Effect of a dietary supplement on the reduction of lymphedema-progression in mouse tail-cut model.

OBJECTIVE: The aim of our study was to evaluate in vivo, in a mouse tail model of lymphedema, the effects of a dietary supplement, Garlive®, based on hydroxytyrosol from olive leaves, spermidine from rice seeds, hesperidin from citrus fruits and vitamin A. Hydroxytyrosol has anti-inflammatory, antioxidant and antimicrobial activities and inhibits leukotriene B4 generation; spermidine is able to inhibit the production of pro-inflammatory cytokines and mediators; hesperidin inhibits the secretion of pro-inflammatory cytokines: IFN-γ, IL-2, IL-4, IL-10; vitamin A deficiency was shown to induce inflammation and aggravate existing inflammatory states, whereas supplementation with vitamin A could ameliorate inflammation. MATERIALS AND METHODS: The active compounds were included in tablets: 250 mg of olive leaf extract titrated in 10% hydroxytyrosol, 200 mg of citrus fruits extract titrated in 60% hesperidin, 10 mg of rice (Oryza sativa) seeds extract titrated in 1% spermidine and 0.8 mg of vitamin A. Mice of an inbred group were randomly selected and divided in the control group and drug-treated group. The wound necessary for lymphedema generation was made on the tail of each mice 1 cm below the base of the trunk. RESULTS: After surgical intervention, there was a gradual increase in the circumference of both ends of the wound. The control group showed higher increase of tail volume than the drug-treated group. The differences in tail swelling between the control group and the drug-treated group were significantly different. The peak of swelling was anticipated to the 6th day in the drug-treated group, whereas in the control group the peak was reached later on. CONCLUSIONS: The tested drug prevented the induction of swelling from day 5th of wound creation and decreased the duration of swelling, favoring the wound healing.

A transgenic reporter under control of an es1 promoter/enhancer marks wound epidermis and apical epithelial cap during tail regeneration in Xenopus laevis tadpole.

Rapid wound healing and subsequent formation of the apical epithelial cap (AEC) are believed to be required for successful appendage regeneration in amphibians. Despite the significant role of AEC in limb regeneration, its role in tail regeneration and the mechanisms that regulate the wound healing and AEC formation are not well understood. We previously identified Xenopus laevis es1, which is preferentially expressed in wounded regions, including the AEC after tail regeneration. In this study we established and characterized transgenic Xenopus laevis lines harboring the enhanced green fluorescent protein (EGFP) gene under control of an es1 gene regulatory sequence (es1:egfp). The EGFP reporter expression was clearly seen in several regions of the embryo and then declined to an undetectable level in larvae, recapitulating the endogenous es1 expression. After amputation of the tadpole tail, EGFP expression was re-activated at the edge of the stump epidermis and then increased in the wound epidermis (WE) covering the amputation surface. As the stump started to regenerate, the EGFP expression became restricted to the most distal epidermal region, including the AEC. EGFP was preferentially expressed in the basal or deep cells but not in the superficial cells of the WE and AEC. We performed a small-scale pharmacological screening for chemicals that affected the expression of EGFP in the stump epidermis after tail amputation. The EGFP expression was attenuated by treatment with an inhibitor for ERK, TGF-ß or reactive oxygen species (ROS) signaling. These treatments also impaired wound closure of the amputation surface, suggesting that the three signaling activities are required for es1 expression in the WE and successful wound healing after tail amputation. These findings showed that es1:egfp Xenopus laevis should be a useful tool to analyze molecular mechanisms regulating wound healing and appendage regeneration.

Effect of hyperbaric oxygen therapy on healing in an experimental model of degloving injury in tails of nicotine-treated rats.

The aim of this study was to investigate the effect of hyperbaric oxygen (HBO) therapy on healing in an experimental model of a degloving injury of the tails of nicotine-treated rats. Thirty-two male Sprague-Dawley rats were randomized to four groups (n = 8): nicotine (group 1); HBO (group 2); nicotine + HBO (group 3); and control (group 4). The mean length of the necrotic parts of the tails at the degloving injury site was significantly higher in group 1 compared with groups 2, 3, and 4, and was significantly lower in group 2 compared with groups 1, 3, and 4. The mean histopathological stage of ulcers at the degloving injury site was statistically significantly higher (more severe) in group 1 compared with groups 2, 3, and 4, and was statistically significantly lower in group 2 compared with groups 1, 3, and 4. It appears that the negative effects of nicotine on wound healing in degloving injuries are negated by the positive effects of immediate HBO therapy.

