Effects of lactation, energetic deficit and remating interval on reproductive performance of primiparous rabbit does.

Female rabbits (n = 151) were assigned at their first parturition (day 0) to one of three treatment groups (A: non-lactating does and ad libitum feeding, R: non-lactating does and restricted feeding, L: lactating does and ad libitum feeding). Additional females (n = 18) were slaughtered at parturition. Experimental females were presented to the male on days 3 (A3, L3 and R3 groups, n = 25, 26 and 26, respectively) or 10 (A10, L10 and R10 groups, n = 25, 25 and 24, respectively) and were slaughtered on day 28 after parturition. Compared to does slaughtered on day 0, adipose tissues were lighter in L (-66%) and R (-32%) females while they were heavier in A females (+40%; P < 0.01). These results suggest that both L and R females were in energy deficit. Receptivity (80% vs. 98%) and conception rate (51% vs. 83%) were lower in L compared to A females (P < 0.01) regardless the day of male presentation. Ovulation rate (-14%) and conception rate (-26%) were lower in R3 than in A3 does (P < 0.05). Ovulation rate was 24% lower in the L10 than in the A10 group (P < 0.01). The uterine contents were lighter (-25%) in L and R than in A does (P < 0.001) regardless the day of male presentation. These results suggest that the energy deficit associated with milk production can partly explain the negative influence of lactation on reproductive performance.     

Influence of metabolic status on oocyte quality and follicular characteristics at different postpartum periods in primiparous rabbit does

Low reproductive performance of high-yield primiparous animals is closely associated with the metabolic stress caused by a simultaneous gestation and lactation. The aim of this work was (1) to analyze body composition and metabolic environment at three time points along lactation (at parturition time; in the lactation period [Day 11 postpartum]; and in the postweaning period [Day 32 postpartum]) of primiparous rabbit does (Oryctolagus cuniculus) and (2) to investigate the ovarian status at insemination time and the possible link with metabolic environment and with their reproductive performance. To this end, does were either submitted to a semi-intensive reproductive rhythm (Group S, inseminated on Day 11 postpartum) or an extensive rhythm (Group E, inseminated on Day 32 postpartum). Body energy (P < 0.05) and protein content (P < 0.001) as well as serum leptin (P < 0.05) and protein concentrations (P < 0.001) increased significantly along the postpartum period. At parturition, body lipid content was significantly lower and serum nonesterified fatty acids concentrations were significantly higher than that on Days 11 postpartum and 32 postpartum. Concerning ovarian status at insemination time, no significant differences were found in mean follicular stages, serum estradiol, progesterone, and prolactin (PRL) concentrations or in prolactin receptor (PRL-R) immunostaining. However, follicles in Group S showed a significantly higher apoptosis index than that of Group E (P < 0.001). The nuclear and cytoplasmic oocyte maturation rates of Group S were also significantly lower than that in Group E. In addition, conception rate and prolificacy were improved in Group E (P < 0.001 and P < 0.05, respectively). In conclusion, in the early postpartum period, metabolic status seems to impact negatively on ovarian follicle and oocyte quality leading to a poor reproductive outcome in primiparous rabbit does.     

Energy balance and reproductive performance in rabbit does

The mortality and the culling rates of females are very high in rabbit breeding. In many cases, the dead or culled females show low reproductive performances and poor body conditions. Energy deficit leads to body mobilization and reduced reproductive performance. Consequently, energy balance and lifespan are closely linked. Two in vivo methods to estimate body composition were recently validated for rabbit does. Ultrasounds and TOBEC methods permitted the evaluation of the changes in energy balance and their relationship to reproductive performance of females. Influence of intrinsic (parity, litter size, genetic line, ...) and extrinsic (reproduction rhythm, temperature, ...) factors on the energetic balance of females are discussed. Energy deficit and depletion of body stores occurring during lactation lead to a decrease in reproductive performance and the physiological mechanisms implicated in these effects were presented. Interest and limitations of several strategies to reduce the body energy deficit in reproducing does and to improve their fertility and thus lifespan are discussed: rearing, feeding, management, genetic selection. A multi-facetted approach, combining these various strategies seems to be required to meet these objectives.     

