Telomere length assessment: biomarker of chronic oxidative stress?

Telomeres are nucleoprotein structures, located at the ends of chromosomes and are subject to shortening at each cycle of cell division. They prevent chromosomal ends from being recognized as double strand breaks and protect them from end to end fusion and degradation. Telomeres consist of stretches of repetitive DNA with a high G-C content and are reported to be highly sensitive to damage induced by oxidative stress. The resulting DNA strand breaks can be formed either directly or as an intermediate step during the repair of oxidative bases. In contrast to the majority of genomic DNA, there is evidence that telomeric DNA is deficient in the repair of single strand breaks. Since chronic oxidative stress plays a major role in the pathophysiology of several chronic inflammatory diseases, it is hypothesized that telomere length is reducing at a faster rate during oxidative stress. Therefore, assessment of telomere length might be a useful biomarker of disease progression. In this review several features of telomere length regulation, their relation with oxidative stress, and the potential application of measurement of telomere length as biomarker of chronic oxidative stress, will be discussed.     

Occurrence of gustducin-immunoreactive cells in von Ebner’s glands of guinea pigs

An immunohistochemical examination of guinea-pig taste buds in vallate papillae revealed gustducin-immunoreactive cells in the area of von Ebner’s glands, minor salivary glands. Since there have been no reports describing those cells in these locations for other species, we investigated these glands in order both to localize the cells and compare their immunoreactive characteristics with corresponding cells in the vallate taste buds. The gustducin-immunoreactive cells coincided with cells containing no secretory granules in the end portion of the glands, which was supported by the electron-microscopic immunocytochemistry. Double immunofluorescence microscopy confirmed these cells to be entirely immunopositive to type III inositol 1,4,5-triphosphate receptor (IP₃R-3), phospholipase Cβ2 (PLCβ2), and villin and also partly immunopositive to neuron-specific enolase (NSE) and calbindin D-28K. The gustducin-immunoreactive cells in the vallate taste buds exhibited completely the same immunoreactivities for these five molecules. Accordingly, the present results give credence to a consideration that the gustducin-immunnoreactive cells in both locations are identical in function(s) e.g., chemo-reception.     

Localized vs. systemic inflammation in guinea pigs: a role for prostaglandins at distinct points of the fever induction pathways?

In guinea pigs, dose-dependent febrile responses were induced by injection of a high (100 microg/kg) or a low (10 microg/kg) dose of bacterial lipopolysaccharide (LPS) into artificial subcutaneously implanted Teflon chambers. Both LPS doses further induced a pronounced formation of prostaglandin E(2) (PGE(2)) at the site of localized subcutaneous inflammation. Administration of diclofenac, a nonselective cyclooxygenase (COX) inhibitor, at different doses (5, 50, 500, or 5,000 microg/kg) attenuated or abrogated LPS-induced fever and inhibited LPS-induced local PGE(2) formation (5 or 500 microg/kg diclofenac). Even the lowest dose of diclofenac (5 microg/kg) attenuated fever in response to 10 microg/kg LPS, but only when administered directly into the subcutaneous chamber, and not into the site contralateral to the chamber. This observation indicated that a localized formation of PGE(2) at the site of inflammation mediated a portion of the febrile response, which was induced by injection of 10 microg/kg LPS into the subcutaneous chamber. Further support for this hypothesis derived from the observation that we failed to detect elevated amounts of COX-2 mRNA in the brain of guinea pigs injected subcutaneously with 10 microg/kg LPS, whereas subcutaneous injections of 100 microg/kg LPS, as well as systemic injections of LPS (intra-arterial or intraperitoneal routes), readily caused expression of the COX-2 gene in the guinea pig brain, as demonstrated by in situ hybridization. Therefore, fever in response to subcutaneous injection of 10 microg/kg LPS may, in part, have been evoked by a neural, rather than a humoral, pathway from the local site of inflammation to the brain.     

Repeat participation among normal healthy research volunteers: professional guinea pigs in clinical trials?

