The neuroprotective effect of human primary astrocytes in multiple sclerosis: In vitro model.

Recent studies highlighted the role of astrocytes in neuroinflammatory diseases, particularly multiple sclerosis, interacting closely with other CNS components but also with the immune cells. However, due to the difficulty in obtaining human astrocytes, their role in these pathologies is still unclear. In this study we develop an astrocyte in vitro model to evaluate their role in multiple sclerosis after being treated with CSF isolated from both healthy and MS diagnosed patients. Gene expression and ELISA assays reveal that several pro-inflammatory markers IL-1ß, TNF-α and IL-6, were significantly downregulated in astrocytes treated with MS-CSF. In contrast, neurotrophic survival, and growth factors, and GFAP, BDNF, GDNF and VEGF, were markedly elevated upon the same treatment. In summary, this study supports the notion of the astrocyte involvement in MS. The results reveal the neuroprotective role of astrocyte in MS pathogenicity by suppressing excessive inflammation and increasing the expression of tropic factors.

Cisplatin Provokes Peripheral Nociception and Neuronal Features of Therapy-Induced Senescence and Calcium Dysregulation in Rats.

Therapy-Induced Senescence (TIS) is a form of senescence that is typically described in malignant cells in response to the exposure of cancer chemotherapy or radiation but can also be precipitated in non-malignant cells. TIS has been shown to contribute to the development of several cancer therapy-related adverse effects; however, evidence on its role in mediating chemotherapy-induced neurotoxicity, such as Chemotherapy-induced Peripheral Neuropathy (CIPN), is limited. We here show that cisplatin treatment over two cycles (cumulative dose of 23 mg/kg) provoked mechanical allodynia and thermal hyperalgesia in Sprague-Dawley rats. Isolation of dorsal root ganglia (DRG) from the cisplatin-treated rats demonstrated robust SA-ß-gal upregulation at both day 8 (after the first cycle) and day 18 (after the second cycle), decreased lmnb1 expression, increased expression of cdkn1a and cdkn2a, and of several factors of the Senescence-associated Secretory Phenotype (SASP) (Il6, Il1b, and mmp9). Moreover, single-cell calcium imaging of cultured DRGs revealed a significant increase in terms of the magnitude of KCl-evoked calcium responses in cisplatin-treated rats compared to vehicle-treated rats. No significant change was observed in terms of the magnitude of capsaicin-evoked calcium responses in cisplatin-treated rats compared to vehicle-treated rats but with decreased area under the curve of the responses in cisplatin-treated rats. Further evidence to support the contribution of TIS to therapy adverse effects is required but should encourage the use of senescence-modulating agents (senotherapeutics) as novel palliative approaches to mitigate chemotherapy-induced neurotoxicity.

Targeting therapy-induced senescence as a novel strategy to combat chemotherapy-induced peripheral neuropathy.

Chemotherapy-induced peripheral neuropathy (CIPN) is a treatment-limiting adverse effect of anticancer therapy that complicates the lifestyle of many cancer survivors. There is currently no gold-standard for the assessment or management of CIPN. Subsequently, understanding the underlying mechanisms that lead to the development of CIPN is essential for finding better pharmacological therapy. Therapy-induced senescence (TIS) is a form of senescence that is triggered in malignant and non-malignant cells in response to the exposure to chemotherapy. Recent evidence has also suggested that TIS develops in the dorsal root ganglia of rodent models of CIPN. Interestingly, several components of the senescent phenotype are commensurate with the currently established primary processes implicated in the pathogenesis of CIPN including mitochondrial dysfunction, oxidative stress, and neuroinflammation. In this article, we review the literature that supports the hypothesis that TIS could serve as a holistic mechanism leading to CIPN, and we propose the potential for investigating senotherapeutics as means to mitigate CIPN in cancer survivors.

Role of bradykinin and prostaglandin EP4 receptors in regulating TRPV1 channel sensitization in rat dorsal root ganglion neurons.

