Menstrual status and thermoregulatory responses of active adolescents during exercise in a cold environment

This study examined the interactions between the reproductive status and the thermoregulatory responses during exercise in the cold in girls involved in competitive sports. Four girls with established menstrual cycles comprised the eumenorrheic menarcheal group (EM) and 5 non-menstruating girls comprised the pre-menarcheal group (PM). During the first visit maximal oxygen consumption, height, weight and percent body fat (%BF) were measured. The second visit involved: a determination of metabolic rate in thermoneutrality (21°C) involving 10-min rest and 20-min cycling (30% of VCL max), and a cold stress test (5°C, 40% humidity, <0.3 m/s air velocity) involving 20-min rest and 40-min cycling (30% of VCL max.). Subjects in the EM group were tested twice in the chamber during the follicular and luteal phases. Pre-menarcheal subjects were found to have significantly (p<0.05) lower core temperatures during the final stages of cold exposure. Overall, body fat was not significantly correlated with core temperature in the cold, however there was a significant surface-to-mass ratio difference between the groups. While in the follicular phase, EM girls had a higher core temperature during cold exposure. Therefore, reproductive hormonal status seems to be an important factor in terms of cold tolerance in females during adolescence.

Neuromuscular adaptations in endurance-trained boys and men

Competitive sports participation in youth is becoming increasingly more common in the Western world. It is widely accepted that sports participation, specifically endurance training, is beneficial for physical, psychomotor, and social development of children. The research on the effect of endurance training in children has focused mainly on healthrelated benefits and physiological adaptations, particularly on maximal oxygen uptake. However, corresponding research on neuromuscular adaptations to endurance training and the latter's possible effects on muscle strength in youth is lacking. In children and adults, resistance training can enhance strength and mcrease muscle activation. However, data on the effect of endurance training on strength and neuromuscular adaptations are limited. While some evidence exists demonstrating increased muscle activation and possibly increased strength in endurance athletes compared with untrained adults, the neuromuscular adaptations to endurance training in children have not been examined. Thus, the purpose of this study was to examine maximal isometric torque and rate of torque development (RID), along with the pattern of muscle activation during elbow and knee flexion and extension in muscle-endurancetrained and untrained men and boys. Subjects included 65 males: untrained boys (n=18), endurance-trained boys (n=12), untrained men (n=20) and endurance-trained men (n=15). Maximal isometric torque and rate of torque development were measured using an isokinetic dynamometer (Biodex III), and neuromuscular activation was assessed using surface electromyography (SEMG). Muscle strength and activation were assessed in the dominant arm and leg, in a cross-balanced fashion during elbow and knee flexion and extension. The main variables included peak torque (T), RTD, rate of muscle activation (Q30), Electro-mechanical delay (EMD), time to peak RTD and co-activation index. Age differences in T, RTD, electro-mechanical delay (EMD) and rate of muscle activation (Q30) were consistently observed in the four contractions tested. Additionally, Q30, nonnalized for peak EMG amplitude, was consistently higher in the endurancetrained men compared with untrained men. Co-activation index was generally low in all contractions. For example, during maximal voluntary isometric knee extension, men were stronger, had higher RTD and Q30, whether absolute or nonnalized values were used. Moreover, boys exhibited longer EMD (64.8 ± 18.5 ms vs. 56.6 ± 15.3 ms, for boys and men respectively) and time to peak RTD (112.4 ± 33.4 ms vs. 100.8 ± 39.1 ms for boys and men, respectively). In addition, endurance-trained men had lower T compared with untrained men, yet they also exhibited significantly higher nonnalized Q30 (1.9 ± 1.2 vs. 1.1 ± 0.7 for endurance-trained men and untrained men, respectively). No training effect was apparent in the boys. In conclusion, the findings demonstrate muscle strength and activation to be lower in children compared with adults, regardless of training status. The higher Q30 of the endurance-trained men suggests neural adaptations, similar to those expected in response to resistance training. The lower peak torque may su9gest a higher relative involvement oftype I muscle fibres in the endurance-trained athletes. Future research is required to better understand the effect of growth and development on muscle strength and activation patterns during dynamic and sub-maximal isometric contractions. Furthennore, training intervention studies could reveal the effects of endurance training during different developmental stages, as well as in different muscle groups.

