Paws Of Animals Research Articles

Analysis of the causes of the occurrence and measures of prevention of pododermatitis in rabbits

Analysis of the causes of the occurrence and measures of prevention of pododermatitis in rabbits

Вариабельность биохимических показателей крови и установление референтных интервалов в доклинических исследованиях. Сообщение 12: обыкновенные игрунки

The common marmosets (Callithrix jacchus) is currently widely used in preclinical studies. These animals are used in preclinical studies aimed at studying pathological conditions associated with the central nervous system, the immune response, the study of infectious and viral diseases, as well as metabolic disorders. The wide involvement of common marmosets in preclinical studies indicates the need for control and periodic monitoring of health. The aim of this study was to establish reference intervals of biochemical blood parameters of healthy mature common marmosets, which can be used as the basic values of animals in preclinical studies. The data obtained from 24 males and 24 females were analyzed. The age of males and females of common marmosets was 12 months, the average body weight was 350 g. Determination of biochemical parameters of all animals was carried out under the same conditions using generally accepted analytical methods. For the study, blood was used, which was taken from the lateral vena of the hind paw of animals. The following parameters were recorded in the blood serum of animals: creatinine, urea, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, cholesterol, triglycerides, total protein, albumin, glucose, creatine kinase, lactate dehydrogenase and total bilirubin. The obtained values were analyzed with other reference intervals found in the scientific literature. In general, the ranges of the conside­red indicators of both males and females were similar, but there were also differences. Thus, the upper limits of the calculated reference intervals of urea, aspartate aminotransferase, alanine aminotransferase, total bilirubin, creatine kinase and triglycerides were lower than similar literary values. According to such obtained ranges of values as total protein, albumin, glucose and lactate dehydrogenase, both similar and higher literary values were found. Differences in the obtained intervals of normal values with similar ranges from other publications may be associated with preanalytical, analytical and postanalytical features of the formulation of the analysis. Seasonality, conditions of keeping and nutrition of the marmosets are also important factors. That is why a serious aspect in preclinical research is the creation of reference intervals of healthy laboratory animals in each scientific center. The obtained intervals can help to avoid erroneous conclusions in the course of studying the safety of new therapeutic agents or modeling a pathological state.

Magnesium sulphate activates the L-arginine/NO/cGMP pathway to induce peripheral antinociception in mice

In the present study, we investigated whether magnesium sulphate activates the L-arginine/NO/cGMP pathway and elicits peripheral antinociception. The male Swiss mice paw pressure test was performed with hyperalgesia induced by intraplantar injection of prostaglandin E2. All drugs were administered locally into the right hind paw of animals. Magnesium sulphate (20, 40, 80 and 160 μg/paw) induced an antinociceptive effect. The dose of 80 μg/paw elicited a local antinociceptive effect that was antagonized by the non-selective NOS inhibitor, L-NOArg, and by the selective neuronal NOS inhibitor, L-NPA. The inhibitors, L-NIO and L-NIL, selectively inhibited endothelial and inducible NOS, respectively, but were ineffective regarding peripheral magnesium sulphate injection. The soluble guanylyl cyclase inhibitor, ODQ, blocked the action of magnesium sulphate, and the cGMP-phosphodiesterase inhibitor, zaprinast, enhanced the antinociceptive effects of intermediate dose of magnesium sulphate. Our results suggest that magnesium sulphate stimulates the NO/cGMP pathway via neuronal NO synthase to induce peripheral antinociceptive effects.

Gelsemium effect against nerve injury-induced mechanical allodynia and hyperalgesia

