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Optical Projection Tomography Imaging Offers New Approach for Breast Cancer Research

By MedImaging International staff writers
Posted on 19 Jan 2012
Image: Instances of irregular MDA-MB-231 nuclear morphologies (scale bar = 5 microns). Panel (A) depicts a multilobular nucleus and panel (B) illustrates a cell with micronuclei. Left images show nuclear surface in blue and cytoplasm in gray, middle images show surface-shaded renderings of the nuclear volume and right images depict volume renderings through the nuclear volume. Increasing nuclear density is color coded from green to red (Photo courtesy of the Biodesign Institute / University of Arizona).
Image: Instances of irregular MDA-MB-231 nuclear morphologies (scale bar = 5 microns). Panel (A) depicts a multilobular nucleus and panel (B) illustrates a cell with micronuclei. Left images show nuclear surface in blue and cytoplasm in gray, middle images show surface-shaded renderings of the nuclear volume and right images depict volume renderings through the nuclear volume. Increasing nuclear density is color coded from green to red (Photo courtesy of the Biodesign Institute / University of Arizona).
A new imaging technique being developed could help researchers pinpoint slight abnormalities in cell nuclear structure, the molecular biosignature of breast cancer, thereby considerably improving diagnostic accuracy and prognosis by providing early detection of the disease.

The team, led by Prof. Deirdre Meldrum, senior scientist and director of the Arizona State University (ASU) Center for Biosignatures Discovery Automation at Biodesign (Tempe, USA), has assessed normal, benign, and malignant cells, using the first research Cell-CT, developed by VisionGate, Inc. (Phoenix, AZ, USA)--a customized instrument capable of imaging cells in precise three-dimensional (3D) with true isotropic resolution. The technology allows the examination of subtle cellular details inaccessible with more conventional forms of microscopy that are inherently 2D.

The group’s findings appeared January 5, 2012, in the journal PLoS ONE. The 3D movie images of cells observed in the study reveal numerous characteristic traces of their condition as normal or aberrant. Prof. Meldrum commented, “There are numerous quantitative morphological parameters that are indicative of disease and may be used as biosignatures for disease staging and diagnosis. For example, a cancerous cell typically has an enlarged nucleus, nuclear invaginations, chromosome mutations, and unique nuclear shape changes.”

Currently, the definitive clinical diagnosis of breast cancer malignancy relies on careful examination of the nuclear structure of cells that have been prepared by histological staining and subjected to bright field microscopy. According to Vivek Nandakumar, lead author of the current study, pathologists qualitatively examine cell features including nuclear size, shape, nucleus-to-cytoplasm ratio, and the texture of cell chromatin. However, these observations do not involve quantitative measurements that would promote a more accurate analysis.

Prof. Meldrum agreed, as to the failings of conventional pathology. As director of the Microscale Life Sciences Center, she has devoted much of her career to the close study of cell heterogeneity, and the manner in which individual cells can go awry as they transition to diseased states. “In our analysis of live single cells we can quantify significant variation from cell to cell under the same conditions,” said Prof. Meldrum.

The scientists utilized Cell-CT to examine 150 cells in each of three specific categories: normal, benign fibrocystic and malignant breast epithelial. Controversy remains as to whether breast fibrosis, which may result from hormonal alterations, is a normal condition or an early indication of malignancy. The condition occurs when ligaments, scars, supportive tissue or other fibrous tissue become more prominent in the breast than fatty tissue.

Cell-CT is a new type of microscope, able to image cells in three-dimensions, using a technique called optical projection tomography. Cell-CT operates similar to a normal CT scanner, though it uses visible photons of light, instead of X-rays. Cells prepared for observation are not placed on slides, but are instead suspended in gel and injected through a microcapillary tube that permits multiple imaging in 360°.

The scanning process produces hundreds of thin slices through the cell. These sections, or tomographs, are reassembled through computer software, forming a detailed 3D image. Movies of cells seen in rotation brightly reveal shape asymmetries, a particularly useful tool for disease diagnosis.

The three cell types examined in the study fell into four distinct nuclear shape categories. Category 1 cell nuclei were slender, with marked concavity. Category 2 cells had a slight concavity and were bulky. Based on these shapes, the first two categories are termed mushroom cap morphology. (The category 2 mushroom cap morphology was the most common nuclear form seen in all three cell types.) Category 3 nuclei were mostly convex in shape, whereas category 4 nuclei were irregular and distorted in shape.

