Development of arterial and lymphatic collaterals under the action of a UHF magnetic field]

The work performed in 48 experimental and 24 control rats dealt with the development of arterial and lymphatic collaterals of pelvic extremities under effects of magnetic field of ultra high frequency (15 and 30 vt) in terms from 2 up to 6 weeks after surgery and exposure to magnetic field. The methods of investigation were arterio- and lymphoroentgenography and dissection. It has been established that formation of arterial collaterals in experimental rats takes place in all zones during the second week after cutting femoral arteries. The rate of the development of arterial collaterals was accelerated as compared with control animals. The lymph outflow after exposure to the influence of magnetic field of UHF goes mainly along superficial collaterals to groin nodes. The influence of the magnetic field of UHF appears to be accompanied by stable dilatation of super ficial lymph vessels, and due to this the lymph outflows along these vessels.     

Ultrastructural alterations in the initial segments and in the recurrent collateral terminals of Purkinje cells following axotomy

Summary Transection of Purkinje cell axons in adult male rats made 1.5 mm or further from the cell body does not lead to the death of the neuron and results in compensatory structural alterations of the surviving axonal portions of the nerve cell. Near to, and at the emergence of recurrent collaterals of Purkinje cell axons, huge varicosities filled with filaments, granular material, lysosomes and mitochondria develop. Terminals of recurrent axon collaterals also exhibit different degrees of structural changes. Most striking of the morphological alterations is the regular presence of nematosomes in the hypertrophic axonal branches, especially in synaptic terminals. Since nematosomes were shown to contain RNA in other types of neurons, their presence in recurrent collaterals may indicate an enhanced synthetic activity in Purkinje axonal processes and endings after axotomy.     

Metaboreceptor-mediated muscle oxygen saturation during recovery following isometric handgrip exercise

The aim of the present study was to determine whether oxygen supply to non-exercised muscle during recovery following fatiguing exercise is influenced by accumulated metabolites within exercised muscle. Twelve healthy male subjects performed 2-min isometric handgrip exercise at 40% maximal voluntary contraction with their right hand and the exercise was followed by a 3-min recovery period. Muscle oxygen saturation (SmO(2)) determined by near-infrared spatially resolved spectroscopy was used as an index of oxygen supply to non-exercised muscle and was measured in biceps brachii and tibialis anterior muscles on the left side. Compared to the pre-exercise baseline level, SmO(2) in the biceps brachii muscle (SmO(2BB)) increased significantly from 30 sec to 1 min after the start of exercise, while SmO(2) in the tibialis anterior muscle (SmO(2TA)) remained stable during the initial 1 min of exercise. Both SmO(2BB) and SmO(2TA) began to decrease at about 1 min and continued to decrease thereafter. Due to the initial increase in SmO(2BB), only SmO(2TA) showed a significant decrease during exercise. During recovery, SmO(2BB) did not differ significantly from the pre-exercise baseline level, whereas SmO(2TA) remained significantly lower until about 1.5 min of recovery and then it did not differ significantly from the baseline level. In another bout, subjects performed handgrip exercise of the same intensity, but post-exercise arterial occlusion (PEAO) of the exercised muscle was imposed for 2 min immediately after the end of exercise. During PEAO, SmO(2BB) decreased significantly compared to the baseline level, whereas SmO(2TA) remained significantly lower until the end of PEAO. The significant decrease in SmO(2BB) and the prolongation of decrease in SmO(2TA) by PEAO suggests that the recovery of SmO(2) in the non-exercised arm and leg is mediated by muscle metaboreceptors.     

Single collateral reconstructions reveal distinct phases of corticospinal remodeling after spinal cord injury

Background

Injuries to the spinal cord often result in severe functional deficits that, in case of incomplete injuries, can be partially compensated by axonal remodeling. The corticospinal tract (CST), for example, responds to a thoracic transection with the formation of an intraspinal detour circuit. The key step for the formation of the detour circuit is the sprouting of new CST collaterals in the cervical spinal cord that contact local interneurons. How individual collaterals are formed and refined over time is incompletely understood.

