Activation of adult rat CNS endothelial cells by opioid-induced toll-like receptor 4 (TLR4) signaling induces proinflammatory, biochemical, morphological, and behavioral sequelae.

CNS immune signaling contributes to deleterious opioid effects including hyperalgesia, tolerance, reward, and dependence/withdrawal. Such effects are mediated by opioid signaling at toll-like receptor 4 (TLR4), presumptively of glial origin. Whether CNS endothelial cells express TLR4 is controversial. If so, they would be well positioned for activation by blood-borne opioids, contributing to opioid-induced pro-inflammatory responses. These studies examined adult primary rat CNS endothelial cell responses to (-)-morphine or its mu opioid receptor (MOR)-inactive metabolite morphine-3-glucuronide (M3G), both known TLR4 agonists. We demonstrate that adult rat CNS endothelial cells express functional TLR4. M3G activated nuclear factor kappaB (NF-κB), increased tumor necrosis factor-α (TNFα) and cyclooxygenase-2 (COX2) mRNAs, and released prostaglandin E2 (PGE2) from these cells. (-)-Morphine-induced upregulation of TNFα mRNA and PGE2 release were unmasked by pre-treatment with nalmefene, a MOR antagonist without TLR4 activity (unlike CTAP, shown to have both MOR- and TLR4-activity), suggestive of an interplay between MOR and TLR4 co-activation by (-)-morphine. In support, MOR-dependent Protein Kinase A (PKA) opposed TLR4 signaling, as PKA inhibition (H-89) also unmasked (-)-morphine-induced TNFα and COX2 mRNA upregulation. Intrathecal injection of CNS endothelial cells, stimulated in vitro with M3G, produced TLR4-dependent tactile allodynia. Further, cortical suffusion with M3G in vivo induced TLR4-dependent vasodilation. Finally, endothelial cell TLR4 activation by lipopolysaccharide and/or M3G was blocked by the glial inhibitors AV1013 and propentofylline, demonstrating endothelial cells as a new target of such drugs. These data indicate that (-)-morphine and M3G can activate CNS endothelial cells via TLR4, inducing proinflammatory, biochemical, morphological, and behavioral sequelae. CNS endothelial cells may have previously unanticipated roles in opioid-induced effects, in phenomena blocked by presumptive glial inhibitors, as well as TLR4-mediated phenomena more broadly.

Spinal upregulation of glutamate transporter GLT-1 by ceftriaxone: therapeutic efficacy in a range of experimental nervous system disorders.

Glutamate neurotransmission is highly regulated, largely by glutamate transporters. In the spinal cord, the glutamate transporter GLT-1 is primarily responsible for glutamate clearance. Downregulation of GLT-1 can occur in activated astrocytes, and is associated with increased extracellular glutamate and neuroexcitation. Among other conditions, astrocyte activation occurs following repeated opioids and in models of chronic pain. If GLT-1 downregulation occurs in these states, GLT-1 could be a pharmacological target for improving opioid efficacy and controlling chronic pain. The present studies explored whether daily intrathecal treatment of rats with ceftriaxone, a beta-lactam antibiotic that upregulates GLT-1 expression, could prevent development of hyperalgesia and allodynia following repeated morphine, reverse pain arising from central or peripheral neuropathy, and reduce glial activation in these models. Ceftriaxone pre-treatment attenuated the development of hyperalgesia and allodynia in response to repeated morphine, and prevented associated astrocyte activation. In a model of multiple sclerosis (experimental autoimmune encephalomyelitis; EAE), ceftriaxone reversed tactile allodynia and halted the progression of motor weakness and paralysis. Similarly, ceftriaxone reversed tactile allodynia induced by chronic constriction nerve injury (CCI). EAE and CCI each significantly reduced the expression of membrane-bound, dimerized GLT-1 protein in lumbar spinal cord, an effect normalized by ceftriaxone. Lastly, ceftriaxone normalized CCI- and EAE-induced astrocyte activation in lumbar spinal cord. Together, these data indicate that increasing spinal GLT-1 expression attenuates opioid-induced paradoxical pain, alleviates neuropathic pain, and suppresses associated glial activation. GLT-1 therefore may be a therapeutic target that could improve available treatment options for patients with chronic pain.

Altered levels of Reelin and its isoforms in schizophrenia and mood disorders.