Transient receptor potential vanilloid type-1 (TRPV1) channels play key roles in chronic pain conditions and are modulated by different inflammatory mediators to elicit heat sensitisation. Bradykinin is a 9-amino acid peptide chain that promotes inflammation. The aim of present study is to investigate how bradykinin and prostaglandin receptors (EP3 and EP4 ) modulate the sensitisation of TRPV1-mediated responses. Calcium imaging studies of rat dorsal root ganglion (DRG) neurons were employed to investigate the desensitizing responses of TRPV1 ion channels by capsaicin, and the re-sensitization of TRPV1 by bradykinin, then to explore the role EP3 and EP4 receptors in mediating these bradykinin-dependent effects. Immunocytochemistry was used to study the co-expression and distribution of EP4, TRPV1, COX-1 and B2 in rat DRG neurons. Desensitization was seen upon repeated capsaicin application, we show that bradykinin-mediated sensitization of capsaicin-evoked calcium responses in rat DRG neurons occurs is dependent on COX-1 activity and utilizes a pathway that involves EP4 but not EP3 receptors. Immunocytochemical techniques revealed that EP4, TRPV1, COX-1 and B2 proteins are expressed mainly in small diameter (<1000 µm2 ) cell bodies of rat DRG neurons which are typically nociceptors. The present study provides suggestive evidence for a potential signalling pathway through which bradykinin may regulate TRPV1 ion channel function via EP4 receptors. In addition to confirming existing knowledge, the anatomical distribution and colocalization of these proteins in DRG neurons as revealed by this study offer valuable insight.

Adenosine Receptors as Potential Therapeutic Analgesic Targets.

Pain represents an international burden and a major socio-economic public health problem. New findings, detailed in this review, suggest that adenosine plays a significant role in neuropathic and inflammatory pain, by acting on its metabotropic adenosine receptors (A1AR, A2AAR, A2BAR, A3AR). Adenosine receptor ligands have a practical translational potential based on the favorable efficacy and safety profiles that emerged from clinical research on various agonists and antagonists for different pathologies. The present review collects the latest studies on selected adenosine receptor ligands in different pain models. Here, we also covered the many hypothesized pathways and the role of newly synthesized allosteric adenosine receptor modulators. This review aims to present a summary of recent research on adenosine receptors as prospective therapeutic targets for a range of pain-related disorders.

Interaction of the synthetic cannabinoid WIN55212 with tramadol on nociceptive thresholds and core body temperature in a chemotherapy-induced peripheral neuropathy pain model.

Chemotherapy-induced peripheral neuropathy (CIPN) is a significant adverse effect of many anticancer drugs. Current strategies for the management of CIPN pain are still largely unmet. The aim of this study is to investigate the antinociceptive potential of combining tramadol with the synthetic cannabinoid WIN55212, and to evaluate their associated adverse effects, separately or in combination, in a CIPN rat model, and to investigate their ability to modulate the transient receptor potential vanilloid 1 (TRPV1) receptor activity. Von Frey filaments were used to determine the paw withdrawal threshold in adult male Sprague-Dawley rats (200-250 g) following intraperitoneal (i.p) injection of cisplatin. Single cell ratiometric calcium imaging was used to investigate WIN55212/tramadol combination ability to modulate the TRPV1 receptor activity. Both tramadol and WIN55212 produced dose-dependent antinociceptive effect when administered separately. The lower dose of tramadol (1 mg/kg) significantly enhanced the antinociceptive effects of WIN55212 without interfering with core body temperature. Mechanistically, capsaicin (100 nM) produced a robust increase in [Ca 2+ ] i in dorsal root ganglia (DRG) neurons ex vivo . Capsaicin-evoked calcium responses were significantly reduced upon pre-incubation of DRG neurons with only the highest concentration of tramadol (10 µM), but not with WIN55212 at any concentration (0.1, 1 and 10 µM). However, combining sub-effective doses of WIN55212 (1 µM) and tramadol (0.1 µM) produced a significant inhibition of capsaicin-evoked calcium responses. Combining WIN55212 with tramadol shows better antinociceptive effects with no increased risk of hypothermia, and provides a potential pain management strategy for CIPN.

