the technique creates ways to determine the structural characteristics of nearly 25% of known proteins, many of which have been missed because of their inability to stack effectively. The team of researchers led by the US Department of Energy’s (DOE) Pacific Northwest National Laboratory (PNNL; Richland, WA, USA) and Lawrence Livermore National Laboratory (LLNL; Livermore, CA, USA) reported their findings with this innovative type of X-ray diffraction in the March 2014 issue of the International Union of Crystallography Journal. “In this paper, we’re proving it’s possible to use an XFEL to study individual monolayers of protein,” said PNNL microscopist Dr. James Evans. “Just being able to see any diffraction is brand new.”

Dr. Evans is co-leader of the team of two dozen scientists with LLNL physicist Dr. Matthias Frank. The bright, fast X-rays were produced at the Linac Coherent Light Source (LCLS) at the SLAC US National Accelerator Laboratory (Menlo Park, CA, USA), the newest of DOE’s major X-ray light source facilities at the national laboratories. LCLS, currently the world’s most powerful X-ray laser, is an X-ray free-electron laser. It generates beams millions of times brighter than earlier X-ray light sources.

At approximately 0.8 nanometer resolution, the proteins appear somewhat fuzzy but match the predicted view based on earlier studies. Dr. Evans reported that this degree of clarity would allow researchers, in some instances, to visualize how proteins alter their shape as they interact with other proteins or molecules in their environment.

To obtain a better view of protein monolayers using XFEL, the scientists will need to enhance the resolution to 0.1 to 0.3 nanometers, as well as capture images of the proteins at different angles, efforts for which are currently ongoing. Researchers have been using X-ray crystallography for over 60 years to determine the shape and form of proteins that form the inner workings of a living organism’s cells. The conventional technology requires, however, that proteins stack into a large crystal, similar to how oranges stack in a crate. The structure of more than 80,000 proteins have been determined this way, leading to a better understanding of diseases, pathogens, and how organisms grow and develop.

However, many proteins found in natural surroundings do not stack easily. Some protrude from the fatty membranes that cover cells, identifying and interacting with other cells and objects, such as bacteria or viruses, in the surrounding area. These proteins are not used to having others of their kind stack on top. These membrane proteins comprise approximately 25% of all proteins but only 2% of proteins for which researchers have determined structures.

Researchers in the last 10 years have been pursuing the theory that one sheet of proteins could be visualized if the X-rays were bright enough but flashed on and off swiftly enough to limit the damage brought about by the powerful X-rays. Researchers, two years ago, demonstrated they could use XFEL technology on crystals of proteins approximately 15 to 20 sheets thick.

The scientists needed to push this further and worked on a way to create one-sheet-thick crystals of two different proteins: a protein called streptavidin and the membrane protein bacteriorhodopsin. The structures of both proteins are well known to scientists, which gave them something with which to compare their findings. The scientists shined the super-bright X-rays for about 30 femtoseconds on the protein crystals. They created so much data in the process that it took them more than one year to study all of it.

The resulting images look like the known structures, confirming that this technology works. Next, the researchers will try to capture proteins changing shape as they engage in a chemical reaction. For this, even shorter flashes of X-rays might be needed to see the action clearly. If effective, shorter flashes of XFEL might mean longer lines at SLAC.

Related Links:
Pacific Northwest National Laboratory
Lawrence Livermore National Laboratory

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52 (116.10984802246): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

53 (55.074691772461): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

54 (477.07557678223): SELECT name FROM jos_deeppockets WHERE id=294750856 LIMIT 1

55 (370.02563476562): SELECT name FROM jos_deeppockets_archive WHERE id=294750856 LIMIT 1

56 (82.015991210938): SELECT a.* FROM jos_components AS a WHERE ( a.admin_menu_link = 'option=com_syndicate' OR a.admin_menu_link = 'option=com_syndicate&hidemainmenu=1' ) AND a.option = 'com_syndicate'

57 (104.90417480469): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.cat=10 AND dp.id=294750856 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

58 (663.04206848145): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets_archive as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.cat=10 AND dp.id=294750856 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

59 (!!!!! 1404.0470123291): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

60 (113.96408081055): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

61 (233.17337036133): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

62 (73.19450378418): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

63 (181.91337585449): select cover from jos_number where journalid in (5) and cover is not null order by left(issue,2)+right(issue,2)*12 desc limit 1

64 (788.92707824707): SELECT fid FROM jos_deeppockets WHERE id=294750856 AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE id=294750856 AND published='1' LIMIT 1

