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text:
Aging is associated with a reduced capacity to cope with physiological stress. To study the molecular mechanisms associated with the decline in stress tolerance that accompanies aging, differences in gene expression between young and old Fischer 344 rats under euthermic control conditions or in response to hyperthermic challenge were evaluated using a cDNA array containing 207 stress-related genes. In the nonstressed control condition, aging resulted in selective upregulation of stress protein genes and transcripts involved in cell growth, death, and signaling, along with a downregulation of genes involved in antioxidant defenses and drug metabolism. Heat stress resulted in a broad induction of genes in the antioxidant and drug metabolism categories and transcripts involved in DNA, RNA, and protein synthesis for both age groups. Old animals had a robust upregulation of genes involved in cell growth, death, and signaling after heat challenge, along with a blunted expression of stress-response genes. In contrast, young animals had a strong induction of stress-response genes after hyperthermic challenge. Changes in expression of selected genes were confirmed by RT-PCR analysis. These findings suggest that aging results in altered gene expression in response to heat stress that is indicative of decreased stress protein transcription and increased expression of oxidative stress-related genes. Thus our findings support the postulate that transcriptional changes in response to a physiological challenge such as hyperthermia contribute to the loss of stress tolerance in older organisms.
- heat stress
- DNA array
- antioxidant enzymes
- stress proteins
- senescence
- DNA microarray
- gene regulation
aging is associated with a progressive decline in physiological function that is manifested at the molecular, cellular, and organ system levels. This decline in function is usually accompanied by a diminished ability to cope with environmental stress (2-4, 8, 17, 20, 25). However, the molecular mechanisms underlying age-related changes in stress responses are not well defined. Previous studies investigating alterations in gene expression have generally been confined to small sets of genes and have been focused on discrete cellular pathways. Specific areas of focus have included modifications in transcriptional regulation of stress-inducible genes (25), blunted synthesis of acute-phase proteins such as heat-shock proteins (5,20), accumulative damage to proteins and DNA and resultant alterations in gene expression patterns (3, 4, 6, 25), and increased oxidative stress and subsequent biomolecular damage resulting from an increased rate of reactive oxygen species (ROS) generation (2).
The aim of this study was to examine the transcriptional response pattern of a wide array of stress-response genes to aging and environmental heating. We postulated that a comparison of gene expression profiles between young and aged animals responding to a physiologically relevant stress such as hyperthermia would provide insight into molecular mechanisms underlying age-related alterations in stress responses. To accomplish this aim, a cDNA expression array system was utilized. This recently available technique is advantageous because the expression profile of a large number of genes can be assessed in a defined set of physiological conditions (26). In addition, this strategy has the potential for identifying specific genes, expression patterns, or cellular pathways associated with both stress and aging that have not been previously identified.
In the present study, oligonucleotide-based arrays containing 207 stress-related genes were utilized to simultaneously compare gene expression profiles in the livers of young and senescent rats at a selected time point after hyperthermic challenge. Alterations in the expression of selected genes on the array were also confirmed with RT-PCR techniques. We focused on the liver because it shows age-dependent evidence of increased cellular ROS production (33) and is a prime target of tissue injury in physiological challenges such as heat stress (11, 18) and ischemia-reperfusion (10). Experimental animals were heat stressed on two consecutive days, separated by 24 h. This design was based on our observations that older rats are markedly less thermotolerant and have a much greater degree of cellular injury than their younger counterparts after repeated heat challenge (11, 12, 21). Thus this experimental design provides an excellent in vivo model for the high morbidity and mortality rates observed in older humans with a stress such as hyperthermia (23,27).
Young (6-mo-old; n = 6; 300–400 g) and old (24-mo-old; n = 6; 350–450 g) male Fischer 344 rats (National Institute on Aging) were used in these experiments. Rats were housed in The University of Iowa Animal Care Facility, and all experimental procedures conformed to institutional animal care guidelines. Animals were maintained at 22–24°C on a 12:12-h light-dark cycle and were provided with food (standard rat chow) and water ad libitum. Rats were randomly assigned to four experimental groups with three rats per group: young control, young heat-stressed, old control, and old heat-stressed groups.