The role of adenosine receptor ligands on inflammatory pain: possible modulation of TRPV1 receptor function.

Chronic pain has a debilitating consequences on health and lifestyle. The currently available analgesics are often ineffective and accompanied by undesirable adverse effects. Although adenosine receptors (AR) activation can affect nociceptive, inflammatory, and neuropathic pain states, the specific regulatory functions of its subtypes (A1, A2A, A2B and A3 ARs) are not fully understood. The aim of this study was to investigate the role of different AR ligands on inflammatory pain. The von Frey filament test was used to assess the anti-nociceptive effects of adenosine ligands on Complete Freund's Adjuvant (CFA)-induced mechanical allodynia in (180-220 g) adult male Sprague Dawley rats (expressed as paw withdrawal threshold, PWT). Neither the A2AAR selective agonist CGS 21680 hydrochloride (0.1, 0.32 and 1 mg/kg) nor the A2BAR selective agonist BAY 60-6583 (0.1, 0.32 and 1 mg/kg) produced any significant reversal of the PWT. However, the A1AR selective agonist ( ±)-5'-Chloro-5'-deoxy-ENBA, the A3AR selective agonist 2-Cl-IB-MECA, the A2AAR selective antagonist ZM 241385 and the A2BAR selective antagonist PSB 603 produced a significant reversal of the PWT at the highest dose of 1 mg/kg. Co-administration of the selective antagonists of A1AR and A3AR PSB36 (1 mg/ml) and MRS-3777 (1 mg/ml); respectively, significantly reversed the anti-nociceptive effects of their corresponding agonists. Furthermore, calcium imaging studies reveled that the effective AR ligands in the behavioral assay also significantly inhibit capsaicin-evoked calcium responses in cultured rat dorsal root ganglia (DRG) neurons. In conclusion, modulating the activity of the transient receptor potential vanilloid 1 (TRPV1) receptor by ARs ligands could explain their anti-nociceptive effects observed in vivo. Therefore, the cross talk between ARs and TRPV1 receptor may represent a promising targets for the treatment of inflammatory pain conditions.

Preclinical comparison of antinociceptive effects between ibuprofen, diclofenac, naproxen, and acetaminophen on acid-stimulated body stretching and acid-depressed feeding behaviors in rats.

Pain is a major problem that burdens the health and economy of societies worldwide. Nonsteroidal anti-inflammatory drugs (NSAIDs) are over-the-counter medications that are widely indicated for mild to moderate pain conditions. Clinically, the selection of a medication among this class is mainly based according to both patient's and doctor's previous experiences. Herein, we studied differences in therapeutic efficacies among the most commonly prescribed NSAIDs and acetaminophen in inflammatory pain rat model. Body stretching and food consumption behaviors were assessed after intraperitoneal administration of lactic acid. Initially, different concentrations of lactic acid were evaluated in adult male rats in both behavioral models. Acid concentrations of 1.8 and 3.2% were selected to assess the effects of ibuprofen, diclofenac, naproxen, and acetaminophen in body stretching and feeding behaviors, respectively. In the feeding study, food restriction for 1-24 h prior to feeding studies was assessed at first, and 24 h was selected for further tests. Acetaminophen (100 mg/kg), diclofenac (10 mg/kg), ibuprofen (10-32 mg/kg), and naproxen (3.2-10 mg/kg) significantly decreased acid-stimulated body stretching. Likewise, acetaminophen (100 mg/kg), diclofenac (10 mg/kg), and ibuprofen (32 mg/kg) increased food consumption significantly after 3.2% lactic acid. There were no significant differences between different test drugs efficacies in both stretching and feeding behaviors. In conclusion, feeding behavior provides a good appraisal for pain and analgesic drugs in preclinical studies. There were comparable efficacies between all tested medications in both lactic acid-stimulated body stretching and -depressed feeding behaviors.

