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X-Ray Diffraction Microscope Designed to Reveal 3D Internal Structure of Whole Cell

By MedImaging International staff writers
Posted on 29 Jul 2010

Three-dimensional (3D) imaging is greatly expanding the ability of researchers to examine biologic specimens, enabling a glance into their internal structures. Moreover, recent developments in X-ray diffraction methods have helped extend the limit of this approach.

While significant progress has been made in optical microscopy to break the diffraction barrier, such techniques rely on fluorescent labeling technologies, which prohibit the quantitative 3D imaging of the entire contents of cells. Cryoelectron microscopy can image structures at a resolution of 3-5 nm, but this only works with thin or sectioned specimens.

Furthermore, although X-ray protein crystallography is currently the primary method used for determining the 3D structure of protein molecules, many biological specimens--such as whole cells, cellular organelles, some viruses and many important protein molecules--are difficult or impossible to crystallize, making their structures inaccessible. Overcoming these limitations requires the employment of different techniques.

Now, in an article published May 31, 2010, in the journal Proceedings of [U.S.] National Academy of Sciences (PNAS), scientists from the University of California, Los Angeles (UCLA; USA) and their collaborators demonstrate the use of a unique X-ray diffraction microscope that enabled them to reveal the internal structure of yeast spores. The team reports the quantitative 3D imaging of a whole, unstained cell at a resolution of 50 nm - 60 nm using X-ray diffraction microscopy, also known as lensless imaging.

Researchers identified the 3D morphology and structure of cellular organelles, including the cell wall, vacuole, endoplasmic reticulum, mitochondria, granules, and nucleolus. The research may open a way to identifying the individual protein molecules inside whole cells using labeling technologies.

The lead authors on the study are Dr. Huaidong Jiang, a UCLA assistant researcher in physics and astronomy, and Dr. John Miao, a UCLA professor of physics and astronomy. The work is a culmination of a collaboration started three years ago with Dr. Fuyu Tamanoi, UCLA professor of microbiology, immunology, and molecular genetics. Dr. Miao and Dr. Tamanoi are both researchers at UCLA's California NanoSystems Institute. Other collaborators include teams at Riken Spring 8 (Hyogo, Japan) and the Institute of Physics, Academia Sinica (Taipei, Taiwan).

"This is the first time that people have been able to peek into the 3D internal structure of a biological specimen, without cutting it into sections, using X-ray diffraction microscopy, Dr. Miao noted. "By avoiding use of X-ray lenses, the resolution of X-ray diffraction microscopy is ultimately limited by radiation damage to biological specimens. Using cryogenic technologies, 3D imaging of whole biological cells at a resolution of 5 nm - 10 nm should be achievable. Our work hence paves a way for quantitative 3D imaging of a wide range of biological specimens at nanometer-scale resolutions that are too thick for electron microscopy.”

Dr. Tamanoi prepared the yeast spore samples analyzed in this study. Spores are specialized cells that are formed when they are placed under nutrient-starved conditions. Cells use this survival strategy to deal with harsh conditions. "Biologists wanted to examine internal structures of the spore, but previous microscopic studies provided information on only the surface features. We are very excited to be able to view the spore in 3D,” Dr. Tamanoi said. "We can now look into the structure of other spores, such as Anthrax spores and many other fungal spores. It is also important to point out that yeast spores are of similar size to many intracellular organelles in human cells. These can be examined in the future.”

Since its first experimental demonstration by Dr. Miao and collaborators in 1999, coherent diffraction microscopy has been applied to imaging a wide range of materials science and biologic specimens, such as nanoparticles, nanocrystals, biomaterials, cells, cellular organelles, viruses, and carbon nanotubes using X-ray, electron, and laser facilities worldwide. Until now, however, the radiation-damage problem and the difficulty of acquiring high-quality 3D diffraction patterns from individual whole cells have prevented the successful high-resolution 3D imaging of biologic cells by X-ray diffraction.

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54 (41.007995605469): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

55 (80.108642578125): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

56 (47.922134399414): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

57 (296.83113098145): SELECT name FROM jos_deeppockets WHERE id=294730475 LIMIT 1

58 (232.93495178223): SELECT name FROM jos_deeppockets_archive WHERE id=294730475 LIMIT 1

59 (49.829483032227): 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'

60 (59.127807617188): 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=294730475 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

61 (498.05641174316): 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=294730475 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

62 (!!!!! 1208.0669403076): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

63 (62.942504882812): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

64 (98.943710327148): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

65 (47.922134399414): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

66 (85.115432739258): 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

67 (622.03407287598): SELECT fid FROM jos_deeppockets WHERE id=294730475 AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE id=294730475 AND published='1' LIMIT 1

68 (46.968460083008): 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`

69 (42.915344238281): select jos_twitters.link from jos_twitters where cat=10

70 (29.087066650391): select jos_facebook.link from jos_facebook where cat=10

71 (30.040740966797): select jos_google_plus.link from jos_google_plus where cat=10

72 (30.040740966797): select jos_linkedin.link from jos_linkedin where cat=10

73 (31.948089599609): SELECT fid FROM jos_deeppockets WHERE id=294730475 AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE id=294730475 AND published='1' LIMIT 1

74 (30.994415283203): 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`

75 (382.90023803711): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294730475 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

76 (225.06713867188): SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294730475 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

