Repeated muscle damage blunts the increase in heat strain during subsequent exercise heat stress.

PURPOSE: Exercise-induced muscle damage (EIMD) has recently been shown to increase heat strain during exercise heat stress (HS), and represents a risk factor for exertional heat illness (EHI). We hypothesised that a repeated bout of EIMD blunts the increase in rectal temperature (T re) during subsequent endurance exercise in the heat. METHODS: Sixteen non-heat-acclimated males were randomly allocated to EIMD (n = 9) or control (CON, n = 7). EIMD performed a downhill running treatment at -10 % gradient for 60 min at 65 % [Formula: see text]O2max in 20 °C, 40 % RH. CON participants performed the same treatment but at +1 % gradient. Following treatment, participants rested for 30 min, then performed HS (+1 % gradient running for 40 min at 65 % [Formula: see text]O2max in 33 °C, 50 % RH) during which thermoregulatory measures were assessed. Both groups repeated the treatment and subsequent HS 14 days later. Isometric quadriceps strength was assessed at baseline, and 48 h post-treatment. RESULTS: The decrease in leg strength 48 h post-EIMD trial 1 (-7.5 %) was absent 48 h post-EIMD trial 2 (+2.9 %) demonstrating a repeated bout effect. Final T re during HS was lower following EIMD trial 2 (39.25 ± 0.47 °C) compared with EIMD trial 1 (39.59 ± 0.49 °C, P < 0.01), with CON showing no difference. Thermal sensation and the T re threshold for sweating onset were also lower during HS on EIMD trial 2. CONCLUSION: The repeated bout effect blunted the increase in heat strain during HS conducted after EIMD. Incorporating a muscle-damaging bout into training could be a strategy to reduce the risk of EHI and improve endurance performance in individuals undertaking heavy exercise with an eccentric component in the heat.

Dispersal, patch leaving, and distribution of Homalodisca vitripennis (Hemiptera: Cicadellidae).

Homalodisca vitripennis (Germar) and related species have caused millions of dollars in damage to southern California vineyards in recent years through the vectoring of Pierce's disease. However, the effects of surrounding vegetation on the dispersal and distribution of H. vitripennis are poorly understood. Therefore, the relationship between dispersal rates and patch quality was tested, as well as the basic predictions of the marginal value theorem. Additional experiments were conducted to compare the H. vitripennis distribution in an isolated crape myrtle (Lagerstroemia indica) patch and a L. indica patch bordering two alternative host patches. In mark-release-recapture tests, H. vitripennis dispersed farther from the release point in a patch of low-quality host plants (Prunus persica) than in patches of high-quality host plants (L. indica). In addition, H. vitripennis remained in L. indica patches longer than in P. persica patches and adjusted patch residence times in P. persica in correlation with known changes in plant physiology. These data suggest that H. vitripennis follows the basic predictions of marginal value theorem. In distribution tests, H. vitripennis were more abundant in the patch center than patch edges in the isolated L. indica patch, but in a patch bordering cottonwood (Populus sp.) and peach (P. persica), H. vitripennis numbers were generally higher along the edges of the patch. These data suggest that alternate hosts bordering cropping systems may be important to the spatial dynamics of H. vitripennis. Implications of these spatial observations on the biology of H. vitripennis and potential control methods are discussed.

Spatiotemporal patterns and dispersal of stink bugs (Heteroptera: Pentatomidae) in peanut-cotton farmscapes.