Brain gene expression differences are associated with abnormal tail biting behavior in pigs.

Knowledge about gene expression in animals involved in abnormal behaviors can contribute to the understanding of underlying biological mechanisms. This study aimed to explore the motivational background to tail biting, an abnormal injurious behavior and severe welfare problem in pig production. Affymetrix microarrays were used to investigate gene expression differences in the hypothalamus and prefrontal cortex of pigs performing tail biting, pigs receiving bites to the tail and neutral pigs who were not involved in the behavior. In the hypothalamus, 32 transcripts were differentially expressed (P < 0.05) when tail biters were compared with neutral pigs, 130 when comparing receiver pigs with neutrals, and two when tail biters were compared with receivers. In the prefrontal cortex, seven transcripts were differently expressed in tail biters when compared with neutrals, seven in receivers vs. neutrals and none in the tail biters vs. receivers. In total, 19 genes showed a different expression pattern in neutral pigs when compared with both performers and receivers. This implies that the functions of these may provide knowledge about why the neutral pigs are not involved in tail biting behavior as performers or receivers. Among these 19 transcripts were genes associated with production traits in pigs (PDK4), sociality in humans and mice (GTF2I) and novelty seeking in humans (EGF). These are in line with hypotheses linking tail biting with reduced back fat thickness and explorative behavior.

Analgesic and antipyretic activities of Drymaria cordata (Linn.) Willd (Caryophyllaceae) extract.

Drymaria cordata (Linn.) Willd (Caryophyllaceae) is an herbaceous plant widely used in traditional African medicine (TAM) for the treatment of diverse ailments including painful and febrile conditions. This study was conducted to investigate the analgesic and antipyretic properties of the whole plant extract of D. cordata. The acetic acid-induced writhing, formalin, and tail clip tests were used to evaluate analgesic activity while the 2,4-dinitrophenol (DNP)-, d-amphetamine-, and yeast-induced hyperthermia tests were used to investigate antipyretic activity in rodents. D. cordata (100, 200, and 400 mg kg(-1), p.o) produced significant (p<0.05) analgesic activity in the mouse writhing, formalin (second phase), and tail clip tests. The effects of D. cordata were generally comparable to those of acetylsalicylic acid (ASA, 100 mg kg(-1), p.o) and morphine (2 mg kg(-1), s.c). Also, D. cordata produced significant (p<0.05) dose-dependent inhibition of temperature elevation in the 2,4-DNP and yeast-induced hyperthermia models with peak effects produced at the dose of 400 mg kg(-1). The effect at this dose was comparable to that of ASA in the two models. In the d-amphetamine method, D. cordata produced significant (p<0.05) dose- and time-dependent reduction of temperature elevation with peak effect produced at the dose of 200 mg kg(-1). The effect of the extract at this dose was greater than that of ASA. The results obtained in this study demonstrate that the aqueous whole plant extract of Drymaria cordata possesses analgesic and antipyretic properties mediated through peripheral and central mechanisms.

Cloning and expression of a retinoic acid receptor ß2 subtype from the adult newt: evidence for an early role in tail and caudal spinal cord regeneration.

Retinoic acid receptor beta 2 (RARß2) has been proposed as an important receptor mediating retinoid-induced axonal growth and regeneration in developing mammalian spinal cord and brain. In urodele amphibians, organisms capable of extensive central nervous system (CNS) regeneration as adults, this receptor had not been isolated, nor had its function been characterized. We have cloned a full-length RARß2 cDNA from adult newt CNS. This receptor, NvRARß2, is expressed in various adult organs capable of regeneration, including the spinal cord. Interestingly, both the NvRARß2 mRNA and protein are up-regulated during the first 2 weeks after amputation of the tail, primarily in the ependymoglial and meningeal tissues near the rostral cut surface of the cord. Treatment with LE135, a RARß-selective antagonist, caused a significant inhibition of ependymal outgrowth and a decrease in tail regenerate length. These data support an early role for this receptor in caudal spinal cord and tail regeneration in this amphibian.

Effect of age on the behavioral and physiological responses of piglets to tail docking and ear notching.