77 (40.054321289062): 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`

78 (!!!!! 1088.1423950195): SELECT show_live FROM jos_com_show_setting LIMIT 1

79 (41.961669921875): SELECT show_live FROM jos_com_show_setting LIMIT 1

80 (28.848648071289): SELECT show_live FROM jos_com_show_setting LIMIT 1

81 (27.894973754883): SELECT show_live FROM jos_com_show_setting LIMIT 1

82 (28.848648071289): SELECT show_live FROM jos_com_show_setting LIMIT 1

83 (27.894973754883): SELECT show_live FROM jos_com_show_setting LIMIT 1

84 (52.928924560547): SELECT show_live FROM jos_com_show_setting LIMIT 1

85 (30.040740966797): SELECT show_live FROM jos_com_show_setting LIMIT 1

86 (28.848648071289): SELECT show_live FROM jos_com_show_setting LIMIT 1

87 (30.040740966797): SELECT show_live FROM jos_com_show_setting LIMIT 1

88 (27.894973754883): SELECT show_live FROM jos_com_show_setting LIMIT 1

89 (30.040740966797): SELECT show_live FROM jos_com_show_setting LIMIT 1

90 (27.894973754883): SELECT show_live FROM jos_com_show_setting LIMIT 1

91 (29.087066650391): SELECT show_live FROM jos_com_show_setting LIMIT 1

92 (50.067901611328): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294730475 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

93 (30.994415283203): SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294730475 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

94 (33.140182495117): 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`

95 (74.863433837891): SELECT show_live FROM jos_com_show_setting LIMIT 1

96 (203.84788513184): SELECT show_live FROM jos_com_show_setting LIMIT 1

97 (366.9261932373): SELECT show_live FROM jos_com_show_setting LIMIT 1

98 (60.796737670898): SELECT show_live FROM jos_com_show_setting LIMIT 1

99 (49.114227294922): SELECT show_live FROM jos_com_show_setting LIMIT 1

100 (41.007995605469): SELECT show_live FROM jos_com_show_setting LIMIT 1

101 (429.86869812012): SELECT show_live FROM jos_com_show_setting LIMIT 1

102 (230.07392883301): SELECT show_live FROM jos_com_show_setting LIMIT 1

103 (54.121017456055): SELECT show_live FROM jos_com_show_setting LIMIT 1

104 (41.961669921875): SELECT show_live FROM jos_com_show_setting LIMIT 1

105 (281.81076049805): SELECT show_live FROM jos_com_show_setting LIMIT 1

106 (194.07272338867): SELECT show_live FROM jos_com_show_setting LIMIT 1

107 (51.975250244141): SELECT show_live FROM jos_com_show_setting LIMIT 1

108 (39.815902709961): SELECT show_live FROM jos_com_show_setting LIMIT 1

109 (277.99606323242): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

110 (157.83309936523): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

111 (283.95652770996): 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=294730475 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

112 (121.8318939209): 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=294730475 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

113 (!!!!! 5259.0370178223): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

114 (319.9577331543): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

115 (731.9450378418): 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=294730475 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

116 (319.00405883789): 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=294730475 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

117 (53.167343139648): 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=294730474 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

118 (283.00285339355): 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=294730474 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

119 (612.02049255371): 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

120 (411.9873046875): 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

121 (!!!!! 1169.2047119141): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

122 (56.982040405273): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

123 (51.021575927734): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

124 (39.100646972656): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

125 (43.869018554688): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

126 (36.001205444336): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

127 (46.968460083008): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

128 (39.100646972656): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

129 (326.87187194824): SELECT * FROM trending_articles WHERE `journal_id` = 10 AND `timestamp` <= NOW() + INTERVAL 7 DAY ORDER BY `order`

130 (!!!!! 38266.897201538): 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

131 (522.85194396973): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294730475 AND type='deeppockets_item' AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294730475 AND type='deeppockets_item' AND published='1' LIMIT 1

132 (!!!!! 15560.865402222): ( 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

133 (64.13459777832): ( 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

134 (41.961669921875): 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

135 (31.948089599609): 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

136 (42.915344238281): 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

137 (30.994415283203): 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

138 (306.84471130371): 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

139 (45.061111450195): 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

140 (36.001205444336): 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

141 (31.948089599609): 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

142 (460.86311340332): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

143 (314.95094299316): SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

144 (36.954879760742): SELECT name FROM jos_deeppockets WHERE id=211

145 (30.040740966797): SELECT name FROM jos_deeppockets WHERE id=211

146 (580.0724029541): 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

147 (458.95576477051): 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

148 (!!!!! 1535.8924865723): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

149 (69.141387939453): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

150 (!!!!! 13059.854507446): TRUNCATE TABLE jos_trademed_module

151 (102.04315185547): 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=294730475 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

152 (!!!!! 3461.8377685547): 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=294730475 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1

153 (646.82960510254): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

154 (73.909759521484): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1

155 (72.002410888672): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

156 (52.213668823242): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

157 (!!!!! 1178.0261993408): 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

158 (74.148178100586): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

159 (!!!!! 3303.050994873): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

160 (103.95050048828): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294730475 AND type='deeppockets_item' AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294730475 AND type='deeppockets_item' AND published='1' LIMIT 1

161 (40.054321289062): 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)

162 (34.093856811523): 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)

163 (108.00361633301): SELECT livesite FROM jos_dp_categories WHERE id=10 AND published='1' LIMIT 1

164 (403.88107299805): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

165 (75.101852416992): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294730475 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294730475 AND published='1' LIMIT 1

166 (56.982040405273): select jos_twitters.link from jos_twitters where cat=10

167 (30.040740966797): select jos_facebook.link from jos_facebook where cat=10

168 (30.040740966797): select jos_google_plus.link from jos_google_plus where cat=10

169 (31.948089599609): select jos_linkedin.link from jos_linkedin where cat=10

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