Experimental Procedures
All rats were handled daily and familiarized to a colonic temperature (Tco) probe during the week before experimentation. The heating protocol utilized has been described previously (11). On the day of an experiment, rats were fitted with a thermistor temperature probe (Yellow Springs Instruments) inserted 6–7 cm into the colon; rats were then placed, conscious and unrestrained, in plastic cages (45 × 25 × 20 cm). Tco was monitored constantly on a digital display. A baseline Tco (37.0–38.0°C for both age groups) was established over a 30-min control period. During the heating protocol, an infrared lamp, positioned ∼40 cm above the rats, was either raised or lowered to obtain an ambient temperature (Ta) of 38–40°C. Movement of the lamp permitted a constant heating rate (∼0.06°C/min) to be attained. Heating was stopped when Tco reached 41°C; however, heating was resumed at appropriate times to maintain Tco at 41°C for 30 min. At the end of this period, the thermistor probe was removed and rats were allowed to passively cool in a cage at room temperature. Animals were subsequently subjected to a second heating protocol 24 h after the first stress. Sham-heated control rats were handled identically to experimental rats, with the exception that Ta was maintained at 22–24°C. At the 2-h time point after the second heating, rats were administered an overdose of pentobarbital sodium (80 mg/kg ip). Liver biopsies were collected, rinsed in phosphate-buffered saline, and then immediately frozen in liquid nitrogen.
RNA isolation.
Total liver RNA was isolated by the guanidinium acid-phenol-chloroform extraction method (7). First, liver tissues from three animals were ground in liquid nitrogen and then pooled. One gram of ground liver tissue was homogenized in a denaturing buffer containing guanidinium acid. Liver protein and DNA were separated from total RNA by three rounds of phenol-chloroform extraction. Total liver RNA was then precipitated by isopropanol. The concentration and quality of the RNA samples were determined with both a spectrophotometric method (a ratio of 260 nm-to-280 nm) and agarose gel techniques.
cDNA array hybridization.
Rat stress-related expression cDNA arrays from Clontech Laboratory (Palo Alto, CA) were used for gene expression profiling. In these arrays, 207 stress-related genes were immobilized in duplicate on a nylon membrane. A total of four membranes were used, and all procedures were followed as recommended by the manufacturer. Liver mRNA was first purified from DNase-treated total RNA with oligo(dT)-linked Oligotex resin (Qiagen). One microgram of mRNA was used to generate [32P]dATP-labeled double-strand cDNA fragments by RT. The labeled fragments were then hybridized onto the DNA array membranes overnight at 68°C. Membranes were subjected to stringent washing three times at 68°C and then exposed at room temperature overnight to a phosphorimaging filter. Images were generated using a phosphorimaging system (Molecular Dynamics Typhoon 8600 variable mode imager, Amersham Pharmacia Biotech).
Microarray phosphorimage assessments.
To ensure accuracy in comparisons among arrays and to avoid saturation of the microarray images, the saturation level of the phosphorimager was first tuned to a low setting so that the genes most abundantly expressed were compared. The majority of the genes were then compared with a normal saturation level, with a few very abundant genes showing oversaturation levels. Very weakly expressed genes were also compared at a setting at which ∼50% of genes were oversaturated. However, this condition did not provide additional information compared with values obtained at the second setting. Instead, the third setting served as a confirmation of the results obtained from the second setting for the genes that were weakly expressed.
To confirm results of the cDNA array, RT-PCR was performed for four genes that exhibited distinct changes in expression with aging and after heating. The genes examined were heat shock protein 27 (HSP27), liver catalase, glutathione transferase subunit P, and c-Jun NH2-terminal kinase 1. Two micrograms of pooled total liver RNA were first converted to cDNA by Moloney murine leukemia virus RT and oligo(dT) according to a protocol provided by Clontech. DNA fragments corresponding to the four genes of interest were synthesized with the use of primers specific to these genes (Clontech). The PCR reaction mixture contained 1 unit of Taq DNA polymerse, 1× reaction buffer with 1.5 mM MgCl, 0.4 μM each of forward and reverse primers, 0.2 mM each of dATP, dTTP, dGTP, and dCTP, 10 μl of RT reaction product, and 1 μl each of sense and anti-sense primers (10 pmol/μl). The primers for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were added simultaneously for each of the four genes being assessed. The PCR cycle was started with a denaturing period (94°C for 3 min), followed by 38 cycles of 1 min at 94°C, 1 min at 55°C, and 1.5 min at 72°C. The PCR cycle was finished with a 7-min extension time at 72°C. DNA fragments were stored at 4°C until they were separated on a 2.5% agarose gel. The separated DNA fragments were visualized under an ultraviolet light with ethidium bromide, and images were obtained using an AlphaImager digital camera (Alpha-Innotech, San Leandro, CA). RT-PCR was repeated three to four times for each gene.
