Chemical activation of thin-fiber phrenic afferents: respiratory responses

In supine chloralose-anesthetized and mechanically ventilated dogs, we assessed the effects of group III and IV thin-fiber phrenic afferents on cardiorespiratory control by injecting capsaicin into the phrenic artery of an in situ isolated and innervated left diaphragm. Inspiratory motor drive was assessed by measuring the electromyogram of left and right diaphragm, left parasternal, and mylohyoid muscles in five protocols. 1) Three boluses (2 ml) of capsaicin (1, 10, and 50 micrograms/ml) were injected 30 min apart. Only the 50-micrograms/ml injection elicited a significant increase in arterial pressure, heart rate, and inspiratory motor drive. 2) Repeated doses of capsaicin were tested. The pressor and hyperpneic responses were weakened. 3) High doses of capsaicin (100 and 500 micrograms/ml) were given. Hyperpneic and pressor responses were similar to those elicited by the 50-micrograms/ml dose. 4) When the left phrenic nerve was sectioned, the pressor and hyperpneic responses to the 50-micrograms/ml injection were abolished. 5) Capsaicin (50 micrograms/ml) was infused into the arterial supply of the in situ vascularly isolated and innervated gastrocnemius. Arterial pressure, breathing frequency, and inspiratory motor drive to all inspiratory muscles increased significantly and to a greater degree than in the diaphragm. In conclusion, diaphragmatic thin-fiber afferents have an excitatory effect on the inspiratory motor drive and arterial pressure that is similar to that seen in limb muscles.     

Titration of botulinum antitoxin of low levels by the score method

Botulinum antitoxin is commonly titrated by injecting a mixture of toxin and antitoxin into mice and by utilizing deaths as a marker to measure the amount of unneutralized toxin. We attempted to titrate antitoxin by converting the severity of symptoms (notably palsy) and time-to-death in days into scores. In neutralization tests with toxin levels at 5.9 LD50 and 23.5 LD50, a linear relationship was obtained for antitoxin dose in a range between 0.03 to 0.003 IU/ml. Statistical analysis showed that homogeneity of variance or slope was not denied for the scores obtained on any day from the first to the fourth days after injection, demonstrating that this method can titrate accurately antitoxin of such a low level as 0.003 IU/ml within 4 days after injection.     

Parathyroid hormone, cAMP, electrolytes and uric acid after high dose CaCl2 in patients with idiopathic stone formation

High dose levels of calcium chloride significantly decrease serum level of parathyroid hormone (PTH) in both patients with history of idiopathic stone formation and healthy controls. Suppressibility of parathyroid glands is obtained by injection of 10 ml 10% calcium chloride without any restriction in calcium diet. Serum levels of calcium measured 14 mg/dl 10 min. after high dose calcium chloride injection and distinctly affect cAMP in urine in patients when compared to healthy controls. The elimination of electrolytes and uric acid is elevated 40 min. after calcium application in both patients and controls and returns to normal values 120 min. later. No stone formation has been observed in an eighteen mouth follow up of all subjects studied.     

Production of antibody to homologous -fetoprotein in rabbits, rats and horses by immunization with human -fetoprotein

The production of antibody to homologous alpha fetoprotein (AFP) in rabbits, rats, and horses by immunication with human AFP is reported. The antigens were administered subcutaneously 5 times at intervals of 7-10 days. Rabbits and dogs received 1 mg of human AFP/ml of the homologous pooled newborn serum with each injection while the rats received 1/2 of the dose. The horses received 5 mg/ml/injection. 2 weeks after the last injection, antisera were collected and immunologic assays were performed by the Ouchterlony method and the reversed version of the Mancini method. High titered antibodies were produced in all animals except in the dog. The rabbit, rat, and horse antibodies crossreacted with their own homologous AFP. Attempts to produce antibody with homologous AFP in rabbits, rats, and dogs were unsuccessful.     

Saponin Adjuvants. IV. Evaluation of The Adjuvant Quil a in the Vaccination of Cattle Against Foot-And-Mouth Disease

The saponin adjuvant Quil A has been investigated in the vaccination of cattle against foot-and-mouth disease. Using a Frenkel type vaccine a dose-response relationship has been established between Quil A and neutralizing antibody titres. Ten ml of vaccine was combined with 0, 50, 200, 800, and 3200 µg of Quil A. The combinations were each injected into 4 animals. The local reaction on the site of injection produced by injection of the vaccine alone and in combination with different doses of Quil A has been estimated. On this basis a therapeutical dose at 1 mg of Quil A has been estimated to combine maximum adjuvant effect with a minimum of adverse reactions. This dose has been tested in the vaccination of cattle with FMD vaccines derived from BHK suspension cell virus of type O and A respectively. The vaccines were tested in 10 ml and 5 ml doses with or without Quil A, and each in 4 animals. It is concluded that Quil A is a valuable adjuvant for use in the induction of neutralizing antibodies against foot-and-mouth disease in cattle.     

