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Biomedical subjects

Mark R Hutchinson

Publications and source records attributed to Mark R Hutchinson.

17 recordsLinked to original sources

The effects of a single exposure to uncontrollable stress on the subsequent conditioned place preference responses to oxycodone, cocaine, and ethanol in rats.

RATIONALE: Acute stress has been shown to facilitate the rewarding effects of a number of commonly abused drugs, although the stressor typically must be administered either immediately before or during drug administration and often in the same environment. We have previously reported that a single session of an uncontrollable (inescapable tailshock, IS), but not controllable (escapable tailshock, ES), stressor can enhance the conditioned place preference (CPP) response to morphine, even when stressor and drug administration are separated temporally and spatially. However, this persistent, trans-situational enhancement did not occur to amphetamine CPP. OBJECTIVES: The following experiments were conducted to determine whether the long-term effects of IS on drug reward are specific to opioids. MATERIALS AND METHODS: Adult, male Sprague-Dawley rats were exposed to a single session of IS or remained in their home cages (HC). Twenty-four hours later, using an unbiased procedure, CPP conditioning was conducted with either oxycodone (0, 2, or 5 mg/kg, sc), cocaine (0, 1, 5, or 10 mg/kg, ip), or ethanol (0.3, 1, or 2 g/kg, ip). Another group of rats were exposed to IS, ES, or HC treatment and conditioned with oxycodone (5 mg/kg, sc) 24 h later. RESULTS: IS enhanced the subsequent CPP response to oxycodone, but not cocaine or ethanol. This enhancement was dependent on the controllability of the stressor, as ES did not affect oxycodone CPP. CONCLUSIONS: These results indicate that the long-term, trans-situational enhancing effect of uncontrollable stress on drug reward is specific to opioids.

Analgesics, Opioid↗

Norman Cousins Lecture. Glia as the "bad guys": implications for improving clinical pain control and the clinical utility of opioids.

Within the past decade, there has been increasing recognition that glia are far more than simply "housekeepers" for neurons. This review explores two recently recognized roles of glia (microglia and astrocytes) in: (a) creating and maintaining enhanced pain states such as neuropathic pain, and (b) compromising the efficacy of morphine and other opioids for pain control. While glia have little-to-no role in pain under basal conditions, pain is amplified when glia become activated, inducing the release of proinflammatory products, especially proinflammatory cytokines. How glia are triggered to become activated is a key issue, and appears to involve a number of neuron-to-glia signals including neuronal chemokines, neurotransmitters, and substances released by damaged, dying and dead neurons. In addition, glia become increasingly activated in response to repeated administration of opioids. Products of activated glia increase neuronal excitability via numerous mechanisms, including direct receptor-mediated actions, upregulation of excitatory amino acid receptor function, downregulation of GABA receptor function, and so on. These downstream effects of glial activation amplify pain, suppress acute opioid analgesia, contribute to the apparent loss of opioid analgesia upon repeated opioid administration (tolerance), and contribute to the development of opioid dependence. The potential implications of such glial regulation of pain and opioid actions are vast, suggestive that targeting glia and their proinflammatory products may provide a novel and effective therapy for controlling clinical pain syndromes and increasing the clinical utility of analgesic drugs.

Analgesia↗

Glia: novel counter-regulators of opioid analgesia.

Development of analgesic tolerance and withdrawal-induced pain enhancement present serious difficulties for the use of opioids for pain control. Although neuronal mechanisms to account for these phenomena have been sought for many decades, their bases remain unresolved. Within the past four years, a novel non-neuronal candidate has been uncovered that opposes acute opioid analgesia and contributes to development of opioid tolerance and tolerance-associated pain enhancement. This novel candidate is spinal cord glia. Glia are important contributors to the creation of enhanced pain states via the release of neuroexcitatory substances. New data suggest that glia also release neuroexcitatory substances in response to morphine, thereby opposing its effects. Controlling glial activation could therefore increase the clinical utility of analgesic drugs.

Analgesics, Opioid↗

Characterisation of the in vitro modulation of splenocyte proliferation by non-4,5-epoxymorphinan opioids.