Comparison of Montreal cognitive assessment and Mattis dementia rating scale in the preoperative evaluation of subthalamic stimulation in Parkinson's disease.

INTRODUCTION: The preoperative evaluation of Parkinson's Disease (PD) patients for subthalamic nucleus deep brain stimulation (STN-DBS) includes the assessment of the neuropsychological status of the patient. A widely used preoperative test is the Mattis Dementia rating scale (MDRS). However, the Montreal cognitive assessment (MoCA) has also been proven to be a sensitive, time-sparing tool with high diagnostic validity in PD. We evaluate the utility of the MoCA as a preoperative screening test for PD patients undergoing bilateral STN-DBS. METHODS: In this single-centre, retrospective study, we analysed pre- and postoperative assessments of MoCA, MDRS, Movement disorder society-Unified PD Rating Scale-motor examination, PD Questionnaire-39 and levodopa equivalent daily dose. Longitudinal outcome changes were analysed using paired t-test, Pearson's correlation coefficient, linear regression and CHAID (chi-square automatic interaction detector) regression tree model. RESULTS: Clinical motor and cognitive scores of 59 patients (61.05±7.73 years, 24 females) were analysed. The MoCA, but not the MDRS, identified significant postoperative cognitive decline in PD patients undergoing STN-DBS. The preoperative MoCA score correlated with postoperative quality of life improvement, whereas the MDRS did not. PD patients with a MoCA score ≤ 23 points had a significant decline of quality of life after DBS surgery compared to patients > 23 points. CONCLUSION: This study identifies the MoCA as an alternative test within the preoperative evaluation of PD patients for the detection of neuropsychological deficits and prediction of the postoperative improvement of quality of life.

The Expression of the Senescence-Associated Biomarker Lamin B1 in Human Breast Cancer.

Senescence is a major response to cancer chemotherapy and has been linked to unfavorable therapy outcomes. Lamin B1 is a component of the nuclear lamina that plays a pivotal role in chromatin stability. Downregulation of lamin B1 represents an established biomarker for cellular senescence. However, the protein expression level of lamin B1 in malignant tissue, particularly of the breast, has not been previously described. In this work, we investigated lamin B1 protein expression in normal breast epithelium, malignant breast tissue (including adjacent non-malignant tissue) and in malignant tissue exposed to neoadjuvant chemotherapy (NAC) using immunohistochemistry (IHC) in three patient groups (n = 15, n = 87, and n = 43, respectively). Our results indicate that lamin B1 mean positive expression was 93% in normal breast epithelium and 88% in malignant breast cells, but significantly decreased (mean: 55%, p < 0.001) in malignant breast tissue after exposure to NAC, suggestive of senescence induction. No significant association between lamin B1 expression and other clinicopathological characteristics or survival of breast cancer patients was recorded. To our knowledge, this is the first report that established the baseline protein expression level of lamin B1 in normal and malignant breast tissue, and its reduction following exposure to chemotherapy. In conclusion, lamin B1 downregulation can be used reliably as a component of multiple biomarker batteries to identify therapy-induced senescence (TIS) in clinical cancer.

Estimation of sex based on metrics of the sternum in a contemporary Jordanian population: A computed tomographic study.