Differences in the reproductive endocrine status of ewes in the early post-partum period and during seasonal anoestrus

More (P less than 0.05) post-partum acyclic ewes (8/9) showed evidence of pulsatile LH release than did seasonally anoestrous ewes (2/8). Mean plasma prolactin concentrations were higher (P less than 0.05) in the post-partum ewes than in the seasonally anoestrous ewes. Hypothalamic and pituitary cytosol oestrogen receptor number, median eminence GnRH content and pituitary LH, FSH and prolactin contents were similar in the two groups of ewes. The number of nuclear-bound oestrogen receptors was greater (P less than 0.01) in pituitaries from seasonally anoestrous ewes than in pituitaries from post-partum ewes. These data suggest that the basis of acyclicity in seasonally anoestrous ewes and in post-partum ewes is probably due to their inability to generate LH pulse frequencies similar to that in the follicular phase of the oestrous cycle. The higher LH pulse frequency in post-partum ewes, compared to that in seasonally anoestrous ewes, suggests that pregnancy may partly negate the reduction in LH secretion that is associated with photoperiod-induced acyclicity. The lower number of nuclear-bound oestrogen receptors in post-partum ewes suggests that there may be lower plasma oestrogen levels and less ovarian follicular growth than in non-suckling anoestrous ewes.     

Exercise distributed across day and night does not alter circadian period in humans

In rodents, increased activity due to running-wheel access is associated with a change in observed circadian period. In humans, exposure to exercise has failed to demonstrate similar effects on period. Methodological issues with prior studies such as light exposure during exercise, length of study, and method of measuring period confounded those evaluations of the effect of exercise on period in humans. In the present experiment, the authors examined the effect of exercise on period in 8 subjects using a 44-day within-subjects inpatient study. They used a 20-h forced desynchrony protocol, in which subjects were exposed to exercise across circadian phases under dim light conditions. Exercise consisted of three 45-min sessions per wake period on an ergometer. Target exercise intensity was ~65% of maximal heart rate. Intrinsic circadian period was measured using both core body temperature and hourly plasma melatonin samples. Consistent with previous reports, the authors find no effect of exercise on endogenous circadian period as measured by either core body temperature or melatonin. Exercise distributed across biological day and night does not appear to affect circadian period.     

Temperature change does not affect force between single actin filaments and HMM from rabbit muscles

The temperature dependence of sliding force, velocity, and unbinding force was studied on actin filaments when they were placed on heavy meromyosin (HMM) attached to a glass surface. A fluorescently labeled actin filament was attached to the gelsolin-coated surface of a 1-microm polystyrene bead. The bead was trapped by optical tweezers, and HMM-actin interaction was performed at 20-35 degrees C to examine whether force is altered by the temperature change. Our experiments demonstrate that sliding force increased moderately with temperature (Q(10) = 1.6 +/- 0.2, +/-SEM, n = 9), whereas the velocity increased significantly (Q(10) = 2.9 +/- 0.4, n = 10). The moderate increase in force is caused by the increased number of available cross-bridges for actin interaction, because the cross-bridge number similarly increased with temperature (Q(10) = 1. 5 +/- 0.2, n = 3) when measured during rigor induction. We further found that unbinding force measured during the rigor condition did not differ with temperature. These results indicate that the amount of force each cross-bridge generates is fixed, and it does not change with temperature. We found that the above generalization was not modified in the presence of 1 mM MgADP or 8 mM phosphate.     

Daunorubicin does not induce immunohistochemically detectable endothelial dysfunction in rabbit aorta and femoral artery

Anthracyclines are one of the most effective anticancer drugs ever developed, but their clinical use has been hampered by the risk of severe cardiotoxicity. In this study, we investigated whether rabbits exposed to a different cumulative dose of anthracycline suffer from immunohistochemically detectable vascular toxicity and endothelial dysfunction. Daunorubicin (3 mg/kg, i.e. 50 mg/m2) was administered i.v. to rabbits once weekly for 1-10 weeks to reach different cumulative doses of the drug (50-500 mg/m2), while control rabbits received saline. The rabbits were sacrificed either 24 hours or 7 days after reaching each particular cumulative dose, and aortas and right femoral arteries were collected for immunohistochemical analysis. Immunohistochemical analysis showed ICAM-1 staining in many aortas from both saline and daunorubicin-treated rabbits without any relationship to the anthracycline treatment. On the contrary, unlike in the lipopolysaccharide-treated or hypercholesterolemic rabbits, no distinct immunoreactivity for other markers of inflammation, oxidative and nitrosative stress (VCAM-1, 4-HNE, iNOS and nitrotyrosine) were detected in aortas and femoral arteries from either control or daunorubicin-treated animals. No relationship to the cumulative dose of the drug or post-expose set up of harvesting was found. In this study, we have demonstrated that daunorubicin does not induce gross histopathological changes in the studied arteries and it fails to induce immunohistochemically detectable endothelial dysfunction. Thus, we propose that endothelial cells are much less susceptible to anthracycline toxicity than cardiac myocytes. In addition, our data suggest that vascular toxicity of anthracyclines plays rather a minor role in the cardiovascular complications of anthracycline chemotherapy.     