Opioids, such as morphine, can directly alter immune function via receptors expressed on immunocompetent cells. However, several studies have questioned the classical opioid nature of this change in immune response. Therefore, it is unclear how opioids that are not from the same structural class as morphine (4,5-epoxymorphinan), will modulate the immune system, if they do not behave in a classical opioid manner. Therefore, the aim of this study was to investigate the in vitro modulatory effects of a range of non-4,5-epoxymorphinan opioids on splenocyte proliferation and compare the response characteristics to their central opioid characteristics. The modulation of concanavalin A stimulated mouse splenocyte proliferation by non-4,5-epoxymorphinan opioids resulted in three types of responses: an inhibitory concentration-response curve (e.g. methadone, inhibitory EC(50)=79.4 microM), an inverted bell shaped curve (e.g. fentanyl, inhibitory EC(50)=0.06 microM) and an induction concentration response curve (e.g. nor-binaltorphimine, induction EC(50)=0.16 microM). Non-stereoselectivity, naloxone-insensitivity, naloxone-sensitivity and non-classical opioid rank order of effect were all observed. These data support the non-classical opioid nature of direct opioid modulation of splenocyte proliferation.

Animals↗

Relationship between 4,5-epoxymorphinan structure and in vitro modulation of cell proliferation.

Morphine belongs to the class of compounds known as 4,5-epoxymorphinans, which can alter immune function directly via receptors expressed by immune cells. However, the opioid characteristics of these receptors are not clear. Therefore, the aim of this study was to investigate the in vitro immunomodulatory effects of 24 structurally related 4,5-epoxymorphinans to allow further characterisation of the receptor that mediates the immunomodulation and to ascertain if there is any structure-effect relationship. The immunomodulation of 4,5-epoxymorphinans using isolated mouse splenocytes stimulated with concanavalin A resulted in five types of responses: an inverted bell shaped curve (oxycodone, inhibitory EC(50)=1.6 nM), an inhibitory concentration response curve (buprenorphine, inhibitory EC(50)=12.6 microM), an inverted bell-shaped curve with induction (morphine, induction EC(50)=1.7 microM), an induction concentration response curve (oxymorphone, induction EC(50)=20 nM), and the lack of any response (e.g. noroxycodone). Non-stereoselectivity, naloxone-insensitivity, naloxone-sensitivity and non-classical opioid rank order of effect were all observed. A structure-effect relationship was developed and significant evidence for non-classical opioid receptor function on immune cells was concluded.

Animals↗

Biomechanics and development of the elbow in the young throwing athlete.

This article reviews basic concepts in fundamental elbow biomechanics, particularly how they relate to the skeletally immature elbow in throwing athletes. Adult phases of throwing and the forces generated in each phase are compared with the developmental phases of throwing in children. The impact of elevated forces, poor mechanics, and poor coordination are emphasized as they relate to potential areas of injury in the skeletally immature. Finally, concepts in prevention focused on biomechanics are offered.

Adolescent↗

(S)-(+)-methadone is more immunosuppressive than the potent analgesic (R)-(--)-methadone.

Methadone is a widely used synthetic opioid which is administered as a racemic mixture of (R)-(--)- and (S)-(+)-enantiomers, with only (R)-(--)-methadone possessing mu opioid receptor agonist activity. Methadone inhibits numerous immune functions in vitro at concentrations above 10 microM in a nonstereoselective and naloxone-insensitive fashion, suggesting the presence of nonclassical opioid receptors on immune cells. No in vivo data on the effects of methadone's enantiomers on immune function are available. Therefore, the stereoselectivity of methadone's analgesia (hot plate latency) in vivo and immune suppression ex vivo (splenocyte proliferation) was investigated in groups of Balb/c mice. Significant analgesia was observed in animals that received racemic methadone (P=0.0012, 52% MPE) and (R)-(--)-methadone (P=0.0002, 70% MPE) when compared to saline-treated controls, while (S)-(+)-methadone was devoid of any such effect (-4% MPE). In vivo (R)-(--)- and racemic methadone caused significant inhibition (P<0.001, greater than -70%) of basal proliferation compared to saline control. In stark contrast to analgesia, in vivo (S)-(+)-methadone caused significantly greater inhibition of basal proliferation (P<0.001, -130%) than (R)-(--)- and racemic methadone. The immune suppression caused by methadone is not purely a classical opioid response but involves nonclassical opioid receptors located at the central level, which have yet to be characterised. Moreover, the dose at which immune suppression occurred could be achieved clinically.