Mechanisms of endothelin-1 induced reactive oxygen species production in vascular adventitial fibroblasts

With the relationship between endothelin-1 (ET-1) stimulation and reactive oxygen species (ROS) production unknown in adventitial fibroblasts, I examined the ROS response to ET-1 and angiotensin (Ang II). ET-1 -induced ROS peaked following 4 hrs of ET-1 stimulation and was inhibited by an ETA receptor antagonist (BQ 123, 1 uM) an extracellular signal-regulated kinase (ERK) 1/2 inhibitor (PD98059, 10 uM), and by both a specific, apocynin (10 uM), and non-specific, diphenyleneiodonium (10 uM), NAD(P)H oxidase inhibitor. NOX2 knockout fibroblasts did not produce an ET-1 induced change in ROS levels. Ang II treatment increased ROS levels in a biphasic manner, with the second peak occurring 6 hrs following stimulation. The secondary phase of Ang II induced ROS was inhibited by an ATi receptor antagonist, Losartan (100 uM) and BQ 123. In conclusion, ET-1 induces ROS production primarily through an ETA-ERKl/2 NOX2 pathway, additionally, Ang II-induced ROS production also involves an ETa pathway.

Lifespan, body size and oxygen : aerobic metabolism and oxidative stress in cultured fibroblasts /

The allometric scaling relationship observed between metabolic rate (MR) and species body mass can be partially explained by differences in cellular MR (Porter & Brand, 1995). Here, I studied cultured cell lines derived from ten mammalian species to determine whether cells propagated in an identical environment exhibited MR scaling. Oxidative and anaerobic metabolic parameters did not scale significantly with donor body mass in cultured cells, indicating the absence of an intrinsic MR setpoint. The rate of oxygen delivery has been proposed to limit cellular metabolic rates in larger organisms (West et al., 2002). As such cells were cultured under a variety of physiologically relevant oxygen tensions to investigate the effect of oxygen on cellular metabolic rates. Exposure to higher medium oxygen tensions resulted in increased metabolic rates in all cells. Higher MRs have the potential to produce more reactive oxygen species (ROS) which could cause genomic instability and thus reduced lifespan. Longer-lived species are more resistant to oxidative stress (Kapahi et al, 1999), which may be due to greater antioxidant and/or DNA repair capacities. This hypothesis was addressed by culturing primary dermal fibroblasts from eight mammalian species ranging in maximum lifespan from 5 to 120 years. Only the antioxidant manganese superoxide dismutases (MnSOD) positively scaled with species lifespan (p<0.01). Oxidative damage to DNA is primarily repaired by the base excision repair (BER) pathway. BER enzyme activities showed either no correlation or as in the case of polymerase p correlated, negatively with donor species (p<0.01 ). Typically, mammalian cells are cultured in a 20% O2 (atmospheric) environment, which is several-fold higher than cells experience in vivo. Therefore, the secondary aim of this study was to determine the effect of culturing mammalian cells at a more physiological oxygen tension (3%) on BER, and antioxidant, enzyme activities. Consistently, standard culture conditions induce higher antioxidant and DNA ba.se excision repair activities than are present under a more physiological oxygen concentration. Therefore, standard culture conditions are inappropriate for studies of oxidative stress-induced activities and species differences in fibroblast DNA BER repair capacities may represent differences in ability to respond to oxidative stress. An interesting outcome firom this study was that some inherent cellular properties are maintained in culture (i.e. stress responses) while others are not (i.e. MR).

