Acute exposure to microgravity does not influence the H-reflex with or without whole body vibration and does not cause vibration-specific changes in muscular activity

PurposeMany potential countermeasures for muscle and bone loss caused by exposure to microgravity require an uncompromised stretch reflex system. This is especially true for whole body vibration (WBV), as the main source of the neuromuscular activity during WBV has been attributed to stretch reflexes. A priori, it cannot be assumed that reflexes and Ia afferent transmission in particular have the same characteristics in microgravity as in normal gravity (NG). Therefore, the purpose of the study was to compare Ia afferent transmission in microgravity and NG and to assess how microgravity affects muscle activity during WBV.MethodsIn 14 participants, electromyographic activity of four leg muscles as well as Hoffmann-reflexes were recorded during NG and microgravity induced by parabolic flights.ResultsThe size of the Hoffmann-reflex was reduced during WBV, but did not differ during acute exposure to microgravity compared to NG. The influence of the gravity conditions on the electromyographic activity did not change depending on the vibration condition.ConclusionsAs far as the electromyographic activity of the recorded leg muscles is concerned, the effect of WBV is the same in microgravity as in NG. Moreover, Ia afferent transmission does not seem to be affected by acute exposure to microgravity when subjects are loaded with body weight and postural sway is minimized.     

Osteocalcin does not influence acute or chronic inflammation in human vascular cells

Some human observational studies have suggested an anti-inflammatory role of osteocalcin (OCN). An inflammatory protocol using interferon-γ and tumor necrosis factor-α (10 ng/ml) was employed to examine the acute (24 hr) and chronic (144 hr) effects of uncarboxylated OCN (ucOCN) in commercial, primary, subcultured human aortic endothelial cells (HAEC), and human smooth muscle cells (HASMCs). The inflammatory protocol increased phosphorylation of intracellular signaling proteins (CREB, JNK, p38, ERK, AKT, STAT3, STAT5) and increased secretion of adhesion markers (vascular cell adhesion molecule-1, intracellular adhesion molecule-1, monocyte chemoattractant protein-1) and proinflammatory cytokines (interleukin-6 [IL-6], IL-8). After acute inflammation, there were no additive or reductive effects of ucOCN in either cell type. Following chronic inflammation, ucOCN did not affect cell responses, nor did it appear to have any pro- or anti-inflammatory effects when administered acutely or chronically on its own in either cell type. Additionally, ucOCN did not affect lipopolysaccharide (LPS)-induced acute inflammation in HAECs or HASMCs. The findings of this study do not support a causal role for OCN within the models of vascular inflammation chosen. Further confirmatory studies are warranted.     

MFGE8 does not influence chorio-retinal homeostasis or choroidal neovascularization in vivo

Purpose

Milk fat globule-epidermal growth factor-factor VIII (MFGE8) is necessary for diurnal outer segment phagocytosis and promotes VEGF-dependent neovascularization. The prevalence of two single nucleotide polymorphisms (SNP) in MFGE8 was studied in two exsudative or “wet” Age-related Macular Degeneration (AMD) groups and two corresponding control groups. We studied the effect of MFGE8 deficiency on retinal homeostasis with age and on choroidal neovascularization (CNV) in mice.

Methods

The distribution of the SNP (rs4945 and rs1878326) of MFGE8 was analyzed in two groups of patients with “wet” AMD and their age-matched controls from Germany and France. MFGE8-expressing cells were identified in Mfge8 ^+/− mice expressing ß-galactosidase. Aged Mfge8 ^+/− and Mfge8 ^−/− mice were studied by funduscopy, histology, electron microscopy, scanning electron microscopy of vascular corrosion casts of the choroid, and after laser-induced CNV.

Results

rs1878326 was associated with AMD in the French and German group. The Mfge8 promoter is highly active in photoreceptors but not in retinal pigment epithelium cells. Mfge8^−/− mice did not differ from controls in terms of fundus appearance, photoreceptor cell layers, choroidal architecture or laser-induced CNV. In contrast, the Bruch''s membrane (BM) was slightly but significantly thicker in Mfge8^−/− mice as compared to controls.