Data Analysis
Images from cDNA arrays were analyzed by using computer software developed by Adryan et al. (1). To analyze gene expression profiles between ages and after heating, detected hybridization signals were first subtracted from the background and then normalized to the signals of the two housekeeping genes: tubulin α-1 and GAPDH (Clontech). A change in hybridization signal was considered significant if there was greater than a twofold difference in intensity between groups (e.g., young vs. old, control vs. heated), which is consistent with previous studies that used similar experimental approaches (22, 24, 31). PCR band densities were analyzed by National Institutes of Health Image software. The density of a band corresponding to gene expression level was normalized to the band of GAPDH, which was analyzed simultaneously for the four genes being assessed. GAPDH was used to ensure that PCR was the same for all groups, to affirm that the same amount of PCR product was loaded on the agarose gel, and to normalize the gene of interest (e.g., HSP27) to a stable gene that should not change expression levels. Changes in gene expression between age groups and after heating are expressed as multiples of differences (fold).
DNA Arrays
Rat cDNA arrays containing 207 stress-related genes were utilized to evaluate gene expression profiles in liver samples from young and old rats in control conditions and at the 2-h time point in recovery from hyperthermic challenge. Representative phosphorimages are presented in Fig. 1. The phosphorimages from microarray experiments were analyzed to generate three pair-wise comparisons between ages (i.e., young vs. old for control) and at the 2-h time point after heat stress (i.e., control vs. heated for each age).
Validation of DNA Array Data
RT-PCR was performed on liver samples from young and old rats subjected to control and heat stress treatments to confirm the cDNA array results. Among the majority comparisons, the mean ratios obtained from RT-PCR were very similar to those from microarray experiments, although only two of the four genes evaluated in comparisons between young and old control animals showed similar trends (Table1). A representative RT-PCR gel for each of the four genes evaluated and the ratio of comparisons for age groups and treatment conditions between individual RT-PCR experiments are shown in Fig. 2.
Global Changes in Gene Expression With Age and Heat Stress
Hybridization signals from the phosphorimages were counted as total number of genes detected. In young animals, 64 genes were detected in the control condition compared with 73 genes at 2 h after stress with the DNA array technique. Old animals had fewer genes detected in the control condition (53 genes detected) but a more robust stress response (88 genes detected) at 2-h postheating than their young counterparts. Surprisingly, hybridization signals for heat shock protein 70 and heat shock factor were not detected on arrays at any time point with the different saturation settings on the phosphorimager.
Both aging and heat stress produced changes in gene expression (induction or suppression) in the liver. Table2 shows the percentage of genes that had at least twofold changes in gene expression with aging and after heat stress. In comparisons between age groups, 9.7% of the 207 genes measured had changes in expression, with 5.3% of the genes induced and 4.3% of genes suppressed in old compared with young animals. After heat stress, changes in gene expression of 12.1 and 15.9% were obtained in the young and old groups, respectively. In both groups, a greater number of genes were induced than suppressed with heat stress.
These genes were further grouped into four categories according to the proposed functions of the proteins they encode. These categories included 1) proteins in cell growth, death, and signaling pathways; 2) stress-response proteins; 3) antioxidant and drug metabolism proteins; and 4) DNA, RNA, and protein synthesis-related enzymes. An overview of the changes in gene expression with aging and heat stress is shown in Table3. Aging per se induced marked alterations in gene expression. These alterations included a greater basal expression of genes for cell growth, death, and signaling, as well as genes involved in stress responses (e.g., heat shock proteins). However, a striking suppression of genes for antioxidant and drug metabolism enzymes was noted in senescent livers. In addition, both young and old animals responded to heat stress with changes in their gene expression profiles. The most striking distinction between young and old animals was the difference in expression of stress-response genes associated with heat stress. After being subjected to heating, stress-response genes were strongly induced in young animals, whereas transcription levels for these stress-response genes were suppressed in the old animals.
