Shock waves as treatment of mouse myofascial trigger points.

INTRODUCTION: An abnormal increase in spontaneous neurotransmission can induce subsynaptic knots in the myocyte called myofascial trigger points. The treatment of choice is to destroy these trigger points by inserting needles. However, 10% of the population has a phobia of needles, blood, or injuries. Therefore, the objective of this study is to verify the usefulness of shock waves in the treatment of myofascial trigger points. METHODS: Two groups of mice have been developed for this: healthy muscles treated with shock waves; trigger points affected muscles artificially generated with neostigmine and subsequently treated with shock waves. Muscles were stained with methylene blue, PAS-Alcian Blue, and labeling the axons with fluorescein and the acetylcholine receptors with rhodamine. Using intracellular recording the frequency of miniature endplate potentials (mEPPs) was recorded and endplate noise was recorded with electromyography. RESULTS: No healthy muscles treated with shock waves showed injury. Twitch knots in mice previously treated with neostigmine disappeared after shock wave treatment. Several motor axonal branches were retracted. On the other hand, shock wave treatment reduces the frequency of mEPPs and the number of areas with endplate noise. DISCUSSION: Shock waves seem to be a suitable treatment for myofascial trigger points. In the present study, with a single session of shock waves, very relevant results have been obtained, both functional (normalization of spontaneous neurotransmission) and morphological (disappearance of myofascial trigger points). Patients with a phobia of needles, blood, or injuries who cannot benefit from dry needling may turn to noninvasive radial shock wave treatment.

Behavioral Biomarkers of Schizophrenia in High Drinker Rats: A Potential Endophenotype of Compulsive Neuropsychiatric Disorders.

Psychogenic polydipsia, which is compulsive, non-regulatory fluid consumption, is present in 6%-20% of chronic psychiatric patients and frequently associated with the schizophrenia diagnosis. In the present study, we investigated the relation between schizophrenia-like symptoms and biomarkers with a compulsive drinking behavior phenotype in rats. Rats that were selected for low drinking vs high drinking behavior following schedule-induced polydipsia (SIP) were assessed in a latent inhibition (LI) paradigm using tone and electrical foot shock and in a spatial reversal learning task to evaluate behavioral inflexibility. We also analyzed the myelin basic protein in different brain areas of high drinker (HD) and low drinker (LD) rats. The HD rats, which were characterized by a compulsive drinking behavior on SIP, had a reduced level of LI effect and increased behavioral inflexibility in the spatial reversal learning task in comparison to the LD group. Moreover, HD rats showed less myelination in the center of the corpus callosum, striatum, and amygdala in comparison to LD rats. These findings strengthen the validity of HD rats that were selected by SIP as a possible phenotype of compulsive neuropsychiatric disorders, as evidenced by the existence of behaviors and biological markers that are related to schizophrenia and obsessive-compulsive disorder, including a reduced LI effect, behavioral inflexibility and reduced brain myelination. Future studies could contribute to the elucidation of the mechanisms underlying the compulsive phenotype of HD rats and its relation to vulnerability to schizophrenia.

Automatic counting and positioning of 5-bromo-2-deoxyuridine (BrdU) positive cells in cortical layers of rat brain slices.

5-Bromo-2-deoxyuridine (BrdU) staining is often used to evaluate cortical layer formation during mammalian brain development. This method allows the quantification of newly generated cells and therefore the study of the effects of xenobiotics or genetic factors on proliferation, cell death and migration behavior in a quantitative manner. However, these endpoints are generally assessed by time-consuming manual evaluation. In the present work, we introduce a novel procedure to identify and quantify BrdU(+) cells within cortical layers, using the commercially available vHCS-Scan V.6.3.1 software to identify BrdU(+) cell coordinates and the novel program 'BrdeLuxe' to define cortical layers and quantitatively assign BrdU(+) cells to them. This procedure is compared to BrdU(+) cell counting with the freeware 'ImageJ' in respect to the manual evaluation, all by two different researchers. BrdeLuxe shows high accuracy and precision for the determination of total number of BrdU(+) cells compared to the manual counting, while ImageJ does not reach such results. Accuracy and precision are also higher for employing the BrdeLuxe program to evaluate the percentage of BrdU(+) cells per brain layer compared to ImageJ. In terms of running time, BrdeLuxe is the fastest method of the three making it more suitable for multiple brain slices analyses.

Oral silicon supplementation: an effective therapy for preventing oral aluminum absorption and retention in mammals.