The recent death of a normal healthy volunteer, as well as the increased use of normal volunteers as research subjects, has heightened the attention given to the participation of normal volunteers in clinical research. An overlooked sub-population of normal healthy volunteers are repeat, veteran volunteers. This essay discusses ethical and methodological issues associated with the use of repeat volunteers in research, along with existing guidelines regarding the use of repeat healthy volunteers, and concludes with recommendations for safeguarding repeat volunteers and ideas supporting centralized recruiting.     

Latissimus dorsi cardiomyoplasty: a chronic experimental porcine model. Feasibility study of cardiomyoplasty in Danish Landrace pigs and Göttingen minipigs

Cardiomyoplasty is an experimental treatment for end-stage heart failure. We hypothesized that the porcine latissimus dorsi muscle (LDM) in an experimental porcine model is a suitable surrogate for a clinically relevant evaluation of this concept. Fourteen Danish Landrace (DL) pigs and six G?ttingen minipigs (GM) were studied. The LDM was evaluated immediately after surgical dissection and in various phases: phase 1 (n = 4)--outcome of a partial vascular isolation (vascular delay), 2 to 3 weeks prior to heart wrapping in DL pigs; phase 2 (n = 6)--long-term flap survival of nonstimulated LDM cardiomyoplasty in DL pigs; phase 3 (n = 6)--outcome of nonstimulated cardiomyoplasty in GM; phase 4--one DL pig had dynamic cardiomyoplasty performed and was subjected to low-intensity LDM stimulation for 8 months. Isolation of the LDM of DL pigs and GM as a pedicled graft had no acute deleterious impact on the global blood supply. In phase 1a, partial vascular isolation and in situ recovery of the LDM resulted in a muscle encapsulated in fibrotic tissue, which hampered a later heart wrap. In phase 1b, a less extensive dissection diminished fibrosis and allowed subsequent wrapping. In phase 2, after 6 weeks of nonstimulated LDM cardiomyoplasty, the LDM of DL pigs was viable, with excellent heart-muscle integration. In phase 3, the same procedure applied in GM yielded the same result as that in DL pigs, but with a higher success rate owing to the learning phase.     

The yield of synaptosomes from the cerebral cortex of guinea pigs estimated by a polystyrene bead “tagging” procedure

Summary The number of synaptosomes (pinched-off nerve endings) produced/g guinea pig cortex on homogenizing this tissue under defined conditions is estimated to be in the region of 4×10¹¹ using two different polystyrene bead tagging procedures. This is the same order of magnitude as the number of nerve endings/g cortex calculated from histological estimates given in the literature of the number of neurones in the cortex and the extent of their cortical connexions.This investigation was supported by U.S. Public Health Service Grant no. NB 03928 from the National Institute of Neurological Diseases and Blindness. Dr. Sheridan was a Postdoctoral fellow of the U.S. Public Health Service during the period of his participation in this investigation (1963/64). The electron microscope facilities were provided by the Wellcome Trust. Mr. T.F.J. Hobson, of the ARC Statistics group, Cambridge, kindly advised on statistical aspects of the work. We are most grateful to him, to Miss L. Swales and Mr. G.H.C. Dowe for their skilled technical assistance and to Dr. E.G. Gray for helpful discussions.     

Brainstem neuronal populations activated in the model of ovalbumine induced allergic rhinitis in guinea pigs — the c-Fos study

Central neuronal interactions may contribute to up regulation of cough in subjects with rhinitis. Previously we have shown that noxious stimulation of the nose induces considerable Fos-like immunoreactivity (FLI) in the solitary nuclei and the region of ventral respiratory group and these neurons are involved in the cough pattern generator. Recent study addressed the question, which additional groups are activated in model of trigeminal hyperresponsiveness and whether some of them might also cooperate with cough pattern gating areas. 24 guinea pigs were sensitized with intraperitoneal ovalbumin (OVA) and later were once weekly challenged with intranasal OVA to develop the neural hyperresponsiveness, 12 animals were left unsensitized. The nasal symptom score was evaluated after each challenge. Finally, animals were anaesthetized and the latest challenges with nasal OVA, capsaicin and saline were applied to induce c-Fos expression in designed groups. Following the survival time animals were deeply anaesthetized, exsanguinated, and transcardially perfused with heparinised saline (200 ml) and paraformaldehyde fixation (200 ml). The brainstems were removed, postfixed, and brainstem slices were processed immunohistochemically (c-Fos, Calbiochem, SR). FLI at the level of obex and areas relative to the obex was analyzed. In all groups (excluding the saline group) the FLI was detected bilaterally in the trigeminal complex, nuclei of solitary tract, lateral reticular and nucleus ambiguus. There were no differences between the OVA and capsaicin groups. Count of Fos-positive neurons within the trigeminal complex does not correlate with the magnitude of clinical symptoms, which gradually increased each week in OVA induced model of hyperresponsiveness. Whereas trigeminal hyperresponsiveness contributes to the up-regulation of cough in animal models, it does not induce any additional neuronal FLI at the middle medulla than observed in naive animals.     