Changes in Gene Expression With Age
Age-related changes in gene expression for nonstressed control animals are shown in Table 4. Only genes with at least a twofold change in detectable hybridization are presented. Twenty of the 207 genes examined displayed a change in gene expression as a function of age in the liver, with 11 genes upregulated and 9 genes downregulated in old compared with young animals. Older animals experienced induction of genes related to cell growth, death, and signaling pathways (6 of 9 genes), as well as induction of the majority of stress protein-related genes (4 of 5; Table 4). However, the expression of genes encoding antioxidant and drug metabolism enzymes was greatly suppressed with aging (6 of 6).
Changes in Gene Expression With Heat Stress
Hyperthermia can induce or suppress a wide variety of genes. To identify the gene expression profiles in young and old animals after heat stress, the transcription levels of 207 stress-related genes were compared at 2 h after heating for each age group. For the young cohort, hybridization levels changed in 25 of the 207 genes surveyed (Table 5). Among these changes, 18 genes were induced and 7 were suppressed. Old animals had a robust response to hyperthermic challenge. The expression of 33 of the 207 stress-related genes was changed in old heat-stressed animals compared with their age-matched controls. Among these genes, 23 were induced and 10 were suppressed at the 2-h recovery point.
When these genes were grouped into functional categories similar to Table 3, old animals had a larger number of changes in gene expression for transcripts in the cell growth, death, and signaling category than young animals after heating (10 vs. 6 genes; Table 5). Within this category, 4 of 6 genes in young animals and 6 of 10 genes in old animals were induced. For stress-response genes, aged animals exhibited a general downregulation in gene expression (5 of 7), whereas young animals had a strong pattern of induction (4 of 5). In the category of genes encoding antioxidant and drug metabolism enzymes, both young and aged animals showed a broad induction in gene expression (6 of 7 for young and 10 of 10 for old). Many genes in the DNA, RNA, and protein synthesis category were upregulated in both groups (4 of 6 for young; 5 of 6 for old) in response to heat stress.
The objective of this study was to examine the transcriptional response pattern of an array of stress-response genes to environmental heating and evaluate the influence of biological aging on these responses. The cDNA array technique provides a sensitive and rapid method for comparing the expression of a large number of genes among a variety of physiological conditions. Information obtained with the use of this strategy also provides insight into the selection of specific genes to evaluate in greater detail and allows for the development of a mechanistic framework regarding the molecular changes associated with the decline in stress tolerance in aged organisms. In our study, use of cDNA expression arrays specific for stress-related genes produced a broad profile of changes in gene expression in the liver after hyperthermic challenge in young and old rats. We focused on the liver because previous studies from our laboratory have demonstrated that this organ plays an important role in an organism's ability to cope with an environmental stress such as hyperthermia (11, 12,19).
In one of the first studies to use oligonucleotide-based arrays to address changes in gene expression with aging, Lee et al. (22) reported that 113 of the 6,347 genes surveyed (2%) had greater than a twofold difference in gene expression in skeletal muscle samples from young compared with old mice (22). In the present study, a higher percentage (∼10%) of genes surveyed showed differential expression with aging. These results appear quite reasonable because all 207 genes contained on the cDNA array utilized are thought to be indicators of stress responses. In addition, the higher rate of differences in expression of stress genes between young and old animals noted in the present study is consistent with the general concept that biological aging is characterized by reduced stress tolerance (2, 20, 25). In support of this concept, our results demonstrate that the older animals had less detectable gene expression signals (53 genes detected) than young animals (66 genes detected) in control conditions but a larger induction of genes after heating (15.9 vs. 12.1%).
Aging is associated with elevated morbidity and mortality rates in response to a variety of challenges (2, 13, 20, 25, 27), thus suggesting a decreased tolerance to stress with advancing age. In studies that compared young and old rats, we previously observed that aged animals are less thermotolerant, have a blunted stress protein response, and manifest extensive cellular injury in tissues such as the liver when heat challenged (11, 20, 21). Therefore, we hypothesized that the pattern of gene expression in old animals would be significantly different from that in young animals. Our results indicate that there is a blunted gene expression profile in old compared with young rats in the nonstressed control condition but a greater response to heat stress with advanced age.