Background Gelsemium dilutions are prescribed for their anxiolytic and its analgesic effects. Investigations using alkaloids isolated from Gelsemium support the hypothesis of Gelsemium-induced analgesia. Aims We evaluated the antinociceptive effect of Gelsemium dilutions 3, 5 and 9C in neuropathic models. Methodology To investigate the potential effect of Gelsemium dilutions 3, 5 and 9C to prevent or to correct sciatic nerve chronic constriction (CCI) injury, we used the von-Frey hair behavioral test. CCI-induced neuropathic pain in rats represents characteristic painful behaviors like hyperalgesia and allodynia, thus validating its suitability for the evaluation of anti-neuropathic drugs. To test the corrective effect, Gelsemium 3, 5 or 9 C or placebo were administered from Day 14 after sciatic nerve ligation, i.e., when neuropathic symptoms were observed. In the preventive strategy, Gelsemium dilutions and placebo began immediately after the sciatic nerve ligation (D0). Results and discussion We observed that in placebo groups, the mechanical-sensitivity thresholds remained unchanged all treatment days whereas Gelsemium dilutions 3C and 5C exerted a strong or moderate antinociceptive effect, respectively. In CCI-rats, the contralateral paw of placebo-treated animals did not respond to 4g non-noxious stimulation but the ipsilateral responses increased progressively (0%-D0, 52%-D14, 72%-D21 and 88%-D28), confirming the occurrence of allodynia. Gelsemium dilution 3C corrective and preventive treatments efficiently reduced the allodynic responses (72-to-38%-D21 and 88-to-22%-D28). Gelsemium 5C also exhibited significant anti-allodynic action while Gelsemium 9C was ineffective. Similarly, one week treatment with Gelsemium 3 or 5C efficiently reduced 15g or 26g-evoked hyperalgesia and the beneficial action improved after 2 weeks. Chronic therapies with current analgesics induce several side-effects. Conclusion The present work, suggest that Gelsemium dilutions devoid of toxicity and used as anxiolytic in humans induced analgesia, opens interesting/alternative perspectives for long-term management of pain without undesirable effects.

Comparative antinociceptive effect of aspirin and aspirin nano particles in semisolid formulae in mice

Aspirin are commonly used analgesic, anti-inflammatory, and anti-pyretic drug in medicine, oral route is the most common one for drug administration as a result it will produce different adverse effects like peptic ulcer, nephropathy, and thrombocytopenia even with low and continuous therapeutic dose, so the alternative topical route is preferable with minimal adverse effects and effective concentration.Therefore, in the present study was to investigate whether the antinociceptive property of aspirin would enhance if used aspirin as nanoparticles after preparing it in several forms (gel, cream and ointment). Thirty-two healthy male mice weighing 30-35 gm. were used in the present study. The animals were divided as a randomized design. Each mouse was treated topically. All drug concentration of aspirin was prepared using gel, cream and ointment as vehicle and topically application on fore and hind paw of experimental animals. Pain was induced by application of hot plate for assessment of latency of pain stimulus. Time from placement to jumping or hind paw licking was recorded as latency of response. The result showed that the median effective concentration (EC50) for analgesic effect of aspirin (gel, cream, and ointment) were 0.848, 0.958 and 1.00% respectively while these EC50s were decrease when used nanoparticles aspirin (gel, cream and ointment) to 0.72, 0.657, and 0.701% respectively. In conclusion, topical applied of aspirin will produce effective therapeutic antinociceptive effects in mice although gel preparation produce a better response followed by cream, then ointment due to pharmacokinetic properties. Also nanoparticle preparation will produce superior response in all forms, whether Nano aspirin is prepared in gel form, cream or ointment.

Effect of Ultra-Micronized-Palmitoylethanolamide and Acetyl-l-Carnitine on Experimental Model of Inflammatory Pain.

Palmitoylethanolamide (PEA), a fatty acid amide, has been widely investigated for its analgesic and anti-inflammatory properties. The ultra-micronized formulation of PEA (um-PEA), that has an enhanced rate of dissolution, is extensively used. Acetyl-l-carnitine (LAC), employed for the treatment of neuropathic pain in humans, is able to cause analgesia by up-regulating type-2 metabotropic glutamate (mGlu2) receptors. In the present study, we tested different associations of um-PEA, LAC and non-micronized PEA (non-m-PEA) in a rat model of carrageenan (CAR)-induced paw edema. Intraplantar injection of CAR into the hind paw of animals caused edema, thermal hyperalgesia, accumulation of infiltrating inflammatory cells and augmented myeloperoxidase (MPO) activity. All these parameters were decreased in a significantly manner by oral administration of a compound constituted by a mixture of um-PEA and LAC in relation 1:1 (5 mg/kg), but not with the association of single compounds administered one after the other. These findings showed the superior anti-inflammatory and anti-nociceptive action displayed by oral administration of um-PEA and LAC versus LAC plus, separate but consecutive, um-PEA in the rat paw CAR model of inflammatory pain.

