Prenatal stress and increased fluctuating asymmetry in the parietal bones of neonatal rats

Recent studies have reported increased fluctuating dental and long bone asymmetry in neonates as a function of prenatal stress. The present study was designed to assess the effects of prenatal stress on a third calcium-dependent system, membranous bone. Pregnant rats were exposed to cold, heat, or noise from conception through parturition. Bilateral parietal bone lengths were measured in the term neonates. Levels of fluctuating asymmetry were found to be significantly increased (p less than .001) in all three stressed groups compared to unstressed controls. Results support the concept of a generalized stress response, and suggestions are offered for human osteological application.     

Toxoplasma gondii: cold stress-induced modulation of antibody responses.

Physical or psychological stressors have been shown to have significant consequences in the immune function and the outcome of disease in human and animal models. Recent work has demonstrated that products released during stress, such as glucocorticoids and catecholamines, can profoundly influence the in vitro growth of pathogens by modulating immune responses. The present study examined the effects of a physical stressor (cold stress) on antigens of Toxoplasma gondii that elicits an antibody-mediated immune response during the acute and chronic phases of infection. Sera obtained from different groups of mice subjected to cold stress during the acute and chronic phases of T. gondii infection were used to measure the levels of antibodies and to localize by Western blot the dominant antigens eliciting IgG and IgM antibody responses. Serum antibodies collected from stressed and infected mice recognized antigens different from those recognized by infected mice without stress. During the acute phase, a stronger IgM antibody response against antigens of 30, 42, 54, and 60 kDa was detected in stressed animals at 3 weeks postinfection. In addition, a 5-kDa antigen was specifically detected in mice subjected to stress during the acute and chronic phases of infection. Levels of specific IgG were increased in infected and in infected and stressed animals that underwent stress in the chronic phase. IgM production did not increase following cold stress in the chronic phase. These results suggest that the strong antibody response in stressed animals is associated with longer parasite persistence in circulation. Stress modulated not only the host immune response but also the ability of parasite antigens to elicit specific antibody responses by the host.     

Cold shock response in sporulating Bacillus subtilis and its effect on spore heat resistance

Cold shock and ethanol and puromycin stress responses in sporulating Bacillus subtilis cells have been investigated. We show that a total of 13 proteins are strongly induced after a short cold shock treatment of sporulating cells. The cold shock pretreatment affected the heat resistance of the spores formed subsequently, with spores heat killed at 85 or 90 degrees C being more heat resistant than the control spores while they were more heat sensitive than controls that were heat treated at 95 or 100 degrees C. However, B. subtilis spores with mutations in the main cold shock proteins, CspB, -C, and -D, did not display decreased heat resistance compared to controls, indicating that these proteins are not directly responsible for the increased heat resistance of the spores. The disappearance of the stress proteins later in sporulation suggests that they cannot be involved in repairing heat damage during spore germination and outgrowth but must alter spore structure in a way which increases or decreases heat resistance. Since heat, ethanol, and puromycin stress produce similar proteins and similar changes in spore heat resistance while cold shock is different in both respects, these alterations appear to be very specific.     

Neonatally-induced thermotolerance: physiological responses

1. Broiler cockerels exposed to neonatal heat (35-37.8 degrees C for 24 hr) at 5 days of age experienced significantly lower mortality upon exposure to elevated temperatures (35-37.8 degrees C) at 43 days of age than did cockerels not given neonatal heat exposure. 2. No differences were found between neonatally stressed and neonatal control groups in water consumption, core and surface temperature, plasma T3 and T4, protein or glucose concentration when exposed to juvenile heat stress. 3. Heterophil to lymphocyte ratios were significantly lower in the neonatally stressed than in the neonatal control group when exposed to heat at 44 days of age. 4. The results indicate that lower mortality during periods of high environmental temperature in the neonatally stressed chicks may involve homeostatic mechanisms different from those utilized during acclimation to high environmental temperature.     

The stress of Lymnaea truncatula just before miracidial exposure with Fasciola hepatica increased the prevalence of infection.