To further evaluate the expression pattern of young and old animals, genes that had at least a twofold change in hybridization were grouped into four functional categories (Table 3). A distinct response pattern was observed in two of the four categories. For example, the old group exhibited an upregulation of several stress-response genes in control conditions. However, these older animals appeared unable to respond to heat stress properly, as demonstrated by the number of stress-response genes that were suppressed after hyperthermic challenge. In contrast, there was a strong induction of these stress-response genes in young animals. These results are in agreement with previous observations from our laboratory, indicating that aging is associated with decreased stress tolerance (11, 12, 19, 20). Our gene profile analysis also gives insight into specific genes that may be responsible for the age-related decrease in stress tolerance. Further studies on the regulation and expression of these genes, as well as the functional characteristics of their proteins, will advance our understanding of the mechanisms underlying age-related alterations in stress tolerance.
One proposed mechanism for aging is the oxidative stress hypothesis, which states that the loss of functional capacity associated with senescence is due to the progressive and irreversible accumulation of oxidative damage (2, 29, 32). It is postulated that the increase in oxidative stress and subsequent damage associated with aging are due to an increased rate of ROS generation, decreased removal of ROS, and/or a greater susceptibility of tissues to oxidative damage. Besides their damaging effects related to aging, ROS can also modulate gene expression, which certainly expands their role in the aging process (15). For example, it has been suggested that the increased oxidative injury that accompanies aging can result from a decrease in antioxidant enzyme defenses (9). The observation in the present study that old animals had a markedly reduced gene expression pattern in the antioxidant enzyme and drug metabolism category supports the basic tenets of the oxidative stress theory of aging.
Transcriptional levels of several genes encoding antioxidant enzymes were altered in response to heat stress. These observations, along with previous studies from our laboratory (11), indicate that ROS generation may play an important role in the liver pathology that accompanies heat stress. A potential source of superoxide radical generation could be from the leakage of mitochondrial electron transport chain reactions (14). Increased production of superoxide radicals by xanthine and/or xanthine oxidase after heat shock has also been reported (28). These superoxide radicals can be further converted to hydrogen peroxide by superoxide dismutases. Excess amounts of hydrogen peroxide, superoxide, and their by-product, hydroxyl radicals, which are produced through Haber-Weiss reactions, can either cause direct damage to cellular macromolecules or act as secondary messengers to produce intracellular oxidative stress. Thus our data implicate the involvement of an oxidative stress mechanism in the decline in stress tolerance that accompanies aging.
When evaluating the current set of data, there are potential limitations impacting the interpretation of the results that warrant discussion. The first involves the use of a single time point after heating (2 h) to evaluate differences in gene expression between age groups. It is possible that the time required to express genes could be altered with age. For example, the observation that there was no age-related difference in the expression of a particular gene at the 2-h time point after heat stress does not preclude the possibility that a difference in gene responses could be manifested at other time points poststress. However, preliminary experiments associated with the present study suggest that the general pattern of alterations in stress gene expression observed between age groups was similar at 2, 12, and 24 h after heat challenge. Additional studies addressing the temporal pattern of gene expression in response to a stress are certainly warranted. A second consideration involves the analysis of pooled liver samples from three animals for each microarray experiment. An experimental design in which a sample from a single animal was analyzed on a microarray would have allowed some insight into the consistency or repeatability of the gene expression patterns observed with aging and heat stress. However, several laboratories have used pooled samples in their gene expression studies (16, 24,30), in part because this approach can reduce interanimal variability and increase the reliability of results.
Taken together, the DNA array results suggest that older animals do not respond to thermal challenges with an appropriate pattern of gene induction. Thus the ability of older animals to maintain cellular integrity and stability may be compromised. Our data also indicate that a physiological challenge such as heat stress generates a gene expression profile in the liver indicative of decreased stress protein transcription and increased expression of oxidative stress-related genes. These findings support the postulate that changes in transcriptional responses to physiological challenges such as hyperthermia contribute to the reduction in stress tolerance in older organisms. The use of the cDNA expression array approach will likely assist in delineating specific genes and molecular pathways associated with aging and the integrated cellular responses to physiologically relevant stressors.
We acknowledge the technical support of Linjung Xu and Joan Seye.
This research was supported by National Institute on Aging Grants AG-12350 and AG-14687.