The cortisol stress response in male round goby (Neogobius melanostomus): effects of living in polluted environments?

Acute exposure to contaminants frequently induces stress, but prolonged exposures, such as those experienced by individuals in wild populations, can impair the capacity to mount a stress response. Using the round goby (Neogobius melanostomus), a small benthic fish and potential sentinel species for habitat contamination in the Great Lakes, we explored the impacts of living in highly polluted areas. Round goby were collected from highly contaminated and less contaminated areas of Hamilton Harbour (a well-known site of polycyclic aromatic hydrocarbon (PAH) and heavy metal contamination). The cortisol stress responses of fish from sites of high and low contamination were compared using an EIA assay on blood collected (n?=?112) either prior to or 0, 10, 30, 60, 240 min and 24 h following the application of a 4-min confined air exposure stressor. Plasma cortisol levels were elevated at 10 and 30 min post-stressor (100.3 and 87.5 ng/mL), and returned to levels similar to baseline (22.3 ng/mL) 1 h after the stressor. In contrast to predictions, round goby from areas of high and low contamination had similar cortisol levels at all timepoints. We also monitored stress responses immediately after a chasing stressor (n?=?19). During the chasing stressor, contaminated-site round goby exhausted faster than individuals from a less contaminated site, although they had similar levels of plasma cortisol (23.5 versus 20.9 ng/mL) and lactate (2.53 versus 1.98 mmol/L). Our results indicate that not all fishes may demonstrate impaired stress responses, even in highly contaminated habitats; however, such animals may still have increased vulnerability to predation.     

Proline accumulation: A parameter for evaluation of sensitivity of tomato varieties to drought stress?

The pattern of proline accumulation and the growth response were followed in several tomato ( Lycopersicon esculentum Mill.) varieties which were exposed to 7 days of drought stress followed by a 15-day period of rewatering. During dehydration, water potential and leaf elongation rates decreased more in var. 'Hosen' and 'S-5' than in 'LX-11', '1970', 'Pakmor', 'Faculty-16', 'Alcobaca' and '475'. Proline accumulation during stress was greatest in the first two varieties. In 'Hosen' and 'S-5' rewatering resulted in a decrease of proline to control levels, whereas in the other varieties accumulation of proline continued long after turgor had been regained. The extent of this continued accumulation was not correlated with the degree to which each variety was dehydrated. Upon rewatering of the plants the rate of leaf elongation was increased, but the final leaf size as well as whole shoot and root fresh weight of the recovered plants were not colated with the degree of "suffering" that each variety experienced during the drought period. Incubation of detached young tomato leaves in polyethylene glycol solution for 48 h resulted in a substantial accumulation of proline. The varietal differences observed under these conditions were reminiscent of the differential responses in proline accumulation obtained in the intact plants. It is concluded that proline accumulation at the time of dehydration signals drought stress in tomato plants but does not correlate with the overall varietal sensitivity to transient dehydration in recovered plants.     

Inhibition of keratinocyte apoptosis by IL-15: a new parameter in the pathogenesis of psoriasis?

