Mark Romanick

Effects of Growth Hormone and Insulin-Like Growth Factor-1 on Hepatocyte Antioxidative Enzymes

The physiological decline that occurs in aging is thought to result, in part, from accumulation of oxidative damage generated by reactive oxygen species during normal metabolic processes. Elevated levels of antioxidative enzymes in liver tissues are present in the Ames dwarf, a growth hormone (GH)-deficient mouse that lives more than 1 year longer than wild-type mice from the same line. In contrast, transgenic mice that overexpress GH exhibit depressed hepatic levels of catalase and have significantly shortened life spans. In this study, we evaluated the in vitro effects of GH and Insulin-like growth factor 1 (IGF-1) on antioxidative enzymes in mouse hepatocytes. Hepatocytes were isolated from wild-type mice following perfusion of livers with a collagenase-based buffer. Dispersed cells were plated on Matrlgel and treated with rat GH (0.1, 1.0, or 10 μg/ml) or IGF-1 (0.5, 5.0, or 50 nM) for 24 hr. Hepatocytes were recovered and protein was extracted for immunoblotting and enzyme activity assays of catalase (CAT), glutathione peroxidase (GPX), and manganese superoxide dismutase (MnSOD). A 41% and 27% decrease in catalase activity was detected in cells treated with GH, whereas IGF-1 reduced CAT activity levels to a greater extent than GH (P < 0.0001). The activity and protein levels of GPX were also significantly depressed In cells treated with GH, whereas activity alone was decreased in cells treated with IGF-1 (P < 0.04). GH significantly suppressed MnSOD levels by 40% and 66% in 1.0 and 0.1 μg/ml concentrations, respectively. Similarly, IGF-1 decreased MnSOD protein levels (5 nM; P < 0.05). These results suggest that GH and IGF-1 may decrease the ability of hepatocytes to counter oxidative stress. In addition, these experiments provide an explanation for the differing antioxidative defense capacity of GH-deficient versus GH-overexpressing mice, and they suggest that GH is directly involved in antioxidant regulation and the aging process.

Mitochondrial oxidant generation and oxidative damage in Ames dwarf and GH transgenic mice

Aging is associated with an accumulation of oxidative damage to proteins, lipids and DNA. Cellular mechanisms designed to prevent oxidative damage decline with aging and in diseases associated with aging. A long-lived mouse, the Ames dwarf, exhibits growth hormone deficiency and heightened antioxidative defenses. In contrast, animals that over express GH have suppressed antioxidative capacity and live half as long as wild type mice. In this study, we examined the generation of H2O2 from liver mitochondria of Ames dwarf and wild type mice and determined the level of oxidative damage to proteins, lipids and DNA in various tissues of these animals. Dwarf liver mitochondria (24 months) produced less H2O2 than normal liver in the presence of succinate (p<0.03) and ADP (p<0.003). Levels of oxidative DNA damage (8ÕHdG) were variable and dependent on tissue and age in dwarf and normal mice. Forty-seven percent fewer protein carbonyls were detected in 24-month old dwarf liver tissue compared to controls (p<0.04). Forty percent more (p<0.04) protein carbonyls were detected in liver tissue (3-month old) of GH transgenic mice compared to wild types while 12 month old brain tissue had 53% more protein carbonyls compared to controls (p<0.005). Levels of liver malonaldehyde (lipid peroxidation) were not different at 3 and 12 months of age but were greater in Ames dwarf mice at 24 months compared to normal mice. Previous studies indicate a strong negative correlation between plasma GH levels and antioxidative defense. Taken together, these studies show that altered GH-signaling may contribute to differences in the generation of reactive oxygen species, the ability to counter oxidative stress and life span.