ABSTRACT: There is a paucity of osteometric standards for sex estimation from unknown skeletal remains in Jordan and the sexual dimorphism of the sternum has not yet been investigated. The aim of this study was to evaluate the sexual dimorphism in sternal measurements using 3D multidetector computed tomography (MDCT), and to assess their reliability for sex estimation in a Jordanian population. A total of 600 MDCT scans (300 males and 300 females) were used and a total of 8 sternal measurements were studied (manubrium length, sternal body length, combined length of manubrium and body, corpus sterni width at 1st and 3rd sternebrae, sternal index and area). Sexual dimorphism was evaluated by means of discriminant function analyses. Significant sexual dimorphism was found mainly in middle-aged and older adults. Including all subjects, multivariate, and stepwise functions gave an overall accuracy of 83.0% and 84.0%, respectively. Additionally, multivariate and stepwise analyses were conducted separately for each age group. The accuracy of sex estimation in multivariate analysis (all variables) varied from 63.2% in the young, and 83.7% in the middle adults to 84.9% for older adults. In stepwise analysis, the highest accuracy rates were provided by only sternal area in young adults (81.6%), and sternal area combined with sternal body length in middle-aged and older adults (84.2% and 85.3%, respectively). The best sex discriminator using univariate analysis (single variable) was sternal area followed by sternal body length (84.0% and 80.8% respectively). Notably, univariate analyses for most variables gave relatively higher classification accuracies in females but were poor at predicting males in the sample (sex bias ranged between -6.4% and -20%). Our data suggest that dimorphism in the human sternum increases with advancing age and separate discriminant functions are needed for each age group in Jordanians. In addition, multivariate and stepwise analyses using sternum gave higher classification accuracies with comparatively lower sex biases compared to univariate analyses.

Age of the donor affects the nature of in vitro cultured human dental pulp stem cells.

OBJECTIVES: The dental pulp stem cells (DPSCs) of six donors (three young donors aged < 19 years and three adult donors aged > 25 and < 30 years) were characterized for their stem cell marker expression and differentiation potential to study the effect of donor age on DPSCs in vitro. METHODS: DPSCs were cultured in αMEM supplemented with 20% fetal calf serum (conventional conditions) or on fibronectin-coated flasks with neurobasal medium supplemented with B27, bFGF and EGF (alternative conditions). DPSCs were characterized by immunofluorescence staining to detect the neural crest/mesenchymal stem cells markers P75 and CD146, respectively. The differentiation potential was tested by the induction of DPSCs into osteogenic, adipogenic and glial lineages and then by detecting the corresponding markers osteocalcin, lipidtox and S100ß, respectively. RESULTS: The DPSCs of the young donors expressed CD146 only under the conventional conditions and expressed P75 regardless of the culture conditions. However, the DPSCs of adult donors expressed CD146 only under the alternative conditions and expressed P75 only under conventional conditions. Only the DPSCs of the young donors differentiated into the glial linage. The DPSCs of the adult donors differentiated more efficiently into the adipogenic linage. Osteogenic differentiation was comparable. CONCLUSION: Donor age affects the expression of stem cell markers and differentiation potential of DPSCs. Moreover, the effect of culture conditions on DPSCs is age dependent.

Accessory Ossicles in the Region of the Foot and Ankle: An Epidemiologic Survey in a Jordanian Population.

Background and Objectives: The incidence of accessory bones in the region of foot and ankle is quite variable between studies and are often confused with avulsion fractures in trauma patients with musculoskeletal injuries. The aim of this study was to assess the incidence of accessory ossicles of the foot and ankle according to gender, side and coexistence, and to determine how frequently accessory ossicles were misdiagnosed as avulsion fractures. Materials and Methods: Oblique and/or lateral foot radiographs of 1000 adult patients referred from emergency departments to foot and ankle clinic were retrospectively reviewed for the presence of accessory ossicles. The Kappa statistic was used in order to assess the validity of radiographic interpretation for the presence of these bones. Results: Accessory ossicles were detected in 40.2% of the radiographs. The incidence rates for the accessory ossicles in order of frequency were: Os trigonum (15.4%), accessory navicular (13.7%), os peroneum (11.5%), os vesalianum (1.1%), os supranaviculare (0.7%), os subfibulare (0.6%), os talotibiale (0.4%), os calcaneus secundarius (0.3%), os supratalare (0.3%), os infranaviculare (0.3%), os intermetatarseum (0.2%), and os subtibiale (0.1%). Coexistence of two or three ossicles in the same foot was observed in 4.4% of the cases, mostly coexistence with os peroneum (2.9%), followed by accessory navicular (1.6%). 2.7% of accessory ossicles were initially misdiagnosed as avulsion fractures at emergency departments. Interrater agreement over identification of different accessory ossicles was found to be reasonably reliable, with a Kappa greater than 0.80 for all assessed bones. Conclusions: In clinical practice, a thorough knowledge of normal anatomical variants is essential to facilitate appropriate diagnosis and treatment and can help to prevent diagnostic errors.