Localization of the mechanism of pH-dependent non- photochemical fluorescence quenching in thylakoid membranes

Higher plants have evolved a well-conserved set of photoprotective mechanisms, collectively designated Non-Photochemical Quenching of chlorophyll fluorescence (qN), to deal with the inhibitory absorption of excess light energy by the photosystems. Their main contribution originates from safe thermal deactivation of excited states promoted by a highly-energized thylakoid membrane, detected via lumen acidification. The precise origins of this energy- or LlpH-dependent quenching (qE), arising from either decreased energy transfer efficiency in PSII antennae (~ Young & Frank, 1996; Gilmore & Yamamoto, 1992; Ruban et aI., 1992), from alternative electron transfer pathways in PSII reaction centres (~ Schreiber & Neubauer, 1990; Thompson &Brudvig, 1988; Klimov et aI., 1977), or from both (Wagner et aI., 1996; Walters & Horton, 1993), are a source of considerable controversy. In this study, the origins of qE were investigated in spinach thylakoids using a combination of fluorescence spectroscopic techniques: Pulse Amplitude Modulated (PAM) fluorimetry, pump-probe fluorimetry for the measurement of PSII absorption crosssections, and picosecond fluorescence decay curves fit to a kinetic model for PSII. Quenching by qE (,..,600/0 of maximal fluorescence, Fm) was light-induced in circulating samples and the resulting pH gradient maintained during a dark delay by the lumenacidifying capabilities of thylakoid membrane H+ ATPases. Results for qE were compared to those for the addition of a known antenna quencher, 5-hydroxy-1,4naphthoquinone (5-0H-NQ), titrated to achieve the same degree of Fm quenching as for qE. Quenching of the minimal fluorescence yield, F0' was clear (8 to 130/0) during formation of qE, indicative of classical antenna quenching (Butler, 1984), although the degree was significantly less than that achieved by addition of 5-0H-NQ. Although qE induction resulted in an overall increase in absorption cross-section, unlike the decrease expected for antenna quenchers like the quinone, a larger increase in crosssection was observed when qE induction was attempted in thylakoids with collapsed pH gradients (uncoupled by nigericin), in the absence of xanthophyll cycle operation (inhibited by DTT), or in the absence of quenching (LlpH not maintained in the dark due to omission of ATP). Fluorescence decay curves exhibited a similar disparity between qE-quenched and 5-0H-NQ-quenched thylakoids, although both sets showed accelerated kinetics in the fastest decay components at both F0 and Fm. In addition, the kinetics of dark-adapted thylakoids were nearly identical to those in qEquenched samples at F0' both accelerated in comparison with thylakoids in which the redox poise of the Oxygen-Evolving Complex was randomized by exposure to low levels of background light (which allowed appropriate comparison with F0 yields from quenched samples). When modelled with the Reversible Radical Pair model for PSII (Schatz et aI., 1988), quinone quenching could be sufficiently described by increasing only the rate constant for decay in the antenna (as in Vasil'ev et aI., 1998), whereas modelling of data from qE-quenched thylakoids required changes in both the antenna rate constant and in rate constants for the reaction centre. The clear differences between qE and 5-0H-NQ quenching demonstrated that qE could not have its origins in the antenna alone, but is rather accompanied by reaction centre quenching. Defined mechanisms of reaction centre quenching are discussed, also in relation to the observed post-quenching depression in Fm associated with photoinhibition.

Response rainbow trout (Salmon gairdneri) blood gas transport system to temperature, oxygen availability and photoperiod

Hematological status in rainbow trout, Salmo gairdneri, was examined in relation to eight combinations of three environmental fa ctors; temperature (5°, 20°C), oxygen availability «35%, >70% saturation) and photoperiod (16L:8D, 8L:16D) and evaluated by 3-factor analysis of variance. Hemog l obin and hematocrit , indicators of oxygenc arrying capacity increased significantly at the higher temperature, following exposure to hypoxia and in relation to reduced light period. Significant variations in mean corpuscular hemoglobin concentration were not detected. The effects of temperature and oxygen availability were more pronounced than that of photoperiod which was generally masked. Although oxygen availability and photoperiod did not interact with temperature, the interaction of the former fac tors was significant. Elec trophoresis revealed twelve hemoglobin isomorphs. Relative concentration changes were found in re lation to the factors c onsidered with temperature>hypoxia>photoperiod. Howeve r , in terms of absolute concentration, effects were hypoxia>temperature>photoperiod. Photoperiod effects were again masked by temperature and (or) hypoxia. Red cell +2 l eve ls of [CI ] and [Mg ], critical elements in the hemoglobin-oxygen affinity regulating system, were also significantly altered. Red cell CI +2 was influenced only by temperature ; Mg by temper ature and oxygen. No photoperiod influence on either ions was observed. Under nominal 'summer' conditions, these changes point to the likelihood of increases in oxygen-c arrying c apac ity coupled with low Hb-02 affinity adjustments which would be expected to increase oxygen delivery rates to their more rapidly metabolising tissues.