Long-lived Ames dwarf mouse exhibits increased antioxidant defense in skeletal muscle

Resting and exercised (both acute and chronic) hindlimb skeletal muscle from long-lived Ames dwarf and wild type mice at 3, 12, 18, and 24 months of age was tested for antioxidant enzyme activity and protein, non-enzymatic antioxidant ratios, mitochondrial hydrogen peroxide concentration, and plasma lactate levels. Differences were observed in GPX enzyme activity between mouse genotypes at all physical activity levels, with dwarf mice exhibiting depressed levels at younger ages (3 months: P = 0.09 [non-swim], P = 0.03 [acute swim], P = 0.04 [chronic swim]) and comparatively higher levels than wild type mice at older ages (18-24 months: P = 0.05 [acute swim], P = 0.07 [chronic swim]). Catalase enzyme activity and the GSH system rarely demonstrated significant differences between genotypes, regardless of age or activity. However, the chronic exercise group displayed a difference in GSH:GSSG ratios between mouse genotypes (P = 0.005). Plasma lactate concentrations were elevated in the wild type mice compared to the dwarf mice at all ages in all activity groups. These results suggest there are biological differences with regard to antioxidant defense that favor the Ames dwarf mouse in active and resting skeletal muscle when compared to wild type mice.

Murine models of atrophy, cachexia, and sarcopenia in skeletal muscle.

With the extension of life span over the past several decades, the age-related loss of muscle mass and strength that characterizes sarcopenia is becoming more evident and thus, has a more significant impact on society. To determine ways to intervene and delay, or even arrest the physical frailty and dependence that accompany sarcopenia, it is necessary to identify those biochemical pathways that define this process. Animal models that mimic one or more of the physiological pathways involved with this phenomenon are very beneficial in providing an understanding of the cellular processes at work in sarcopenia. The ability to influence pathways through genetic manipulation gives insight into cellular responses and their impact on the physical expression of sarcopenia. This review evaluates several murine models that have the potential to elucidate biochemical processes integral to sarcopenia. Identifying animal models that reflect sarcopenia or its component pathways will enable researchers to better understand those pathways that contribute to age-related skeletal muscle mass loss, and in turn, develop interventions that will prevent, retard, arrest, or reverse this phenomenon. This article is part of a Special Issue entitled: Animal Models of Disease.

A prospective study of injury affecting competitive collegiate swimmers.

The purposes of this study were to determine the incidence and distribution of injuries affecting collegiate competitive swimmers and to test possible injury risk factors. A prospective cohort design was used to follow 34 swimmers (16M, 18F) from an NCAA Division I Midwest University over one academic year. Exposure-based injury rates were determined for both practice and competition. Risk of injury was assessed relative to gender, years swimming, and history of injury. Twenty of 34 swimmers sustained 31 injuries with overall injury rates of 5.55 injuries per 1000 athlete exposures and 3.04 injuries per 1000 hours exposure. Practice injury rates for female swimmers were higher than for womens practice injury rates in other NCAA sports. The most common injury locations were the shoulder, back, and knee. Risk factors that remained significant in the multivariate analysis were history of injury to the same anatomical location and history of injury at other anatomical sites.

Injury Type and Incidence Among Elite Level Curlers During World Championship Competition

Our objective was to investigate the incidence of musculoskeletal injuries sustained by elite level curling athletes during international competition. This study was conducted during the 2008 World Mens Curling Championships. All registered athletes and the tournament medical team were given report forms for documenting injuries that occurred during the tournament. Report form information included demographics, area injured, types of injuries sustained, and curling-specific aggravating conditions. During the competition five injuries were reported, resulting in an injury rate of .07 injuries per game. Only one reported injury resulted in missed competition (.014 injuries per game). All reported injuries involved increased pain during curling-specific activities. At the elite international competitive level, injury incidence in curling was found to be low. Future exploration over the course of a season may be beneficial to identify risk factors and to assist with formulating training strategies to decrease injury risk.