Single-miracidium infections of Lymnaea truncatula with Fasciola hepatica were carried out under laboratory conditions to determine whether the stress of snails just before miracidial exposure had any influence on the prevalence of Fasciola infection, redial burden, and cercarial shedding. Three methods, i.e., the fasting of L. truncatula for 3 days in water filtered through a Millipore membrane, the effect of 6-8 degrees C water for 15 min, or the immersion of L. truncatula in a detergent solution at low concentration for 15 min, were used to stress snails. Enhanced susceptibility of snails to F. hepatica infection was noted in stressed groups (93-96% vs 48-50% in controls). The number of free rediae did not show any variation in controls as well as in stressed groups, except for fasted snails in which free rediae were significantly fewer. No differences in cercarial production between controls and the cold group were noted. Fasting, cold shock, or detergent exposure prior to exposure to F. hepatica miracidia might have weakened the snails so that they were not as efficient in avoiding miracidial penetration, thus leading to higher infection rates.     

Prenatal Ethanol Exposure: Changes in Regional Brain Catecholamine Content Following Stress

Abstract: Previous studies have shown that fetal ethanol exposure (FEE) may have long-term effects on the function of catecholaminergic neurons in different regions of the CNS. The present study is the first to examine the effects of FEE on regional brain catecholamine responses following acute stress (a single 60-min restraint stress), repeated stress (single periods of restraint stress on 1, 5, or 10 consecutive days), and recovery from stress (recovery for up to 60 min in the home cage following a single 60-min period of restraint stress). Both male and female offspring from FEE, pair-fed (PF), and ad libitum-fed control (C) groups were tested in adulthood to determine catecholamine content in the cortex, hypothalamus, and hippocampus. A single period of restraint reduced cortical norepinephrine (NE) content in FEE and PF animals compared with that in the cortex of C animals, and reduced hypothalamic NE content in FEE female offspring below that found in animals in all other groups. In contrast, hippo-campal NE content was higher in FEE than in C animals following a single period of restraint; PF animals had intermediate levels of hippocampus NE and did not differ significantly from either FEE or C animals. Following repeated periods of restraint, cortical NE content was lower in FEE than in C animals; PF animals once again had intermediate levels of NE. Importantly, basal (non stressed) NE content did not differ among groups in any brain area examined. In addition, several significant changes in regional brain catecholaminergic responses to acute stress were observed in animals across all treatment groups. Females generally had significantly lower cortical NE levels than males following both single and multiple exposures to restraint. In addition, the cortical NE content decreased below non-stressed levels in all groups following a single restraint period, and remained significantly below basal levels during the 60-min recovery period, whereas the hypothalamic NE content was significantly decreased immediately following the restraint period but showed some recovery toward basal levels by 60 min. There were no significant changes over time in hippocampal NE level or in cortical or hypothalamic dopamine (DA) content following a single restraint stress. Following multiple periods of restraint, hippocampal NE levels were significantly increased and hypothalamic DA levels were significantly decreased in all animals compared with basal levels. These data suggest that the brain noradrenergic response to acute stress is particularly sensitive to the effects of FEE, and that with regard to the hypothalamus, male and female offspring were differentially affected. Furthermore, nutritional effects appear to play some role in mediating the changes in regional brain catecholamine content that are observed. In addition, stress effects on brain catecholamine content across all treatment groups were found to be both region and sex specific.     

The effects of cold-stress,hibernation, and prolonged inactivity on bone dynamics in the golden hamster,Mesocricetus auratus

The effects of cold-stress and hibernation on bone dynamics in the femurs of hamsters were investigated using histometric analyses. Control animals were maintained at 27° C for 90 days; experimental animals were kept at 5° C and hibernated for 7, 15, 21, 50, or 90 days. Histometric analyses of cross sections indicated that bone diameter and cortical thickness at the femoral midshaft increased after 83 days of extreme cold and 7 days of hibernation but decreased significantly after 69 days of cold stress and 21 days of hibernation. Osteoporosis was evident although the number of osteons per unit area of bone increased during hibernation. An initial decrease in the number of non-Haversian longitudinal vessels per unit area of bone was seen in experimental animals which was apparently related to a corresponding reduction in cortical thickness. Lacunar area increased in these animals, suggesting that osteocytic osteolysis may be a significant mechanism for calcium regulation during hibernation.     