In the southeast United States, a field of peanuts, Arachis hypogaea L., is often closely associated with a field of cotton, Gossypium hirsutum L. The objective of this 4-yr on-farm study was to examine and compare the spatiotemporal patterns and dispersal of the southern green stink bug, Nezara viridula L., and the brown stink bug, Euschistus servus (Say), in six of these peanut-cotton farmscapes. GS(+) Version 9 was used to generate interpolated estimates of stink bug density by inverse distance weighting. Interpolated stink bug population raster maps were constructed using ArcMap Version 9.2. This technique was used to show any change in distribution of stink bugs in the farmscape over time. SADIE (spatial analysis by distance indices) methodology was used to examine spatial aggregation of individual stink bug species and spatial association of the two stink bug species in the individual crops. Altogether, the spatiotemporal analyses for the farmscapes showed that some N. viridula and E. servus nymphs and adults that develop in peanuts disperse into cotton. When these stink bugs disperse from peanuts into cotton, they aggregate in cotton at the interface, or common boundary, of the two crops while feeding on cotton bolls. Therefore, there is a pronounced edge effect observed in the distribution of stink bugs as they colonize the new crop, cotton. The driving force for the spatiotemporal distribution and dispersal of both stink bug species in peanut-cotton farmscapes seems to be availability of food in time and space mitigated by landscape structure. Thus, an understanding of farmscape ecology of stink bugs and their natural enemies is necessary to strategically place, in time and space, biologically based management strategies that control stink bug populations while conserving natural enemies and the environment and reducing off-farm inputs.

REVIEW paper: mare reproductive loss syndrome.

An epidemic of early fetal loss (EFL), late fetal loss (LFL), fibrinous pericarditis, and unilateral uveitis which occurred during the spring of 2001, are together now known as the mare reproductive loss syndrome (MRLS). A similar epidemic with less intensity was reported during the same period of time from southern Ohio, West Virginia, and Tennessee. The same syndrome with lesser intensity recurred in 2002. The estimated economic loss from the syndrome in 2001 and 2002 together was approximately $500 million. Both EFL and LFL were characterized by the absence of specific clinical signs in aborting mares. Nonhemolytic Streptococcus spp. and Actinobacillus spp. accounted for 65% of the organisms isolated from fetuses submitted for a postmortem during the MRLS period in 2001 and 2002. The pathologic findings in fetoplacental units of LFL included bronchopneumonia and funisitis, and there were no findings in EFL. Epidemiologic studies conducted in 2001 suggested an association between the presences of eastern tent caterpillars (ETC) in pastures with MRLS. Experimental studies in pregnant mares by exposure to ETC, or administration by stomach tube or with feed material, reproduced EFL and LFL. Similar experimental studies in mouse, rats, and goats with ETC were unsuccessful. Currently, 2 hypotheses are proposed for MRLS. One hypothesis proposes that an ETC-related toxin with secondary opportunistic bacterial invasion of the fetus leads to MRLS. The second hypothesis suggests that a breach of gastrointestinal mucosal integrity by hairs of ETC leads to a bacteremia and results in MRLS. In 2004, a similar equine abortion storm was reported from Australia and caterpillar exposure was identified as a risk factor for the abortion. In 2006, the syndrome was observed in Florida and New Jersey.

Hormone profiles of mares affected by the mare reproductive loss syndrome.

While searching for the cause of the Mare Reproductive Loss syndrome (MRLS), we postulated that 1 of 3 tissues in 40-120 D pregnant mares was the likely primary target of the noxious factor that caused early abortions: The corpora lutea (CL), the endometrium or the fetus and/or its membranes. At this stage of gestation, progesterone (P4) is solely produced by luteal tissue, eCG by endometrial cups in the endometrium and oestrogens by the feto-placental unit. We determined whether concentrations of P4, eCG and/or total conjugated oestrogens (CE) would indicate which tissue was targeted during the MRLS. P4, eCG and CE were measured in single serum samples collected from 216 mares, 60-110 D after ovulation during the 2001 MRLS outbreak. All mares had previously been confirmed pregnant by ultrasonography. The following data was obtained from each mare: Interval from ovulation, pregnancy status and normalcy of fetal fluids at the time of sampling, and pregnancy status 3 weeks after sampling and at term. There were no meaningful differences in hormone concentrations between pregnant mares that had normal and excessively echogenic fetal fluids at the time of sampling. CE were lower (p < 0.05) in mares that aborted after sample collection than in mares the carried to term. In 8 mares from which multiple samples were obtained, CE consistently decreased prior to any decreases in P4 or eCG. Arguments are presented that lead to the hypothesis that the fetal trophoblast was the primary target of the MRLS agent.

