The Fourth National Institutes of Health Symposium on the Functional Genomics of Critical Injury: Surviving stress from organ systems to molecules.

Recent strides in computational biology and high-throughput technologies have generated considerable interest in understanding complex biological systems. The application of these technologies to critical illness and injury offers the potential to define adaptive and maladaptive programs of gene expression induced by infection, shock, trauma, or other inflammatory triggers, and to detect biomarkers and genetic polymorphisms linked to these responses and outcome. A systems biology approach is timely because despite substantial effort, treatment approaches directed at a single mediator or inflammatory pathway have met with little success in altering outcomes of critically ill or injured patients. Highlights from the Fourth National Institute of Health Functional Genomics of Critical Illness and Injury Symposium are described herein, in addition to deliverables for the field identified during panel discussions. Next steps for the community and suggestions for future research are presented.

Physiological responses of over-wintering common carp (Cyprinus carpio) to disturbance by Eurasian otter (Lutra lutra).

Using a tame animal, the impact of otter (Lutra lutra) disturbance on over-wintering carp (Cyprinus carpio) was monitored in two experiments, 133 and 140 days, respectively, over two consecutive winters (November-April). The level of stress in over-wintering carp exposed to various intensities of disturbance by otters was quantified using biological indicators of stress (cortisol, cortisone, indices of nitrogen, carbohydrate, lipid and mineral metabolism and activity of basic blood plasma enzymes) taken from blood plasma of stocked carp at the end of the winter seasons (when the photoperiod was 12 light:12 dark, respectively, 13L:10D). Moreover, condition (Fulton's coefficient of condition and fat content in muscles) and mortality rate of that carp were measured after over-wintering and also after the subsequent vegetation period. The analysis of blood and tissue samples of experimental fish showed changes in nitrogen, carbohydrate and mineral metabolism as well as levels of hormones and fat reserves. Higher response to stress in metabolism of carp with lower intensity of disturbance by otter suggests that high level of disturbance can lead to metabolic adaptation of carp to stress. The effect of stress on the mortality rate of carp during the over-wintering is not clear. Nevertheless, the negative effect of stress on survival, condition and growth rate of carp in the subsequent vegetation period was not observed.

Frequency of the HAL-1843 mutation of the ryanodine receptor gene in dead and nonambulatory-noninjured pigs on arrival at the packing plant.

Four Midwestern packing plants (designated as plants A, B, C, and D) were visited on 53 occasions, and tissue samples were collected postmortem from a total of 2,019 pigs to determine the frequency of the HAL-1843 mutation of the ryanodine receptor gene in dead (DOA), nonambulatory-noninjured (NANI), and normal animals. The sampled pigs came from approximately 130,000 animals from 454 farms and were transported on 861 trailer loads, with an average of 152 pigs/load and an average pig live BW/load of 125 (SD 7.02) kg/pig. Frequency of animals with the HAL-1843 mutation was low, with only 2.7% of the pigs being either homozygous recessives (nn, 0.45%) or carriers (Nn, 2.3%) for the mutation and 97.3% of the pigs being homozygous for the normal allele (NN). The mutation was present in all 3 classes of pig, with 1.8% of normal, 1.8% of NANI, and 4.7% of DOA animals having at least 1 copy. Two of the plants (A and C) had a greater frequency (P < 0.05) of carrier (3.7 and 3.5 vs. 1.1 and 1.0 for plants A and C vs. B and D, respectively) and homozygous recessive (1.0 and 0.9 vs. 0.0 and 0.0, respectively) animals than the others (plants B and D). There was a greater frequency (P < 0.05) of carriers in DOA animals than in the normal or NANI pigs (3.7 vs. 1.7 and 1.5 for DOA vs. normal and NANI, respectively). The 55 pigs that had at least 1 copy of the mutation came from 53 farms; therefore, the mutation was relatively widespread, being present in approximately 11% of the farms sampled. Although the HAL-1843 mutation is still present in commercial pig populations in the United States, its low frequency in DOA and NANI pigs suggests that it is not a major cause of these transport losses.

Warm weather transport of broiler chickens in Manitoba. I. Farm management factors associated with death loss in transit to slaughter.

This observational study was conducted to identify flock of origin level factors associated with mortality during transport to slaughter of 1 090 733 Manitoba broiler chickens in spring and early summer. The death loss in transit was 0.346% (3778 birds). Death loss on the farm of origin during the growing phase of production and bird body weight at slaughter were associated with increased mortality in-transit. Death loss during production significantly exceeded, while crowding of growing birds was significantly less than, European proposed animal welfare standards.

Diminished lifespan and acute stress-induced death in DNA-PKcs-deficient mice with limiting telomeres.