Training distribution and the acquisition of maximal isometric elbow flexion strength

Twenty-six sedentary, college-aged females were matched and randomly assigned to one of two groups. The massed group (n=13) completed 15 maximal isometric elbow flexion strength trials in one session, while the distributed group (n=13) performed five such contractions on three successive days. After a two-week and three month rest interval, both groups returned to perfonn another five maximal isometric elbow flexion strength trials to assess retention of any potential strength gains. Elbow flexion torque and surface electromyography (SEMG) of the biceps and triceps were monitored concurrently. There was a significant (P < 0.05) increase in strength in both groups from block one (first five contractions) to block four (first retest) and from block one to block five (second retest). Both groups exhibited a similar linear increasing (P < 0.05) trend in biceps root-mean-square (RMS) SEMG amplitude. A significant (P < 0.05) decrease in triceps RMS SEMG amplitude was found between block one and block four for the distributed group. However, a significant (P < 0.05) increase was then found between block one and five for the massed group, and between blocks four and five for distributed group. These results suggest that there is flexibility in resistive exercise schedules. An increase in neural drive to the agonist muscle continued throughout testing. This was accompanied by a reduction in antagonist co activation that was a short-tenn (two weeks) training effect, dissipated over the longer rest interval (three months).

CCK, dietary variety, and food intake in rats /

Among the environmental factors that can affect food intake is the extent of dietary variety available in the environment. Numerous studies have demonstrated that variety in a meal can increase the amount of food consumed in humans, rats, and other species. A physiological mechanism that has been demonstrated to affect food intake is the gut peptide cholecystokinin (CCK) which is released from the upper small intestine during the ingestion of food. Peripherally administered CCK has a robust inhibitory effect on the intake of a single-food meal. Thus, dietary variety and CCK both affect meal size, with dietary variety increasing intake and CCK decreasing intake. This raises the question ofhow dietary variety and CCK might interact to affect meal size. Previous studies of CCK's effects have focused on situations in which only one food was available for consumption. However, in an animal's natural environment it would frequently occur that the animal would come across a number of foods either simultaneously or in quick succession, thus providing the animal access to a variety of foods during a meal. Accordingly, the effect ofCCK on food intake in single-food and multiple-food meals was examined. It was found that food intake was greater in multiple-food than in single-food meals provided that foods in the multiplefood meal were presented either simultaneously or in increasing order of preference. When foods in the multiple-food meal were presented in decreasing order of preference, intake was similar to that observed in single-food meals. In addition, it was found that CCK inhibited food intake in a dose-dependent manner, and that its effects on food intake were similar regardless of meal type. Therefore, the inhibitory effects ofCCK were not diminished when a variety of foods were available for consumption. Furthermore, the finding that CCK did not differentially affect the intake of the two types of meals does not provide support for the recent-foods hypothesis which postulates that CCK decreases food intake by reducing the palatability of only recently consumed foods. However, it is consistent with the all-foods hypothesis, which predicts that CCK reduces food intake by decreasing the palatability of all foods. The 600 ng/kg dose of the CCK^-antagonist lorglumide significantly antagonized the inhibitory effect of exogenous CCK on food intake, and the magnitude of this effect was similar for both types of meal. These results suggest that exogenous CCK inhibits food intake through the activation ofCCK^ receptors. However, when administered by itself, the 600^ig/kg dose of lorglumide did not increase food intake in either single-food or multiple-food meals, suggesting that peripheral endogenous CCK may not play a major role in the control of food intake.