HIV protease inhibitor induces fatty acid and sterol biosynthesis in liver and adipose tissues due to the accumulation of activated sterol regulatory element-binding proteins in the nucleus.

The mechanism by which human immunodeficiency virus (HIV) protease inhibitor therapy adversely induces lipodystrophy and hyperlipidemia has not been defined. This study explored the mechanism associated with the adverse effects of the prototype protease inhibitor ritonavir in mice. Ritonavir treatment increased plasma triglyceride and cholesterol levels through increased fatty acid and cholesterol biosynthesis in adipose and liver. Ritonavir treatment also resulted in hepatic steatosis and hepatomegaly. These abnormalities, which were especially pronounced after feeding a Western type high fat diet, were due to ritonavir-induced accumulation of the activated forms of sterol regulatory binding protein (SREBP)-1 and -2 in the nucleus of liver and adipose, resulting in elevated expression of lipid metabolism genes. Interestingly, protease inhibitor treatment did not alter SREBP mRNA levels in these tissues. Thus, the adverse lipid abnormalities associated with protease inhibitor therapy are caused by the constitutive induction of lipid biosynthesis in liver and adipose tissues due to the accumulation of activated SREBP in the nucleus.

Stomachs of mice lacking the gastric H,K-ATPase alpha -subunit have achlorhydria, abnormal parietal cells, and ciliated metaplasia.

The H,K-ATPase of the gastric parietal cell is the most critical component of the ion transport system mediating acid secretion in the stomach. To study the requirement of this enzyme in the development, maintenance, and function of the gastric mucosa, we used gene targeting to prepare mice lacking the alpha-subunit. Homozygous mutant (Atp4a(-/-)) mice appeared healthy and exhibited normal systemic electrolyte and acid-base status but were achlorhydric and hypergastrinemic. Immunocytochemical, histological, and ultrastructural analyses of Atp4a(-/-) stomachs revealed the presence of chief cells, demonstrating that the lack of acid secretion does not interfere with their differentiation. Parietal cells were also present in normal numbers, and despite the absence of alpha-subunit mRNA and protein, the beta-subunit was expressed. However, Atp4a(-/-) parietal cells had dilated canaliculi and lacked typical canalicular microvilli and tubulovesicles, and subsets of these cells contained abnormal mitochondria and/or massive glycogen stores. Stomachs of adult Atp4a(-/-) mice exhibited metaplasia, which included the presence of ciliated cells. We conclude that ablation of the H,K-ATPase alpha-subunit causes achlorhydria and hypergastrinemia, severe perturbations in the secretory membranes of the parietal cell, and metaplasia of the gastric mucosa; however, the absence of the pump appears not to perturb parietal cell viability or chief cell differentiation.

Impaired cardiac performance in heterozygous mice with a null mutation in the sarco(endo)plasmic reticulum Ca2+-ATPase isoform 2 (SERCA2) gene.

The sarco(endo)plasmic reticulum Ca2+-ATPase isoform 2 (SERCA2) gene encodes both SERCA2a, the cardiac sarcoplasmic reticulum Ca2+ pump, and SERCA2b, which is expressed in all tissues. To gain a better understanding of the physiological functions of SERCA2, we used gene targeting to develop a mouse in which the promoter and 5' end of the gene were eliminated. Mating of heterozygous mutant mice yielded wild-type and heterozygous offspring; homozygous mutants were not observed. RNase protection, Western blotting, and biochemical analysis of heart samples showed that SERCA2 mRNA was reduced by approximately 45% in heterozygous mutant hearts and that SERCA2 protein and maximal velocity of Ca2+ uptake into the sarcoplasmic reticulum were reduced by approximately 35%. Measurements of cardiovascular performance via transducers in the left ventricle and right femoral artery of the anesthetized mouse revealed reductions in mean arterial pressure, systolic ventricular pressure, and the absolute values of both positive and negative dP/dt in heterozygous mutants. These results demonstrate that two functional copies of the SERCA2 gene are required to maintain normal levels of SERCA2 mRNA, protein, and Ca2+ sequestering activity, and that the deficit in Ca2+ sequestering activity due to the loss of one copy of the SERCA2 gene impairs cardiac contractility and relaxation.