An adequate and appropriate response to physiological and pathophysiological stresses is critical for long-term homeostasis and viability of the aging organism. Previous work has pointed to the immune system, telomeres and DNA repair pathways as important and distinct determinants of a normal healthy lifespan. In this study, we explored the genetic interactions of telomeres and DNA-PKcs, a protein involved in non-homologous end-joining (NHEJ) and immune responses, in the context of a key aspect of aging and lifespan--the capacity to mount an acute and appropriate immune-mediated stress response. We observed that the combination of DNA-PKcs deficiency and telomere dysfunction resulted in a shortened lifespan that was reduced further following viral infection or experimental activation of the innate immune response. Analysis of the innate immune response in the DNA-PKcs-deficient mice with short dysfunctional telomeres revealed high basal serum levels of tumor necrosis factor alpha (TNFalpha) and hyper-active cytokine responses upon challenge with polyinosinic-polycytidylic acid (poly-IC). We further show that serum cytokine levels become elevated in telomere dysfunctional mice as a function of age. These results raise speculation that these genetic factors may contribute to misdirected immune responses of the aged under conditions of acute and chronic stress.

Effects of travel distance and the season of the year on death rates of broilers transported to poultry processing plants.

Death losses among broilers transported to processing plants are caused by poor welfare. The number of birds dying during transport and in processing plants shortly after arrival there may serve as an indicator of the quality of welfare during loading and transport. In the Czech Republic, shipments of broilers to processing plants were monitored from 1997 to 2004. It was found that the mortality of broilers during transport was 0.247%, but it varied according to the transport distance to the processing plant from 0.146% (50 km maximum) to 0.862% (>300 km). The broiler mortality in transit was also influenced by the season of the year. The highest mortality was found in summer months, especially in June, July, and August, and in winter months, especially in December, January, and February. A comparison between period 1 (1997 to 2000) and period 2 (2001 to 2004) showed a long-term adverse trend in the number of broilers dying during transport to processing plants for all of the distances monitored (except transport distances <50 km), because the overall number of dead birds on arrival to processing plants increased from 0.224 to 0.265% (index of 1.18). This difference is statistically significant (P = 0.000). The stress caused to broilers by transport to processing plants is reflected in higher transport-related mortality of the birds. Long-term trends point to an increase in death losses of broilers. Longer transport distances and transportation in summer and winter months have led to an increase in death losses among broilers transported to processing plants.

Effect of floor space during transport of market-weight pigs on the incidence of transport losses at the packing plant and the relationships between transport conditions and losses.

Data on 74 trailer loads of finishing pigs (mean BW = 129.0, SEM = 0.63 kg) from wean-to-finish buildings on 2 farms within 1 production system were collected to investigate the effect of amount of floor space on the trailer (0.39 or 0.48 m2/pig) during transport on the incidence of losses (dead and nonambulatory pigs) at the packing plant and to study the relationships between transport conditions and losses. Pigs were loaded using standard commercial procedures for pig handling and transportation. Two designs of flat-deck trailers with 2 decks were used. Floor space treatments were compared in 2 similarly sized compartments on each deck of each trailer type. Differences in floor space were created by varying the number of pigs in each compartment. The incidence of nonambulatory pigs at the farm during loading and at the plant after unloading, average load weight, load number within each day, event times, and temperature and relative humidity in the trailer from loading to unloading were recorded. Of the 12,511 pigs transported, 0.26% were non-ambulatory at the farm, 0.23% were dead on arrival, and 0.85% were nonambulatory at the plant. Increasing transport floor space from 0.39 to 0.48 m2/pig reduced the percentage of total nonambulatory pigs (0.62 vs. 0.27 +/- 0.13%, respectively; P < 0.05), nonambulatory, noninjured pigs (0.52 vs. 0.15 +/- 0.11%, respectively; P < 0.01), and total losses (dead and nonambulatory pigs) at the plant (0.88 vs. 0.36 +/- 0.16%, respectively; P < 0.05) and tended to reduce dead pigs (0.27 vs. 0.08 +/- 0.08%, respectively; P = 0.06). However, transport floor space did not affect the percentage of nonambulatory, injured pigs at the plant. Nonambulatory pigs at the farm were positively correlated with relative humidity during loading and load number within the day (r = 0.46 and 0.25, respectively; P < 0.05). The percentage of total losses at the plant was positively correlated to waiting time at the plant, unloading time, and total time from loading to unloading (r = 0.24, 0.51, and 0.36, respectively; P < 0.05). Average temperature during loading, waiting at the farm, transport, waiting at the plant, unloading, and average pig weight on the trailer were not correlated to losses. These results suggest that floor space per pig on the trailer and transport conditions can affect transport losses.