Address for reprint requests and other correspondence: K. C. Kregel, Integrative Physiology Laboratory, 532 FH, The Univ. of Iowa, Iowa City, IA 52242 (E-mail:kevin-kregel{at}uiowa.edu).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
First published November 2, 2001;10.1152/japplphysiol.00733.2001
- Copyright © 2002 the American Physiological Society

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Uzo Aduba stars as Suzanne "Crazy Eyes" Warren on the Netflix's Orange Is the New Black and she recently sat down for a SAG Conversation to chat about her career, OitNB, acting and more.
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← Previous Story Watch: SAG Conversations with the Cast of 'Life in Pieces'
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William H. Macy: "This may sound pretentious, but I am getting better at what I do every day"
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Photographs, news ads and articles, excerpts from student handbooks, and nearly 60 clothing items tell the story of student life as reflected in a century of style. Continue reading
Pioneering female photographers helped launch the historic preservation movement in North Carolina. Continue reading
During the 1930s, photographers Bayard Wootten and Frances Benjamin Johnston documented the early architecture of the American South. Continue reading
Dr. David Moore will discuss excavations at the site near Morganton, N.C. It was the earliest European settlement in what is now the United States. Continue reading
North Carolina Digital Heritage Center received an "Award for Excellence in Web Presence" on Oct. 3 from the North Carolina Genealogical Society. Continue reading
Program with Don Tate, author and illustrator of a new children's biography about Chatham County slave and poet George Moses Horton. Continue reading
Cookbooks from the North Carolina Collection showcases the range and history of cooking in the Tar Heel State in an exhibition on view in Wilson Library. Continue reading
Areas of the Wilson Special Collections Library will be closed to the public on June 19. We apologize for the inconvenience. Continue reading
Sarah Carrier joined the staff of the Wilson Special Collections Library at UNC on June 1. Continue reading
A blog that allowed readers to follow the Civil War in the words of those who lived it reached its end on April 26, after 1,450 posts. Continue reading


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- //Home
- //Music
- //Ninjasonik
No Swords Or Masks EP
US: 6 Mar 2012
UK: Import
Reverend McFly and Telli Gramz, two parts of one Ninjasonik, are known to stir up a solid ruckus live, but whatever wizardry they take to the stage is either impotent or absent in the studio: It's hard to see what could possibly get anyone excited about their flaccid alloy of electro and hip-hop. Absent the rapidfire spitwads of Bushwick basement touchstones (Pabst, tattoos, skinny jeans) that no doubt helped endear their debut Art School Girls to the Street Carnage hype machine, the duo and a bevy of hired guns double their track lengths on No Swords or Masks to accomodate terrible singing and longer verses extolling the virtues of bathroom sex and themselves, among other things. Flashes of vocal dexterity show some growth in Telli's MCing, but the Reverend remains mostly a fashion icon, and together they're still dumb as nails, which the redemption raps of "Around the World" and "Gave It Everything" make painfully clear.
Goodwood Atoms "Into the Bay" (audio) (premiere) // Music
Barnaby Carter "Evening Redness" (audio) (premiere) // Music
Apollo Brown and Skyzoo "A Couple Dollars (feat. Joell Ortiz)" // Music
How Röyksopp's 'Melody A.M.' Brought Electronica Into the Mainstream
// Sound Affects
"With their debut, the Norwegian duo essentially provided the everyman's guide to electronic music.READ the article


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2 Held, 1 Sought in West Plains Homicide
WEST PLAINS -- Authorities in south-central Missouri say two people have been arrested and a third is being sought in the killing of a man near West Plains.
The body of 29-year-old Jackie Yarber, of Birch Tree, was discovered Saturday off Howell County Road 69-70. KYTV reports (http://bit.ly/qBz6hw) that sheriff's officers received a tip about the death Friday night. A 24-year-old man and a 21-year-old woman, both from West Plains, were arrested Monday.
Investigators believe Yarber was killed while three people were stealing his truck. Prosecutors say it appears he was run over or stabbed to death.