The characteristics of the anti-inflammatory action of sodium diclofenac in cold and normal environment

Aim. To determine the effect of low temperature and the resulting hypothermia on the anti-inflammatory activity of classical NSAID – sodium diclofenac on the model of carrageenan-induced paw edema in mice.Materials and methods. The model of exudative inflammation – carragenin-induced paw edema was used. Mice were given an injection into the plantar aponeurosis of the left hind paw with 0.05 ml of a 1 % carrageenan solution. Sodium diclofenac was administered as a suspension intragastrically for 30 minutes before carrageenan in an effective frigoprotective dose of 14 mg/kg. The study was performed in three temperature regimes: 1) at a constant temperature +22 °C; 2) during cold exposure of animals (+10 °C) for 3 hours after the carrageenan injection; 3) during cold exposure of animals (+10 °С) for 3 hours before and 3 hours after carrageenan injection. The severity of the inflammatory reaction was evaluated by the increase in the mass of the paws of animals. The anti-inflammatory activity of sodium diclofenac was determined by the ability to reduce the mass of the edematous limb compared to the untreated control. To assess the severity of hypothermia, the body temperature of animals was measured using an infrared thermometer.Results and discussion. At a temperature of 22 °C, carrageenan increased the median paw mass by 84 mg (73 %), sodium diclofenac decreased its growth by 22.5 mg (16.5 %). It indicates the inherent anti-inflammatory effect of this drug. In regime 2 the phlogogenic effect of carrageenan persisted, but its severity was significantly less (p ≤ 0.01) – the increase in paw mass was only 54.5 mg (44 %). Under these conditions, the anti-inflammatory effect of diclofenac was absent – the mass of the edematous paw increased by 51.5 mg (42 %). In regime 3 the pro-inflammatory effect of carrageenan was even weaker: the mass of the edematous paw increased by only 42.5 mg (36 %), sodium diclofenac also did not show the anti-inflammatory effect under these conditions, increasing the median paw mass by 42.5 mg (32 %). Sodium diclofenac significantly reduced the degree of hypothermia in regimes 2 and 3, confirming the presence of frigoprotective properties. The relationship between the degree of hypothermia and the severity of the inflammatory reaction was analyzed.Conclusions. The pro-inflammatory effect of carrageenan decreases with hypothermia. Sodium diclofenac exhibits frigoprotective properties, reducing the severity of hypothermia. The anti-inflammatory effect of the drug on the model of carrageenan-induced paw edema disappears at low temperatures.

Analgesic and anti-inflammatory effects and molecular mechanisms of Kadsura heteroclita stems, an anti-arthritic Chinese Tujia ethnomedicinal herb