Methodology/Principal Findings

We traced the hindlimb corticospinal tract at different timepoints after lesion to show that cervical collateral formation is initiated in the first 10 days. These collaterals can then persist for at least 24 weeks. Interestingly, both major and minor CST components contribute to the formation of persistent CST collaterals. We then developed an approach to label single CST collaterals based on viral gene transfer of the Cre recombinase to a small number of cortical projection neurons in Thy1-STP-YFP or Thy1-Brainbow mice. Reconstruction and analysis of single collaterals for up to 12 weeks after lesion revealed that CST remodeling evolves in 3 phases. Collateral growth is initiated in the first 10 days after lesion. Between 10 days and 3–4 weeks after lesion elongated and highly branched collaterals form in the gray matter, the complexity of which depends on the CST component they originate from. Finally, between 3–4 weeks and 12 weeks after lesion the size of CST collaterals remains largely unchanged, while the pattern of their contacts onto interneurons matures.

Conclusions/Significance

This study provides a comprehensive anatomical analysis of CST reorganization after injury and reveals that CST remodeling occurs in distinct phases. Our results and techniques should facilitate future efforts to unravel the mechanisms that govern CST remodeling and to promote functional recovery after spinal cord injury.     

Desensitization of alpha-1 adrenergic receptor mediated smooth muscle contraction in aorta from endotoxic rats

Desensitization of vascular smooth muscles in endotoxemia was studied using the aorta from intraperitoneally endotoxin-injected rats. The KCl- and phenylephrine-induced contractions were significantly decreased in the endotoxic aorta compared to the control. In the endotoxic aorta the phenylephrine-induced contracture showed a gradual tension decrease after reaching a plateau and was attenuated by prior exposure to high concentration of phenylephrine, while KCl produced a sustained contraction and it was not affected by prior exposure to phenylephrine. The phenylephrine- and KCl-induced contractures of the control aorta showed stable plateaus and were not affected by prior exposure to phenylephrine. Neither diminished contractile force nor in vitro desensitization of phenylephrine contracture of isolated aorta was prevented by pretreatment of endotoxic rats with an alpha-adrenergic antagonist, phentolamine. These findings suggest that the contractile response to phenylephrine is easily desensitized in the endotoxic aorta compared to the control and neither this in vitro desensitization nor the diminution of contractile force is caused by in vivo exposure of aorta to a high concentration of catecholamines during endotoxemia.     

Recovery following exhaustive dynamic exercise in the human biceps muscle

The aim of the study was to investigate the recovery of the maximum voluntary contraction force (MVC), the endurance time and electromyographical (EMG) parameters following exhaustive dynamic exercise of the m. biceps brachii. EMG recordings were made in ten healthy subjects using bipolar surface electrodes placed over the common belly of the left arm biceps muscle. Up to 25 h post-exercise, the maximum contraction force and the EMG signal were recorded alternately at regular intervals. The EMG signal was recorded during 30-s contractions at 40% of the pre-fatigued MVC. Four hours and 25 h post-exercise, the endurance time of a 40% pre-fatigued MVC was recorded. Up to 25 h after the exercise the maximum contraction force, the endurance time and the EMG parameters were significantly different from the pre-exercise values. Nine out of ten subjects complained that muscle soreness had developed. Thus, long-lasting changes are found after exhaustive dynamic exercise, not only in the MVC and the muscle's endurance capacity, but also in the EMG signal.     

Establishment and characterization of porcine aortic endothelial cell cultures with prolonged replicative lifespan by a non-enzymatic method

The aim of this work was the establishment and characterization of porcine aortic endothelial cell cultures with prolonged lifespan. Endothelial cells where isolated from porcine thoracic aorta and established in primary culture; after sub cultivation, the cells showed typical morphology of endothelial phenotype with cobblestone appearance and growth in monolayer; they were positive against anti-CD31 and anti-CD54 immunostaining and Ac-LDL-Dil uptake. The cells were able to migrate in culture and showed a normal growth curve. The phenotype of our in vitro model of endothelial cells was stable through subcultivation; so, it should be a valuable tool for diverse studies of the endothelial response against physiological and pathological stimulus, particularly it could be useful to study interactions of endothelium with pathogenic bacteria causing diseases in pigs, as they are difficult to study in vivo.     

