Leptin and interleukin-6 alter the function of mesolimbic dopamine neurons in a rodent model of prenatal inflammation

Maternal inflammation during critical stages of gestation is thought to underlie the link between prenatal infection and several neurodevelopmental psychiatric disorders in the offspring, including schizophrenia. Increased activity of mesolimbic dopamine (DA) neurons, a hallmark of psychosis, is found in offspring of rodents exposed to a prenatal inflammatory challenge but it is unclear how this effect is elicited. Using an experimental model of localized aseptic inflammation with turpentine oil (TURP) we sought to establish whether circulating interleukin-6 (IL-6) and leptin play a role in the effects of prenatal inflammation on DA neurons. Both mediators are involved in the systemic inflammatory response to immunogens, with IL-6 mediating the early phase, followed by leptin in the late phase of the response. Maternal treatment with TURP at gestational day (GD) 15 enhanced the locomotor response to the DA indirect agonist, amphetamine (AMPH), increased the expression of tyrosine hydroxylase (TH), an enzyme involved in DA synthesis, DA levels and the expression of the post-synaptic protein spinophilin in the nucleus accumbens (NAcc) in the adult offspring. All of these alterations were totally abolished by co-treating the pregnant dams with a neutralizing IL-6 antiserum. Neutralization of maternal leptin prevented the enhanced behavioral sensitization and elevation of DA and spinophilin in the NAcc but spared other changes regulated by IL-6, such as increased NAcc TH levels and acute locomotor response to AMPH. Our results provide novel evidence to suggest that prenatal surges in both maternal circulating IL-6 and leptin contribute to the appearance of sensitized DA function in the adult offspring.     

Preliminary In Vitro Antisickilng Properties of Crude Juice Extracts of Persia Americana,Citrus Sinensis,Carica Papaya and Ciklavit?

The antisickling properties of crude juice extracts of the edible portions of three commonly consumed tropical fruits namely Persia americana, Citrus sinensis, and Carica papaya were investigated in vitro alongside a new drug preparation called Ciklavit® that has antisickling activity. Four different solvent extracts of the crude juice of each fruit including aqueous, acidic, alkaline and alcoholic extracts were prepared and their antisickling effects on sickle cell trait (HbAS) and sickle cell disease (HbSS) blood samples checked alongside Ciklavit®. Blood samples were stabilized using normal saline and the antisickling effects were checked by counting the number of sickle cells remaining after incubation of the blood samples with the crude fruit extracts and Ciklavit® for twenty-four hours. The results showed that Ciklavit® produced a sustained reduction in the number of sickle cells in both HbAS and HbSS blood samples. Also the alkaline and alcoholic extracts of P. americana and C. papaya produced significant reduction in the number of sickle cells.     

Leptin actions on food intake and body temperature are mediated by IL-1

Leptin regulates energy balance through its actions in the brain on appetite and energy expenditure and also shares properties with cytokines such as IL-1. We report here that leptin, injected into rats intracerebroventricularly or peripherally, induces significant dose-dependent increases in core body temperature as well as suppression of appetite. Leptin failed to affect food intake or body temperature in obese (fa/fa) Zucker rats, which posses a defective leptin receptor. Furthermore, injection of leptin increased levels of the proinflammatory cytokine IL-1beta in the hypothalamus of normal Sprague-Dawley rats. Central injection of IL-1 receptor antagonist (IL-1ra) inhibited the suppression of food intake caused by central or peripheral injection of leptin (60 and 84%, respectively) and abolished the leptin-induced increase in body temperature in both cases. Mice lacking (gene knockout) the main IL-1 receptor (80 kDa, R1) responsible for IL-1 actions showed no reduction in food intake in response to leptin. These data indicate that leptin actions in the brain depend on IL-1, and we show further that the effect of leptin on fever, but not food intake, is abolished by a cyclooxygenase inhibitor. Thus, we propose that in addition to its role in body weight regulation, leptin may mediate neuroimmune responses via actions in the brain dependent on release of IL-1 and prostaglandins.     