Conclusions

Despite a reproducible minor increase of rs1878326 in AMD patients and a very modest increase in BM in Mfge8^−/− mice, our data suggests that MFGE8 dysfunction does not play a critical role in the pathogenesis of AMD.     

Parity of female goats does not influence their estrous and ovulatory responses to the male effect

The objective of the present study was to determine whether parity is a factor that influences the estrous and ovulatory responses of female goats when they are stimulated by males that show increased sexual activity. To stimulate sexual activity, four adult male goats were subjected to photoperiodic treatment for 2.5 months comprising long days, with the treatment commencing on 1 November. On 14 April at 1900 h, a group of multiparous females (n = 21) and a group of 16 months-old nulliparous females (n = 19) were exposed to four bucks (two per group) for 15 days. Throughout the study period, the estrous behavior of these female goats was detected twice on a daily basis. Ovulations of the female goats were determined by ecography on days 7 and 18 after exposure to males. The sexual behavior of males was recorded twice every day from 0800 to 0900 h and from 1730 to 1830 h during the first 4 days after introduction in the pen of females. The total cumulative proportion of multiparous females that had ovulations (100%) and displayed estrous behavior (100%) during the 15 days of exposure to males did not differ (P > 0.05) from that of nulliparous females (100% and 95%, respectively). The interval between introduction of males and onset of estrous behavior did not differ (P > 0.05) between multiparous (1.9 ± 0.1 days) and nulliparous (1.7 ± 0.2 days) females. The proportion of females displaying a short estrous cycle was greater (P < 0.05) in multiparous (13/21, 62%) than in nulliparous (5/19, 26%) females. Duration of these shorter than typical estrous cycles did not differ (P > 0.05) between groups (multiparous: 5.2 ± 0.3 days, nulliparous: 4.5 ± 0.1 days). The number of anogenital sniffings was greater (P < 0.001) in males exposed to nulliparous than in those exposed to multiparous females. In contrast, the number of mounting attempts was greater (P < 0.01) in males that were introduced to multiparous than in those that were introduced to nulliparous does. The number of flehmen, nudging, self-marking with urine, and mounts was not different (P > 0.05) between males that were in contact with multiparous and nulliparous females. These results indicate that regardless of parity, female goats respond to male introduction if they are stimulated by males that were previously exposed to artificial long days to increase their sexual behavior.     

Chronic administration of NMU into the paraventricular nucleus stimulates the HPA axis but does not influence food intake or body weight

C-reactive protein (CRP), a predictor of future cardiovascular diseases, has been reported to damage the vascular wall by inducing endothelial dysfunction and inflammation. This proatherogenic CRP was speculated to have a role in attenuating angiogenic functions of human endothelial progenitor cells (EPCs), possibly impairing vascular regeneration and increasing cardiovascular vulnerability to ischemic injury. Herein, we investigated the direct effect of CRP on angiogenic activity and gene expression in human EPCs. Incubation of EPCs with human recombinant CRP significantly inhibited EPC migration in response to vascular endothelial growth factor, possibly by decreasing the expression of endothelial nitric oxide synthase and subsequent nitric oxide production. In addition, CRP-treated EPCs showed the reduced adhesiveness onto an endothelial cell monolayer. When assayed for the gene expression of arteriogenic chemo-cytokines, CRP substantially decreased their expression levels in EPC, in part due to the upregulation of suppressors of cytokine signaling proteins. These results suggest that CRP directly attenuates the angiogenic and possibly arteriogenic functions of EPCs. This CRP-induced EPC dysfunction may impair the vascular regenerative capacity of EPCs, thereby leading to increased risk of cardiovascular diseases.     