More News
COLUMBIA - The holiday season got off to an early start Monday with the beginning of the Almeta Crayton Memorial... More >>
ASHLAND - Candidates for Southern District Boone County Commissioner faced off in a political forum at Southern Boone County High... More >>
BOONE COUNTY - The Missouri State Highway Patrol issued an endangered person advisory for a woman on Monday. According... More >>
SPRINGFIELD (AP) — A nonprofit said drought and population growth may put a strain on water sources in southwest Missouri.... More >>
KANSAS CITY, Mo. (AP) — A Kansas City-area man is accused of duping dozens of women into having sex with... More >>
FULTON – The Brick District Playhouse Theater will be able to install an HVAC heating/cooling system thanks to a Rural... More >>
KANSAS CITY, Mo. (AP) — Missouri said the U.S. Constitution doesn't guarantee a reporter or anyone else a right to... More >>
JEFFERSON CITY – River City Habitat for Humanity celebrated its 99 th home opening Monday night with the dedication of... More >>
COLUMBIA – MoDOT began road work on Business Loop 70 between College Avenue and Illinois Avenue Monday. MoDOT said... More >>
MOBERLY - The Randolph County Health Department confirmed several shigellosis cases in Moberly Public Schools during September. It now reports... More >>
JEFFERSON CITY — Gov. Jay Nixon attended a round table discussion in Jefferson City, which is the first in a... More >>
COLUMBIA - A car ran into the side of a home off of Rainbow Trout Circle, Monday. The driver and... More >>
COLUMBIA - Tracking foot traffic in Columbia might be near impossible right now, but that information could be invaluable to... More >>
COLUMBIA — If you live in Boone County and are looking to buy a home, you have to move fast.... More >>
COLUMBIA — When I meet with new clients, I ask them about their fitness and wellness goals. What are their... More >>
KINGDOM CITY (AP) — Authorities say one person was killed and another injured in an explosion at a central Missouri... More >>
BERKELEY (AP) — A St. Louis newspaper says St. Louis County prosecutors and the FBI are investigating allegations of absentee... More >>
ST. JOSEPH (AP) — Authorities say an 18-year-old has been charged in the death of a woman whose body was... More >>
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Islamabad:Any players wishing to play for Pakistan cannot continue to play in the unsanctioned Indian Cricket League, the Pakistan Cricket Board said on Wednesday, adding that it would decide on what penalties to impose on ICL players once they resign from the competition.
"Players must resign from the ICL if they are to be considered for selection by the PCB," the statement said.
Earlier this month the International Cricket Council rejected an ICL application for official recognition at its board meeting in Dubai, but left it up to the individual cricket boards to decide on what penalties to impose on players who decide to return to officially-sanctioned cricket.
The Board of Control for Cricket in India on Wednesday announced an amnesty for players involved in the ICL if they sever ties with the unsanctioned Twenty20 tournament by the end of next month.
The Indian players can participate in domestic cricket immediately, but will have to wait for a year before they are considered for selection for the Indian national team.
The PCB has already lifted a domestic ban on several Pakistani players competing in the ICL.
"The PCB will decide on a case-to-case basis the penalties to be imposed and the time period for which they will remain banned," the PCB said.
The PCB has imposed a deadline of the end of May for all Pakistani ICL players to decide their future.


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Mara M. Watkins, M.A., LPC-Intern
|Professions: Counseling Psychology, ,
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You are human and as humans we all have our low moments. It helps to get an objective point of view sometimes. Usually when we talk to someone close to us about a problem they might start talking about their experience, or tell you what you should and shouldn't do or feel. I see therapy as more of a luxury because it use to be only accessible to the wealthy and celebrities. Movies and TV shows have created an unrealistic perception of therapy and those seeking therapy. You might feel like the only one but take a moment to google celebrities with mental illness and you will be surprised because you probably never thought of them as flawed. Think of it like watching a movie, you're able to see the whole picture and things the characters in the movie can't see. The characters on the other hand are just going along unaware of the bigger picture and factors affecting their outcome. If you were able to warn the characters in the movie and tell them about the parts they are missing then they could possibly change the ending. Therapy is like having someone view your life movie while giving you insights so you might gain new perspectives and a chance to enhance your experience. It's like having a secret weapon to combat issues in your life because no one will know you're in therapy unless you tell them.