Significantly, cells taken from the cancerous cell line showed the largest fraction of irregular, category 4 and category 2 nuclei and the smallest fraction of nuclei with a category 3 convex shape. The malignant cells also displayed the greatest shape heterogeneity within category 4. The fibrocystic cell sample contained the largest portion of category 3 and the lowest portion of category 1 nuclei. The largest overall shape heterogeneity with respect to the four shape categories occurred in the normal cells.

Cell and nuclear volume were observed to increase as one travels from normal to fibrocystic to malignant cells, though fibrocystic cells had, on average, the largest nucleus-to-cytoplasm volume. Textural differences among cells and arrangement of chromatin were also observed. In all, the scientists computed 42 distinct 3D morphologic and textural descriptors of cellular and nuclear structure. Cell-CT technology is able to resolve cell features down to less than one half micron.

Study coauthor Roger Johnson, research laboratory manager at the Center for Biosignatures Discovery Automation, emphasized that the subtle nuclear differences seen, especially for the cancerous cells, would likely have been missed had the samples been examined with conventional 2D imagery. As a result, the architecture imaged with Cell-CT supersedes the existing nuclear grades established for cancer diagnosis using a microscope.

Although much progress has been made in determining the transformation of cells from normal to diseased states, patient outcomes for many forms of cancer remain discouragingly poor. Many believe a new paradigm for investigating such cancers will need to be established, and the field has drawn interested researches from diverse disciplines. Dr. Paul Davies, another coauthor of the current study is a physicist and cosmologist in ASU’s College of Liberal Arts and Sciences and part of a new US National Cancer Institute-funded consortium, focused on studying the physical science of cancer. “We expect that insights and methods drawn from physical science will lead to radical new ideas for understanding and tackling cancer,” said Dr. Davies.

These findings provide new clues into the variations of nuclear structure that often signal cell malignancy. The unparalleled structural details produced by Cell-CT have the potential to improve dramatically 3D nuclear morphometry, leading to a sensitive and specific nuclear grade classification for breast cancer diagnosis.