The role of learning and growth hormone-releasing factor in the control of protein intake in rats /

Both learning and basic biological mechanisms have been shown to play a role in the control of protein int^e. It has previously been shown that rats can adapt their dietary selection patterns successfully in the face of changing macronutrient requirements and availability. In particular, it has been demonstrated that when access to dietary protein is restricted for a period of time, rats selectively increase their consumption of a proteincontaining diet when it becomes available. Furthermore, it has been shown that animals are able to associate various orosensory cues with a food's nutrient content. In addition to the role that learning plays in food intake, there are also various biological mechanisms that have been shown to be involved in the control of feeding behaviour. Numerous studies have documented that various hormones and neurotransmitter substances mediate food intake. One such hormone is growth hormone-releasing factor (GRF), a peptide that induces the release of growth hormone (GH) from the anterior pituitary gland. Recent research by Vaccarino and Dickson ( 1 994) suggests that GRF may stimulate food intake by acting as a neurotransmitter in the suprachiasmatic nucleus (SCN) and the adjacent medial preoptic area (MPOA). In particular, when GRF is injected directly into the SCN/MPOA, it has been shown to selectively enhance the intake of protein in both fooddeprived and sated rats. Thus, GRF may play a role in activating protein consumption generally, and when animals have a need for protein, GRF may serve to trigger proteinseeking behaviour. Although researchers have separately examined the role of learning and the central mechanisms involved in the control of protein selection, no one has yet attempted to bring together these two lines of study. Thus, the purpose of this study is to join these two parallel lines of research in order to further our understanding of mechanisms controlling protein selection. In order to ascertain the combined effects that GRF and learning have on protein intake several hypothesis were examined. One major hypothesis was that rats would successfully alter their dietary selection patterns in response to protein restriction. It was speculated that rats kept on a nutritionally complete maintenance diet (NCMD) would consume equal amount of the intermittently presented high protein conditioning diet (HPCD) and protein-free conditioning diet (PFCD). However, it was hypothesized that rats kept on a protein-free maintenance diet (PFMD) would selectively increase their intake of the HPCD. Another hypothesis was that rats would learn to associate a distinct marker flavour with the nutritional content of the diets. If an animal is able to make the association between a marker flavour and the nutrient content of the food, then it is hypothesized that they will consume more of a mixed diet (equal portion HPCD and PFCD) with the marker flavour that was previously paired with the HPCD (Mixednp-f) when kept on the PFMD. In addition, it was hypothesized that intracranial injection of GRF into the SCN/MPOA would result in a selective increase in HPCD as well as Mixednp-t consumption. Results demonstrated that rats did in fact selectively increase their consumption of the flavoured HPCD and Mixednp-f when kept on the NCMD. These findings indicate that the rats successfully learned about the nutrient content of the conditioning diets and were able to associate a distinct marker flavour with the nutrient content of the diets. However, the results failed to support previous findings that GRF increases protein intake. In contrast, the administration of GRF significantly reduced consumption of HPCD during the first hour of testing as compared to the no injection condition. In addition, no differences in the intake of the HPCD were found between the GRF and vehicle condition. Because GRF did not selectively increase HPCD consumption, it was not surprising that GRF also did not increase MixedHP-rintake. What was interesting was that administration of GRF and vehicle did not reduc^Mixednp-f consumption as it had decreased HPCD consumption.

Oxygen exchange in galls of Eurosta solidaginis (Diptera: Tephritidae)

Hypoxia in plant tissue should affect animals living within. Gallmakers stimulate their plant hosts to produce the gall they inhabit and feed on, and also influence the gall phenotype for other adaptations, such as defense against predators. The potential for hypoxia in galls of Eurosta solidaginis was studied in the context of potential adaptations to gall oxygen level, using a combination of direct measurement, mathematical modelling, and respirometry on both gallmakers and hosts. Modelling results suggested mild hypoxia tolerable to the larva persists for most of the growth season, whereas more severe hypoxia may occur earlier in fully-grown young galls. Field data from one of the two years studied showed hypoxia more severe than expected, and coincided with adverse weather conditions and high larval mortality. The hypoxia may be related to host response to adverse weather. Whether hypoxia directly caused larval mortality requires further study.