Effect of exposure to diabetic and fasted plasma on glucose oxidation in rat aorta

Glucose metabolism is depressed in aortic intima-media of fasted and diabetic rats. The aim of this study was to elucidate the influence of diabetic and fasted plasma on glucose oxidation in rat aorta. Male Sprague-Dawley rats weighing about 200 g were used. Diabetes was induced by streptozotocin (65 mg/kg) and the rats were used after a diabetes duration of two weeks. Fasted rats were used after food deprivation for 3 days. Aortic intima-media was preincubated in plasma for 120 or 240 min. During a further incubation for 2 hours in Krebs-Henseleit bicarbonate buffer the oxidation of 14C-glucose to 14CO2 was measured. Preincubation of normal aorta in diabetic or fasted rat plasma and diabetic human plasma significantly depressed the subsequently determined glucose oxidation in comparison to aorta preincubated in normal plasma. Preincubation of aorta from diabetic or fasted rats in normal rat plasma enhanced the glucose oxidation compared with the glucose oxidation in aorta of diabetic or fasted rats after preincubation in the corresponding plasma. These results suggest that diabetic and fasted plasma contains factor(s) which in vitro depress glucose oxidation in vascular smooth muscle and, thus, may be of importance for the lowered glucose oxidation found in vascular smooth muscle preparations obtained from diabetic or fasted animals.     

Changes in the expression of calbindin D-28k in the gerbil hippocampus following seizure

Previous studies have reported that calbindin D-28k (CB), a calcium-binding protein, containing neurons in the hippocampus play an important role in hippocampal excitability in epilepsy, because CB modulates the free calcium ion during seizure. Hence, in the present study, we investigated changes of CB expression in the hippocampus and its association in the Mongolian gerbil to identify roles of CB in epileptogenesis. CB immunoreactivity in the hippocampus was significantly lower in the pre-seizure group of seizure sensitive (SS) gerbils as compared with those seen in the seizure resistant (SR) gerbils. The distribution of CB immunoreactivity in the hippocampus showed significant difference after seizure on-set in SS gerbils. CB immunoreactivity in the hippocampal CA1, CA2 areas, and subiculum was lowest at 3h after seizure on-set; thereafter, the immunoreactivity became to increase to 12h after seizure on-set. Mossy fibers, Schaffer collaterals and dentate granule cells showed the highest CB immunoreactivity at 3h after seizure on-set; thereafter, the immunoreactivity became to decrease. In the case of the intrinsic and output connections of the hippocampus, a rapid decrease of CB serves an inhibitory function, which regulates the seizure activity and output signals from the hippocampus.     

Decreased DNA repair capacity of UV-irradiated cells following interferon treatment

The aim of this study was to examine the effect of interferons (IFNs) on the recovery of UV-damaged cells by means of measuring cell viability rates. The influence of the recombinant human interferons IFN-alpha, IFN-beta and IFN-gamma on the repair capacity of the UV-irradiated human cell lines WISH and HeLa was studied. The ability of cells to repair UV-induced damage was determined by the comet assay and both short- and long-term survival assays in proliferating cell cultures. We found that INFs negatively regulated DNA repair in cells damaged by UV light. One day after treatment, in both cell lines tested, IFN-alpha had a stronger inhibitory effect than IFN-gamma. Combined treatment with different IFNs exhibited a stronger inhibitory effect on cell recovery than treatment with each of them. The protein kinase inhibitor wortmanin further aggravated the effect of IFNs on cell survival.     

Time required for the restoration of normal heavy exercise VO2 kinetics following prior heavy exercise.