Intracerebroventricular and septal injections of arginine vasopressin are not antipyretic in the rabbit

Arginine vasopressin (AVP) has been reported to have an antipyretic effect in the ewe and guinea pig near term. Perfusions with AVP of sites in the septal region also reduced fever in non-pregnant sheep. In the present experiments adult rabbits with third cerebral ventricular or septal cannulas were restrained in a 23°C environment, and rectal temperature was recorded every 10 min. Fever induced by IV administration of leukocytic pyrogen was not reduced by AVP (25–100 ng) given intraventricularly 20 min later. Doses of 1–5 μg AVP injected into the septum likewise were not antipyretic but actually caused an increase in fever. This augmentation of the febrile response is consistent with results of previous studies in this laboratory in which AVP increased hyperthermia in a hot environment and enhanced hyperthermic responses to PGE₂. The data from these experiments provide no evidence that central AVP is an endogenous antipyretic in rabbits; rather, it may be that central AVP augments fever in this species.     

To breed or not to breed: Physiological correlates of reproductive status in a facultatively biennial iguanid

It is unusual for seasonal breeders to frequently skip opportunities for reproduction. We investigated the relationship between physiological state and reproductive decision-making in Galápagos marine iguanas (Amblyrhynchus cristatus), a species in which females typically reproduce biennially, although the proportion of breeding individuals varies significantly across years. Nearly all adult-sized females initiated follicular development prior to the lekking period, but 38% of females resorbed all developing follicles 5-15 days before the start of copulations. Receptive and non-receptive females differed in reproductive hormones during the mate choice period. Testosterone peaked in receptive females immediately prior to copulation, indicating that testosterone or its derivative estradiol likely mediates female receptivity in Galápagos marine iguanas. Non-receptive females showed significant peaks in both testosterone and progesterone during follicular atresia, suggesting that these hormones may be involved in inhibiting vitellogenesis. Two to three weeks prior to the period of reproductive decision-making (and the onset of follicular atresia in non-receptive females) receptive females were in higher body condition, were developing larger follicles, and had lower levels of both baseline and stress-induced corticosterone. Reproduction is extremely costly in this long-lived species, and increases the likelihood of mortality in the year following breeding; females could therefore gain significant benefits from being attuned to indicators of reproductive success. We suggest that corticosterone may modulate reproductive decisions by altering individual sensitivity to both internal and external cues of the likelihood of successful reproduction.     

Mutation from guanine to adenine in 25S rRNA at the position equivalent to E. coli A2058 does not confer erythromycin sensitivity in Sacchromyces cerevisae

The macrolide erythromycin binds to the large subunit of the prokaryotic ribosome near the peptidyltransferase center (PTC) and inhibits elongation of new peptide chains beyond a few amino acids. Nucleotides A2058 and A2059 (E. coli numbering) in 23S rRNA play a crucial role in the binding of erythromycin, and mutation of nucleotide A2058 confers erythromycin resistance in both gram-positive and gram-negative bacteria. There are high levels of sequence and structural similarity in the PTC of prokaryotic and eukaryotic ribosomes. However, eukaryotic ribosomes are resistant to erythromycin and the presence of a G at the position equivalent to E. coli nucleotide A2058 is believed to be the reason. To test this hypothesis, we introduced a G to A mutation at this position of the yeast Saccharomyces cerevisiae 25S rRNA and analyzed sensitivity toward erythromycin. Neither growth studies nor erythromycin binding assays on mutated yeast ribosomes indicated any erythromycin sensitivity in mutated yeast strains. These results suggest that the identity of nucleotide 2058 is not the only determinant responsible for the difference in erythromycin sensitivity between yeast and prokaryotes.     

Cold stress does not induce stress proteins SP 25 and SP 72 in rat skeletal muscle.

Cryotherapy is a common treatment for musculoskeletal injuries, yet the mechanism(s) underlying its effects remain unclear. Since cryotherapeutic treatment often involves temperatures that are known to induce the protective stress proteins (SPs), we determined whether SP 25 and SP 72 expression was altered following a 20-min cold stress to the hindlimb muscles of Sprague-Dawley rats. The right hindlimb of anesthetized animals was placed in an ice bath until muscle temperature decreased to either 8.4 +/- 0.4 degrees C or 19.7 +/- 0.3 degrees C for 20 min. After a 24-h recovery, the white and red gastrocnemius, plantaris, soleus, extensor digitorum longus, and tibialis anterior muscles from both legs were removed and rapidly frozen in liquid nitrogen. Portions of the muscles were homogenized and SP 25 and SP 72 content was assessed by SDS-PAGE/Western blot analyses. Quantification of SP 25 and SP 72 by densitometric scanning of blots demonstrated no significant increases in SP 25 or SP 72 content in any of the muscles exposed to either the 8 or the 20 degrees C cold stress compared to muscles from the unstressed contralateral limbs. These results suggest that a 20-min cold stress of 8 degrees C or 20 degrees C does not increase muscle SP 25 or SP 72 content.     