Phenotype resembling Gitelman's syndrome in mice lacking the apical Na+-Cl- cotransporter of the distal convoluted tubule.

Mutations in the gene encoding the thiazide-sensitive Na+-Cl- cotransporter (NCC) of the distal convoluted tubule cause Gitelman's syndrome, an inherited hypokalemic alkalosis with hypomagnesemia and hypocalciuria. These metabolic abnormalities are secondary to the deficit in NaCl reabsorption, but the underlying mechanisms are unclear. To gain a better understanding of the role of NCC in sodium and fluid volume homeostasis and in the pathogenesis of Gitelman's syndrome, we used gene targeting to prepare an NCC-deficient mouse. Null mutant (Ncc-/-) mice appear healthy and are normal with respect to acid-base balance, plasma electrolyte concentrations, serum aldosterone levels, and blood pressure. Ncc-/- mice retain Na+ as well as wild-type mice when fed a Na+-depleted diet; however, after 2 weeks of Na+ depletion the mean arterial blood pressure of Ncc-/- mice was significantly lower than that of wild-type mice. In addition, Ncc-/- mice exhibited increased renin mRNA levels in kidney, hypomagnesemia and hypocalciuria, and morphological changes in the distal convoluted tubule. These data indicate that the loss of NCC activity in the mouse causes only subtle perturbations of sodium and fluid volume homeostasis, but renal handling of Mg2+ and Ca2+ are altered, as observed in Gitelman's syndrome.

Balance and hearing deficits in mice with a null mutation in the gene encoding plasma membrane Ca2+-ATPase isoform 2.

Plasma membrane Ca2+-ATPase isoform 2 (PMCA2) exhibits a highly restricted tissue distribution, suggesting that it serves more specialized physiological functions than some of the other isoforms. A unique role in hearing is indicated by the high levels of PMCA2 expression in cochlear outer hair cells and spiral ganglion cells. To analyze the physiological role of PMCA2 we used gene targeting to produce PMCA2-deficient mice. Breeding of heterozygous mice yielded live homozygous mutant offspring. PMCA2-null mice grow more slowly than heterozygous and wild-type mice and exhibit an unsteady gait and difficulties in maintaining balance. Histological analysis of the cerebellum and inner ear of mutant and wild-type mice revealed that null mutants had slightly increased numbers of Purkinje neurons (in which PMCA2 is highly expressed), a decreased thickness of the molecular layer, an absence of otoconia in the vestibular system, and a range of abnormalities of the organ of Corti. Analysis of auditory evoked brainstem responses revealed that homozygous mutants were deaf and that heterozygous mice had a significant hearing loss. These data demonstrate that PMCA2 is required for both balance and hearing and suggest that it may be a major source of the calcium used in the formation and maintenance of otoconia.

Renal and intestinal absorptive defects in mice lacking the NHE3 Na+/H+ exchanger.

NHE3 is one of five plasma membrane Na+/H+ exchangers and is encoded by the mouse gene Slc9a3. It is expressed on apical membranes of renal proximal tubule and intestinal epithelial cells and is thought to play a major role in NaCl and HCO3- absorption. As the distribution of NHE3 overlaps with that of the NHE2 isoform in kidney and intestine, the function and relative importance of NHE3 in vivo is unclear. To analyse its physiological functions, we generated mice lacking NHE3 function. Homozygous mutant (Slc9a3-/-) mice survive, but they have slight diarrhoea and blood analysis revealed that they are mildly acidotic. HCO3- and fluid absorption are sharply reduced in proximal convoluted tubules, blood pressure is reduced and there is a severe absorptive defect in the intestine. Thus, compensatory mechanisms must limit gross perturbations of electrolyte and acid-base balance. Plasma aldosterone is increased in NHE3-deficient mice, and expression of both renin and the AE1 (Slc4a1) Cl-/HCO3- exchanger mRNAs are induced in kidney. In the colon, epithelial Na+ channel activity is increased and colonic H+,K+-ATPase mRNA is massively induced. These data show that NHE3 is the major absorptive Na+/H+ exchanger in kidney and intestine, and that lack of the exchanger impairs acid-base balance and Na+-fluid volume homeostasis.