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70 (285.86387634277): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294738934 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
71 (291.82434082031): SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294738934 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
72 (53.882598876953): 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`
73 (109.91096496582): SELECT show_live FROM jos_com_show_setting LIMIT 1
74 (35.04753112793): SELECT show_live FROM jos_com_show_setting LIMIT 1
75 (47.922134399414): SELECT show_live FROM jos_com_show_setting LIMIT 1
76 (33.855438232422): SELECT show_live FROM jos_com_show_setting LIMIT 1
77 (30.040740966797): SELECT show_live FROM jos_com_show_setting LIMIT 1
78 (33.140182495117): SELECT show_live FROM jos_com_show_setting LIMIT 1
79 (26.941299438477): SELECT show_live FROM jos_com_show_setting LIMIT 1
80 (28.133392333984): SELECT show_live FROM jos_com_show_setting LIMIT 1
81 (26.941299438477): SELECT show_live FROM jos_com_show_setting LIMIT 1
82 (28.133392333984): SELECT show_live FROM jos_com_show_setting LIMIT 1
83 (28.133392333984): SELECT show_live FROM jos_com_show_setting LIMIT 1
84 (27.894973754883): SELECT show_live FROM jos_com_show_setting LIMIT 1
85 (26.941299438477): SELECT show_live FROM jos_com_show_setting LIMIT 1
86 (48.87580871582): SELECT show_live FROM jos_com_show_setting LIMIT 1
87 (46.968460083008): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294738934 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
88 (30.040740966797): SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294738934 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
89 (37.908554077148): 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`
90 (46.968460083008): SELECT show_live FROM jos_com_show_setting LIMIT 1
91 (30.040740966797): SELECT show_live FROM jos_com_show_setting LIMIT 1
92 (28.133392333984): SELECT show_live FROM jos_com_show_setting LIMIT 1
93 (!!!!! 2465.9633636475): SELECT show_live FROM jos_com_show_setting LIMIT 1
94 (59.127807617188): SELECT show_live FROM jos_com_show_setting LIMIT 1
95 (30.040740966797): SELECT show_live FROM jos_com_show_setting LIMIT 1
96 (31.948089599609): SELECT show_live FROM jos_com_show_setting LIMIT 1
97 (49.114227294922): SELECT show_live FROM jos_com_show_setting LIMIT 1
98 (29.087066650391): SELECT show_live FROM jos_com_show_setting LIMIT 1
99 (28.133392333984): SELECT show_live FROM jos_com_show_setting LIMIT 1
100 (26.941299438477): SELECT show_live FROM jos_com_show_setting LIMIT 1
101 (28.133392333984): SELECT show_live FROM jos_com_show_setting LIMIT 1
102 (26.941299438477): SELECT show_live FROM jos_com_show_setting LIMIT 1
103 (28.848648071289): SELECT show_live FROM jos_com_show_setting LIMIT 1
104 (71.048736572266): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
105 (45.061111450195): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
106 (55.074691772461): 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=294738934 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
107 (406.02684020996): 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=294738934 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
108 (!!!!! 3373.1460571289): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
109 (85.830688476562): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
110 (52.928924560547): 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=294738934 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
111 (473.0224609375): 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=294738934 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
112 (605.82160949707): 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
113 (442.0280456543): 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
114 (82.015991210938): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
115 (72.002410888672): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
116 (61.988830566406): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
117 (51.021575927734): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
118 (57.93571472168): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
119 (52.928924560547): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
120 (54.121017456055): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
121 (40.769577026367): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
122 (258.20732116699): SELECT * FROM trending_articles WHERE `journal_id` = 10 AND `timestamp` <= NOW() + INTERVAL 7 DAY ORDER BY `order`
123 (!!!!! 40771.007537842): 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
124 (524.99771118164): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294738934 AND type='deeppockets_item' AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294738934 AND type='deeppockets_item' AND published='1' LIMIT 1
125 (!!!!! 15852.928161621): ( 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
126 (68.902969360352): ( 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
127 (313.04359436035): 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
128 (40.054321289062): 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
129 (257.96890258789): 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
130 (40.054321289062): 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
131 (257.01522827148): 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
132 (37.908554077148): 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
133 (193.83430480957): 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
134 (35.04753112793): 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
135 (!!!!! 1867.0558929443): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
136 (429.15344238281): SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
137 (269.88983154297): SELECT name FROM jos_deeppockets WHERE id=215
138 (46.014785766602): SELECT name FROM jos_deeppockets WHERE id=215
139 (599.14588928223): 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 '((;)|(^))215((;)|($))' AND catid IN (0,110) ORDER BY publish_up ASC LIMIT 3
140 (446.08116149902): SELECT *, DATE_FORMAT(publish_up,'%d %b %Y') AS date_publish_up, DATE_FORMAT(publish_down,'%d %b %Y') AS date_publish_down FROM jos_events WHERE state=1 AND publish_up > NOW() and extra_info REGEXP '((;)|(^))215((;)|($))' AND catid IN (0,110) ORDER BY publish_up ASC LIMIT 3
141 (92.029571533203): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
142 (56.982040405273): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
143 (!!!!! 19803.047180176): TRUNCATE TABLE jos_trademed_module
144 (571.01249694824): 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=294738934 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
145 (90.122222900391): 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=294738934 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
146 (!!!!! 1453.8764953613): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
147 (71.048736572266): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
148 (646.82960510254): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
149 (118.01719665527): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
150 (!!!!! 1816.0343170166): 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
151 (449.89585876465): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
152 (55.074691772461): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
153 (344.03800964355): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294738934 AND type='deeppockets_item' AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294738934 AND type='deeppockets_item' AND published='1' LIMIT 1
154 (464.91622924805): 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=215 or channel_id is null)
155 (37.908554077148): 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=215 or channel_id is null)
156 (115.87142944336): SELECT livesite FROM jos_dp_categories WHERE id=10 AND published='1' LIMIT 1
157 (576.01928710938): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
158 (397.92060852051): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294738934 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294738934 AND published='1' LIMIT 1
159 (144.00482177734): select jos_twitters.link from jos_twitters where cat=10
160 (31.948089599609): select jos_facebook.link from jos_facebook where cat=10
161 (112.05673217773): select jos_google_plus.link from jos_google_plus where cat=10
162 (36.954879760742): select jos_linkedin.link from jos_linkedin where cat=10
163 (358.81996154785): update jos_bannersmanager_banner set impmade=impmade+1 where id in(14202,14203)
164 (!!!!! 1188.0397796631): insert into jos_banners_delta values (14202,'2025-06-24 03:54:48','216.73.216.120','view'),(14203,'2025-06-24 03:54:48','216.73.216.120','view')