Ethnopharmacological relevanceRheumatoid arthritis (RA) is a chronic autoimmune disease characterized by failure of spontaneous resolution of inflammation. The stem of Kadsura heteroclite (KHS) is a well-known anti-arthritic Tujia ethnomedicinal plant, which named Xuetong in folk, has long been used for the prevention and treatment of rheumatic and arthritic diseases. Aim of the studyThe analgesic and anti-inflammatory effects and the potential mechanisms behind such effects of KHS would be investigated by using different animal models. Materials and methodsThe abdominal writhing episodes of mice induced by intraperitoneal injection of acetic acid and the tail-flick response induced by radiant heat stimulation were used to evaluate the analgesic effect of KHS. The number of abdominal writhing episodes of mice and the latency of tail-flick in rats were measured and recorded. In acute inflammatory models, the ear edema of mice was induced by applying xylene on the ear surface, while the paw edema of male and female rats was induced by subcutaneous injection of carrageenan into the right hind paws of animals. The carrageenan-induced paw swelling in rats were selected as an anti-acute inflammatory mechanism of KHS. Serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor (TNF-α) were measured by ELISA, and protein expression of cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) were detected by Western blot. ResultsThe maximal tolerated single dose of KHS was determined to be 26 g/kg in both sexes of mice. Pharmacological studies showed that KHS at the dose of 200 mg/kg significantly prolonged the reaction time of rats to radiant heat stimulation and suppressed abdominal writhing episodes of mice induced by intraperitoneal injection of acetic acid. KHS at the dose of 200, 400, and 800 mg/kg, showed dose-dependent inhibition of xylene-induced ear swelling in mice. KHS at the dose of 100, 200, 400, and 800 mg/kg demonstrated dose- and time-dependent suppression of paw edema induced by subcutaneous injection of carrageenan in both all rats. Mechanistic studies revealed that the anti-inflammatory effect of KHS was associated with inhibition of the production of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α and effectively decreased the expression of COX and iNOS proteins in the carrageenan-injected rat serum, paw tissues and inflammatory exudates. The positive reference drug, rotundine at a dosage of 100 mg/kg and indomethacin at a dosage of 10 mg/kg were used in both mice and rat models. ConclusionThese results suggested that KHS has significant effects on analgesia and anti-inflammation with decreasing the pro-inflammation cytokines of IL-1β, IL-6, and TNF-α and inhibiting the proteins expression of COX-2 and iNOS.

Kappa opioid signaling in the right central amygdala causes hind paw specific loss of diffuse noxious inhibitory controls in experimental neuropathic pain.

Diffuse noxious inhibitory controls (DNICs) is a pain-inhibits-pain phenomenon demonstrated in humans and animals. Diffuse noxious inhibitory control is diminished in many chronic pain states, including neuropathic pain. The efficiency of DNIC has been suggested to prospectively predict both the likelihood of pain chronification and treatment response. Little is known as to why DNIC is dysfunctional in neuropathic pain. Here, we evaluated DNIC in the rat L5/L6 spinal nerve ligation (SNL) model of chronic pain using both behavioral and electrophysiological outcomes. For behavior, nociceptive thresholds were determined using response to noxious paw pressure on both hind paws as the test stimulus before, and after, injection of a conditioning stimulus of capsaicin into the left forepaw. Functionally, the spike firing of spinal wide-dynamic-range neuronal activity was evaluated before and during noxious ear pinch, while stimulating the ipsilateral paw with von Frey hairs of increased bending force. In both assays, the DNIC response was significantly diminished in the ipsilateral (ie, injured) paw of SNL animals. However, behavioral loss of DNIC was not observed on the contralateral (ie, uninjured) paw. Systemic application of nor-binaltorphimine, a kappa opioid antagonist, did not ameliorate SNL-induced hyperalgesia but reversed loss of the behavioral DNIC response. Microinjection of nor-binaltorphimine into the right central amygdala (RCeA) of SNL rats did not affect baseline thresholds but restored DNIC both behaviorally and electrophysiologically. Cumulatively, these data suggest that net enhanced descending facilitations may be mediated by kappa opioid receptor signaling from the right central amygdala to promote diminished DNIC after neuropathy.

Environmental enrichment alleviates chronic pain in rats following a spared nerve injury to induce neuropathic pain. A preliminary study.

BackgroundIn mice, chronic pain can be alleviated with enriched environments (EEs). The purpose of this preliminary study is to investigate whether pain behaviors in rats with peripheral neuropathy would be altered when keeping these animals in either 1) standard laboratory cages or in 2) a significantly EE.MethodsTwo groups of rats (n=8/group) underwent a spare nerve injury surgery of the right hind leg; one group (n=8) was returned to standard ventilated cages (2 rats/cage), the other (n=8) placed in an EE (8 rats/ferret cage with toys). A third group (n=8) underwent a sham surgery and was used as control. These animals were returned to standard ventilated cages (2 rats per cage). Spare nerve injury surgery consisted of ligation/transection of the tibial and common peroneal branches of the sciatic nerve of the right leg only. Von Frey Filaments were applied to test mechanical sensitivity of both hind paws.ResultsThe right paw of nerve-injured animals was hypersensitive to mechanical stimuli at 2, 4, and 8 weeks following the surgery; however, animals in the EE conditions showed significantly (P<0.05) less mechanical sensitivity than rats left in the standard caging environment (2, 4, and 8 weeks postsurgery: standard environment 2.8±0.5, 2.8±0.7, and 2.6±0.4 and EE 4.7±0.6, 5.8±0.5, and 5.5±0.7). Sham animals were unaffected by the surgery.ConclusionEnvironmental enrichment alleviated mechanically induced chronic pain in a spared nerve injury rat model of neuropathic pain. Findings also suggest that environmental enrichment, as a method to alleviate pain, may be species-specific, motor behaviors being a very important parameter when considering pain modulation.