65 (360.01205444336): SELECT id,name,sponsor_text FROM jos_deeppockets dp left outer join jos_channel_sponsor cs on dp.id=cs.fid WHERE cat=10 AND TYPE='file_folder' AND published=1 AND id=211 ORDER BY `ordering`

66 (!!!!! 6723.8807678223): select jos_twitters.link from jos_twitters where cat=10

67 (134.94491577148): select jos_facebook.link from jos_facebook where cat=10

68 (41.961669921875): select jos_google_plus.link from jos_google_plus where cat=10

69 (30.994415283203): select jos_linkedin.link from jos_linkedin where cat=10

70 (76.055526733398): SELECT fid FROM jos_deeppockets WHERE id=294750856 AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE id=294750856 AND published='1' LIMIT 1

71 (54.836273193359): SELECT id,name,sponsor_text FROM jos_deeppockets dp left outer join jos_channel_sponsor cs on dp.id=cs.fid WHERE cat=10 AND TYPE='file_folder' AND published=1 AND id=211 ORDER BY `ordering`

72 (529.05082702637): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294750856 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

73 (416.04042053223): SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294750856 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

74 (!!!!! 2225.1605987549): SELECT id,name,title FROM jos_deeppockets WHERE cat=10 AND TYPE='file_folder' AND published=1 AND id != 294743300 AND id != 294743301 AND id != 294743302 AND id != 294743303 AND id != 294743304 AND id != 294743305 AND id != 294743306 AND id != 294743307 AND id != 294743308 AND id != 294743309 AND id != 294743310 AND metakey != 'aacc' ORDER BY `ordering`

75 (86.069107055664): SELECT show_live FROM jos_com_show_setting LIMIT 1

76 (34.809112548828): SELECT show_live FROM jos_com_show_setting LIMIT 1

77 (30.040740966797): SELECT show_live FROM jos_com_show_setting LIMIT 1

78 (30.040740966797): SELECT show_live FROM jos_com_show_setting LIMIT 1

79 (29.087066650391): SELECT show_live FROM jos_com_show_setting LIMIT 1

80 (28.848648071289): SELECT show_live FROM jos_com_show_setting LIMIT 1

81 (70.810317993164): SELECT show_live FROM jos_com_show_setting LIMIT 1

82 (31.948089599609): SELECT show_live FROM jos_com_show_setting LIMIT 1

83 (34.093856811523): SELECT show_live FROM jos_com_show_setting LIMIT 1

84 (36.001205444336): SELECT show_live FROM jos_com_show_setting LIMIT 1

85 (40.054321289062): SELECT show_live FROM jos_com_show_setting LIMIT 1

86 (36.001205444336): SELECT show_live FROM jos_com_show_setting LIMIT 1

87 (30.040740966797): SELECT show_live FROM jos_com_show_setting LIMIT 1

88 (29.087066650391): SELECT show_live FROM jos_com_show_setting LIMIT 1

89 (78.916549682617): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294750856 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

90 (41.961669921875): SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294750856 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

91 (50.783157348633): SELECT id,name,title FROM jos_deeppockets WHERE cat=10 AND TYPE='file_folder' AND published=1 AND id != 294743300 AND id != 294743301 AND id != 294743302 AND id != 294743303 AND id != 294743304 AND id != 294743305 AND id != 294743306 AND id != 294743307 AND id != 294743308 AND id != 294743309 AND id != 294743310 AND metakey != 'aacc' ORDER BY `ordering`

92 (85.115432739258): SELECT show_live FROM jos_com_show_setting LIMIT 1

93 (55.074691772461): SELECT show_live FROM jos_com_show_setting LIMIT 1

94 (66.041946411133): SELECT show_live FROM jos_com_show_setting LIMIT 1

95 (41.961669921875): SELECT show_live FROM jos_com_show_setting LIMIT 1

96 (41.961669921875): SELECT show_live FROM jos_com_show_setting LIMIT 1

97 (30.994415283203): SELECT show_live FROM jos_com_show_setting LIMIT 1

98 (28.848648071289): SELECT show_live FROM jos_com_show_setting LIMIT 1

99 (38.14697265625): SELECT show_live FROM jos_com_show_setting LIMIT 1

100 (33.140182495117): SELECT show_live FROM jos_com_show_setting LIMIT 1

101 (29.087066650391): SELECT show_live FROM jos_com_show_setting LIMIT 1

102 (28.848648071289): SELECT show_live FROM jos_com_show_setting LIMIT 1

103 (29.087066650391): SELECT show_live FROM jos_com_show_setting LIMIT 1

104 (41.007995605469): SELECT show_live FROM jos_com_show_setting LIMIT 1

105 (40.054321289062): SELECT show_live FROM jos_com_show_setting LIMIT 1

106 (167.1314239502): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

107 (68.902969360352): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

108 (89.883804321289): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.cat=10 AND dp.id=294750856 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