Prior heavy exercise markedly alters the O2 uptake (VO2) response to subsequent heavy exercise. However, the time required for VO2 to return to its normal profile following prior heavy exercise is not known. Therefore, we examined the VO2 responses to repeated bouts of heavy exercise separated by five different recovery durations. On separate occasions, nine male subjects completed two 6-min bouts of heavy cycle exercise separated by 10, 20, 30, 45, or 60 min of passive recovery. The second-by-second VO2 responses were modeled using nonlinear regression. Prior heavy exercise had no effect on the primary VO2 time constant (from 25.9 +/- 4.7 s to 23.9 +/- 8.8 s after 10 min of recovery; P = 0.338), but it increased the primary VO2 amplitude (from 2.42 +/- 0.39 to 2.53 +/- 0.41 l/min after 10 min of recovery; P = 0.001) and reduced the VO2 slow component (from 0.44 +/- 0.13 to 0.21 +/- 0.12 l/min after 10 min of recovery; P < 0.001). The increased primary amplitude was also evident after 20-45 min, but not after 60 min, of recovery. The increase in the primary VO2 amplitude was accompanied by an increased baseline blood lactate concentration (to 5.1 +/- 1.0 mM after 10 min of recovery; P < 0.001). Baseline blood lactate concentration was still elevated after 20-60 min of recovery. The priming effect of prior heavy exercise on the VO2 response persists for at least 45 min, although the mechanism underpinning the effect remains obscure.     

The amphibian microbiome exhibits poor resilience following pathogen-induced disturbance

Infectious pathogens can disrupt the microbiome in addition to directly affecting the host. Impacts of disease may be dependent on the ability of the microbiome to recover from such disturbance, yet remarkably little is known about microbiome recovery after disease, particularly in nonhuman animals. We assessed the resilience of the amphibian skin microbial community after disturbance by the pathogen, Batrachochytrium dendrobatidis (Bd). Skin microbial communities of laboratory-reared mountain yellow-legged frogs were tracked through three experimental phases: prior to Bd infection, after Bd infection (disturbance), and after clearing Bd infection (recovery period). Bd infection disturbed microbiome composition and altered the relative abundances of several dominant bacterial taxa. After Bd infection, frogs were treated with an antifungal drug that cleared Bd infection, but this did not lead to recovery of microbiome composition (measured as Unifrac distance) or relative abundances of dominant bacterial groups. These results indicate that Bd infection can lead to an alternate stable state in the microbiome of sensitive amphibians, or that microbiome recovery is extremely slow—in either case resilience is low. Furthermore, antifungal treatment and clearance of Bd infection had the additional effect of reducing microbial community variability, which we hypothesize results from similarity across frogs in the taxa that colonize community vacancies resulting from the removal of Bd. Our results indicate that the skin microbiota of mountain yellow-legged frogs has low resilience following Bd-induced disturbance and is further altered by the process of clearing Bd infection, which may have implications for the conservation of this endangered amphibian.Subject terms: Microbial ecology, Community ecology     

Effect of autologous marrow transplantation on bone marrow recovery following large-field fractionated irradiation of dogs

After large-field fractionated irradiation of dogs at a cumulative dose of 54 Gy, a stable bone marrow depletion occurs persisting for a year following irradiation. The automyelotransplantation after the end of the exposure elicits a transient recovery of the exposed bone marrow, 1.5-2 months after the beginning of irradiation, followed by a secondary depletion of the exposed haemopoietic sites. The control and the automyelotransplanted animals exhibited bone marrow recovery one year and six months after irradiation, respectively, the cellularity being maintained at a high level for 3 years of observation.     

Aorta,pulmonary artery,and blood flows on them in chickens in the second half of embryogenesis and after hatching

The aim of the study was to determine changes in the blood flow in arterial trunks (coming out of the heart of chickens) by changes of the lumen of these arteries during embryogenesis (on the 10th, 15th, and 19th days) and 6 days after the hatching. For this purpose, posthumous morphometry of aorta, pulmonary arteries, and arterial (Botallo’s) ducts (AD) from their exit from the heart until final extraorgan branching was conducted. It was demonstrated that, in this period, (1) initial lumens of aorta and pulmonary arteries are equal to each other and are equally increased (with temporary stop in last quarter of embryogenesis) with an increase of the body weight (BW); (2) the portion of the right ventricle in a total blood circulation minute volume (BCMV) is somewhat smaller than the portion of the left ventricle, but it approaches equality to it by the end of embryogenesis; (3) with the growth of embryos, the portion of total BCMV flowing through the anterior (before the inflow of AD into the aorta) part of the body decreases (from 41 to 33%); that in the average part increases (from 17 to 31%); that in the posterior part (after bifurcation of aorta), where chorioallantoic membrane (CAM) is located, remains almost unchanged; (4) after the hatching (and disappearance of CAM), BCMV of the left ventricle multiply increases due to the junction of two blood flows from the heart (through the ascending aorta and AD) into a single flow, which flows sequentially by lesser and greater circulations, resulting in multiple increase in the organ blood flow.     