Orexin-A does not stimulate food intake in old rats

Aging is associated with a progressive decrease in appetite and food intake. Both A and B orexins, expressed in specific neurons of the lateral hypothalamic area, have been implicated in the regulation of sleep and feeding. In this study, the stimulatory effect of intracerebroventricular administration of the orexins on food intake was compared between young (4-mo-old) and old (25- to 27-mo-old) male Wistar rats. A stainless steel cannula was implanted stereotactically into the left lateral ventricle. After a 7-day recovery period, different doses (0-30 nmol) of orexins were injected into the left lateral ventricle without anesthesia. Food and water consumptions were measured at 1, 2, and 4 h after injection. The protein levels of orexin receptors, a specific receptor for orexin-A (OX1R) and a receptor for both orexin-A and -B (OX2R), in the hypothalamus were determined by Western blot analysis and compared between young and old rats. Intracerebroventricular administration of orexin-A stimulated food intake in a dose-dependent manner in young rats. However, no effects were observed at any dose in old rats. The protein level of OX1R in the hypothalamus was significantly lower in old rats than in young rats, although the protein level of OX2R was comparable between groups. Results of the present study indicate that the function of the orexin system is diminished in old rats. The decrease in the OX1R protein level in the hypothalamus could be responsible for orexin-A's lack of stimulation of food intake in old rats.     

Infection with Wolbachia does not influence crossing-over in Drosophila melanogaster

The influence of endosymbiotic bacterium Wolbachia on the recombination processes in the sex chromosome of Drosophila melanogaster on the region between white and cut genes has been studied. The analysis of the crossing-over frequency in various variants of strain crossing, infected and noninfected, with bacteria has been carried out. The results have shown the absence of the influence of infection with Wolbachia on the frequency of crossing-over in the studied region of the X chromosome of D. melanogaster.     

Herbivory damage does not indirectly influence the composition or excretion of aphid honeydew

Recent research has shown that extrafloral nectar secretion by plants, which also attracts ants, is a defense inducible by herbivory damage. Our research addresses the question of whether plants can manipulate the honeydew secretions of homopterans as an inducible defense in response to herbivory in the same manner as extrafloral nectaries. We investigated changes in honeydew composition and excretion rate by the facultatively ant-attended aphid Chaitophorus saliniger Shinji (Homoptera: Aphididae) depending on the presence or absence of herbivory by caterpillars (Clostera anastomosis L.; Lepidoptera: Notodontidae) on their host plant, the willow Salix gilgiana Seemen. Our results found no evidence to suggest that herbivory damage to the aphids host plant causes significant changes in aphid population growth, honeydew droplet volume, volume of honeydew excreted per aphid per hour, or composition of three abundant sugars in aphid honeydew as a result of herbivory damage to the aphids host plant, suggesting that, in this particular system, the plant is not manipulating the aphids honeydew output for its own benefit.     

High calcium intake does not prevent stress-salt hypertension in dogs

Avoidance conditioning sessions and isotonic saline (1.3 L/day) were administered to dogs for 12 days under conditions of a low (0.1%) or high (1.5%) calcium diet. Twenty-four-hour mean arterial pressure increased comparably during the stress-salt conditioning periods on both the low (systolic: +16 +/- 5 mm Hg; diastolic: +6 +/- 2 mm Hg) and high (systolic: +17 +/- 4 mm Hg; diastolic: +11 +/- 4 mm Hg) calcium diets. Urine volume, sodium excretion, and serum calcium levels on the high calcium diet were not significantly different from those on the low calcium diet. In a second experiment, calcium was infused continuously for six days into the arterial circulation of normotensive or stress-salt hypertensive dogs at a rate of 0.12-0.23 mEq/min. Although serum calcium levels increased by up to 50% under these conditions, there were no significant effects on 24-hour levels of arterial pressure. In contrast to the protective effect of augmented potassium intake, these findings indicate that calcium intake does not influence the development of stress-salt hypertension in dogs.     

Beta-carotene does not influence fertility in beef heifers

Eighty-four 18-month-old crossbred beef heifers, 3 to 4 months pregnant, were assigned by stratified randomization to either a high or low (control) beta-carotene (B-car) diet to determine the effect of long-term supplementation of B-car on reproductive performance. The heifers were followed through pregnancy, calving and subsequent breeding. The basal diet consisted of barley, canola meal and barley straw. Heifers supplemented by B-car received 625 mg B-car per day in the concentrate. Vitamin A and D complex injections were given monthly to all heifers. Heifers were bred by artificial insemination after Day 60 postpartum. Throughout the study heifers fed the B-car supplement had higher levels of B-car in plasma (> 300 ug/dl) (P < 0.01) than the heifers fed the control diet (< 50 ug/dl). Vitamin A status was satisfactory in all heifers throughout the study. Birth weight of calves, weight gain, and incidence of mortality were not influenced by B-car. For the control and B-car treatments, days postpartum to first normal luteal phase were 67.5 and 62.6 days; days postpartum to first detected estrus were 70.1 and 65.3, and services per conception were 1.24 and 1.29, respectively. Long-term supplementation of B-car increased prepartum plasma progesterone but had no effect on postpartum fertility.     