Dog walking brings Toxocara eggs to people's homes

To find out the transmission routes for Toxocara infection, the possibility of transfer of Toxocara eggs from the soil on the paws of animals and on the shoes of people was explored. For this purpose, a study was conducted to find helminth eggs in washings from the paws of dogs after walking, from the shoes of their owners, as well as non-dog owners. Toxocara eggs were detected in 19.4% of the dogs’ paws washings and in 11.4% of washings from the shoes of their owners. The number of eggs found on the paws was about twice as high as on the shoes. The mean number of eggs in the sample was 2.9 in washings from the paws and 1.8 from the shoes. In the samples, Toxocara cati eggs prevailed both in occurrence and in abundance. Out of the total number of positive samples, the eggs of T. cati were found in 83%, and T. canis in 42%. 79% of the found eggs belonged to T. cati and 21% to T. canis. In the washings from shoes of people that do not own dogs, the eggs of parasites were not found. This study demonstrates that the helminth eggs can be transferred from contaminated soil to people’s homes on the paws and shoe soles. Even animals without a patent infection may take part in propagation of infection causing risks of human toxocariasis. In dogs, in addition, the transferred on paws T. canis eggs can serve as a causative agent of permanent, cumulative subclinical infection with a deferred manifestation in posterity. It is supposed that infestation “through the paws” is one of the probable routes of transmission of toxocariasis in dogs.

Automated Assessment of Loss of Consciousness Using Whisker And Paw Movements During Anesthetic Dosing in Head-Fixed Rodents.

The precise identification of loss of consciousness (LOC) is key to studying the effects of anesthetic drugs in neural systems. The standard behavioral assay for identifying LOC in rodents is the Loss of Righting Reflex (LORR), assessed by placing the animal in the supine position every minute until it fails to right itself. However, this assay cannot be used when the rodents are head-fixed, which limits the use of powerful techniques such as multi-electrode recordings, in vivo patch clamp, and neuronal imaging. In these situations, an alternative way to assess LOC is needed. We propose that loss of movement (LOM) in whiskers and paws of head-fixed animals can be used as an alternative behavioral assay in head-fixed animals. Unlike LORR, LOM in whiskers and paws is much harder to detect by visual inspection. Therefore, we developed a method to automatically assess for LOM of whiskers and paws in head fixed rodents during in vivo patch clamp recordings. Our method uses an algorithm based on optical flow and point-process filtering which can be run on images acquired on regular cameras at low frame-rates. We show that the algorithm can achieve at least comparable accuracy in detecting LOC when compared with consensus among human observers, as well as improved precision when compared with individual observers. In the future, we aim to to expand the method to detect more behavioral end-points during anesthesia such as paradoxical excitation. Eventually, we hope to enable multi-modal anesthesia studies, which incorporates behavioral and neurophysiological data.

Pharmacological and biochemical studies on protective effects of mangiferin and its interaction with nitric oxide (NO) modulators in adjuvant-induced changes in arthritic parameters, inflammatory, and oxidative biomarkers in rats.