Cellular Mechanisms of Resveratrol's Interaction with Mitochondrial Reactive Oxygen Species Metabolism

Resveratrol, a polyphenol found naturally in red wines, has attracted great interest in both the scientific community and the general public for its reported ability to protect against many of the diseases facing Western society today. While the purported health effects of resveratrol are well characterized, details of the cellular mechanisms that give rise to these observations are unclear. Here, the mitochondrial antioxidant enzyme Mn superoxide dismutase (MnSOD) was identified as a proximal target of resveratrol in vitro and in vivo. MnSOD protein and activity levels increase significantly in cultured cells treated with resveratrol, and in the brain tissue of mice given resveratrol in a high fat diet. Preventing the increase in MnSOD levels eliminates two of resveratrol’s more interesting effects in the context of human health: inhibition of proliferative cell growth and cytoprotection. Thus, the induction of MnSOD is a critical step in the molecular mechanism of resveratrol. Mitochondrial morphology is a malleable property that is capable of impeding cell cycle progression and conferring resistance against stress induced cell death. Using confocal microscopy and a novel ‘cell free’ fusion assay it was determined that concurrent with changes in MnSOD protein levels, resveratrol treatment leads to a more fused mitochondrial reticulum. This observation may be important to resveratrol’s ability to slow proliferative cell growth and confer cytoprotection. Resveratrol's biological activities, including the ability to increase MnSOD levels, are strikingly similar to what is observed with estrogen treatment. Resveratrol fails to increase MnSOD levels, slow proliferative cell growth and confer cytoprotection in the presence of an estrogen receptor antagonist. Resveratrol's effects can be replicated with the specific estrogen receptor beta agonist diarylpropionitrile, and are absent in myoblasts lacking estrogen receptor beta. Four compounds that are structurally similar to resveratrol and seven phytoestrogens predicted to bind to estrogen receptor beta were screened for their effects on MnSOD, proliferative growth rates and stress resistance in cultured mammalian cells. Several of these compounds were able to mimic the effects of resveratrol on MnSOD levels, proliferative cell growth and stress resistance in vitro. Thus, I hypothesize that resveratrol interacts with estrogen receptor beta to induce the upregulation of MnSOD, which in turn affects cell cycle progression and stress resistance. These results have important implications for the understanding of RES’s biological activities and potential applications to human health.

Effect of increased milk intake combined with endurance exercise training on body composition, blood-lipids and inflammatory markers in overweight young males

Consuming low-fat milk (LFM) after resistance training leads to improvements in body composition. Habitual aerobic exercise and dairy intake are relatively easy lifestyle modifications that could benefit a population at risk for becoming obese. Thus, the purpose of this study was to investigate combining increased LFM intake with endurance exercise on body composition, blood-lipid profile and metabolic markers. 40 young males were randomized into four groups: one ingesting 750mL LFM immediately post-exercise, the other 6hrs post-exercise; and two isocaloric carbohydrate groups ingesting at the two different times. Participants completed a 12 week endurance-training program (cycling 1 hour/day at ~60%VO2peak, 5 days/week). 23 participants completed the study. Increases in lean mass (p < 0.05), and decreases in anti-inflammatory marker adiponectin (p < 0.05) were seen in all groups. No other significant changes were observed. Future analyses should focus on longer duration exercise and include a larger sample.

The Association of Energy Intake with Body Mass in Children With and Without Probable Developmental Coordination Disorder

Objective To determine if there is an association between energy intake (EI) and overweight or obesity status (OWOB) in children with and without probable developmental coordination disorder (p-DCD). Methods 1905 children were included. The Bruininks-Oseretsky Test of Motor Proficiency was used to assess p-DCD, body mass index for OWOB, and the Harvard Food Frequency Questionnaire for EI. Comparative tests and logistic regressions were performed. Results Reported EI was similar between p-DCD and non-DCD children among boys (2291 vs. 2281 kcal/day, p=0.917), but much lower in p-DCD compared to non-DCD girls (1745 vs.. 2068 kcal/day, p=0.007). EI was negatively associated with OWOB in girls only (OR: 0.82 (0.68, 0.98)). Conclusions Girls with p-DCD have a lower reported EI compared to their non-DCD peers. EI is negatively associated with OWOB in girls with p-DCD. Future research is needed to assess longitudinally the potential impact of EI on OWOB in this population.