A comprehensive study of the anatomical variations of the posterolateral tubercle of talus / Un estudio integral de las variaciones anatómicas del tubérculo posterolateral del talo

SUMMARY: The aim of our study was to determine the prevalence and the anatomical variations of the posterolateral tubercle of talus in relation to sex on CT imaging. A total of 1478 ankle CT scans was retrospectively reviewed for the different anatomical variants of the posterolateral tubercle of talus, the type and size of os trigonum. Normal sized lateral tubercle was found in 46.1 %, an enlarged posterolateral tubercle (Stieda's process) in 26.1 %, os trigonum in 20.5 % and almost absent tubercle in 7.3 %. A statistically higher prevalence of Stieda's process was found in males while os trigonum was higher in females (p0.05). The posterolateral tubercle of talus and its accessory ossicle, the os trigonum, could vary morphologically. The data of this study could be helpful in understanding the clinical problems that could be associated with some of these variants.

RESUMEN: El objetivo de nuestro estudio fue determinar la prevalencia y las variaciones anatómicas del tubérculo posterolateral del talo en relación con el sexo en la imagen de TC. Se revisaron retrospectivamente un total de 1478 TC de tobillo para las diferentes variantes anatómicas del tubérculo posterolateral del talo, el tipo y tamaño del os trigonum. Se encontró tubérculo lateral de tamaño normal en 46,1 %, tubérculo posterolateral (proceso de Stieda) en 26,1 %, os trigonum en 20,5% y tubérculo casi ausente en 7,3 %. Se encontró una prevalencia estadísticamente más alta del proceso de Stieda en los hombres, mientras que el os trigonum fue mayor en las mujeres (p 0,05) no se observaron diferencias significativas. El tubérculo posterolateral del talo y su osículo accesorio, el os trigonum, podrían originar problemas clínicos que podrían estar asociados con algunas de estas variantes.

Unique Variant Spectrum in a Jordanian Cohort with Inherited Retinal Dystrophies.

Whole Exome Sequencing (WES) is a powerful approach for detecting sequence variations in the human genome. The aim of this study was to investigate the genetic defects in Jordanian patients with inherited retinal dystrophies (IRDs) using WES. WES was performed on proband patients' DNA samples from 55 Jordanian families. Sanger sequencing was used for validation and segregation analysis of the detected, potential disease-causing variants (DCVs). Thirty-five putatively causative variants (6 novel and 29 known) in 21 IRD-associated genes were identified in 71% of probands (39 of the 55 families). Three families showed phenotypes different from the typically reported clinical findings associated with the causative genes. To our knowledge, this is the largest genetic analysis of IRDs in the Jordanian population to date. Our study also confirms that WES is a powerful tool for the molecular diagnosis of IRDs in large patient cohorts.

Estimation of sex from measurements of foramen magnum region in a contemporary Jordanian population: A computed tomographic study.