Early life stress increases testosterone and corticosterone and alters stress physiology in zebra finches

Early life stress has enduring effects on behavior and physiology. However, the effects on hormones and stress physiology remain poorly understood. In the present study, parents of zebra finches of both sexes were exposed to an increased foraging paradigm from 3 to 33 days post hatching. Plasma and brains were collected from chicks at 3 developmental time points: post hatching days 25, 60 and adulthood. Plasma was assayed for testosterone (T), estradiol (E2), and corticosterone (CORT). The paraventricular nucleus of the hypothalamus was assessed for corticotrophin releasing factor (CRH) and glucocorticoid receptor (GR) expression. As expected, body mass was lower in nutritionally stressed animals compared to controls at multiple ages. Nutritionally stressed animals overall had higher levels of CORT than did control and this was particularly apparent in females at post hatching day 25. Nutritionally stressed animals also had a higher number of cells expressing CRH and GR in the paraventricular nucleus of the hypothalamus than did controls. There was an interaction, such that both measures were higher in control animals at PHD 25, but higher in NS animals by adulthood. Females, regardless of treatment, had higher circulating CORT and a higher number of cells expressing CRH than did males. Nutritionally stressed animals also had higher levels of T than did control animals, and this difference was greatest for males at post hatching day 60. There were no effects of nutritional stress on E2. These findings suggest that nutritional stress during development has long-lasting effects on testosterone and stress physiology.     

Impaired bone formation in transgenic mice resulting from altered integrin function in osteoblasts

To determine the role of integrins in mature osteoblasts in vivo, we expressed in transgenic mice a dominant-negative integrin subunit (beta1-DN) consisting of the beta1 subunit cytoplasmic and transmembrane domains, driven by the osteoblast-specific osteocalcin promoter. Immature osteoblasts isolated from transgenic animals differentiated normally in vitro until the osteocalcin promoter became active; thereafter they detached from the substratum, suggesting that beta1-DN was impairing adhesion in mature osteoblasts. Transgenic animals had reduced bone mass, with increased cortical porosity in long bones and thinner flat bones in the skull. At 35 days, the rate of bone formation was reduced in cortical bone, and the parietal bones were 45% thinner than in wild-type animals. Active osteoblasts were less polar and had larger areas of cytoplasm with intracellular stores of matrix molecules. Osteocyte lacunae appeared normal around the cell body but did not have normal canilicular structures. At 90 days, the parietal bone of transgenic males was of normal width, suggesting that the original defect in matrix deposition had been repaired or compensated for. In contrast, transgenic females still had decreased bone mass in the parietal bone at 90 days. The decreased bone mass in TG females was accompanied by increased staining for osteoclast activity, suggesting that there was a sex-specific defect in mature animals.     

Chronic-intermittent cold stress in rats induces selective ovarian insulin resistance

In rat ovary chronic cold stress increases sympathetic nerve activity, modifies follicular development, and initiates a polycystic condition. To see whether there is a relationship between the previously described changes in follicular development and metabolic changes similar to those in women with polycystic ovary, we have studied the effect of chronic cold stress (4 degrees C for 3 h/day, Monday to Friday, for 4 wk) on insulin sensitivity and the effect of insulin on sympathetic ovarian activity. Although cold-stressed rats ate more than the controls, they did not gain more weight. Insulin sensitivity, determined by hyperinsulinemic-euglycemic clamp, was significantly increased in the stressed animals. Insulin in vitro increased the basal release of norepinephrine from the ovaries of control rats but not from those of stressed rats, suggesting a local neural resistance to insulin in stressed rats. The levels of mRNA and protein for IRS1 and SLC2A4 (also known as GLUT4), molecules involved in insulin signaling, decreased significantly in the ovaries but not in the muscle of stressed rats. This decrease was preferentially located in theca-interstitial cells compared with granulosa cells, indicating that theca cells (the only cells directly innervated by sympathetic nerves) are responsible for the ovarian insulin resistance found in stressed rats. These findings suggest that ovarian insulin resistance produced by chronic stress could be in part responsible for the development of the polycystic condition induced by stress.     