Effect of ethanol and probenecid on the excretory function of the cerebral choroid plexus in rats

Probenecid at a dose 100 and 200 mg/kg, i.v. has been found to decrease in a dose-dependent manner the level of radioactivity of cerebrospinal fluid (CSF) measured at 1, 15, 30 and 60 min. after the intravenous injection of 14C-tyrosine, 14C-tryptophan and 14C-DOPA. Ethanol at a dose 2 and 4 g/kg, i. p. has not changed the level of radioactivity of the CSF. It is suggested that mentioned in the literature an increased accumulation of the labeled tyrosine, tryptophan and DOPA in the brain structures after their intravenous injection is not related to the inhibitory effect of ethanol on the excretory function of the choroid plexus of the brain. On the other hand, it is concluded that probenecid is able to inhibit the excretion from the brain of some acid compounds including tyrosine, tryptophan and DOPA.     

Characteristics of the disorder in the composition of the internal medium and kidney function of vertebrate animals after cisplatin administration

In rats, 5 days after injection of cisplatin (5 mg/kg of body weight) the urea content of the blood serum significantly increased; in pigeons, elevation of the uric acid was observed. No significant changes were found in the blood serum of the frog Rana temporaria even after injection of 40 mg/kg of cisplatin. In the black sculpin Myoxocephalus scorpius, injections of 50 mg/kg of cisplatin resulted in hypermagnesemia. Swelling of the kidney and changes in its electrolyte content were observed in rats, pigeons, and frogs, the wet weight of the kidney increased in rats and pigeons. In all the animals, accumulation of platinum in the kidney was observed, its content being dependent on the injected dose. The data obtained reveal lower sensitivity of the kidney in cold blood vertebrates to toxic effect of cisplatin and demonstrate that the pattern of disturbances in composition of the blood serum is related to the extent of the excretory function of the kidney within the species.     

Inhibiting effect of cholera vibrio neuraminidase in Rauscher mouse leukemia]

A study was made of the inhibitory effect of neuraminidase of cholera vibrio in mouse Rauscher leukemia. It was shown that the processing of cells of the spleen of mice suffering from leukemia with neuraminidase (in a dose of 50 units/ml and more) significantly inhibited the capacity of these cells to induce leukemia in their injection to BALB/c mice. In the mentioned concentration neuraminidase injected repeatedly parenterally produced no therapeutic effect in Rauscher leukemia.     

Kinetics of phenazepam-14C excretion from the body of white rats in the single and prolonged administration of the preparation]

During 5 days after intraperitoneal injection of 14C-phenazepam into albino rats, about 77% of the total radioactivity was excreted with urine and feces in both intact animals and in those premedicated with phenazepam for 15 days. The excretory processes are described by the first order equations. The rates of phenazepam total excretion are identical in single and repeated injections. At the same time, phenazepam injected into the animals at a single dose is predominantly excreted with urine, while in multiple administration it is excreted with feces. Excretion of phenazepam with urine acquires the biexponential features, provided it is injected in multiple doses.     

Effects of varying doses of methionine sulfoximine on liver glutamine synthetase activity and time courses of blood and urinary nitrogenous compounds in the chicken (Gallus domesticus)

1. The activity of liver glutamine synthetase was inhibited to 7-12% of the control activity by an intracardiac injection with methionine sulfoximine (MSM) at dosages of 20, 50, 75 and 100 mg/kg body wt. 2. Plasma glutamine concentrations in all the MSM treatments decreased sharply, then reached steady-state levels within 0.5-2.5 hr, which were almost proportional to a dose of MSM. 3. Blood ammonia concentration sharply increased to a steady-state level attained at 4.5 hr, which was proportional to a dose of MSM. The excretion rate of urinary ammonia augmented linearly up to the dose dependent maximum rates within 2-5 hr. 4. Plasma uric acid concentration dropped linearly by about 6.4 mg/100 ml at doses of 50, 75 and 100 mg MSM and by 3.7 mg/100 ml at a dose of 20 mg MSM within 2.5 hr, then recovered a little. 5. The decreases in excretion rates of urinary uric acid for the first 4 hr were almost the same at doses of 50 mg and larger, being twice as large as that of the control chicken. 6. Any doses of MSM affected neither the time course of excretion rate of total urinary nitrogen nor its total amounts for 7 hr after MSM treatment.     