Electro-cortical signs of early neuronal damage following transient global cerebral ischemia in rat

During recovery after a transient global cerebral ischemia (TGCI), rat electrocorticogram (ECoG) shows epochs of synchronized activity (SA) alternating with epochs of low amplitude background activity (BA). The aim of this study was to compare the changes in these electrical activities during a 30-min recovery period that followed either a noninjuring (3 minutes, N=10) or an injuring (10 minutes, N=10) TGCI. During TGCI there was a 3 fold reduction in amplitudes of both SA and BA but no changes in frequency. During reperfusion following a 3 minutes TGCI, the amplitudes of both SA and BA recovered to about 70%. During the reperfusion that followed a 10 minutes TGCI, BA showed no recovery, whereas SA recovered to about 40%. During the 30 min reperfusion, there was a timedependent decrease in the frequency of SA, but independent on the duration of TGCI. In contrast, the frequency of the BA did not change during reperfusion. Our data indicate that following cerebral ischemia the recovery of SA can take place independently of BA. The lack of recovery in BA may indicate early subcortical neuronal damage.     

De- and regeneration of the bulbospinal serotonin neurons in the rat following 5,6-or 5,7-Dihydroxytryptamine treatment

Summary In an attempt to determine the conditions which permit central 5-HT neurons to respond to a chemical injury of their axons by sprouting and regeneration, the pattern and time-course of recovery of 5-HT concentrations and regrowth of bulbospinal 5-HT axons were evaluated in rats subjected to intraventricular treatment with either 75 g 5,6- or 150 g 5,7-DHT. While 5,6-DHT treatment is followed by a significant recovery of 5-HT concentrations in the telodiencephalon, brainstem and upper part of the spinal cord within 3 months, there is no significant restoration of the severely depleted 5-HT levels in the telodiencephalon and spinal cord, and only limited recovery in 5-HT content of the brainstem preparation after 5,7-DHT.These differences conform to the observation of widespread and effective regrowth and regeneration of the bulbospinal 5-HT neurons in the 5,6-DHT treated lower brainstem and upper spinal cord but restricted and localized sprouting efforts in the 5,7-DHT treated lower medulla oblongata. This could be explained by a cell body near lesion of the non-terminal indoleamine axons by 5,7-DHT which results in a late retrograde, irreversible degeneration of most of the indoleamine pericarya from group B1 and many of group B3.It is concluded that the preservation of a critical length of the main axon and part of its collaterals is necessary for the neuron's survival, and that the individual pattern of the neuropil architecture of brain centres which are invaded by the axonal sprouts may significantly influence their growth characteristics and thus either favour or impede their chance to reestablish connections with their original effector. Aberrant, localized, intense sprouting of drug-damaged axons may in itself reflect the need of the neuron—deprived of most of its axonal tree—to reestablish its original total axonal length by multiple branching.Supported by grants from the Deutsche Forschungsgemeinschaft. The authors are indebted to Rolf Franck for his technical assistance.Supported by grants from the Swedish Medical Research Council (No. 04 X-3874 and 04 X-56).     

Radiosensitivity of vascular tissue. II. Differential radiosensitivity of aortic cells in vitro

The cellular outgrowths from three layers of rabbit and monkey aorta were used as primary cultures. Irradiation of the tissue fragments at the time of explanation resulted in a reduction in outgrowth of 50% with a dose of 200 rad, and in a reduction of over 90% with doses of 300 rad and above. When comparable cultures were irradiated after 2 months in vitro as a mature actively metabolizing but slowly proliferating cell population, radioresistance was increased. Subcultures of medial smooth muscle cells irradiated during their logarithmic growth phase showed a linear dose response in the cell number parameter up to 150 rad. A dose of 250 rad resulted in complete flattening of the growth curve, with a reduction in labeling index, after a 3-hr terminal [3H]TdR pulse. On the other hand, the labeling index indicated some recovery 3 days after irradiation in cultures receiving less than 250 rad. Under the same experimental conditions, cells derived from the intima of the same aorta showed no recovery when increase in cell numbers over time, or the number of labeled cells per area, were used as parameters. Cells derived from adventitia showed a relative increase in the number of labeled cells per area 4 and 7 days after irradiation following an initial decrease on Day 1.     