Variation in the mortality rate of turkeys during transport to the slaughterhouse with travel distance and month.

Failure to comply with animal welfare requirements during the transport of turkeys to the slaughterhouse increases stress in animals, which is manifested by increased mortality rate during transport. The numbers of turkeys that died during transport or soon after arrival may serve as an important parameter to indicate the level of animal welfare during transport of turkeys. The number of turkeys that died during transport to slaughterhouses in the Czech Republic in the period from 1997 to 2004 was investigated. The mortality rate found was 0.28% +/- 0.06% but varied with travel distance. The lowest mortality rate was found in case of travel distance below 50 km (0.18% +/- 0.08%) while long travel distances resulted in considerable increase in the mortality rates of turkeys (between 0.28% +/- 0.07 and 0.37% +/- 0.10%). The mortality rate of transported turkeys was also affected by the particular month of the year. Thus, the highest overall mortality rate occurred at long travel distances during winter months, i.e. in December (0.34% +/- 0.18%), January (0.32% +/- 0.06%), and February (0.36% +/- 0.07%). The comparison of individual years has shown a long-term trend towards a decrease in turkeys' mortality during transportation to slaughterhouses from 0.32% in 1998 to 0.20% in 2004. The decrease was statistically significant (Spearman's rank correlation coefficient r = -0.86, p < 0.01). This trend can be evaluated as positive.

Is incomplete recovery from work a risk marker of cardiovascular death? Prospective evidence from industrial employees.

OBJECTIVE: A chronic lack of recovery from work during leisure time is hypothesized to indicate a health risk among employees. We examined whether incomplete recovery from work predicted cardiovascular mortality. METHODS: This prospective cohort study involved 788 industrial employees (534 men, 254 women, mean age 37.3, SD = 12.0) who were initially free from cardiovascular diseases. The baseline examination in 1973 determined cases of cardiovascular disease, cardiovascular risk factors, and the extent of recovery from work. Data on mortality in 1973 to 2000 were derived from the national mortality register. RESULTS: Sixty-seven cardiovascular deaths and 102 deaths from noncardiovascular causes occurred during the mean follow-up of 25.6 years. Employees who seldom recovered from work during free weekends had an elevated risk of cardiovascular death (p = .007) but not of other mortality (p = .82). The association between incomplete recovery and cardiovascular death remained after adjustment for age, sex, and 16 conventional risk factors, including occupational background, cholesterol, systolic pressure, body mass index, smoking, alcohol consumption, physical inactivity, depressive symptoms, fatigue, lack of energy, and job stress. The association was not explained by deaths that occurred close to the assessment of recovery from work. CONCLUSIONS: This study suggests that incomplete recovery from work is an aspect of the overall risk profile of cardiovascular disease mortality among employees.

MCP-1 is up-regulated in unstressed and stressed HO-1 knockout mice: Pathophysiologic correlates.

BACKGROUND: Up-regulation of heme oxygenase-1 (HO-1) occurs in, and often confers protection to, the injured kidney. Up-regulation of monocyte chemoattractant protein-1 (MCP-1) promotes not only acute and chronic nephritides but also acute ischemic and nephrotoxic injury. The present study was stimulated by the hypothesis that expression of MCP-1 is suppressed by HO-1, and analyzed the effect of HO-1 on the expression of MCP-1 in stressed and unstressed conditions. METHODS: Expression of MCP-1 and pathophysiologic correlates were examined in HO-1 knockout (HO-1-/-) and wild-type (HO-1+/+) mice in the unstressed state in young and aged mice, and following nephrotoxic and ischemic insults. RESULTS: In unstressed HO-1-/- mice, plasma levels of MCP-1 protein were elevated, and MCP-1 mRNA expression was increased in circulating leukocytes and in the kidney. Such early and heightened up-regulation of MCP-1 was eventually accompanied by phenotypic changes in the aged kidney consistent with MCP-1, namely, proliferative changes in glomeruli, tubulointerstitial disease, and up-regulation of transforming growth factor-beta1 (TGF-beta1) and collagens I, III, and IV. In response to a nephrotoxic insult such as hemoglobin, MCP-1 mRNA was up-regulated in a markedly sustained manner in HO-1-/- mice. In response to a duration of ischemia that exerted little effect in HO-1+/+ mice, HO-1-/- mice exhibited higher expression of MCP-1 mRNA, enhanced activation of nuclear factor-kappaB (NF-kappaB) (the transcription factor that regulates MCP-1), markedly greater functional and structural renal injury, increased caspase-3 expression, and increased mortality. CONCLUSION: In the absence of HO-1, expression of MCP-1 is significantly and consistently enhanced in unstressed and stressed conditions. We speculate that the protective effects of HO-1 in injured tissue may involve, at least in part, the capacity of HO-1 to restrain up-regulation of MCP-1.