Bone modeling response to voluntary exercise in the hindlimb of mice

The functional adaptation of juvenile mammalian limb bone to mechanical loading is necessary to maintain bone strength. Diaphyseal size and shape are modified during growth through the process of bone modeling. Although bone modeling is a well-documented response to increased mechanical stress on growing diaphyseal bone, the effect of proximodistal location on bone modeling remains unclear. Distal limb elements in cursorial mammals are longer and thinner, most likely to conserve energy during locomotion because they require less energy to move. Therefore, distal elements are hypothesized to experience greater mechanical loading during locomotion and may be expected to exhibit a greater modeling response to exercise. In this study, histomorphometric comparisons are made between femora and tibiae of mice treated with voluntary exercise and a control group (N = 20). We find that femora of exercised mice exhibit both greater bone growth rates and growth areas than do controls (P < 0.05). The femora of exercised mice also have significantly greater cortical area, bending rigidity, and torsional rigidity (P < 0.05), although bending and torsional rigidity are comparable when standardized by bone length. Histomorphometric and cross-section geometric properties of the tibial midshaft of exercised and control mice did not differ significantly, although tibial length was significantly greater in exercised mice (P < 0.05). Femora of exercised mice were able to adapt to increased mechanical loading through increases in compressive, bending, and torsional rigidity. No such adaptations were found in the tibia. It is unclear if this is a biomechanical adaptation to greater stress in proximal elements or if distal elements are ontogenetically constrained in a tradeoff of bone strength of distal elements for bioenergetic efficiency during locomotion.     

Effect of misoprostol in preventing stress-induced intestinal fluid secretion in rats

Psychological stress may alter gastrointestinal function by central nervous system controlled alteration of local intestinal mediators. Prostaglandins have been shown to prevent epithelial damage to various noxious stimuli. The purpose of this study was to determine (a) if wrap restraint stress altered in vivo intestinal fluid absorption in rats, and (b) if the prostaglandin E1 analogue, misoprostol, could correct observed fluid malabsorption. In vivo loop studies demonstrated net fluid secretion in the ileum and colon of cold wrap restraint stressed rats. In cold wrap restraint stressed rats, misoprostol reversed net secretion to absorption, but it had no effect on fluid absorption in controls. Mild wrap restraint stress did not alter in vivo fluid absorption. We conclude that cold wrap restraint stress is accompanied by net intestinal fluid secretion that can be effectively reversed with misoprostol.     

Effect of drinking earthen pot water on physiological response and behavior of sheep under heat stress

Heat stress is a major limiting factor for animal welfare and sheep production. Traditionally in India, the villagers used to keep their drinking water in the earthen pot to make it cold during summer. The cold drinking water (24–28 °C) during summer gives a feeling of relief from the heat. Therefore, the present study was carried out to assess the effect of drinking earthen pot water on physiological response and behavior of sheep under heat stress for one month. For this purpose, eighteen Avishaan rams were selected from the experimental animal flock and they were equally divided into three groups; viz., control (CON), heat stress (HS) and heat stress with earthen pot water (HSC). The animals of HS and HSC were exposed to higher ambient temperatures to induce heat stress inside the psychometric chamber. The animals of CON and HS were provided with ad-libitum water of their ambient temperature whereas; HSC groups were provided with ad-libitum cold water (24–28 °C) earthen pot water. All the animals were offered with 400 gm concentrate mixture and ad-libitum Cenchrus hay. The bodyweight of HS rams was significantly reduced (P < 0.05) at the end of the experimental period as compared to their initial body weight. The total roughage and dry matter intake was significantly higher (P < 0.01) in HSC rams as compared to HS rams. The plasma thyroxine concentration was significantly lower (P < 0.05) in HSC as compared with HS group. The rumination time significantly reduced (P < 0.05) in HSC group. However, The blood biochemical did not differ among the groups. Therefore, it may be concluded that Avishaan rams have the ability to adapt to heat stress. Nevertheless, the availability of earthen pot cold drinking water under heat stress reduced their body weight loss, improves their metabolic activity and ultimately improves their welfare.     