Several studies have shown variability in basicranial measurements between populations. Therefore, each population should have specific standards to optimize the accuracy of identification. The aim of this study was to evaluate the sexual dimorphism in foramen magnum and occipital condyles measurements using 3D multidetector computed tomography (MDCT), and to assess their utility and reliability for sex estimation in a Jordanian population by means of discriminant function analyses. A total of 500 MDCT scans (288 males and 212 females) were used and a total of 8 basicranial measurements were studied (3 measurements were derived from foramen magnum, and 5 measurements were derived from occipital condyles). Significant sexual dimorphism was found in all basicranial measurements. The most dimorphic variables were length of occipital condyle and maximum bicondylar distance. Including all variables, multivariate and stepwise functions gave an overall accuracy of 77.8% and 78.6%, respectively. However, the multivariate analyses conducted separately for measurements derived from foramen magnum and occipital condyles gave lower overall accuracy of 68.6% and 70.0%, respectively. Basicranial measurements derived from foramen magnum alone predicted males with relatively higher accuracy but were poor at predicting females in the sample (82.6% were males, 49.5% were females, sex bias 33.1%). Adding occipital condyles measurements to the multivariate analysis increased the percentage of correct sexing in females and reduced considerably the sex bias (78.8% male, 76.4% female, sex bias 1.4%). Discriminant function analysis using basicranial measurements derived from both foramen magnum and occipital condyles measurements can be utilized to estimate sex in our population.

Alkaptonuria with extensive ochronotic degeneration of the Achilles tendon and its surgical treatment: a case report and literature review.

Background: Alkaptonuria is a rare genetic metabolic disorder due to deficiency of homogentisate 1,2-dioxygenase (HGD), an enzyme catalyzing the conversion of homogentisate to 4-maleylacetoacetate in the pathway for the catabolism of phenylalanine and tyrosine. HGD deficiency results in accumulation of homogentisic acid and its pigmented polymer. Ochronosis is a bluish-black discoloration due to the deposition of the polymer in collagenous tissues. Extensive ochronotic involvement of the Achilles tendon in alkaptonuria and its surgical treatment is rarely reported. Case report: A 43-year-old man presented to our clinic in March 2019 with sudden onset of left Achilles tendon pain with no history of prior trauma. Surgical exploration revealed a complete disruption of the tendon at its attachment to the calcaneus. Black pigmentation was extensive and reached the calcaneal tuberosity, extending about 7 cm from the insertion. Discussion: Achilles reconstruction was performed using flexor hallucis longus tendon transfer. The patient experienced uncomplicated healing with satisfactory functional results. Conclusion: Orthopedic surgeons should be aware of the progressive nature of alkaptonuria. Extensive degenerative changes of the ruptured tendon should be suspected so that physicians can plan tendon repair and facilitate prompt surgical intervention.

Impairment in locomotor activity as an objective measure of pain and analgesia in a rat model of osteoarthritis.

A major problem with current animal models of pain is their lack of face validity and their vulnerability for false positive results. The present study evaluated the efficacy of the open field locomotor system, as an objective measure of pain-related behavior and analgesic efficacy in rodents. Adult, male, Sprague-Dawley rats (180-250 g) received intra-articular injections of monoiodoacetate (MIA; 1 mg) in the left knee joint. Mechanical allodynia using von Frey filaments, the weight bearing difference test and the open field locomotor activity test were performed every other day for 21 days, following the MIA injection. The antinociceptive effects of ibuprofen (50 and 100 mg/kg) on the MIA-induced nociception were also evaluated. MIA induced a significant reduction in the paw withdrawal threshold (PWT) and a significant alteration in the weight bearing difference compared with control rats. Similarly, MIA induced a significant reduction in locomotor activity, with respect to X total counts, that represent the overall locomotor activity in the horizontal plane, and X ambulatory counts, which in turn represent small scale movements, such as scratching and grooming, and lastly, Z total counts, that represent rearing or standing. Both doses of ibuprofen resulted in a significant reversal of the MIA-induced alterations in PWT and weight bearing difference. Furthermore, the two doses of ibuprofen resulted in a significant reversal of the MIA-induced reduction in locomotor activity, with respect to X ambulatory counts, but not Z total counts. Only the higher dose of ibuprofen reversed the X total counts. The open field locomotor system may successfully be used to predict the analgesic efficacy of compounds in models of joint inflammation and osteoarthritis.