Species variations in the threshold molecular-weight factor for the biliary excretion of organic anions

1. The excretion in the bile and urine after intravenous injection of 16 organic anions having molecular weights between 355 and 752 was studied in female rats, guinea pigs and rabbits. 2. These compounds were mostly excreted unchanged, except for three of them, which were metabolized to a slight extent (<7% of dose). 3. The rat excreted all the compounds extensively (22-90% of dose) in the bile. 4. In guinea pigs four of the compounds with mol.wt. 355-403 were excreted in the bile to the extent of 7-16% of the dose, four with mol.wt. 407-465 to the extent of 25-44% and eight compounds with mol.wt. 479-752 to the extent of 44-100%. 5. In rabbits four compounds with mol.wt. 355-465 were excreted in the bile to the extent of 1-8% of the dose, two compounds with mol.wt. 479 and 495 to the extent of 24 and 22%, and six compounds with mol.wt. 505-752 to the extent of 31-94%. 6. These results, together with those of other investigations from this laboratory, are discussed and the conclusion is reached that there is a threshold molecular weight for appreciable biliary excretion (i.e. more than 10% of dose) of anions, which varies with species: about 325+/-50 for the rat, 400+/-50 for the guinea pig and 475+/-50 for the rabbit. 7. Anions with molecular weights greater than about 500 are extensively excreted in the bile of all three species. 8. That proportion of the dose of these compounds which is not excreted in the bile is excreted in the urine, and in the three species, bile and urine are complementary excretory pathways, urinary excretion being greatest for the compounds of lowest molecular weight and tending to decrease with increasing molecular weight. 9. Some implications of this interspecies variation in the molecular-weight requirement for extensive biliary excretion are discussed.     

Central effects of leptin on cardiovascular and neurohormonal responses in conscious rabbits

We determined the cardiovascular and neurohormonal responses to intracerebroventricular injection of leptin in conscious rabbits. Intracerebroventricular injection of leptin elicited dose-related increases in mean arterial pressure and renal sympathetic nerve activity while producing no consistent, significant increases in heart rate. Peak values of mean arterial pressure and renal sympathetic nerve activity induced by intracerebroventricular injection of 50 microgram of leptin (+17.3 +/- 1.2 mmHg and +47.9 +/- 12.0%) were obtained at 10 and 20 min after injection, respectively. Plasma catecholamine concentrations significantly increased at 60 min after intracerebroventricular injection of leptin (control vs. 60 min; epinephrine: 33 +/- 12 vs. 97 +/- 27 pg/ml, P < 0.05; norepinephrine: 298 +/- 39 vs. 503 +/- 86 pg/ml, P < 0.05). Intracerebroventricular injection of leptin also caused significant increases in plasma vasopressin and glucose levels. However, pretreatment with intravenous injection of pentolinium (5 mg/kg), a ganglion blocking agent, abolished these cardiovascular and neurohormonal responses. On the other hand, intravenous injection of the same dose of leptin (50 microgram) as used in the intracerebroventricular experiment failed to cause any cardiovascular and renal sympathetic nerve responses. These results suggest that intracerebroventricular leptin acts in the central nervous system and activates sympathoadrenal outflow, resulting in increases in arterial pressure and plasma glucose levels in conscious rabbits.     

Comparison of the pharmacokinetics of subcutaneous and intravenous administration of a phosphorothioate oligodeoxynucleotide in cynomolgus monkeys

The pharmacokinetics of subcutaneous (s.c.) administration of a phosphorothioate oligodeoxynucleotide (PS-ODN) was evaluated in cynomolgus monkeys. In a single dose study, monkeys were injected s.c. or intravenously (i.v.) with doses of either 1 or 5 mg/kg ISIS 2302. The bioavailability of s.c. injection ranged from 26% to 55% and appeared to be dependent on the concentration of the dosing solution rather than the dose. The bioavailability of a subcutaneously administered 5 mg/kg dose of ISIS 2302 was 55% using a 50 mg/ml dosing solution and only 26% using a 10 mg/ml dosing solution. Slow absorption from the s.c. injection site significantly blunted the maximal concentration (Cmax) compared with i.v. administration. The time to peak plasma concentration (Tmax) increased slightly with increasing dose, from 0.5 to 1 hour for the 1 mg/kg dose to 1 to 2.5 hours for the 5 mg/kg dose. Plasma half-lives were prolonged after s.c. administration, indicating more dependence on absorption than elimination. The half-lives after s.c. administration averaged 3 hours, whereas after i.v. administration, the half-lives were <1 hour. Metabolism of the ISIS 2302 after s.c. injection was consistent with exonucleolytic cleavage, as previously observed after i.v. administration. In summary, s.c. administration of PS-ODN resulted in prolonged and extensive absorption of the ODN.     