Relationship between maximum daily temperature and mortality of broiler chickens during transport and lairage.

1. Based on records of all (n = 59 171 843) broiler chickens slaughtered over three years at one processing plant, the overall mortality of birds in transit was 0.126%. 2. There was a pronounced seasonal effect with increased mortality in the summer months, particularly June, July and August. Mortality increased when the maximum daily temperature (measured in the shade) rose above about 17 degrees C at the start of the summer. 3. Between 17.0 and 19.9 degrees C, mortality was 30% higher than at lower temperatures. Between 20.0 and 22.9 degrees C it increased 2.6-fold, and at temperatures of above 23 degrees C 6.6-fold. There was no evidence of birds dying from hypothermia at very low ambient temperatures (down to -1 degrees C). 4. The implication from the results is that above a maximum daily temperature of 17 degrees C steps may need to be taken to ameliorate the damaging effects of transport on bird welfare.

E-PASS (The Estimation of Physiologic Ability and Surgical Stress) scoring system helps the prediction of postoperative morbidity and mortality in thoracic surgery.

OBJECTIVE: When a new scoring system, 'E-PASS', standing for the Estimation of Physiologic Ability and Surgical Stress that predicts the postoperative surgical risk by quantification of the patient's reserve and surgical stress applied to a population of general thoracic surgery patients, it should be investigated if this system could help us or not. METHODS: The comprehensive risk score (CRS) of the E-PASS and the clinical course were evaluated retrospectively in 282 consecutive patients with primary lung cancer (group A), and in 458 patients who underwent elective thoracic operations (group B). RESULTS: The morbidity and mortality rates in both group A and group B increased as the CRS increased. The CRS correlated significantly with the morbidity score, length of stay and cost of hospitalization. CONCLUSIONS: E-PASS scoring system may be useful in surgical decision-making and evaluating quality of care in patients who are tolerable for lung resection.

Sociodemographic and clinical predictors of mortality in geriatric depression.

OBJECTIVE: It is well documented that depression and early mortality are related, and current research suggests that depression may influence vascular causes of death. The authors report on prospectively observed mortality in elderly depressed patients, comparing differences in sex and vascular risk. METHODS: The subjects were 338 patients with unipolar major depression; patients with neurological or other psychiatric disorders were excluded. All subjects received a clinical evaluation and a standardized interview to establish DSM-III depression diagnosis. The cohort had a mean age of 67.2 years and was followed up at approximately 10 years from last contact. RESULTS: Mortality was associated with older age, older age at depression onset, and being male. Also, in men, mortality was associated with higher baseline CES-D scores, and, in women, with having a higher vascular risk rating or late age at depression onset. CONCLUSION: This is the first study demonstrating a gender-by-vascular condition interaction effect on mortality. Women with vascular risk factors may require closer follow-up to control vascular conditions. The relationship between greater depression and increased mortality risk among men is interesting, and further studies will be required to replicate and understand it.

Detection of hormesis effect in longevity: simulation approach for heterogeneous population.

Manifestation of hormesis in longevity was modelled by modification of the mortality rate during and after the period of a stress factor action. In heterogeneous population this can lead to observation of unchanged mortality during action of the stress and decrease in mortality after stress period. Stochastic simulations were made to investigate the possibility of detecting the hormesis effect on the basis of the stress-control longitudinal data. The goal of the stochastic simulation was to investigate the role in the hormesis detection of control and stressed group size, of population heterogeneity variance value, of stress and hormesis attributable risks as well as the role of a prior information about the survival in the control group. It was demonstrated that if the attributable risks for stress and hormesis effects are approximately equal, then in both 'high' and 'low' heterogeneous populations the hormesis phenomenon is detected with probability higher than 75% even in relatively 'small' groups of 50 subjects. In case of 'weak' effect the hormesis phenomenon is not detected in a 'highly heterogeneous' population even in a group composed of 1000 subjects. In a 'low heterogeneous' population the hormesis phenomenon is detected with probability higher than 70% when the group size is not less than 200 subjects. Information about the survival in control group did not play a critical role in all experiments and exact survival curve may be replaced by the traditional Kaplan-Meier estimate.