Current study was designed to evaluate protective effect of mangiferin and its interaction with low dose of nitric oxide (NO) modulators in complete Freund's adjuvant (CFA) inoculated rats. Male wistar rats (200-300g, n = 8 per group) were used in the study. On day ''0'' of study arthritis was induced in rats by injecting 0.2ml CFA in sub-planter region of right hind paw of animals. Treatment with methotrexate (5mg/kg), mangiferin (10-30mg/kg) alone and in combination with NO modulators was given (i.p.) from days 14 to 28. After 28days, blood and joint synovial fluid was collected for biochemical analysis and rat paws were excised to estimate MDA and SOD in tissue (paw) homogenates. CFA inoculation significantly increases (1) arthritic index, (2) ankle diameter, (3) paw volume, and (4) serum TNF-α, IL-6, IL-1β, and synovial TNF-α levels (p < 0.001). The serum Th1 (IFN-γ) and Th2 (IL-4) cytokine levels, MDA levels in rat paw tissue homogenates and serum NF-κB levels were also found significantly increased. Significant decrease in serum IL-10 levels and SOD activity was found after CFA inoculation. These CFA-induced arthritic changes, cytokine profile, and oxidative stress markers were significantly reversed by mangiferin (10-30mg/kg) treatment alone and in combination with L-arginine and L-NAME nitric oxide modulators (p < 0.05). Treatment with methotrexate (5mg/kg) also significantly reversed these adjuvant changes (p < 0.05). However, effect of methotrexate was less marked as compared to mangiferin (30mg/kg) alone and in combination with L-NAME (10mg/kg), but was comparable or slightly better than mangiferin (10 and 20mg/kg). Thus, on the basis of our findings, we can suggest that interaction of mangiferin with nitric oxide modulators may have therapeutic value for chronic inflammatory disease such as RA.

Serotonin induces peripheral antinociception via the opioidergic system

PurposeStudies conducted since 1969 have shown that the release of serotonin (5-HT) in the dorsal horn of the spinal cord contributes to opioid analgesia. In the present study, the participation of the opioidergic system in antinociceptive effect serotonin at the peripheral level was examined. MethodsThe paw pressure test was used with mice (Swiss, males from 35 g) which had increased pain sensitivity by intraplantar injection of PGE2 (2 μg). Serotonin (250 ng), administered locally to the right paw of animals, produces antinociception in this model. ResultsThe selective antagonists for mu, delta and kappa opioid receptors, clocinnamox clocinnamox (40 μg), naltrindole (60 μg) and nor-binaltorfimina (200 μg), respectively, inhibited the antinociceptive effect induced by serotonin. Additionally, bestatin (400 μg), an inhibitor of enkephalinases that degrade peptides opioids, enhanced the antinociceptive effect induced by serotonin (low dose of 62.5 ng). ConclusionsThese results suggest that serotonin possibly induce peripheral antinociception through the release of endogenous opioid peptides, possible from immune cells or keratinocytes.

Effectiveness of bis(phenylimidazoselenazolyl) diselenide on a mouse model of inflammatory nociception

The injection of complete Freund’s adjuvant (CFA) in the hindpaw of rodents induces tissue inflammation and nociceptive hypersensitivity. In addition, it has been reported that organoselenium compounds have antinociceptive properties in animal models. The purpose of this study was to investigate the potential antinociceptive effect of bis(phenylimidazoselenazolyl) diselenide (BPIS) in the inflammatory nociception model in mice and its possible mechanism of action. C57BL/6 mice received CFA intraplantar in right hindpaw and the inflammatory response was verified 24h after injection as well as the antinociceptive effect of BPIS. The CFA-induced mechanical allodynia was reversed by BPIS treatment (1mg/kg, p.o.) observed through the von Frey hair test. Additionally, L-arginine (600mg/kg; i.p.), administered before BPIS treatment, blocked its antinociceptive effect. Regarding myeloperoxidase activity, NOx and 3-nitrotyrosine levels, BPIS administration did not reverse alterations observed in the paw of animals injected with CFA. BPIS reversed the increase in spinal NOx content induced by CFA. In the spinal cord, it was also found that CFA induced an increase in malondialdehyde content and a decrease in glutamate uptake, and these alterations were reversed by BPIS. Moreover, BPIS treatment induced an increase in non-protein thiol levels in spinal cord of animals that received CFA injection. No alterations were found in glutathione peroxidase, reductase and S-transferase activities of experimental groups. The obtained data reinforce the relevance of BPIS as an antinociceptive agent as well as highlight the importance of the nitric oxide pathway in the spinal cord and its antioxidant potential for its mechanism of action.