Susceptibility of zebra fish Danio rerio to infection by Flavobacterium columnare and F. johnsoniae

Flavobacterium columnare is a serious pathogen in a wide range of fish species. F. johnsoniae is an opportunistic pathogen of certain fish. Both are gliding bacteria. These species were tested for their ability to infect the zebra fish Danio rerio. Both injection and bath infection methods were tested. The results indicate that F. johnsoniae is not an effective pathogen in D. rerio, but that F. columnare is an effective pathogen. F. johnsoniae did not cause increased death rates following bath infection, but did cause increased death rates following injection, with an LD50 (mean lethal dose) of approximately 3 x 10(10) cfu (colony-forming units). Non-motile mutants of F. johnsoniae produced a similar LD50. F. columnare caused increased death rates following both injection and bath infections. There was considerable strain variation in LD50, with the most lethal strain tested producing an LD50 of 3.2 x 10(6) cfu injected and 1.1 x 10(6) cfu ml(-1) in bath experiments, including skin damage. The LD50 of F. columnare in zebra fish without skin damage was > 1 x 10(8), indicating an important effect of skin damage.     

Anticonvulsant Activity and Toxicity of Essential Oil and Methanolic Extract of Zhumeria majdae Rech, a Unique Iranian Plant in Mice

Drug-resistance and adverse effects of current drugs are the most obstacles in the treatment of epilepsy. In a plan for finding new natural anticonvulsant agents, we studied the anticonvulsant effects of essential oil (ZMEO) and methanolic extract (ZMME) of Zhumeria majdae in pentylene tetrazol (PTZ) and maximal electro-shock (MES) models in mice. Mice received different doses of ZMEO and ZMME, 30?min before induction of chemical and electrical convulsions. Neurotoxicity (movement toxicity and sedation) was evaluated using rota-rod test. The mortality was determined after 24?h following injection of different doses of the ZMEO and ZMME. The obtained results show that ZMEO dose-dependently protected mice from tonic convulsions induced by PTZ and MES with effective doses (ED₅₀) of 0.26 (0.13?C0.39) and 0.27 (0.17?C0.37)?ml/kg respectively. Toxic doses (TD₅₀) in rota-rod test for ZMEO was 0.55 (0.42?C0.70)?ml/kg. ZMME at dose of 2?g/kg decreased tonic convulsions as much as 40?%. For ZMEO, TD₅₀ of 0.55 (0.45?C0.69)?ml/kg was obtained. ZMME significantly decreased the walking time in rota-rod test at dose of 2?g/kg. Lethal dose (LD₅₀) of ZMEO was determined as 2.35 (1.98?C2.65)?ml/kg. ZMME showed about 34?% death of the animals at dose 5?g/kg. The essential oil of Z. majdae could be a good candidate for further anticonvulsive studies.     

Direct hypophysial inhibition of luteinizing hormone release by dopamine in the rabbit.

The role and site of action of dopamine in regulating gonadotropin secretion remain unclear. In the present study, we investigated the possibility that dopamine regulates LH secretion by acting directly on the pituitary gland of the rabbit. The effect of dopamine infusion on LHRH-evoked LH release was determined in intact and pituitary stalk sectioned animals. Intravenous injection of LHRH (1 μg) in intact and acutely stalk sectioned rabbits increased peripheral plasma LH levels from a resting value of 0.2 ng/ml to maximal values of 12–14 ng/ml within 10–20 min. When dopamine was infused iv at a dose of 6.6 μg/min/kg BW from 30 min before LHRH injection until 120 min after, the rise in plasma LH levels in intact and stalk sectioned animals was decreased by 50–70%. However, dopamine infused at a lower dose (0.66 μg/min/kg BW) or at a higher dose (66.0 μg/min/kg BW), did not affect the LHRH-induced secretion of LH. These results suggest that dopamine can exert a direct hypophysial inhibitory effect on release of LH. They also demonstrate that dopamine is inhibitory only within a restricted dose-range, extending to the pituitary an established property of dopamine in the cardiovascular system.     