[Nitrogen oxide storage as a factor of adaptive defence]. / Deponirovanie oksida azota kak faktor adaptivnoi zashchity]

Adaptation to environmental factors possesses multiple NO-dependent protective effects and stimulates the NO storage. An adaptation to a mild stress was shown to reduce the death rate in rats from 57% to 8% and to prevent a heat shock-induced hypotension and endothelial overactivation. Treatment of the rats with the NO-synthase inhibitor L-NNA interfered with the NO storage and formation of protective effects, while the NO donor dinitrosyl iron complex facilitated the NO storage and simulated the adaptive defence. The data obtained suggest an important role of the NO storage in adaptive defence of the organism.

Winter mortality in relation to climate.

We report further details of the Eurowinter survey of cold related mortalities and protective measures against cold in seven regions of Europe, and review these with other evidence on the relationship of winter mortality to climate. Data for the oldest subject group studied, aged 65-74, showed that in this vulnerable group, high levels of protection against indoor and outdoor cold at given outdoor temperatures were found mainly in countries with cold winters, and were associated with low levels of excess mortality at a given level of outdoor cold. Regions such as London that had poor protection against cold and/or high baseline mortalities had higher levels of winter excess mortality than expected for the coldness of their winters.

Effects of prenatal exposure to a mild chronic variable stress on body weight, preweaning mortality and rat behavior.

Early stimulation has been shown to produce long-lasting effects in many species. Prenatal exposure to some strong stressors may affect development of the nervous system leading to behavioral impairment in adult life. The purpose of the present work was to study the postnatal harmful effects of exposure to variable mild stresses in rats during pregnancy. Female Holtzman rats were submitted daily to one session of a chronic variable stress (CVS) during pregnancy (prenatal stress; PS group). Control pregnant rats (C group) were undisturbed. The pups of PS and C dams were weighed and separated into two groups 48 h after delivery. One group was maintained with their own dams (PS group, N = 70; C group, N = 36) while the other PS pups were cross-fostered with C dams (PSF group, N = 47) and the other C pups were cross-fostered with PS dams (CF group, N = 58). Pups were undisturbed until weaning (postnatal day 28). The male offspring underwent motor activity tests (day 28), enriched environment tests (day 37) and social interaction tests (day 42) in an animal activity monitor. Body weight was recorded on days 2, 28 and 60. The PS pups showed lower birth weight than C pups (Duncan's test, P<0.05). The PS pups suckling with their stressed mothers displayed greater preweaning mortality (C: 23%, PS: 60%; chi2 test, P<0.05) and lower body weight than controls at days 28 and 60 (Duncan's test, P<0.05 and P<0.01, respectively). The PS, PSF and CF groups showed lower motor activity scores than controls when tested at day 28 (Duncan's test, P<0.01 for PS group and P<0.05 for CF and PSF groups). In the enriched environment test performed on day 37, between-group differences in total motor activity were not detected; however, the PS, CF and PSF groups displayed less exploration time than controls (Duncan's test, P<0.05). Only the PS group showed impaired motor activity and impaired social behavior at day 42 (Duncan's test, P<0.05). In fact, CVS treatment during gestation plus suckling with a previously stressed mother caused long-lasting physical and behavioral changes in rats. Cross-fostering PS-exposed pups to a dam which was not submitted to stress counteracted most of the harmful effects of the treatment. It is probable that prenatal stress plus suckling from a previously stressed mother can induce long-lasting changes in the neurotransmitter systems involved in emotional regulation. Further experiments using neurochemical and pharmacological approaches would be interesting in this model.

Less-invasive surgery for gastric cancer prolongs survival in patients over 80 years of age.

This study was undertaken to determine the most appropriate form of surgery for elderly patients with gastric cancer in relation to postoperative complications and long-term survival. A total of 72 consecutive patients over 80 years of age who underwent partial or total gastrectomy were evaluated using an E-PASS scoring system. This system is comprised of a preoperative risk score (PRS), a surgical stress score (SSS), and a comprehensive risk score (CRS) determined by both the PRS and SSS. Patients with a CRS > or = 0.5 had significantly higher rates of morbidity and mortality at 45.0% and 20.0%, respectively, than those with CRS < or = 0.5, at 17.0% and 2.1%, respectively. A Cox regression analysis of long-term survival, including death from other causes, identified five significant prognostic factors, namely: stage, curability, SSS, CRS, and allogeneic blood transfusion. Among the patients without any apparent residual cancer, a significantly better survival was seen in those who underwent less invasive surgery (SSS < 0.25), those with a CRS < or = 0.5, and those who had not been given a blood transfusion. These results suggest that less invasive surgery not requiring a blood transfusion is advisable for patients over 80 years of age with gastric cancer. Furthermore, gastrectomy with a CRS > or = 0.5 may have a poor therapeutic effect on both early and long-term outcome.