Reelin is a secreted extracellular matrix protein approximately 410 kDa mol. wt that is reduced in brains of patients with schizophrenia, autism, bipolar disorder and major depression. Recent reports also indicate its near absence in sera of some patients with an autosomal recessive form of lissencephaly. Moreover, Reelin is involved not only in normal cortical lamination of the brain during mammalian embryogenesis but is also implicated in cell signaling systems subserving cognition in adult brain. Here, we show that blood levels of Reelin and its isoforms are altered in three psychiatric disorders, namely, schizophrenia, bipolar disorder and major depression. The changes include significant increases in 410 kDa Reelin moiety of 49% in schizophrenic patients (p < 0.022) of four ethnic compositions (Caucasian, Vietnamese, Hmong and Laotian) and non-significant increases in depressed patients by 34% vs control blood. In contrast, 410 kDa Reelin levels decreased by 33% in bipolar blood, albeit non-signficantly, vs. controls. There was a significant increase of 90% (p < 0.0061) in 330 kDa Reelin in Caucasian schizophrenics; the depressed value was elevated by 30% vs. control but non-significantly. Again, in contrast, bipolar 330 kDa value decreased by 31% vs control (p < 0.0480). Finally, all 180 kDa Reelin values varied minimally in schizophrenics vs controls. In contrast, the 180 kDa Reelin values dropped significantly by 49% (p < 0.0117) and 29% (p < 0.0424) in bipolar and depressed patients, respectively, compared with controls. The alterations in blood Reelin values appear to be specific since levels of two other blood proteins, ceruloplasmin and albumin did not vary significantly between all psychiatric subjects and controls. These findings suggest that blood Reelin levels and its isoforms may be used as potential peripheral markers to diagnose presence of several psychiatric disorders and may also serve as targets for future therapeutic interventions.

Ultrasound and endocrine evaluation of the ovarian cycle and early pregnancy in the Sumatran rhinoceros, Dicerorhinus sumatrensis.

Longitudinal ultrasound and endocrine evaluations were conducted in two adult female Sumatran rhinoceroses (Dicerorhinus sumatrensis) over a period of 12-22 months to learn more about their reproductive physiology. Rectal ultrasonography was conducted to monitor ovarian activity. Blood samples were collected and analysed for progesterone and LH, and faecal samples were analysed for progestin metabolites. One female showed cyclic ovarian activity during the study period, whereas the other female showed no evidence of ovarian activity. The cyclic Sumatran rhinoceros appeared to be an induced ovulator, the first of its kind reported within the Perrisodactyla. Ultrasound examinations of the ovaries revealed the formation of anovulatory haemorrhagic follicles when the animal was not mated. These follicles appeared to undergo varied degrees of luteinization that resulted in irregular faecal progestin profiles. When allowed to mate, the female showed a 21 day reproductive cycle that was reflected in both faecal progestin and serum progesterone profiles. The concentration of serum LH was baseline before mating, increased approximately 30-fold within 1-2 h of intromission and returned to baseline within 22 h. Ovulation occurred within 46 h of copulation. The female conceived three times during the study. Pregnancy was detected using ultrasonography 14-16 days after mating, and the concentration of both serum progesterone and faecal progestins remained high. Early embryogenesis appeared to be similar to that in horses. However, each pregnancy terminated unexpectedly within the first 3 months of gestation. This study demonstrates the important role that basic research and reproductive technology can play in developing a natural breeding programme for an endangered animal in captivity.

[Internal ureter splinting, a promising therapy for iatrogenic ureteral injuries and ureteral stenoses in the small pelvis]. / Die interne Harnleiterschienung, eine erfolgversprechende Therapie bei iatrogenen Ureterverletzungen und Ureterstenosen im kleinen Becken.

The technique of endoscopically placed PVC indwelling splints into the lesioned or obstructed ureter is described. With the method of consecutive expanding of the ureter and following closed splinting, good results in 8 of 8 patients were achieved. The splints have been in place for 2,5 to 42 months. Indications, technique, complications and results are herein described.

Self-destructive behavior on an inpatient ward.

Suicidal and self-destructive behavior on a psychiatric inpatient service are said to be related to the degree of staff demoralization and dissension. Staff factors that may permit or encourage self-destructive acts include poor communications, staff disagreements, scapegoating of patients, poor staff judgment, staff self-preoccupation, and reversal of staff-patient roles. However, it is also possible that a major contributory factor is not individual patient or staff psychopathology, but rather the destruction of the underlying traditions and values of the ward which occurs at times of major change. The thesis is presented that the ritualization of ward values, when operative, provides a coherent world-view which renders the therapeutic activities, and life itself, meaningful, and that such ritualization can provide a framework of stability in times of critical staff turnover. Without such tradition and ritualization, however, therapeutic activities become hollow and meaningless, and fail to provide self-destructive patients a reason to view their life more positively.