Purinoceptor desensitization impairs but does not abolish the non-cholinergic motor transmission in rat isolated urinary bladder

In order to determine the effectiveness of purinoceptor desensitization in blocking the non-cholinergic motor transmission in rat detrusor, isolated preparations of urinary bladder, pretreated with atropine 3 microM + indomethacin 10 microM were contracted by alpha, beta-methylene-ATP, a stable analogue of ATP and by electrical stimulation at frequencies of 1, 2, 4, 8 and 16 Hz. The mean maximum tension generated by alpha, beta-methylene-ATP was 43% of that by electrical field stimulation. Desensitization to the contractile action of alpha, beta-methylene-ATP was achieved by administration of alpha, beta-methylene-ATP 10 microM, three times at 10 min intervals. Responses to electrical field stimulation were reduced following induction of alpha, beta-methylene-ATP desensitization but a sizeable proportion of the response persisted (51% at 1 Hz; 29% at 8 Hz). Tetrodotoxin 0.5 microM abolished the responses to electrical field stimulation, persisting after alpha, beta-methylene-ATP desensitization. It is concluded that ATP is unlikely to be the sole non-cholinergic motor transmitter in the rat detrusor.     

Sites of action of IL-1 in the development of fever and cytokine responses to tissue inflammation in the rat

1. The objective of the present study was to determine the sites of action of the cytokine, interleukin-1 (IL-1), in the febrile response to local inflammation in the rat, by comparing the importance of IL-1 in the local tissues, the circulation and the brain. This was achieved by injecting lipopolysaccharide (LPS, 100 μg kg⁻¹) into a subcutaneous air pouch and testing the effects of blocking IL-1 action with the human recombinant interleukin-1 receptor antagonist (IL-1ra) injected either into the air pouch, intraperitoneally (1 mg kg⁻¹, 0+1 h, i.p.), or intracerebroventricularly (200 μg/rat, 0+1 h, i.c.v.).
2. To investigate the effect of IL-1ra on fever and the induction of local and circulating cytokines (IL-1 and IL-6), separate experiments were performed in which groups of animals were killed 1.5, 3 or 5 h after LPS injection. Plasma and pouch fluid samples were collected for bioassay of IL-1 and IL-6.
3. Injection of LPS into the air pouch significantly increased (1.5°C) body temperature, local (air pouch) concentrations of bioactive IL-1 and IL-6, and circulating bioactive IL-6, compared to saline-treated controls.
4. Injection of IL-1ra into the pouch significantly attenuated LPS fever (P<0.001). This decrease in body temperature was associated with significant inhibition of local IL-1 bioactivity 1.5 (96%), 3 (84%) and 5 h (72%), and in bioactive IL-6 in pouch lavage fluid 1.5 (45%) and 5 h (35%), after LPS injection. The concentration of bioactive IL-6 in the plasma was significantly reduced (39%) at 3 h, when temperature was approaching the maximal value.
5. Both systemic (i.p.) and central (i.c.v.) administration of IL-1ra significantly attentuated LPS fever (P<0.05). However, it had no effect on either local concentrations of bioactive IL-1 or IL-6, or circulating IL-6, at any of the sample points.
6. These data suggest that IL-1 is released locally, at the site of tissue inflammation and that it is an important mediator of the febrile response to local inflammation. The results also indicate that IL-1 produced locally may contribute to the production of IL-6 which is released into the circulation, and that IL-1 has important actions in the generation of fever at other sites, including the brain.

     

Subacute and chronic bone infections: diagnosis using In-111, Ga-67 and Tc-99m MDP bone scintigraphy, and radiography

The usefulness of indium-111 white blood cell scintigraphy in the diagnosis of subacute or chronic bone infection was examined in 21 orthopedic patients. In-111 WBC imaging was compared with gallium-67 and technetium-99m methylene diphosphonate skeletal scintigraphy and bone radiography, all studies being performed within 1 week. In-111 WBC scintigraphy showed no definite advantage over Ga-67 scintigraphy in the identification of chronic bone infection. The two tests had the same sensitivity (80%) and similar specificity (In-111 WBC 75%, Ga-67 83%; difference not significant). Bone radiography had a sensitivity of 60% and a specificity of 67%. A negative Tc-99m MDP bone scintigram ruled out infection (sensitivity 100%), but because of low specificity (25%), final evaluation required performance of Ga-67 or In-111 WBC scintigraphy.