Treatment with Sulforaphane Produces Antinociception and Improves Morphine Effects during Inflammatory Pain in Mice.

The activation of nuclear factor erythroid 2-related factor 2 (Nrf2) exerts potent antioxidative and anti-inflammatory effects; however, its participation in the modulation of chronic inflammatory pain and on the antinociceptive effects of μ-opioid receptor (MOR) agonists has not been evaluated. We investigated whether the induction of Nrf2 could alleviate chronic inflammatory pain and augment the analgesic effects of morphine and mechanisms implicated. In male C57BL/6 mice with inflammatory pain induced by complete Freund's adjuvant (CFA) subplantarly administered, we assessed: 1) antinociceptive actions of the administration of 5 and 10 mg/kg of a Nrf2 activator, sulforaphane (SFN); and 2) effects of SFN on the antinociceptive actions of morphine and on protein levels of Nrf2, heme oxygenase 1 (HO-1), and NAD(P)H: quinone oxidoreductase 1 (NQO1) enzymes, microglial activation and inducible nitric oxide synthase (NOS2) overexpression, as well as on mitogen-activated protein kinase (MAPK) and MOR expression in the spinal cord and paw of animals with inflammatory pain. Results showed that treatment with SFN inhibited allodynia and hyperalgesia induced by CFA and increased the local antinociceptive actions of morphine. This treatment also augmented the expression of Nrf2, HO-1, NQO1, and MOR, and inhibited NOS2 and CD11b/c overexpression and MAPK phosphorylation induced by inflammation. Thus, this study shows that the induction of Nrf2 might inhibit inflammatory pain and enhance the analgesic effects of morphine by inhibiting oxidative stress and inflammatory responses induced by peripheral inflammation. This study suggests the administration of SFN alone and in combination with morphine are potential new ways of treating chronic inflammatory pain.

Anti-Inflammatory Subfractions Separated from Acidified Chloroform Fraction of Fenugreek Seeds (Trigonella foenum-graecum L.)

ABSTRACTConsidering the side effects of current anti-inflammatory drugs, novel therapeutic agents are desired. We have succeeded in separating flavonoid-rich fractions with anti-inflammatory effect from fenugreek seeds (Trigonella foenum-graecum L.). In this work, we aimed to carry out further fractionation to find active anti-inflammatory subfractions. Trigonelline content of the plant was determined by spectrophotometric method. Fenugreek seeds were extracted consecutively with petroleum ether, acidified chloroform (ACC), alkaline chloroform (AKC), methanol, and water. ACC fraction, which had exhibited the highest anti-inflammatory effect, was further fractionated using column chromatography. Obtained subfractions were evaluated using carrageenan-induced paw edema (CIPE) method. Animals were pretreated by test compounds, and after 30 minutes edema was induced by subcutaneous injection of 100 µl of 1% w/v carrageenan into the right paw of animals. Volume difference of both paws was measured at different times after carrageenan injection. The concentration of trigonelline was determined as 16.2%. ACC fraction inhibited paw edema significantly in comparison to control (p < .05). Four subfractions (dry weight percentage basis) were selected for pharmacological study. F3 subfraction exhibited the greatest inhibition at 15 mg/kg (p < .001). ACC fraction and F4 significantly inhibited paw edema at doses of 5, 10, and 15 mg/kg (p < .001). Phytochemical studies indicated the presence of flavonoids in ACC and active subfractions. Further separation can lead to finding active components from active subfractions, which probably belong to flavonoid phytochemicals. Considering the gastroprotective effect of fenugreek, we hope the separated fractions also would be free of gastrointestinal side effects.

Pharmacological studies on the anti-inflammatory and immunomodulatory role of pentoxifylline and its interaction with nitric oxide (NO) in experimental arthritis in rats.