Recovery of yeast cells following the combined action of hyperthermia and ionizing radiation

Consecutive action of elevated temperature (50 degrees C) and gamma-irradiation on yeast cells Saccharomyces cerevisiae was studied. It was shown that yeast cells can recover from lethal thermal and radiation lesions after the combined action of the two factors. The efficiency of recovery does not depend upon the sequence of treatments. Heating (50 degrees C) before or after gamma-irradiation increases the radiation response of yeast when plating the cells on a nutrient agar containing 1.5 M KCl. The synergistic effect decreases with yeast cells kept in water at 28 degrees C before plating. The influence of one factor on the effectiveness of recovery from damages induced by the other was estimated.     

Genotoxicity and repair capability of Mus musculus DNA following the oral exposure to Tramadol

Tramadol is an analgesic and psychoactive drug that acts primarily upon the central nervous system where it alters brain function, resulting in temporary changes in perception, mood, consciousness and behavior. The aim of present study was to analyze the genotoxicity and repair capability of DNA after Tramadol exposure in albino mice (Mus musculus). For this purpose, forty mice were divided equally into four groups as; a control group (without drug) and three treatment groups that were treated with three doses of Tramadol as minimum dose group, Intermediate dose group and maximum dose group, corresponding to 25 mg/kg, 50 mg/kg and 75 mg/kg of body weight respectively. The dose was given orally for 15 days. After 15 days peripheral blood was drawn from half mice of each group and subjected to comet assay. While the remaining half mice were given a recovery period of 15 days and same procedure was used for blood collection and comet assay. Significant difference in various comet parameters was observed among control and exposed groups. Maximum damage was observed at highest concentration 75 mg/kg of Tramadol and minimum damage was observed at dose 25 mg/kg of Tramadol, while results of repaired mice group showed that repair capability of Tramadol was minor and recovery of Tramadol required a lot of time. It can be concluded that Tramadol cause genotoxicity that is dose dependent and has low repair capability.     

Regulation of CCL5 expression in smooth muscle cells following arterial injury

Chemokines play a crucial role in inflammation and in the pathophysiology of atherosclerosis by recruiting inflammatory immune cells to the endothelium. Chemokine CCL5 has been shown to be involved in atherosclerosis progression. However, little is known about how CCL5 is regulated in vascular smooth muscle cells. In this study we report that CCL5 mRNA expression was induced and peaked in aorta at day 7 and then declined after balloon artery injury, whereas IP-10 and MCP-1 mRNA expression were induced and peaked at day 3 and then rapidly declined.The expression of CCL5 receptors (CCR1, 3 & 5) were also rapidly induced and then declined except CCR5 which expression was still relatively high at day 14 after balloon injury. In rat smooth muscle cells (SMCs), similar as in aorta CCL5 mRNA expression was induced and kept increasing after LPS plus IFN-gamma stimulation, whereas IP-10 mRNA expression was rapidly induced and then declined. Our data further indicate that induction of CCL5 expression in SMCs was mediated by IRF-1 via binding to the IRF-1 response element in CCL5 promoter. Moreover, p38 MAPK was involved in suppression of CCL5 and IP-10 expression in SMCs through common upstream molecule MKK3. The downstream molecule MK2 was required for p38-mediated CCL5 but not IP-10 inhibition. Our findings indicate that CCL5 induction in aorta and SMCs is mediated by IRF-1 while activation of p38 MAPK signaling inhibits CCL5 and IP-10 expression. Methods targeting MK2 expression could be used to selectively regulate CCL5 but not IP-10 expression in SMCs.