To breed or not to breed: past reproductive status and environmental cues drive current breeding decisions in a long-lived amphibian

Iteroparity is an adaptive response to uncertainty in reproductive success. However, spreading reproductive success over multiple reproduction events during a lifetime is constrained by adult mortality and the stochasticity associated with interactions between external factors and physiological states. The acquisition of information about environmental conditions during the growth of progeny and sufficient resources during the non-reproductive period are key factors for breeding success. Consequently, we hypothesized that long-lived animals may skip a breeding opportunity when information about unfavourable environmental conditions is available. In addition, nutritional constraints could prevent an animal from replenishing its reserves sufficiently to invest in the current breeding period. We investigated these questions using capture–recapture data from a 5-year study on a large population of yellow-bellied toads in a forest in north-eastern France. We took advantage of various advances in multi-state capture–recapture models (e.g. unobservable states and mixture models) to test our hypotheses. Our results show that the combined effects of rainfall deficit and the breeding/non-breeding state of individuals during the past breeding season affect breeding probability during the following breeding opportunity. We also found that females breed less frequently than males, suggesting that the overall energy cost of reproduction differs between genders. Finally, the results indicate that toad survival appears to be negatively influenced by rainfall deficits. We discuss the yellow-bellied toad’s reproductive behaviour in term of bet-hedging strategy and life history trait evolution.     

Dietary protein restriction during lactation in primiparous sows with different live weights at farrowing: II. Consequences on reproductive performance and interactions with metabolic status

The hypothesis that the restriction of dietary protein during lactation has different impacts on reproductive performance in light and heavy sows at farrowing was investigated, as well as the relationships between reproductive parameters and sow metabolic data. At farrowing, 38 primiparous sows were assigned to one of three groups: sows weighing 180 kg not restricted in dietary protein during lactation (180CP); sows weighing 180 or 240 kg restricted in protein (180LP and 240LP). Twenty-four sows were catheterized and serial blood samples were collected 1 d before and 1 d after weaning. The sows were inseminated at the first estrus after weaning and slaughtered at d 30 of gestation. Protein restriction reduced the proportion of sows that returned to estrus within 8 d after weaning in the 180LP sows (P < 0.03), but not in the 240LP sows. It also induced a reduction in ovulation rate in the 180LP sows (P < 0.05) and, to a lesser extent, in the 240LP sows (P = 0.12). When the sows were categorized according to return to estrus (WOI < or = 8 or > 8 d), basal and mean concentrations of LH increased after weaning only in sows with a short WOI. Sows with a delayed estrus exhibited a higher ratio of plasma tyrosine to large neutral amino acids (AA, P < 0.01). In conclusion, large body reserves at farrowing buffer, at least in part, the detrimental effect of a strongly negative nitrogen balance on reproduction. We suggest that the alteration of AA profiles induced by dietary protein restriction and body protein loss alters LH secretion via modifications of the neurotransmitters involved in GnRH secretion.     

Passive immunization to outer membrane proteins MLP and PAL does not protect mice from sepsis

Multiple older studies report that immunoglobulin directed to rough mutant bacteria, such as E. coli J5, provides broad protection against challenge with heterologous strains of Gram-negative bacteria. This protection was initially believed to occur through binding of immunoglobulin to bacterial lipopolysaccharide (LPS). However, hundreds of millions of dollars have been invested in attempting to develop clinically-effective anti-LPS monoclonal antibodies without success, and no study has shown that IgG from this antiserum binds LPS. Identification of the protective mechanism would facilitate development of broadly protective human monoclonal antibodies for treating sepsis. IgG from this antiserum binds 2 bacterial outer membrane proteins: murein lipoprotein (MLP) and peptidoglycan-associated lipoprotein (PAL). Both of these outer membrane proteins are highly conserved, have lipid domains that are anchored in the bacterial membrane, are shed from bacteria in blebs together with LPS, and activate cells through Toll-like receptor 2. Our goal in the current work was to determine if passive immunization directed to MLP and PAL protects mice from Gram-negative sepsis. Neither monoclonal nor polyclonal IgG directed to MLP or PAL conferred survival protection in 3 different models of sepsis: cecal ligation and puncture, an infected burn model, and an infected fibrin clot model mimicking peritonitis. Our results are not supportive of the hypothesis that either anti-MLP or anti-PAL IgG are the protective antibodies in the previously described anti-rough mutant bacterial antisera. These studies suggest that a different mechanism of protection is involved.     