Neutrophil depletion does not prevent lung edema after endotoxin infusion in goats

Neutropenia was produced in goats by injection of either nitrogen mustard, (1.5 mg/kg) or hydroxyurea (200 mg X kg-1 X day-1). A nitrogen mustard (M + E) group (n = 6), a hydroxyurea (H + E) group (n = 5), and a control (E) group (n = 7) were given 1-h infusions of endotoxin (5 micrograms/kg total dose), then monitored for up to 5 h. Postmortem extravascular lung water (EVLW) was significantly higher in the M + E group (14.2 +/- 4.4 ml/kg) and the E group (11.9 +/- 3.9 ml/kg) when compared with a normal control (6.6 +/- 1.3 ml/kg) group that did not receive endotoxin. EVLW in a group made neutropenic with nitrogen mustard (6.7 +/- 1.3 ml/kg) and the H + E (7.9 +/- 1.5 ml/kg) groups were not statistically different from each other or from normal controls. Circulating neutrophil counts averaged 32 +/- 42 cells/microliter in the M + E group and 180 +/- 210 cells/microliter in the H + E group. Only minimal histological changes were seen in the H + E group, but the E and M + E lungs had severe pulmonary edema. We conclude that neutrophils are not required for increased EVLW and decreased arterial O2 partial pressure after endotoxin infusion, and hydroxyurea prevents at least part of the pulmonary edema after endotoxin by a mechanism that is not neutrophil dependent.     

Prepartum peripherally-induced hyposmia does not reduce postpartum anoestrus duration in nursing goats

Parturient goats rapidly develop exclusive nursing of their own litter that relies on olfactory recognition of the young. They also show a period of postpartum anoestrus whose duration depends on the presence of the kid. In cattle, maternal selectivity is one of the factors that delays the recovery of sexual activity. To investigate the possible influence of maternal selectivity on the duration of postpartum anoestrus in goats, we compared the recovery of estrus behavior by daily estrus detection with an active buck in intact and selective nursing goats (n = 24) with that of dams rendered non-selective by peripheral hyposmia with ZnSO4 (n = 18). Postpartum anoestrus duration was shorter in intact (68+/-7 days) than in hyposmic mothers (93+/-7 days; P < 0.05). However, the cycles of normal duration were less frequent in intact goats (P = 0.03). We conclude that in nursing goats, preventing the establishment of selective nursing by prepartum peripheral hyposmia does not reduce postpartum anoestrus duration. Our results suggest that daily exposure to the buck may result in an earlier recovery of ovarian activity in intact mothers.     

Seeing group members eating a familiar food enhances the acceptance of novel foods in capuchin monkeys

In primates, social context is one of the factors that increases the acceptance of novel foods. Previous experiments showed that tufted capuchins, Cebus apella, eat significantly more of novel foods when in the presence of group members eating the same novel foods. Several processes may have led to these results. The mere presence of group members may reduce the individual's stress of being alone, or its neophobic response and, consequently, may increase its food consumption. The individual may be influenced by what group members do, and local/stimulus enhancement and/or social facilitation may occur. To investigate the above processes, we assessed whether an individual capuchin monkey's consumption of novel foods is lower when (1) the individual is alone with nobody in the nearby cage than when (2) group members are present in the nearby cage with no food or when (3) they are present and eating a familiar food. We tested 15 subjects with three novel foods, each presented in one condition. In both social conditions, the more group members there were by the food box the more the experimental subject ate. In addition, when group members were present and eating food, there was a significant increase in the acceptance of the three foods, regardless of what group members were eating. We argue that social facilitation of eating is a quicker way to overcome neophobia and only social facilitation of eating what the others are eating can be considered a safe way to learn about a safe diet. Copyright 2000 The Association for the Study of Animal Behaviour.     