Silicon is an essential element for some lower forms of life. However, it is not generally considered an essential nutrient for mammals and the mechanisms underlying its potential essentiality remain partially unknown. In recent years, a possible association between the aluminum and silicon levels in drinking water and Alzheimer's disease (AD) has been suggested. It has been reported that silicon might have a protective effect for limiting oral aluminum absorption. This review is focused primarily on the potential role of silicon in preventing oral aluminum absorption and retention in mammals. The results of a number of studies suggest that dietary silicon supplementation could be of therapeutic value for preventing chronic aluminum accumulation in the brain, and hence, be a potential therapy for AD. However, it must be noted that controversy remains about whether aluminum accumulation in the brain is a cause or a consequence of AD. It is suggested that further investigation of this issue is warranted.

Combined effects of perfluorooctane sulfonate (PFOS) and maternal restraint stress on hypothalamus adrenal axis (HPA) function in the offspring of mice.

Although it is known that prenatal exposure to perfluorooctane sulfonate (PFOS) can cause developmental adverse effects in mammals, the disruptive effects of this compound on hormonal systems are still controversial. Information concerning the effects of PFOS on hypothalamus adrenal (HPA) axis response to stress and corticosterone levels is not currently available. On the other hand, it is well established that stress can enhance the developmental toxicity of some chemicals. In the present study, we assessed the combined effects of maternal restraint stress and PFOS on HPA axis function in the offspring of mice. Twenty plug-positive female mice were divided in two groups. Animals were given by gavage 0 and 6 mg PFOS/kg/day on gestation days 12-18. One half of the animals in each group were also subjected to restraint stress (30 min/session, 3 sessions/day) during the same period. Five plug-positive females were also included as non-manipulated controls. At 3 months of age, activity in an open-field and the stress response were evaluated in male and female mice by exposing them to 30 min of restraint stress. Male and female offspring were subsequently sacrificed and blood samples were collected to measure changes in corticosterone levels at four different moments related to stress exposure conditions: before stress exposure, immediately after 30 min of stress exposure, and recuperation levels at 60 and 90 min after stress exposure. Results indicate corticosterone levels were lower in mice prenatally exposed to restraint. In general terms, PFOS exposure decreased corticosterone levels, although this effect was only significant in females. The recuperation pattern of corticosterone was mainly affected by prenatal stress. Interactive effects between PFOS and maternal stress were sex dependent. The current results suggest that prenatal PFOS exposure induced long-lasting effects in mice.

Behavioral effects in adult mice exposed to perfluorooctane sulfonate (PFOS).

Nowadays, very little information concerning the effects on behavior in mammals of perfluorooctane sulfonate (PFOS), a widely distributed persistent environmental pollutant, is available. In the present study, we assessed the behavioral effects of PFOS on 3 months old mice after 1 month of exposure to this pollutant. Thirty adult mice were divided into three groups. Animals were given by gavage 0, 3, and 6 mg PFOS/kg/day for four consecutive weeks. After the treatment period, mice were evaluated for several skills by testing motor and sensory function by means of a functional observation battery (FOB), general activity and exploratory behavior in an open-field, and learning and memory in a water maze task. One week after behavioral testing, serum was collected for corticosterone analyses. No adverse effects were observed in the FOB. In general terms, activity in the open-field was similar in all groups being the only observed differences limited to the group given PFOS at 3mg/kg/day (spent less time in the center) and the group exposed to 6 mg PFOS/kg/day) (reduced rate of vertical activity). Concerning the effects of PFOS in the water maze, although all animals learned the task, no effect of the dose was observed during the acquisition. In the retention test, a deleterious effect of PFOS was noted. These results indicate that PFOS exposure induced only slight behavioral effects in adult male mice.

Influence of maternal restraint stress on the long-lasting effects induced by prenatal exposure to perfluorooctane sulfonate (PFOS) in mice.

The behavioral effects of concurrent maternal exposure to restraint stress and perfluorooctane sulfonate (PFOS) were assessed in the offspring of mice at 3 months of age. Plug positive females were divided into two groups. Animals were given by gavage 0 and 6mg PFOS/kg/day on gestation days 12-18. One-half of the animals in each group were subjected to restraint stress (30min/session, three sessions per day) during the same period. At 3 months, mice were evaluated for general activity in an open-field, and for learning and memory in a water maze task. The group prenatally exposed to PFOS and restraint presented a reduced mobility in the open-field. In the water maze, an interaction between sex and restraint was observed. Delayed task learning was also detected in females prenatally exposed to PFOS and restraint. An overall effect of restraint was observed in mice on retention of the task, suggesting a better retention in restrained animals. On the other hand, corticosterone levels were lower in animals prenatally subjected to restraint stress. The current results suggest interactive effects between PFOS and maternal stress.

Interactions in developmental toxicology: concurrent exposure to perfluorooctane sulfonate (PFOS) and stress in pregnant mice.