Keratinocytes (KC) are important source of and targets for several cytokines. Although KC express IL-15 mRNA, the functional effects of IL-15 on these epithelial cells remain to be dissected. Investigating primary human foreskin KC and HaCaT cells, we show here by semiquantitative RT-PCR and flow cytometric analysis that both translate IL-15 and IL-15R mRNA and express IL-15 and IL-15Ralpha protein on the cell surface, suggesting that human KC can employ IL-15 for juxtacrine signaling. While IL-15 exerted no significant effect on KC proliferation and IL-6 or IL-8 secretion, IL-15 inhibited both anti-Fas and methylcellulose-induced KC apoptosis in vitro. This is in line with the recognized potent anti-apoptotic effects of IL-15. IL-2, whose receptor shares two components with the IL-15R, failed to inhibit KC apoptosis. Together with the role of IL-15 in sustaining chronic immune reactions, this invited the question of whether a reduction of KC apoptosis by IL-15 may be involved in the pathogenesis of psoriasis, a chronic hyperproliferative inflammatory skin disease characterized by abnormally low KC apoptosis in the epidermis. Remarkably, compared with nonlesional psoriatic skin and skin of healthy volunteers, lesional psoriatic epidermis showed high IL-15 protein expression in the epidermis and enhanced binding activity for IL-15. Therefore, antagonizing the inhibitory effects of IL-15 on KC apoptosis deserves exploration as a novel therapeutic strategy in psoriasis management.     

How do frugivores track resources? Insights from spatial analyses of bird foraging in a tropical forest

Frugivores often track ripe fruit abundance closely across local areas despite the ephemeral and typically patchy distributions of this resource. We use spatial auto- and cross-correlation analyses to quantify spatial patterns of fruit abundance and avian frugivory across a 4-month period within a forested 4.05-ha study grid in Puerto Rico. Analyses focused on two tanager species, Spindalis portoricensis and Nesospingus speculiferus, and their principal food plants. Three broad questions are addressed: (1) at what spatial scales is fruit abundance and frugivory patchy; (2) at what spatial scales do frugivores respond to fruit abundance; and (3) to what extent do spatial patterns of frugivory overlap between bird species? Fruit patch size, species composition, and heterogeneity was variable among months, despite fruit patch locations remaining relatively consistent between months. Positive correlations between frugivory and fruit abundance suggested tanagers successfully tracked fruit abundance. Frugivory was, however, more localized than fruit abundance. Scales of spatial overlap in frugivory and monthly variation in the foraging locations of the two tanager species suggested that interspecific facilitation may have been important in determining bird foraging locations. In particular, S. portoricensis, a specialist frugivore, may have relied on the loud calls of the gregarious generalist, N. speculiferus, to find new foraging areas. Such a mechanism could help explain the formation of mixed species feeding flocks and highlights the potential importance of facilitation between species that share resources.     

Memory in the grip of emotions and stress: a necessarily harmful impact?

While intense negative events are vividly recalled, information learned during stressful situations is poorly remembered. These differential effects of emotions and stress on memory have been attributed to the physiological manifestations generated during those affective states. Intense emotional and stressful events trigger the secretion of catecholamines and of glucocorticoids, in particular. These hormones would be modulatory agents of memory functions. In the first part of this paper, we review the specific effects emotions and stress have on memory. We then summarize the psychological and biological determinants responsible for these effects. Finally, we discuss different methodological issues that could explain the discrepancy found between the impact of emotions and stress on memory. Defining more precisely the effects emotion and stress have on memory will lead to a better comprehension of the cognitive problems that characterize patients dealing with emotional turmoil, such as patients suffering from depression or post-traumatic stress disorder.     

Oxidation of uric acid in rheumatoid arthritis: is allantoin a marker of oxidative stress?

Free radicals are implicated in many diseases including atherosclerosis, cancer and also in rheumatoid arthritis. Reaction of uric acid with free radicals, such as hydroxyl radical and hypochlorous acid (HOCl) results in allantoin production. In this study, we measured the serum allantoin levels, oxidation products of uric acid, as a marker of free radical generation in rheumatoid arthritis. Fasting blood samples were obtained from 21 rheumatoid patients and 15 healthy controls. In this study, the serum allantoin and uric acid levels were measured by a gas chromatography-mass spectrometry method and the ratios were calculated. The mean allantoin and uric acid levels and ratios in the patient group were 22.1±11.3, 280.5±65.0 and 8.0±3.7 μM, while in the control group they were 13.6±6.3, 278.3±53.6 and 4.9±2.1 μM, respectively. The effects of gender, age, menopausal status, duration of disease and medications on serum allantoin and uric acid levels of the patient and control groups were studied. Our results suggest that uric acid acts as a free radical scavenger and thus is converted to allantoin. Increased allantoin levels suggest the possible involvement of free radicals in rheumatoid arthritis.     