Leptospiral abortion and leptospiruria in horses from the same farm.

Leptospirosis was documented as the cause of abortion in a 5-year-old mare. Leptospires were detected in tissue specimens from fetal kidneys and from placenta by histologic evaluation of silver-stained sections. Antibodies against Leptospira interrogans serovar pomona were detected in fetal serum at a titer of 1,600 by use of a microscopic agglutination test. The mare had serum titers of 6,400; 0; 400; 800; 3,200; and 6,400 to L interrogans serovars bratislava, canicola, grippotyphosa, hardjo, icterohaemorrhagiae, and pomona, respectively. A serologic survey identified titers of at least 6,400 against serovars bratislava and pomona in 5 other horses on the farm. Titers of at least 100 against serovar bratislava were detected in 53% of the horses on the farm. Leptospires were detected by direct fluorescent-antibody testing in urine samples from the mare that aborted and from 2 of the other 5 horses.

Coupling between oxidative metabolism and active transport in the midgut of tobacco hornworm.

Active K transport (Isc) in the midgut of tobacco hornworm Manduca sexta has been shown to be highly dependent on oxidative metabolism. However, the oxygen consumption rate (rO2) was not altered by conditions that drastically affect Isc. Respiration was normally maximal, inasmuch as uncouplers did not increase rO2. This rate could be maintained without any added substrate probably by oxidation of endogenous substrates. Additional succinate increased rO2 by 17%. Simultaneous monitoring of Isc and the redox level of the respiratory chain components demonstrated that 1) succinate (5 mM) reduced all the respiratory enzymes while increasing Isc by 17%; 2) sesamol (5 mM), a mitochondrial uncoupler, reoxidized all respiratory enzymes and inhibited Isc by about 50%; 3) cyanide (1 mM) fully reduced the cytochromes and completely inhibited Isc. These redox responses indicate that the mitochondria in this tissue are normally coupled, even if respiration is maximal and is not modulated by active transport. Mitochondria isolated from the midgut show coupling and respiratory control by ADP, appearing to behave like mitochondria from other tissues. Therefore, a cytoplasmic constraint must exist in this tissue that continually elicits an unmodulated maximal respiratory rate.

Role of ATP in respiratory control and active transport in tobacco hornworm midgut.

The intracellular ATP, ADP, AMP, and orthophosphate (Pi) levels were measured in the midgut of Manduca sexta. The nucleotide levels were identical in tissues either "fresh" frozen or equilibrate in regular (32 mM) K or low (8 mM) K solutions. The calculated [ATP]/[ADP][Pi]ratio was approximately 300 M-1, which is low compared to other tissues. Given the ability of this ratio to control the respiratory rate, it is speculated that this low value may cause the maximal uncontrolled respiration normally observed in the midgut. The kinetics to anoxia of active transport (Isc) and the redox level of the mitochondrial cytochromes were measured simultaneously in the midgut. The cytochromes became reduced with a time constant of 0.75 +/- 0.15 min, whereas that for Isc inhibition was 2.1 +/- 0.15 min after a delay of 0.25 min. The difference between these two kinetic rates indicates that an intermediate form of energy exists in this tissue to energize active K transport. Measurements of ATP levels during the transition to anoxia indicate that its decay kinetics are sufficiently slow for ATP to be the immediate energy source for active transport in this tissue.

Dilantin-calcium interaction and active Na transport in frog skin.

The effects of diphenylhydantoin (DPH) and calcium were simultaneously studied on solute transport in frog skin. In the presence of external Ca, DPH elicited a significant increase in active Na transport; in its absence, DPH had no significant effect on active transport. DPH increased passive permeability, but this action was independent of Ca. It appears that the increase in active transport elicited by DPH is due to a specific interaction with Ca at the transport site.