109 (623.94142150879): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets_archive as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.cat=10 AND dp.id=294750856 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

110 (!!!!! 4685.1634979248): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

111 (158.07151794434): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

112 (698.08959960938): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.id=294750856 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

113 (742.91229248047): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets_archive as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.id=294750856 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

114 (82.969665527344): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.id=294750855 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

115 (672.81723022461): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets_archive as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.id=294750855 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

116 (!!!!! 1058.1016540527): SELECT dp.name, dp.id, dp2.name as channel, ex.name as author_name, ex.specialization, ex.images FROM jos_deeppockets as dp LEFT JOIN jos_experts as ex ON ex.id = dp.author LEFT JOIN jos_deeppockets as dp2 ON dp2.id = dp.fid WHERE dp.author != 0 AND dp.cat = 10 AND dp.published = 1 AND ( (dp.fid = 294743300) OR (dp.fid = 294743301) OR (dp.fid = 294743302) OR (dp.fid = 294743303) OR (dp.fid = 294743304) OR (dp.fid = 294743305) OR (dp.fid = 294743306) OR (dp.fid = 294743307) OR (dp.fid = 294743308) OR (dp.fid = 294743309) OR (dp.fid = 294743310) ) ORDER BY RAND() LIMIT 1

117 (!!!!! 1775.9799957275): SELECT dp.name, dp.id, dp2.name as channel, ex.name as author_name, ex.specialization, ex.images FROM jos_deeppockets as dp LEFT JOIN jos_experts as ex ON ex.id = dp.author LEFT JOIN jos_deeppockets as dp2 ON dp2.id = dp.fid WHERE dp.author != 0 AND dp.cat = 10 AND dp.published = 1 AND ( (dp.fid = 294743300) OR (dp.fid = 294743301) OR (dp.fid = 294743302) OR (dp.fid = 294743303) OR (dp.fid = 294743304) OR (dp.fid = 294743305) OR (dp.fid = 294743306) OR (dp.fid = 294743307) OR (dp.fid = 294743308) OR (dp.fid = 294743309) OR (dp.fid = 294743310) ) ORDER BY RAND() LIMIT 1

118 (160.93254089355): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

119 (77.009201049805): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

120 (74.863433837891): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

121 (72.002410888672): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

122 (66.995620727539): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

123 (67.949295043945): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

124 (64.13459777832): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

125 (427.0076751709): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

126 (542.87910461426): SELECT * FROM trending_articles WHERE `journal_id` = 10 AND `timestamp` <= NOW() + INTERVAL 7 DAY ORDER BY `order`

127 (!!!!! 58478.116989136): SELECT * FROM jos_deeppockets WHERE jos_deeppockets.cat = 10 AND jos_deeppockets.type = 'deeppockets_item' AND jos_deeppockets.published = 1 AND jos_deeppockets.created + INTERVAL 14 DAY >= NOW() ORDER BY jos_deeppockets.hits DESC LIMIT 7

128 (!!!!! 1220.9415435791): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294750856 AND type='deeppockets_item' AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294750856 AND type='deeppockets_item' AND published='1' LIMIT 1

129 (!!!!! 26702.880859375): ( SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.video != '' AND dp.video_source != '' AND dp.cat = 10 AND dp.published = 1 AND dp.type != 'deeppockets_item_link' ORDER BY created DESC LIMIT 3 ) UNION ( SELECT dpa.id, dpa.name, dpa.title, dpa.video, dpa.video_source, dpa.created FROM jos_deeppockets_archive AS dpa WHERE dpa.video != '' AND dpa.video_source != '' AND dpa.cat = 10 AND dpa.published = 1 AND dpa.type != 'deeppockets_item_link' ORDER BY created DESC LIMIT 3 ) ORDER BY created DESC LIMIT 3

130 (139.95170593262): ( SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.video != '' AND dp.video_source != '' AND dp.cat = 10 AND dp.published = 1 AND dp.type != 'deeppockets_item_link' ORDER BY created DESC LIMIT 3 ) UNION ( SELECT dpa.id, dpa.name, dpa.title, dpa.video, dpa.video_source, dpa.created FROM jos_deeppockets_archive AS dpa WHERE dpa.video != '' AND dpa.video_source != '' AND dpa.cat = 10 AND dpa.published = 1 AND dpa.type != 'deeppockets_item_link' ORDER BY created DESC LIMIT 3 ) ORDER BY created DESC LIMIT 3