Analysis of DNA damage induced by aflatoxin B1 in Dunkin–Hartley guinea pigs

Aflatoxin B1 (AFB1) is among the most potent naturally occurring carcinogens and classified as a group I carcinogen. Since the ingestion of aflatoxin-contaminated food is associated with several liver diseases, the aim of the present study was to evaluate the effect of 2, 20, and 200 ppb of AFB1 on DNA damage in peripheral blood lymphocytes and liver cells in Dunkin-Hartley guinea pigs. The animals were divided into four groups according to the given diet. After the treatment the lymphocytes and liver cells were isolated and DNA damage determined by Comet assay. The levels of DNA damage in lymphocytes were higher animals treated with 200 ppb of AFB1-enriched diet (P = 0.02). In the liver cells there were a relationship between the levels of DNA damage and the consumption of AFB1 in all studied groups. These results suggest that Comet assay performed on lymphocytes is a valuable genotoxic marker for high levels of exposure to AFB1 in guinea pig. Additionally our results indicate that the exposure to this toxin increases significantly and increases the level of DNA damage in liver cells, which is a key step on liver cancer development. We also suggest that the Comet assay is an useful tool for monitoring the genotoxicity of AFB1 in liver.     

Are characiform Fishes Gondwanan in Origin? Insights from a Time-Scaled Molecular Phylogeny of the Citharinoidei (Ostariophysi: Characiformes)

Fishes of the order Characiformes are a diverse and economically important teleost clade whose extant members are found exclusively in African and Neotropical freshwaters. Although their transatlantic distribution has been primarily attributed to the Early Cretaceous fragmentation of western Gondwana, vicariance has not been tested with temporal information beyond that contained in their fragmentary fossil record and a recent time-scaled phylogeny focused on the African family Alestidae. Because members of the suborder Citharinoidei constitute the sister lineage to the entire remaining Afro-Neotropical characiform radiation, we inferred a time-calibrated molecular phylogeny of citharinoids using a popular Bayesian approach to molecular dating in order to assess the adequacy of current vicariance hypotheses and shed light on the early biogeographic history of characiform fishes. Given that the only comprehensive phylogenetic treatment of the Citharinoidei has been a morphology-based analysis published over three decades ago, the present study also provided an opportunity to further investigate citharinoid relationships and update the evolutionary framework that has laid the foundations for the current classification of the group. The inferred chronogram is robust to changes in calibration priors and suggests that the origins of citharinoids date back to the Turonian (ca 90 Ma) of the Late Cretaceous. Most modern citharinoid genera, however, appear to have originated and diversified much more recently, mainly during the Miocene. By reconciling molecular-clock- with fossil-based estimates for the origins of the Characiformes, our results provide further support for the hypothesis that attributes the disjunct distribution of the order to the opening of the South Atlantic Ocean. The striking overlap in tempo of diversification and biogeographic patterns between citharinoids and the African-endemic family Alestidae suggests that their evolutionary histories could have been strongly and similarly influenced by Miocene geotectonic events that modified the landscape and produced the drainage pattern of Central Africa seen today.     

Review: Insect meal: a future source of protein feed for pigs?

Are insects the farm animal of the future? A key agenda for agricultural production systems is the development of sustainable practices whereby food and feed can be produced in an environmentally efficient manner. These goals require novel approaches to complex problems and demand collaboration between scientists, producers, consumers, government and the general population. The provision of feed for animals is a major contributor to land and water use and greenhouse gas (GHG) emissions. Further, overfishing and a reduction in available land and water resources on which crops can be grown has led to an increase in price of protein ingredients such as fish meals and oils and soybean meals. Determination of novel solutions to meet the feed protein requirements of production animals is key to the development of sustainable farming practices. The Australian pork industry aims to develop production systems that efficiently use available resources (such as feed and energy) and limit the production of emissions (such as manure waste and GHGs). Invertebrates (insects e.g. black soldier flies) are naturally consumed by monogastric and aquatic species, yet the large-scale production of insects for feed (or food) is yet to be exploited. Most insects are low producers of GHGs and have low land and water requirements. The large-scale production of insects can contribute to a circular economy whereby food and feed waste (and potentially manure) are reduced or ideally eliminated via bioconversion. While the concept of farm-scale production of insects as domestic animal feed has been explored for decades, significant production and replacement of traditional protein sources has yet to be achieved. This review will focus on the potential role of insect-derived protein as a feed source for the Australian pig production industry.     