Quantitative RT-PCR analysis and immunohistochemical localization of HSP70 in sea bass Dicentrarchus labrax exposed to transport stress

In aquaculture, fish are exposed to stressful conditions, which cause an increased synthesis of heat shock proteins (HSPs) at the cellular level. In this work we considered the expression of the constitutive and inducible forms of HSP70 as an indicator of stress caused by transport, during development of the sea bass (Dicentrarchus labrax), a teleost fish of high value for aquaculture. Qualitative RT-PCR analysis revealed expression of inducible HSP70 gene in larvae and fry (25, 40 and 80 days) as well as in adult tissues (liver, brain, muscle, gills, kidney, gonads, heart, spleen and skin) of both control and stressed animals. Expression of inducible HSP70 mRNA examined in different adult tissues by Real-Time PCR, was significantly higher in skin and skeletal muscle of stressed animals than in controls. Immunolocalization of inducible and constitutive forms of heat shock protein 70 (HSP70 and HSC70), reported here for the first time, demonstrated an ubiquitous distribution of HSC70 protein in several tissues of both stressed and control animals (at all stages), while inducible HSP70 protein was found only in skeletal muscle of stressed animals. In all stressed animals, regardless of their developmental stage, cortisol levels were higher than in control animals.     

Beta3 adrenoceptors substitute the role of M(2) muscarinic receptor in coping with cold stress in the heart: evidence from M(2)KO mice

We investigated the role of beta3-adrenoceptors (AR) in cold stress (1 or 7?days in cold) in animals lacking main cardioinhibitive receptors-M2 muscarinic receptors (M(2)KO). There was no change in receptor number in the right ventricles. In the left ventricles, there was decrease in binding to all cardiostimulative receptors (beta1-, and beta2-AR) and increase in cardiodepressive receptors (beta3-AR) in unstressed KO in comparison to WT. The cold stress in WT animals resulted in decrease in binding to beta1- and beta2-AR (to 37%/35% after 1?day in cold and to 27%/28% after 7?days in cold) while beta3-AR were increased (to 216% of control) when 7?days cold was applied. MR were reduced to 46% and 58%, respectively. Gene expression of M2 MR in WT was not changed due to stress, while M3 was changed. The reaction of beta1- and beta2-AR (binding) to cold was similar in KO and WT animals, and beta3-AR in stressed KO animals did not change. Adenylyl cyclase activity was affected by beta3-agonist CL316243 in cold stressed WT animals but CL316243 had almost no effects on adenylyl cyclase activity in stressed KO. Nitric oxide activity (NOS) was not affected by BRL37344 (beta3-agonist) both in WT and KO animals. Similarly, the stress had no effects on NOS activity in WT animals and in KO animals. We conclude that the function of M2 MR is substituted by beta3-AR and that these effects are mediated via adenylyl cyclase rather than NOS.     