Analgesics, Opioid↗

In vitro opioid induced proliferation of peripheral blood immune cells correlates with in vivo cold pressor pain tolerance in humans: a biological marker of pain tolerance.

There is substantial evidence for bidirectional communication between the immune system and the central nervous system, as the cells and signalling molecules of the immune system influence many central nervous system functions, for instance nociception. Opioids, such as morphine, produce analgesia and numerous other central and peripheral effects including sedation and euphoria, while their effects on the immune system are wide-ranging. There is considerable interindividual variability in basal nociception and response to opioids, however, the physiological and biological mechanisms underlying this are unclear. Therefore, we investigated the relationship between the immune system and basal nociceptive thresholds, using the proliferative response of isolated peripheral blood mononuclear cells and cold pressor pain tolerance. Here we show that the percent increase in proliferation of peripheral immune cells from 13 healthy subjects incubated with morphine ex vivo is highly correlated with the subjects' tolerance to noxious cold stimuli (Pearson r = 0.92, P < 0.0001). These pilot data provide evidence of a novel objective biological marker of pain tolerance in humans, which also links the immune and opioid systems with basal pain tolerance.

Adult↗

CYP2D6 and CYP3A4 involvement in the primary oxidative metabolism of hydrocodone by human liver microsomes.

AIM: To determine the Michaelis-Menten kinetics of hydrocodone metabolism to its O- and N-demethylated products, hydromorphone and norhydrocodone, to determine the individual cytochrome p450 enzymes involved, and to predict the in vivo hepatic intrinsic clearance of hydrocodone via these pathways. METHODS: Liver microsomes from six CYP2D6 extensive metabolizers (EM) and one CYP2D6 poor metabolizer (PM) were used to determine the kinetics of hydromorphone and norhydrocodone formation. Chemical and antibody inhibitors were used to identify the cytochrome p450 isoforms catalyzing these pathways. Expressed recombinant cytochrome p450 enzymes were used to characterize further the metabolism of hydrocodone. RESULTS: Hydromorphone formation in liver microsomes from CYP2D6 EMs was dependent on a high affinity enzyme (Km = 26 microm) contributing 95%, and to a lesser degree a low affinity enzyme (Km = 3.4 mm). In contrast, only a low affinity enzyme (Km = 8.5 mm) formed this metabolite in the liver from the CYP2D6 PM, with significantly decreased hydromorphone formation compared with the livers from the EMs. Norhydrocodone was formed by a single low affinity enzyme (Km = 5.1 mm) in livers from both CYP2D6 EM and PM. Recombinant CYP2D6 and CYP3A4 formed only hydromorphone and only norhydrocodone, respectively. Hydromorphone formation was inhibited by quinidine (a selective inhibitor of CYP2D6 activity), and monoclonal antibodies specific to CYP2D6. Troleandomycin, ketoconazole (both CYP3A4 inhibitors) and monoclonal antibodies specific for CYP3A4 inhibited norhydrocodone formation. Extrapolation of in vitro to in vivo data resulted in a predicted total hepatic clearance of 227 ml x h-1 x kg-1 and 124 ml x h-1 x kg-1 for CYP2D6 EM and PM, respectively. CONCLUSIONS: The O-demethylation of hydrocodone is predominantly catalyzed by CYP2D6 and to a lesser extent by an unknown low affinity cytochrome p450 enzyme. Norhydrocodone formation was attributed to CYP3A4. Comparison of recalculated published clearance data for hydrocodone, with those predicted in the present work, indicate that about 40% of the clearance of hydrocodone is via non-CYP pathways. Our data also suggest that the genetic polymorphisms of CYP2D6 may influence hydrocodone metabolism and its therapeutic efficacy.