Protandim does not influence alveolar epithelial permeability or intrapulmonary oxidative stress in human subjects with alcohol use disorders

Alcohol use disorders (AUDs), including alcohol abuse and dependence, have been linked to the development of acute lung injury (ALI). Prior clinical investigations suggested an association between AUDs and abnormal alveolar epithelial permeability mediated through pulmonary oxidative stress that may partially explain this relationship. We sought to determine if correcting pulmonary oxidative stress in the setting of AUDs would normalize alveolar epithelial permeability in a double-blinded, randomized, placebo-controlled trial of Protandim, a nutraceutical reported to enhance antioxidant activity. We randomized 30 otherwise healthy AUD subjects to receive directly observed inpatient oral therapy with either Protandim (1,350 mg/day) or placebo. Subjects underwent bronchoalveolar lavage (BAL) and blood sampling before study drug administration and after 7 days of therapy; all AUD subjects completed the study protocol without adverse events. BAL total protein was measured at each timepoint as an indicator of alveolar epithelial permeability. In subjects with AUDs, before study drug initiation, BAL total protein values were not significantly higher than in 11 concurrently enrolled controls (P = 0.07). Over the 7-day study period, AUD subjects did not exhibit a significant change in BAL total protein, regardless of their randomization to Protandim {n = 14, -2% [intraquartile range (IQR), -56-146%]} or to placebo [n = 16, 77% (IQR -20-290%); P = 0.19]. Additionally, among those with AUDs, no significant changes in BAL oxidative stress indexes, epithelial growth factor, fibroblast growth factor, interleukin-1β, or interleukin-10 were observed regardless of drug type received. Plasma thiobarbituric acid reactive substances, a marker of lipid peroxidation, decreased significantly over time among AUD subjects randomized to placebo (P < 0.01). These results suggest that Protandim for 7 days in individuals with AUDs who are newly abstinent does not alter alveolar epithelial permeability. However, our work demonstrates the feasibility of safely conducting clinical trials that include serial bronchoscopies in a vulnerable population at risk for acute lung injury.     

Sulfation of tyrosine residues does not influence secretion of alpha 2-antiplasmin or C4

Sulfation of tyrosine residues is a biosynthetic modification of many secretory proteins. The function of this modification is not known, but it has been proposed that tyrosine sulfate residues may act as sorting signals to direct proteins along the secretory pathway. We tested this hypothesis by examining the effect of sulfation inhibitors on the kinetics of secretion of proteins by HepG2 cells. The inhibitors induced no change in the rate of secretion of alpha 2-antiplasmin and C4 (fourth component of complement), both of which contain tyrosine sulfate residues. Sulfation of tyrosine residues does not contribute to secretion of these proteins.     

A high intake of industrial or ruminant trans fatty acids does not affect the plasma proteome in healthy men

Consumption of industrial trans fat raises the risk of cardiovascular disease, but it is unclear whether cis9,trans11-conjugated linoleic acid (CLA)--a trans fatty acid in dairy products--modulates disease development. We investigated the effects of complete diets providing 7% of energy as industrial trans fat or cis9, trans11 CLA, compared with oleic acid, on regulation of plasma proteins in 12 healthy men. Diets were provided for 3 wk each, in random order. Plasma was collected at the end of each 3 wk intervention period, depleted of its 12 most abundant proteins and analyzed by 2-DE. Principal component analysis of protein spot intensity values revealed that the nature of the dietary intervention did not significantly affect the plasma proteome. The intervention provided in the 1st period produced a significant treatment effect compared with the interventions provided in the other two periods, and there was a significant subject effect. In conclusion, the nature of an extreme dietary intervention, i.e. 7% of energy provided by industrial trans fat or cis9,trans11 CLA, did not markedly affect the plasma proteome. Thus plasma proteomics using 2-DE appears, by and large, an unsuitable approach to detect regulation of plasma proteins due to changes in the diet.