Serum leptin levels in community acquired pneumonia (CAP) are related to nutritional status and not to acute phase reaction

To determine whether leptin in patients with CAP acts as a nutritional or as an inflammatory marker and whether leptin plays any role regarding survival, we included 222 patients diagnosed of CAP, 142 men and 80 women, median age 74 years. We did not find significant differences in serum leptin levels between CAP patients and healthy controls, even after adjusting by BMI. Serum leptin levels were directly related with BMI, body fat and muscle mass and inversely related with inflammatory markers, including pro- and anti-inflammatory cytokines. Patients with positive blood cultures showed lower serum leptin and raised inflammatory markers. Although patients who died showed lower values of serum leptin, multivariate analysis showed that the prognostic value of low serum leptin levels depends on impaired nutritional status. In conclusion, we suggest that in CAP patients, leptin does not act as an inflammatory reactant but as a nutritional marker.     

Prostaglandin translocation from the lumen of the rabbit uterus in vitro in relation to day of pregnancy or pseudopregnancy

Transport of 3H-labeled prostaglandins (PGs) E2 and F2 alpha from the uterine lumen across the uterine wall has been studied in rabbit uteri in vitro in incubations lasting up to 180 min, in relation to sexual state of the rabbit, incubation temperature, intraluminal PG concentration, addition of metabolic inhibitors and time of incubation. PG accumulation by the tissue increased rapidly up to 30 min and then remained relatively constant. By 30 min, radioactivity was found in the external incubation medium, and this increased linearly with time. The translocation of PGF2 alpha was significantly greater in pseudopregnant than in pregnant animals on Day 6, whereas that of PGE2 was significantly higher in pregnant than in pseudopregnant animals on Day 6.8. In pregnant animals, both PGF2 alpha and PGE2 were translocated to the exterior more rapidly on Day 6.8 than on Days 5 or 6. Transport of PGs was reduced by low temperature, unaffected by metabolic inhibitors and only that of PGE2 increased with increased (5 microM) intraluminal concentrations. During incubation, the tissue remained viable as judged by T/M ratios (dpm tissue/dpm medium) for 204 thallium. Transport of [14C] sucrose was much slower than that of [14C] urea, which was greater than the fastest rates exhibited by the PGs. In general, amounts of radioactivity found in antimesometrial, mesometrial and lateral portions of the uterine wall, or in implantation and interimplantation areas did not differ, but more was found in the endometrium than the myometrium. PGF2 alpha was translocated unmetabolized to the external medium, while only two-thirds of the PGE2 was translocated unchanged, and one-third converted to PGF2 alpha. It is concluded that the rabbit uterus shows some selectivity in handling PGs in relation to stage of pregnancy.     

Binding and endocytosis of cluster glycosides by rabbit hepatocytes. Evidence for a short-circuit pathway that does not lead to degradation

Synthetic cluster glycosides containing either one, two, or three galactosyl or lactosyl residues per ligand were used to test the effect of carbohydrate clustering on binding by the rabbit hepatic Gal/GalNAc-binding lectin using either isolated rabbit hepatocytes or the solubilized, affinity-purified lectin. The tris- and bis-glycosides were superior to the mono-glycosides for inhibition of 125I-asialoorosomucoid binding to rabbit hepatocytes at 0 degrees C. The concentrations of the tris-glycosides required for 50% inhibition of 125I-asialoorosomucoid binding (4-8 microM) to hepatocytes were 50-100 times lower than the concentrations of the corresponding mono-glycosides required for 50% inhibition (400-500 microM). The isolated lectin, however, did not effectively discriminate between the mono-, bis-, and tris-glycosides, possibly indicating an organizational difference between the lectin in the cell membrane and the isolated lectin. When the cluster glycosides were labeled with 125I-tyrosine, it was shown that the tris- and bis-galactosides were bound to the hepatocytes at 37 degrees C, and that binding was followed by a step that led to ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid resistance, probably internalization. The process could be specifically inhibited by the neoglycoprotein Gal44-AI-bovine serum albumin, or by IgG specific for the hepatic lectin, but not by preimmune IgG. Internalization of the cluster glycosides did not lead to accumulation of ligand inside the cell, nor to degradation. Instead, the ligands were quickly released from the cells.