Use of the cytokinesis-block micronucleus method in mouse splenocytes

Mouse splenocytes have been used in the cytokinesis-block method for the evaluation of micronuclei induced by mutagenic agents in vitro as well as in vivo. Stimulation with concanavalin A for 48 h followed by 16-24-h treatment with 5 micrograms/ml cytochalasin B was found to be an optimum condition to obtain micronuclei in the binucleated splenocytes after the cells were cultured in vitro. Under the above conditions splenocytes from mice pretreated with a single i.p. injection of cyclophosphamide gave a significant increase in micronucleus production. This increase was dependent on the dose of cyclophosphamide (r = 0.99). A dose of 50 mg/kg resulted in 22% of the binucleated cells producing micronuclei, more than 20 times the level in the untreated control. The increase was also dependent on the time of cyclophosphamide injection before removal of the spleen. A duration of 4-8 h after cyclophosphamide injection gave rather sharp optimum values for the production of micronuclei. When splenocytes from non-treated mice were treated with mitomycin C together with cytochalasin B in the above in vitro condition, there was a significant increase in micronucleus production in the binucleated cells. It was also dependent on the dose of mitomycin C (r = 0.975) and a dose of 0.5 micrograms/ml resulted in a more than 20-fold increase over the untreated control. Thus, the use of mouse splenocytes in the cytokinesis-block micronucleus assay was shown to be sensitive enough for testing mutagenic agents in vivo as well as in vitro.     

Resistance of cats to the diabetogenic effect of alloxan

Experimental diabetes mellitus can be induced chemically in many species of animals with streptozotocin or alloxan. However, the cat is known to be resistant to the diabetogenic effect of streptozotocin. The purpose of this study was to find the optimal dose and rate of injection of alloxan to consistently produce hyperglycemia (blood sugar levels greater than 300 mg/dl) in cats. Alloxan was administered to 22 cats at various concentrations (50, 100 and 150 mg/kg) and different rates of injection (0.5, 1.0 and 1.5 ml/min). No hyperglycemic effect was observed at any of the concentrations or different rates of injection. Cats receiving high concentrations and/or high rates of injection of alloxan died due to kidney damage. The results of this study suggest that the cat is resistant to the diabetogenic effect of alloxan, but is susceptible to its toxic side effects.     

Short-term immobilization of mice by methohexitone

The feasibility of short-term immobilization for <5 min of female mice by methohexitone sodium was studied. In C3H/Neu mice, methohexitone at a dose <40 mg/kg did not result in chemical restraint, doses >50 mg/kg caused considerable lethality. A dose of 44 mg/kg, applied intraperitoneally at a concentration of 6.46 mg/ml, is suitable for immobilization without complications. This concentration was chosen in order to achieve an injection volume of about 0.15 ml for a mouse with an average body weight of 22 g, corresponding to about 1 mg/mouse. Complete immobilization, defined as absence of the righting reflex, was observed within 3.3 +/- 0.8 min (mean +/- SD, n = 10) after the injection and lasted for 1.5 +/- 0.7 min. Recovery of the animals was complete after a total period of 10 to 15 min post-injection. No gross pathomorphological changes were induced when intraperitoneal injections of methohexitone were repeated 10 times within 10 days. In the present study, complete immobilization of the mice was safely achieved after 87 out of 90 injections. In conclusion, immobilization by intraperitoneal injection of methohexitone is a feasible and reliable method in the experimental studies of female mice.     

Free radical scavenging and metal chelation by Tinospora cordifolia,a possible role in radioprotection

Aqueous extract of T. cordifolia inhibited Fenton (FeSO4) reaction and radiation mediated 2-deoxyribose degradation in a dose dependent fashion with an IC50 value of 700 microg/ml for both Fenton and radiation mediated 2-DR degradation. Similarly, it showed a moderate but dose dependent inhibition of chemically generated superoxide anion at 500 microg/ml concentration and above with an IC50 value of 2000 microg/ml. Aqueous extract inhibited the formation of Fe2+-bipiridyl complex and formation of comet tail by chelating Fe2+ ions in a dose dependent manner with an IC50 value of 150 microg/ml for Fe2+-bipirydyl formation and maximally 200 microg/ml for comet tail formation, respectively. The extract inhibited ferrous sulphate mediated lipid peroxidation in a dose-dependent manner with an IC50 value of 1300 microg/ml and maximally (70%) at 2000 microg/ml. The results reveal that the direct and indirect antioxidant actions of T. cordifolia probably act in corroboration to manifest the overall radioprotective effects.