Present study was designed to evaluate protective effects of pentoxifylline and its potentiation with low dose of nitric oxide (NO) modulators in adjuvant-induced experimental arthritis in rats. Wistar rats (200-300g, n=8 per group) of both sexes were used in the study. On day "0" experimental arthritis was induced by injecting 0.2ml of Complete Freund's adjuvant (CFA) in sub-planter region of right hind paw of animals. Pentoxifylline treatment alone and in combination with NO modulators was given (i.p.) from day 14 to 28. Various arthritic parameters were recorded and blood and joint synovial fluid was collectedfor biochemical analysis. CFA inoculation significantly increases (1) arthritic index (2) ankle diameter (3) paw volume (4) histopathology score (5) serum TNF-α, IL-6, IL-1β and synovial TNF-α levels (p<0.001) (6) serum Th1 and Th2 cytokine levels g) MDA levels in rat paw tissue homogenates (7) serum NF-κB levels. Significant decrease in serum IL-10 levels and SOD activity was observed in rats after CFA inoculation. Decrease in body weight and suppressed general quality of life of CFA inoculated rats was also observed. These CFA-induced arthritic changes were significantly reversed by pentoxifylline alone and in combination with low dose of NO modulators (p<0.05). These results are suggestive of protective effects of pentoxifylline and its potentiation in combination with low dose of NO modulators. These results may provide new pharmacological therapy for management of rheumatoid arthritis (RA).

Carcass and non-carcass components of Santa Ines lambs subjected to food restriction

The objective of this study was to evaluate the effect of sex class (castrated; non-castrated) and level of food restriction (0%, 30% and 60% of ad libitum consumption) on the quantitative composition of carcass and non-carcass components of Santa Ines lambs. A completely randomized 3x2 design (restriction level x sex class) was used to evaluate thirty lambs approximately two months of age with an average initial body weight of 13 ± 1.49 kg. When the average body weight of the animals in one of the treatment groups reached 28 kg, all animals were slaughtered. Sex class had effect on body weight at slaughter, empty body weight, hot carcass weight and cold carcass weight. The weight of the leg was greater in non-castrated animals. There was linear decreased effect according to increasing levels of restriction for the carcass cuts, except for leg and rear loin yield. There was statistic difference between sex class for the weights of kidneys, paw, and large intestine of non-castrated animals (P &lt; 0.05). There was a negative linear association between level of food restriction and mass of blood, head, leather, paw, rumen, reticulum, abomasum, small intestine, large intestine; perirenal, omental, mesenteric and heart fat. The quantitative composition of carcass and non-carcass components of Santa Ines lambs is influenced by sex class and food restriction level.

The prokineticin Bv8 sensitizes cutaneous terminals of female mice to heat

Injection of the noxious peptide Bv8 has previously been shown to induce a biphasic thermal hyperalgesia in rodents, the first peak presumably due to peripheral sensitization. This hypothesis has never been directly confirmed. We have assessed whether Bv8 can indeed sensitize peripheral nerve fibres in the mouse to heat. We used recordings from single cutaneous fibres, cutaneous calcitonin gene-related peptide (CGRP) release and immunostaining in nerves and plantar skin to evaluate the Bv8 effects on cutaneous nerves. Application of Bv8 at nanomolar concentrations (30-310nmol/L) to skin preparations significantly increased the heat-induced discharge, the heat-induced afterdischarge and reduced threshold temperature of single unmyelinated polymodal fibres. Furthermore, application of Bv8 to hind-paw skin flaps or trigeminal ganglia significantly elevated their heat-induced CGRP release. Capsaicin-induced and to a lesser extent also KCl-induced CGRP releases were also augmented after Bv8 treatment. Immunohistochemistry revealed co-localization of prokineticin 2 (Bv8 ortholog in rodents) and CGRP in both plantar skin and nerve tissues. These results confirm that Bv8 sensitizes cutaneous nerve endings to heat, partly, although not exclusively through TRPV1 activation. Our results thus support the hypothesis that the first hyperalgesic phase to follow Bv8 injection to hind paws of intact animals is due to peripheral sensitization of nociceptors. WHAT DOES THIS STUDY ADD?: Our data provide mechanistic insights into the effect Bv8 application exerts on afferent nerve endings and into the concomitant development of thermal hyperalgesia.