131 (575.06561279297): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294803337 LIMIT 1

132 (65.088272094727): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294803337 LIMIT 1

133 (!!!!! 1178.0261993408): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294803276 LIMIT 1

134 (51.021575927734): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294803276 LIMIT 1

135 (350.9521484375): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets_archive AS dp WHERE dp.id = 294803276 LIMIT 1

136 (53.882598876953): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets_archive AS dp WHERE dp.id = 294803276 LIMIT 1

137 (732.1834564209): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294802958 LIMIT 1

138 (79.154968261719): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294802958 LIMIT 1

139 (555.99212646484): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

140 (428.19976806641): SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

141 (220.06034851074): SELECT name FROM jos_deeppockets WHERE id=211

142 (74.148178100586): SELECT name FROM jos_deeppockets WHERE id=211

143 (797.03330993652): SELECT *, DATE_FORMAT(publish_up,'%d %b %Y') AS date_publish_up, DATE_FORMAT(publish_down,'%d %b %Y') AS date_publish_down FROM jos_events WHERE state=1 AND publish_up > NOW() and extra_info REGEXP '((;)|(^))211((;)|($))' AND catid IN (0,110) ORDER BY publish_up ASC LIMIT 3

144 (868.08204650879): SELECT *, DATE_FORMAT(publish_up,'%d %b %Y') AS date_publish_up, DATE_FORMAT(publish_down,'%d %b %Y') AS date_publish_down FROM jos_events WHERE state=1 AND publish_up > NOW() and extra_info REGEXP '((;)|(^))211((;)|($))' AND catid IN (0,110) ORDER BY publish_up ASC LIMIT 3

145 (119.20928955078): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

146 (75.817108154297): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

147 (!!!!! 20133.018493652): TRUNCATE TABLE jos_trademed_module

148 (735.04447937012): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.cat=10 AND dp.id=294750856 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

149 (!!!!! 5439.9967193604): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets_archive as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.cat=10 AND dp.id=294750856 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

150 (547.17063903809): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

151 (!!!!! 4586.935043335): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

152 (471.11511230469): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

153 (523.09036254883): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

154 (!!!!! 4935.0261688232): SELECT jos_item.* FROM jos_item INNER JOIN jos_number ON jos_item.numberID=jos_number.id WHERE 1=1 AND (jos_item.imgMini<>'') AND jos_item.type='L' AND jos_item.numberID= ( SELECT jos_number.id FROM jos_number WHERE jos_number.journalID IN (10) and jos_number.id in (select distinct jos_number.id from jos_number join jos_item on jos_item.numberid=jos_number.id where jos_item.type='P') ORDER BY (CONVERT(SUBSTRING(issue,1,2),SIGNED) + (CONVERT(SUBSTRING(issue,3,2),SIGNED)*100)) DESC LIMIT 0,1 ) ORDER BY RAND() DESC LIMIT 6

155 (617.02728271484): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

156 (241.04118347168): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

157 (169.03877258301): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294750856 AND type='deeppockets_item' AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294750856 AND type='deeppockets_item' AND published='1' LIMIT 1

158 (884.05609130859): select distinct links.* from jos_textlinks links join jos_textlinks_placement placement on links.id=placement.link_id where journal_id=(10) and (channel_id=0 or channel_id is null)

159 (!!!!! 5004.8828125): select distinct links.* from jos_textlinks links join jos_textlinks_placement placement on links.id=placement.link_id where journal_id=(10) and (channel_id=0 or channel_id is null)

160 (308.99047851562): SELECT livesite FROM jos_dp_categories WHERE id=10 AND published='1' LIMIT 1

161 (!!!!! 2171.0395812988): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

162 (!!!!! 2605.9150695801): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750856 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750856 AND published='1' LIMIT 1

163 (149.96528625488): select jos_twitters.link from jos_twitters where cat=10

164 (139.95170593262): select jos_facebook.link from jos_facebook where cat=10

165 (173.80714416504): select jos_google_plus.link from jos_google_plus where cat=10

166 (65.088272094727): select jos_linkedin.link from jos_linkedin where cat=10

167 (!!!!! 1665.1153564453): update jos_bannersmanager_banner set impmade=impmade+1 where id in(14615,14202,14203)

168 (!!!!! 3865.0035858154): insert into jos_banners_delta values (14615,'2025-08-19 02:37:00','216.73.216.0','view'),(14202,'2025-08-19 02:37:00','216.73.216.0','view'),(14203,'2025-08-19 02:37:00','216.73.216.0','view')

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