Preliminary results of residual deficits observed in athletes with concussion history: Combined EEG and cognitive study

Assessment, treatment, and management of sport-related concussions are a widely recognized public health issue. Although several neuropsychological and motor assessment tools have been developed and implemented for sports teams at various levels and ages, the sensitivity of these tests has yet to be validated with more objective measures to make return-to-play (RTP) decisions more confidently. The present study sought to analyze the residual effect of concussions on a sample of adolescent athletes who sustained one or more previous concussions compared to those who had no concussion history. For this purpose, a wide variety of assessment tools containing both neurocognitive and electroencephalogram (EEG) elements were used. All clinical testing and EEG were repeated at 8 months, 10 months, and 12 months post-injury for both healthy and concussed athletes. The concussed athletes performed poorer than healthy athletes on processing speed and impulse control subtest of neurocognitive test on month 8, but no alterations were marked in terms of visual and postural stability. EEG analysis revealed significant differences in brain activities of concussed athletes through all three intervals. These long-term neurocognitive and EEG deficits found from this ongoing sport-related concussion study suggest that the post-concussion physiological deficits may last longer than the observed clinical recovery.Assessment, treatment, and management of sport-related concussions are a widely recognized public health issue. Although several neuropsychological and motor assessment tools have been developed and implemented for sports teams at various levels and ages, the sensitivity of these tests has yet to be validated with more objective measures to make return-to-play (RTP) decisions more confidently. The present study sought to analyze the residual effect of concussions on a sample of adolescent athletes who sustained one or more previous concussions compared to those who had no concussion history. For this purpose, a wide variety of assessment tools containing both neurocognitive and electroencephalogram (EEG) elements were used. All clinical testing and EEG were repeated at 8 months, 10 months, and 12 months post-injury for both healthy and concussed athletes. The concussed athletes performed poorer than healthy athletes on processing speed and impulse control subtest of neurocognitive test on month 8, but no alterations were marked in terms of visual and postural stability. EEG analysis revealed significant differences in brain activities of concussed athletes through all three intervals. These long-term neurocognitive and EEG deficits found from this ongoing sport-related concussion study suggest that the post-concussion physiological deficits may last longer than the observed clinical recovery.

Neurocognitive deficits observed on high school football players with history of concussion: A preliminary study

Sport-related concussion diagnosis, monitoring, treatment, and recovery is rightfully recognized as one of the primary health concerns nowadays. Although much clinical and systematic research, involving athletes of different ages playing different games, has been conducted, there is still a paucity of objective measures to examine and monitor the post-concussion recovery of athletes to make a confident Return to Play (RTP) decision. The aim of the present study is to evaluate the residual symptoms and neurocognitive recovery patterns of concussion by comparing adolescent athletes having a history of one or multiple concussions with athletes who have no previous history of concussion. A variety of neurocognitive assessment tools including ImPACT (Immediate Post-Concussion Assessment and Cognitive Testing) for memory, attention and processing speed, K-D (King-Devick) test for visual and suboptimal brain function analysis, and BESS (Balance Error Scoring System) to examine postural stability, were been combined. A significant difference was detected for the ImPACT test during impulse control and processing speed subtests. The analysis also revealed long-term concussion effects on the performance of athletes who had multiple concussion histories. These enduring neurocognitive deficits observed from this study suggest that athletes with concussion history may require more prolonged recovery time compared to healthy athletes.

A Novel EEG Based Spectral Analysis of Persistent Brain Function Alteration in Athletes with Concussion History

The neurocognitive sequelae of a sport-related concussion and its management are poorly defined. Detecting deficits are vital in making a decision about the treatment plan as it can persist one year or more following a brain injury. The reliability of traditional cognitive assessment tools is debatable, and thus attention has turned to assessments based on electroencephalogram (EEG) to evaluate subtle post-concussive alterations. In this study, we calculated neurocognitive deficits combining EEG analysis with three standard post-concussive assessment tools. Data were collected for all testing modalities from 21 adolescent athletes (seven concussive and fourteen healthy) in three different trials. For EEG assessment, along with linear frequency-based features, we introduced a set of time-frequency (Hjorth Parameters) and nonlinear features (approximate entropy and Hurst exponent) for the first time to explore post-concussive deficits. Besides traditional frequency-band analysis, we also presented a new individual frequency-based approach for EEG assessment. While EEG analysis exhibited significant discrepancies between the groups, none of the cognitive assessment resulted in significant deficits. Therefore, the evidence from the study highlights that our proposed EEG analysis and markers are more efficient at deciphering post-concussion residual neurocognitive deficits and thus has a potential clinical utility of proper concussion assessment and management.