Neonatal piglets are often subject to potentially painful processing procedures such as tail docking and ear notching during the first few days after birth. However, these procedures may influence the development of suckling behavior and passive transfer of immunoglobulins, especially if done within the first day postpartum. The objective of this experiment was to compare the effects of processing piglets during the first 24 h versus at 3 d of age on suckling and pain-related behavior, the passive transfer of immunoglobulins, and growth. Six piglets per litter from 20 litters (n = 120 piglets) were used in a 3 x 2 complete block design. Piglets were weighed at birth and assigned to 1 of 3 treatments (balanced by birth weight): control (unmanipulated), sham processed (manually manipulated), and processed (tail docked and ear notched) at 1 of 2 ages (1 or 3 d of age). Vocalizations were recorded during the procedures, and piglets were observed after the procedures for pain-related behavior. Suckling behavior was observed for 6 h on each of d 1 to 4. Colostrum samples were collected after the birth of all piglets (before first suck), and blood samples were collected on d 5 to examine concentrations of immunoglobulins (IgA and IgG) and IGF-I. Body weights were measured at birth and on d 5 and 14. During the procedures, processed piglets, regardless of age, vocalized at a greater frequency (P < 0.001) and produced more high frequency calls (P = 0.016) than sham-processed piglets. All piglets on d 1 produced more high frequency calls than all piglets on d 3 (P = 0.047). Immediately after the procedures, sham-processed and processed piglets spent less time lying and more time standing than control piglets (P < 0.001), whereas processed piglets jammed their tail between their legs more than sham-processed or control piglets (P < 0.001). Lying, standing and tail posture were not influenced by age, nor were there age by treatment interactions. Piglets on d 1 trembled more than piglets on d 3 (P < 0.001), and this tended to be exacerbated by processing (P = 0.076). There was no effect of treatment or age of treatment on suckling behavior. Processed piglets had decreased IgG serum concentrations compared with sham-processed and control piglets (P = 0.029), although there was no interaction between treatment and age of treatment (P = 0.67). Whereas tail docking and ear notching do appear to result in short-term pain and modulated immune status, processing on d 1 appears neither better nor worse than processing on d 3.

Antinociceptive properties of ethanolic extract and fractions of Pterodon pubescens Benth. seeds.

We have previously demonstrated that the hydroalcoholic extract from Pterodon pubescens Benth. seeds (sucupira branca, Leguminosae) exhibits anti-arthritic activity and that its oleaginous extract (OEP) and PF1 fraction exhibit acute and topic anti-edematogenic activities. In this work, we studied the antinociceptive activity of OEP and its fractions on the acetic acid-induced abdominal constriction and formalin assays in SW male mice. OEP was obtained by ethanol extraction and its four fractions by sequential liquid-liquid extraction. PF2 GC/MS profile indicated it contains furane diterpenes derivatives of vouacapan and non-vouacapan compounds. The antinociceptive properties were demonstrated to OEP and predominantly to PF1 and PF2 by the writhing test. In the formalin assay, PF1 inhibited both phases and PF2 inhibited mainly the late one. Then, PF1 and PF2 seemed to present antinociceptive effects by different mechanisms, peripheral and/or central inhibitory ones, and showed maximum antinociceptive properties with very low doses, providing a rationale for its popular use in pain disorders.

The N-Methyl-D-aspartate receptor antagonist dizocilpine inhibits associative antinociceptive tolerance to morphine in mice: relation with memory.

I and coauthor previously reported the memory facilitation effect of morphine. The main purpose of this study was to evaluate the involvement of the N-methyl-D-aspartate (NMDA) receptor in associative tolerance to morphine by a contextual procedure. Antinociceptive response latency was measured by the tail-pinch method during repeated morphine (5 mg/kg, s.c.) injection for four consecutive days with pretreatment by dizocilpine (0.01, 0.05, 0.1 mg/kg, i.p.) at 30 min prior to morphine injection in the training phase and before and after morphine injection in the test phase. The nociceptive response latency was shortened by the single administration of dizocilpine (0.05 to 0.25 mg/kg, i.p.). Pretreatment by dizocilpine at 0.05 or 0.1 mg/kg weakened the antinociception to morphine on Day 1, but decreased the tolerance throughout the training phase. In the test phase, the animals were allocated into the same and different contexts. In the test phase, hyperalgesia before morphine injection in the same context and antinociception after morphine injection in the different context were evident in the saline-pretreated group in the training phase, but they were not observed in those contexts in the dizocilpine-pretreated groups. These results suggest that memory dysfunction with dizocilpine inhibits the recovery of associative tolerance to morphine by contextual change.