Analgesics, Opioid↗

Quantification of the O- and N-demethylated metabolites of hydrocodone and oxycodone in human liver microsomes using liquid chromatography with ultraviolet absorbance detection.

High-performance liquid chromatographic assays for the O- and N-demethylated oxidative metabolites of hydrocodone and oxycodone formed in human liver microsomes are described. A solvent-solvent extraction/re-extraction procedure followed by reversed-phase HPLC with UV detection at 210 nm allows for the quantification of hydromorphone, norhydrocodone, oxymorphone and noroxycodone. Calibration curve concentration ranges were 0.63-400 microM (0.18-114 microg/ml) and 1.25-400 microM (0.36-114 microg/ml) for hydromorphone and norhydrocodone, respectively and 0.13-20 microM (0.04-6.03 microg/ml) and 1-200 microM (0.30-60 microg/ml) for oxymorphone and noroxycodone, respectively. Assay performance was determined by intra- and inter-assay precision and inaccuracies for quality control samples and was <15% for all metabolites at each quality control concentration. These methods provide good precision, accuracy and sensitivity for use in in vitro kinetic studies investigating the oxidative metabolism of hydrocodone and oxycodone in human liver microsomes.

Chromatography, High Pressure Liquid↗

Resident's ridge: assessing the cortical thickness of the lateral wall and roof of the intercondylar notch.

PURPOSE: The purpose of this study was to better delineate the anatomy of "resident's ridge," a term coined by William Clancy Jr., M.D., to describe the raised bony landmark commonly visualized just anterior to the femoral attachment of the anterior cruciate ligament (ACL). This landmark can mislead the novice surgeon into misplacing the femoral tunnel of ACL reconstructions. TYPE OF STUDY: Cadaveric anatomic study. METHODS: Ten human distal femurs harvested from embalmed specimens were fixed, sectioned, and analyzed for the presence and descriptive characteristics of resident's ridge. A single, blinded examiner evaluated slope, cortical thickness at 4 sites, and the presence or absence of a distinct ridge relative to the attachment of the ACL. RESULTS: A defined resident's ridge was present in 9 of 10 specimens. This was directly associated with a change in slope of the intracondylar roof in the same 9 of 10 patients. The mean cortical thickness at the ACL attachment site was 1.6 mm. This was thicker than at resident's ridge (mean, 0.90 mm), the cartilage-intercondylar notch junction (mean, 0.96 mm), and a point midway between the ACL attachment and the cartilage-intercondylar notch junction (mean, 0.90 mm). CONCLUSIONS: The phenomenon of "resident's ridge" is accounted for by a distinctive change in slope of the femoral notch roof that occurs just anterior to the femoral attachment of the ACL. The density change apparent at the time of notchplasty is probably caused by the transition between normal cortical thickness just anterior to the ACL and the cortical thickness of the ACL attachment. No distinctive increased cortical thickness can be identified as "resident's ridge."

Anterior Cruciate Ligament↗

Overuse and throwing injuries in the skeletally immature athlete.

Over 25 million children participate in school-sponsored sports, and an additional 20 million participate in extracurricular organized sports. Over the past decade, increased intensity of training, more pressure for success, new opportunities for structured play, and more organized advanced leagues and traveling teams have led to a corresponding increase in overuse injuries in the skeletally immature athlete. Perhaps the classic sports model for overuse injuries of the upper extremity is baseball. Throwing sports contribute to an increased incidence of elbow and shoulder injuries that might be related to intensity of training, throwing mechanics, and poor conditioning, including core strength. Specific areas of concern regarding overuse injuries in young athletes include such diagnoses as little leaguer's shoulder, little leaguer's elbow, osteochondritis dissecans of the elbow, tennis elbow, and distal radial epiphysitis. Ultimately, overuse injuries, and particularly physeal injuries, should be suspected in any young athlete who has pain in the upper extremity. Comparative bilateral radiographs are the rule in workup.

Adolescent↗

Diacetylmorphine degradation to 6-monoacetylmorphine and morphine in cell culture: implications for in vitro studies.