The maternal and developmental toxicity of combined exposure to restraint stress and perfluorooctane sulfonate (PFOS) was assessed in mice. On gestation Days 6-18, four groups of plug-positive female mice were orally exposed to PFOS at 0, 1.5, 3 and 6 mg/kg/day. Four additional groups of plug-positive animals received the same PFOS doses being restrained during 30 min three times per day. A control group was also included. Cesarean sections were performed on Day 18 of gestation and fetuses were weighed and examined for external, internal and skeletal malformations and variations. Before sacrifice of the dams, blood was collected and serum samples were prepared for thyroid hormones (total and free T3 and T4) and corticosterone analyses. The results of the present study show that both PFOS and restraint stress induced maternal toxicity. In turn, PFOS-induced fetal toxicity was evidenced by increased prenatal mortality. The only effect of restraint on fetal toxicity was a reduction on body weight and an increased prenatal mortality in fetuses concurrently exposed to 1.5 mg/kg of PFOS and restraint. PFOS-induced adverse effects on maternal and fetal toxicity in mice were observed at lower doses than those previously reported.

Concurrent exposure to aluminum and stress during pregnancy in rats: Effects on postnatal development and behavior of the offspring.

The present study was conducted to assess the potential combined influence of maternal restraint stress and aluminum (Al) exposure on postnatal development and behavior in the offspring of exposed rats. Female rats were concurrently exposed to 0 (control group), 50 or 100 mg/kg/day of Al administered as Al nitrate nonahydrate in drinking water with citric acid (355 or 710 mg/kg/day) for a period of 15 days prior to mating with untreated males. Aluminum exposure was maintained throughout the gestational, lactational and post-weaning periods. On days 6-20 of gestation, one-half of the pregnant animals in each group were restrained for 2 h/day. Food consumption and maternal body weight were decreased in the groups exposed to restraint only or combined with the highest Al dose. All of the animals were allowed to deliver and wean their offspring. The pups were evaluated for physical development and neuromotor maturation. Moreover, open-field activity, passive avoidance, and spatial learning in a water maze were also determined on postnatal days 30, 35 and 60, respectively. Body weight of pups treated with 100 mg/kg/day of Al was decreased relative to controls from postnatal day 12 through 21, sexual maturation was delayed in Al treated females and in males exposed to 100 mg/kg/day. Forelimb grip strength was reduced in males exposed to 100 mg/Al/kg/day and in females exposed to this Al dose plus prenatal restraint. Learning in a passive avoidance task indicated facilitated performance for Al treated rats at 100 mg/kg/day combined with prenatal restraint as evidenced by longer avoidance latencies, while learning in a water maze task showed a shorter latency to find the platform on acquisition day 2 for Al treated rats. However, no effects of Al on water maze performance were detected during the retention probe trial in which the only effect noted was an increase in the platform quadrant swim time for the prenatal restraint group. In general terms, the results of the present study did not show a notable influence of maternal restraint on the Al-induced postnatal developmental and behavioral effects in the offspring of prenatally Al-exposed rats.

Metal concentrations in hair and cognitive assessment in an adolescent population.

The objective of this study was to establish the potential relationship between the levels of various metals in hair and cognitive functions in children living in zones of Tarragona (Catalonia, Spain) with different metal pollution levels. Thirty-nine boys and 61 girls (12-14 yr old) from various schools were selected for the study. The concentrations of cadmium (Cd), chromium (Cr), mercury (Hg), lead (Pb), manganese (Mn), nickel (Ni), and tin (Sn) in scalp hair were determined by inductively coupled plasma- mass spectrometry (ICP-MS). Attention, visuospatial capabilities, and abstract reasoning were assessed as indicators of cognitive impairment. Three categories of attention were defined: low, medium, and high. A significant negative correlation (p=0.019) between Pb levels in hair and attention was observed. Significant differences between Pb levels in hair in low- and medium-performance groups and those in the high-performance group were also found. Moreover, a positive correlation (p=0.048) between Hg hair concentrations and visuospatial capabilities was also noted.

Behavioral effects of adult rats concurrently exposed to high doses of oral manganese and restraint stress.

The behavioral effects of concurrent exposure of high doses of manganese (Mn) and restraint stress were assessed in adult rats. Male Sprague-Dawley rats (250-300 g) received 0, 275 and 550 mg/kg/day of Mn in the drinking water for 19 weeks. Each group was divided into two subgroups. Animals in one subgroup were restrained for 2h/day. During the treatment period, food and water intake, and body weight were weekly recorded. At the end of the treatment period, activity levels were monitored in an open-field. Learning was evaluated by a water-maze task during five consecutive days. A trial probe was also conducted to assess the time spent in the platform quadrant. Body weight and food consumption were significantly reduced in the group receiving 550 mg/kg/day of Mn. A two-way analysis of variance (ANOVA) revealed an overall effect of Mn on the total distance traveled. Differences on spatial learning were observed in the acquisition period, in which rats given 550 mg/kg/day of Mn (alone or restrained) were impaired in comparison with the control and the restrained only groups. In the probe trial, there was an impaired retention in the group treated with Mn at 550 mg/kg/day. The results of this investigation in the open-field and water maze suggest that it would be plausible that restraint stress and a high exposure to Mn interact at common neurotransmitter levels but inducing opposite effects.