Anti-nociceptive effect of Arbutus andrachne L. methanolic leaf extract mediated by CB1, TRPV1 and PPARs in mouse pain models.

Arbutus andrachne L. is a medicinal plant that grows in Jordan and has many valuable effects. In the present study, the anti-nociceptive effect of A. andrachne methanolic leaf extract was determined in mice using thermal and chemical tests. Our findings show that different doses of A. andrachne extract reduced the number of writhings significantly compared to control group. The leaf extract also reduced the time of paw licking in the early and late phases of formalin test. In all the conducted tests, 300 mg/kg body wt. was the best effective dose. A peroxisome proliferator-activated receptor alpha (PPARα) antagonist reversed the action of the plant extract in the early phase of formalin test while antagonists of the PPARα, PPAR gamma (PPARγ) and cannabinoid 1 (CB1) receptors were responsible for abolishing its effect in the late phase of this test. Also, the extract administration increased the latency time in hot plate and tail flick, an effect that was reversed by the antagonists of PPARγ, CB1 and transient receptor potential vanilloid 1 (TRPV1). No effect was noticed for α2-adrenergic receptor antagonist in the action of A. andrachne in any of the conducted tests in this study. Furthermore, analysis of the constituents in the methanolic leaf extract using liquid chromatography mass spectrometry (LCMS) showed that the extract is rich in compounds that have anti-nociceptive and/or anti-inflammatory effects such as arbutin, rutin, linalool, linoleic acid, gallic acid, lauric acid, myristic acid, hydroquinone, ß-sitosterol, ursolic acid, isoquercetin, quercetin, (+)-gallocatechin, kaempferol, α-tocopherol, myricetin 3-O-rhamnoside and catechin gallate. In conclusion, A. andrachne showed promising anti-nociceptive effects in thermal and chemical models of pain. These findings can open an avenue for natural pain relief.

Analgesic Effects and Impairment in Locomotor Activity Induced by Cannabinoid/Opioid Combinations in Rat Models of Chronic Pain.

Both opioids and cannabinoids have well-known antinociceptive effects in different animal models of chronic pain. However, unwanted side effects limit their use. The aim of this study is to evaluate the antinociceptive effect of combining synthetic cannabinoids with subtherapeutic doses of opioids, and to evaluate the effects of these drugs/combinations on rat's locomotor activity. Intra-plantar injection of Complete Freund's Adjuvant (CFA) into the left hindpaw and intraperitoneal injection of streptozotocin (STZ) were used to induce inflammatory and diabetic neuropathic pain in adult male Sprague-Dawley rats, respectively. Von Frey filaments were used to assess the antinociceptive effects of opioids (morphine and tramadol) and the synthetic cannabinoids (HU210 and WIN55212) or their combinations on CFA and STZ-induced mechanical allodynia. Open field test was used to evaluate the effect of these drugs or their combinations on locomotion. HU210 and WIN55212 did not produce significant antinociceptive effect on inflammatory pain while only the maximal dose of HU210 (1 mg/kg) was effective in neuropathic pain. Only the maximal doses of morphine (3.2 mg/kg) and tramadol (10 mg/kg) had significant anti-allodynic effects in both models. Tramadol (1 mg/kg) enhanced the antinociceptive effects of WIN55212 but not HU210 in neuropathic pain with no effect on inflammatory pain. However, in open field test, the aforementioned combination did not change tramadol-induced depression of locomotion. Tramadol and WIN55212 combination produces antinociceptive effects in neuropathic but not inflammatory pain at low doses with no additional risk of locomotor impairment, which may be useful in clinical practice.