Diacetylmorphine deacetylates rapidly to 6-monoacetylmorphine and then to morphine. The immunomodulatory effects of diacetylmorphine are under investigation by several groups utilising various methods including in vitro cell culture; however, diacetylmorphine stability under these conditions is unknown. The aim of this study was to quantify diacetylmorphine degradation under cell culture conditions and to determine the mechanism by which this occurs. Diacetylmorphine degradation in a mouse splenocyte mitogenesis assay was investigated. Morphine and 6-monoacetylmorphine were quantified using HPLC with UV detection. After 6 h, approximately 73% of diacetylmorphine had been hydrolysed in the presence of cells. The half-life of diacetylmorphine was 1.4 h in cell media alone and 1.2-2.2 h in incubations containing cells, while the half-life of 6-monoacetylmorphine was 3.1 h in cell media alone and 0.99-1.2 h in incubations containing cells. 6-Monoacetylmorphine and morphine formation were found to be dependent on incubation time and diacetylmorphine concentration, and were not dependent on esterase activity, mitogen concentration, presence of erythrocytes and cell media evaporation. Only morphine formation was dependent on lymphocyte concentration. 6-Monoacetylmorphine formation was independent of cells and appeared to be due to the conditions of the cell culture (pH and temperature), while morphine formation was dependent to a greater extent on cells, but independent of esterase activity. The study highlights the limitations of conclusions made in previous studies which have not recognised diacetylmorphine instability.

Animals↗

The microscopic pathoanatomy of acute anterior shoulder dislocations in a simian model.

PURPOSE: The purpose of this study was to determine the extent of capsular injury that occurs at a microscopic level in association with acute anterior shoulder dislocations. TYPE OF STUDY: The study was an anatomic, cadaveric study using a simian model. The microscopic slide review was blinded regarding the presence or absence of dislocation. METHODS: Three cadaveric monkey shoulder specimens were dissected free of muscle tissue but left undislocated as controls. Eleven cadaveric monkey shoulder specimens were dislocated with an abduction and external rotation force and subsequently evaluated grossly and histologically for the presence and direction of gross and microscopic injury. RESULTS: All dislocated specimens had an associated Bankart lesion. In addition, all dislocated specimens had histologic evidence of capsular microtearing. The tears were in no specific location or direction and were at times located perpendicular to, obliquely to, or between the longitudinal orientation of the fibers. CONCLUSIONS: Based on this model, a Bankart lesion is a common and usual occurrence with acute anterior shoulder dislocations. In these specimens, the Bankart lesion was never an isolated pathology. Microscopic capsular injury was always associated. Additionally, the direction of the capsular injury is highly variable.

Animals↗

Anatomic structures at risk during minimal-incision endoscopically assisted fascial compartment releases in the leg.

BACKGROUND: Although minimal-incision surgical techniques are recommended for treatment of chronic exertional compartment syndrome of the leg, which is an increasing problem among endurance athletes, there is little information about anatomic correlation with structures at risk. HYPOTHESIS: Fascial releases performed with endoscopic assistance are safer than the percutaneous method. STUDY DESIGN: Controlled laboratory study. METHODS: Ten endoscopically assisted and six percutaneous fascial releases were performed on 16 human cadaveric lower limbs. Formal dissection documented length of release and proximity of structures at risk. RESULTS: Endoscopically assisted fascial release led to reduced risk of superficial peroneal nerve injury compared with a blind percutaneous release through a 2- to 3-cm incision. Both techniques had unacceptable rates of saphenous vein injury (30% to 100%), and releases performed percutaneously had greater length. CONCLUSIONS: Risk of superficial peroneal nerve injury was less with single-incision endoscopically assisted fascial release. Risk of injury to the saphenous vein from either technique appeared to be unacceptable. CLINICAL RELEVANCE: Single-incision endoscopically assisted fascial release of the anterior and lateral compartments may be a useful technique with low risk of peroneal nerve injury. Clinical studies will confirm whether this technique proves safer or more effective than those currently used for chronic exertional compartment syndrome of the leg.

Cadaver↗