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Full name Jonothan Vincent Chisholm
Born November 13, 1980, Gladstone, Queensland
Current age 35 years 350 days
Major teams Queensland Country
Batting style Right-hand bat
Bowling style Right-arm fast
This is his first season of representative cricket and began with selection in the North Queensland team for their two matches against Victoria Institute of Sport at Mackay in September. Though his bowling was rather expensive in the match for North Queensland against South Queensland he shows plenty of potential for future development and his selection is seen as a worth while investment. He was educated at Trinity Beach State School, St Augustines College, Cairns and is now a student at TAFE in Cairns. (Warwick Torrens, December 2000)
Club: Barron River, Cairns
Dhoni's 9000 runs: 244 innings, 10109 balls
Almost two-thirds of Dhoni's 9000 ODI runs have come as captain and all of them with the extra responsibility of keeping
Virat Kohli and MS Dhoni dominate the stats highlights from the Mohali ODI
Dhoni revels in freedom to play at No. 4
India's limited-overs captain has wanted to bat up 'for a long time'; when he got the chance in Mohali, he played the big shots unhindered by the pressures of finishing a chase
Rossouw comes in from the cold with eye on summer Tests
A thoughtless remark nearly derailed the South African batsman's career before it began. And then he got five ducks
An all-time Pakistan Test XI chosen by our readers
Recent middle-order mainstays make it to Pakistan's all-time XI selected on the occasion of the country's 400th Test
Test or T20? The choice that changes cricket (96)
Australia's clash of Twenty20 and Test fixtures, with players forced to choose between them, will come to be seen as a tipping point for the international game
Explainer - Why the BCCI changed its stance on DRS (83)
Higher frame rates for cameras and a safeguard to prevent operator error were the key improvements in technology that led to the BCCI agreeing to trial the system
Dhoni revels in freedom to play at No. 4 (71)
India's limited-overs captain has wanted to bat up 'for a long time'; when he got the chance in Mohali, he played the big shots unhindered by the pressures of finishing a chase
Resilient Johnson's ups and downs (52)
Talking points from Mitchell Johnson's new autobiography
Taking 20 wickets against England was a significant step forward for Bangladesh - but work is required, particularly on pace bowling, to maintain their progress


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Discover a selection of top forums tabletop vision on the internet. Come to share your passion on forums tabletop vision and meet other fans of tabletop vision. Then, why don't you create your own forum of tabletop vision ?
Free public forum for advanced and productive discussion of philosophy and related subjects
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Free forum : Bringing the Collection of Thinkers community back together again.
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The place to come to find a group or player for any of your tabletop adventures.
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There's no direct allusion to the former Wisconsin/North Carolina State star this week, but it might be worth noting how difficult it is for a player chosen after the second round to start on Opening Day in his rookie season. The last time a rookie selected after the second round started a season opener was way back in 2005, when Kyle Orton, a fourth-rounder with the Bears and the 106th prospect off the board that year, opened the campaign.
"Believe me, it's really on-the-job training," Orton, now the backup to Tony Romo in Dallas, told The Sports Xchange.
The former Purdue quarterback was 10-5 as a starter his rookie season, including an eight-game winning streak.
There were 46 quarterbacks picked after the first two rounds in the six-year stretch 2006-11, and none started the season opener as a rookie for his respective team.
The undersized Wilson, plucked by the Seahawks in the third round, with the No. 75 overall choice, has overcome long odds most of his career. And the past isn't necessarily a prelude to the future or a predictor of how he'll eventually fare in the league. But the strong endorsement of coach Pete Carroll aside, Wilson will have a rough time overcoming the recent history of passers taken in the third round or later.
So it's no surprise that Carroll has tabbed six-year pro Tarvaris Jackson to take the first-team snaps at the outset of camp. A much bigger surprise will be if it's Wilson at the top of the depth chart for the Seahawks' opener at Arizona on Sept. 9.
"I really do not believe it's a big deal. I just think that they don't, in general, give (short quarterbacks) the opportunities, or stick with them. A guy that's undersized has to prove himself right away. And if you don't have success right away, you're out the door. (It's) similar to a (Tim) Tebow situation. If Tim Tebow had taken over the Broncos last year and lost his first two or three games, he'd never have seen the field again. Instead, he won whatever it was - five, six in a row - and they kept doubting him until next week, until next week, until next week. (But) if he were the 6-4 guy who was a true pocket passer, drafted in the first round and the franchise decided, 'This is our guy,' then he goes out and loses his first three or four starts . . . well, there is a learning curve involved. And they have patience with him. And that's my frustration with it." - former NFL quarterback Doug Flutie, per NFL.com, on the league's perceived bias against short quarterbacks


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