Influence of maternal stress on metal-induced pre- and postnatal effects in mammals: a review.

Studies in rodents have shown that, during pregnancy, maternal stress from restraint, noise, light, and heat among other factors may be associated with adverse effects on embryo/fetal and postnatal development. Moreover, it is also well known that exposure to certain metal levels during gestation can also cause maternal and developmental toxicity. Because potentially, pregnant women may be concurrently exposed to metals and various types of stress, the influence of maternal stress on the metal-induced adverse pre- and postnatal effects has been investigated for a number of elements. This influence is reviewed here. It is concluded that maternal stress enhances the metal-induced embryo/fetal and developmental toxicity only at doses of the metal which are also clearly toxic to the dam.

Interactions of caffeine and restraint stress during pregnancy in mice.

The maternal and developmental toxicity of combined exposure to restraint stress and caffeine was assessed in mice. On gestational Days 0-18, three groups of plug-positive females (n = 13-15) were given by gavage caffeine at 30, 60, and 120 mg/kg/day. Three additional groups received the same caffeine doses and were restrained for 2 hr/day. Control groups included restrained and unrestrained plug-positive mice not exposed to caffeine. All animals in the group concurrently exposed to 120 mg/kg/day of caffeine and restraint died during the experimental period. In the remaining groups, cesarean sections were performed on Day 18 of gestation, and the fetuses were weighed and examined for external, internal, and skeletal malformations and variations. Although maternal and embryo/fetal toxicity were observed at all caffeine doses, the adverse maternal and developmental effects were significantly enhanced in the groups concurrently exposed to caffeine and restraint. It was especially remarkable at 60 and 120 mg/kg/day. The results of this study suggest that maternal and developmental toxic effects might occur if high amounts of caffeine were consumed by women under a notable stress during pregnancy.

Effects of prenatal exposure to manganese on postnatal development and behavior in mice: influence of maternal restraint.

Manganese (Mn) is an essential trace element whose deficiency and excess have been reported to cause central nervous system (CNS) disturbances. On the other hand, during pregnancy, maternal stress has been shown to enhance the developmental toxicity of a number of metals. In this study, the maternal toxicity and developmental effects of a concurrent exposure to Mn and restraint stress were evaluated in mice. Pregnant animals were divided into three groups and received subcutaneous injections of manganese chloride tetrahydrate (MnCl2.4H2O) at 0, 1 and 2 mg/kg/day on Gestation Days 6-18. Each group was divided into two subgroups. Mice in one subgroup were subjected to restraint for 2 h/day on Days 6-18 of gestation. Pregnant mice were allowed to deliver, and pups were evaluated for physical and neuromotor maturation. Subsequently, adult mice were also evaluated for activity and learning. A significant increase in perinatal mortality was observed at 2 mg/kg/day Mn. A delay in some developmental landmarks (eye opening, testes descent) due to Mn exposure (2 mg/kg/day) was also seen in both restrained and unrestrained animals. No differences in motor resistance and coordination, or in learning at the passive avoidance test, were noted in adult mice. At the current Mn doses, combined exposure to Mn and stress during the prenatal period did not produce long-lasting effects on adult mice.

Influence of age on aluminum-induced neurobehavioral effects and morphological changes in rat brain.

Aluminum (Al) is potentially toxic for mammals. In contrast to well documented Al neurotoxicity, neurobehavioral studies of Al in rodents have generally not produced robust or consistent results. In the present study, 16 young male (21 days old) and 16 old male (18 months) rats were exposed to 0 (control group) and 100 mg/kg/day of Al administered as Al nitrate nonahydrate in drinking water concurrently with citric acid (356 mg/kg/day) for a period of 100 days. At the end of the exposure period, rats were evaluated for motor activity in an open-field apparatus and learning in a passive avoidance test. After behavioral testing, rats were sacrificed and brain samples were collected to determine Al concentrations and to study synapses in the left CA1 fields of hippocampal formation. While no significant effects of Al exposure between groups could be detected on behavior, the total number of synapses decreased with age and Al exposure. In turn, the percentage of perforated synapses significantly increased in old Al-loaded rats.