Stress- and cold-induced adrenocortical responses in repetitively immobilized or cold-acclimated rats

To evaluate the role of adrenocortical hormones in stress- or cold-induced nonshivering thermogenesis, plasma corticosterone (CS) and deoxycorticosterone (DOCS) were measured with the aid of HPLC under various conditions. Repetitive immobilization stress (3 h/day, for 1 or 4 weeks) elevated the resting level (24 h after the last immobilization) of CS, but not DOCS. Acute stress (immobilization for 30 min) or cold exposure (-5 degrees C for 15 min) caused marked increases of CS and DOCS in both nonstressed naive controls and repetitively stressed rats. Four weeks, but not 1 week, of repetitive immobilization stress potentiated the responsiveness of CS to both acute stress and cold, and that of DOCS to acute stress, but not to cold. Cold acclimation (5 degrees C, 4 weeks) significantly elevated both corticosteroids but did not affect the resting levels (18 h after being transferred to 25 degrees C) or the responsiveness of both CS and DOCS to either acute stress or cold. These results suggest that repetitive immobilization stress, but not cold acclimation, could enhance nonshivering thermogenesis, at least in part, through an improvement in the responsiveness of adrenocortical hormone secretion to acute stress or cold.     

Activity of Stress-related Antioxidative Enzymes in the Invasive Plant Crofton Weed （Eupatorium adenophorum）

Crofton weed is an invasive weed in southwestern China. The activities of several antioxidative enzymes involved in plant protection against oxidative stress were assayed to determine physiological aspects of the crofton weed that might render the plant vulnerable to environmental stress. Stresses imposed on crofton weed were heat （progressively increasing temperatures： 25 ℃, 30 ℃, 35 ℃, 38℃ and 42 ℃ at 24 h intervals）, cold （progressively decreasing temperatures： 25 ℃, 20 ℃, 15℃, 10 ℃ and 5℃ at 24h intervals）, and drought （without watering up to 4days）. The three stresses induced oxidative damage as evidenced by an increase in lipid peroxidation. The effect varied with the stress imposed and the length of exposure. The activity of superoxide dismutase （SOD） increased in response to all stresses but was not significantly different from the controls （P 〈 0.05） when exposed to cold stress. Catalase （CAT） activity decreased in response to heat and drought stress but increased when exposed to cold conditions. Guaiacol peroxidase （POD） and glutathione reductase （GR） activities increased in response to cold and drought but decreased in response to heat stress. The activity of ascorbata peroxidase （APX） responded differently to all three stresses. Monodehydroascorbate reductase （MDHAR） activity decreased in response to heat and drought, and slightly increased in response to the cold stress but was not significantly different from the controls （P 〈 0.05）. The activity of dehydroascorbata reductase （DHAR） increased in response to all three stresses. Taken together, the co-ordinate increase of the oxygen-detoxifying enzymes might be more effective to protect crofton weed from the accumulation of oxygen radicals at low temperatures rather than at high temperatures.     

Predisposition for and Prevention of Subjective Tinnitus Development

Dysfunction of the inner ear as caused by presbyacusis, injuries or noise traumata may result in subjective tinnitus, but not everyone suffering from one of these diseases develops a tinnitus percept and vice versa. The reasons for these individual differences are still unclear and may explain why different treatments of the disease are beneficial for some patients but not for others. Here we for the first time compare behavioral and neurophysiological data from hearing impaired Mongolian gerbils with (T) and without (NT) a tinnitus percept that may elucidate why some specimen do develop subjective tinnitus after noise trauma while others do not. Although noise trauma induced a similar permanent hearing loss in all animals, tinnitus did develop only in about three quarters of these animals. NT animals showed higher overall cortical and auditory brainstem activity before noise trauma compared to T animals; that is, animals with low overall neuronal activity in the auditory system seem to be prone to develop tinnitus after noise trauma. Furthermore, T animals showed increased activity of cortical neurons representing the tinnitus frequencies after acoustic trauma, whereas NT animals exhibited an activity decrease at moderate sound intensities by that time. Spontaneous activity was generally increased in T but decreased in NT animals. Plastic changes of tonotopic organization were transient, only seen in T animals and vanished by the time the tinnitus percept became chronic. We propose a model for tinnitus prevention that points to a global inhibitory mechanism in auditory cortex that may prevent tinnitus genesis in animals with high overall activity in the auditory system, whereas this mechanism seems not potent enough for tinnitus prevention in animals with low overall activity.