When does stress end? Evidence of a prolonged stress reaction in shiftworking truck drivers

This study aimed to analyze individual cortisol levels in relation to work conditions, sleep, and health parameters among truck drivers working day shifts (n = 21) compared to those working irregular shifts (n = 21). A total of 42 male truck drivers (39.8 (+/-) 6.2 yrs) completed questionnaires about sociodemographics, job content, work environment, health, and lifestyle. Rest-activity profiles were measured using actigraphy, and cardiovascular blood parameters were collected. Salivary cortisol samples were obtained: (i) at waking time, (ii) 30?min after waking, and (iii) at bedtime, during both one workday and one day off from work. Irregular-shift workers, compared to day-shift workers, showed significantly higher waist-hip ratio, very-low-density lipoprotein (VLDL) cholesterol, tiredness after work, years working as a driver, truck vibration, and less job demand (p < .05). High cortisol levels in irregular-shift workers were correlated with certain stressors, such as short sleep duration and low job satisfaction, and to metabolic parameters, such as total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), VLDL, and triglycerides. Day-shift workers had higher cortisol levels collected 30?min after waking (p = .03) and a higher cortisol awakening response (CAR; p = .02) during workdays compared to off days. Irregular-shift workers had higher cortisol levels on their off days compared to day-shift workers (p = .03). In conclusion, for the day-shift workers, a higher cortisol response was observed on workdays compared to off days. Although no direct comparisons could be made between groups for work days, on off days the irregular-shift workers had higher cortisol levels compared to day-shift workers, suggesting a prolonged stress response in the irregular-shift group. In addition, cortisol levels were correlated with stressors and metabolic parameters. Future studies are warranted to investigate further stress responses in the context of irregular work hours.     

Wild genius - domestic fool? Spatial learning abilities of wild and domestic guinea pigs

Background

In social insects, the queen is essential to the functioning and homeostasis of the colony. This influence has been demonstrated to be mediated through pheromone communication. However, the only social insect for which any queen pheromone has been identified is the honey bee (Apis mellifera) with its well-known queen mandibular pheromone (QMP). Although pleiotropic effects on colony regulation are accredited to the QMP, this pheromone does not trigger the full behavioral and physiological response observed in the presence of the queen, suggesting the presence of additional compounds. We tested the hypothesis of a pheromone redundancy in honey bee queens by comparing the influence of queens with and without mandibular glands on worker behavior and physiology.

Results

Demandibulated queens had no detectable (E)-9-oxodec-2-enoic acid (9-ODA), the major compound in QMP, yet they controlled worker behavior (cell construction and queen retinue) and physiology (ovary inhibition) as efficiently as intact queens.

Conclusions

We demonstrated that the queen uses other pheromones as powerful as QMP to control the colony. It follows that queens appear to have multiple active compounds with similar functions in the colony (pheromone redundancy). Our findings support two hypotheses in the biology of social insects: (1) that multiple semiochemicals with synonymous meaning exist in the honey bee, (2) that this extensive semiochemical vocabulary exists because it confers an evolutionary advantage to the colony.     

Is there a gloxylate cycle in the liver of the fetal guinea pig?

Studies on the effect of the inhibitor of fatty acid oxidation (+)-octanoylcarnitine on the perfused liver of the 48–51 days fetal guinea pig indicate that the oxidation of endogenous fatty acids is a major source of carbon for the citric acid cycle and for synthesis of hexose. Consistent with this the liver can convert isocitrate to glyoxylate and glyoxylate to malate and may therefore operate a glyoxylate cycle allowing the net production of sugars from acetyl-CoA.