Pre-dosing Values Research Articles

Effects of intravenous infusion, vest wearing and repeated intravenous blood collection on clinical pathology parameters in cynomolgus monkeys (Macaca fascicularis).

Nonhuman primates are used extensively in a variety of nonclinical safety evaluation studies of new drugs. In those studies, intravenous infusion is a common treatment method, a noninvasive telemetry system is usually used for cardiovascular safety and pharmacology evaluation, and blood samples are repeatedly collected for various analysis. Intravenous infusion, vest wearing, and repeated intravenous blood collection can caused a stress response in cynomolgus monkeys, which may lead to changes in clinical pathology parameters in them. Here, we aimed to test the effects of the above operations on clinical pathology parameters in cynomolgus monkeys. Twenty monkeys (10 male/10 female) were administered 0.9% sodium chloride injections via intravenous infusions on Days 1 and 10. Each animal wore a vest before each dosing, and the vest was removed at 24 h after each dosing. Blood samples were collected before the first dose and at 2 min, 24 h, 48 h, 72 h, and 168 h after each dosing. As compared to values before the first dose (D-1) increases in reticulocytes (percentage and absolute count) and decreases in erythrocytes (red blood cells, hemoglobin, and hematocrit) were noted after dosing. The decrease in erythrocytes and increase in reticulocytes were considered to the related to the repeated intravenous blood collection. Increases in leukocytes (white blood cells and absolute count and percentage of neutrophils) and platelets (mean platelet volume and platelet distribution width) were noted at 2 min or 24 h post dose. Increases in aspartate aminotransferase, direct bilirubin, creatine kinase, C-reactive protein, and human cardiac troponin I and decreases in inorganic phosphate were noted at 2 min to 72 h post dose.

The Anti-Factor XII Antibody AB052 Is Antithrombotic without Hemostatic Impairment in a Primate Model of Extracorporeal Membrane Oxygenation

In standard clinical practice, accumulation of contact initiated thrombi within extracorporeal membrane oxygenation (ECMO) systems is generally controlled by systemic anticoagulation using unfractionated heparin (UFH). However, the efficacy of UFH is limited by dose-restricting hemorrhagic safety concerns, and thus oxygenator replacement and thrombo-embolic complications are frequent during sustained oxygenation. Since coagulation factor XII (FXII) is essential in contact initiated blood coagulation, but has no known contribution to hemostasis, we sought to further explore the roles of FXII in hemostasis and thrombosis in a baboon ECMO model utilizing the anti-FXII monoclonal antibody AB052. In vitro, using human plasma, AB052 is 30-times more potent at inhibiting FXII activity than corn trypsin inhibitor, as assessed by a doubling of the activated partial thromboplastin time (aPTT). In baboon ECMO experiments, a Terumo CAPIOX RX5 baby oxygenator was inserted into a chronic arteriovenous shunt with blood flow regulated to 100 ml/min. An intravenous bolus of AB052 (5 mg/kg), UFH (20 U/kg), or the combination was given between 15 and 30 min prior to each experiment. ACT times immediately prior to experiments were 382±9 sec (AB052), 271±11 sec (UFH), 473±26 sec (combination), compared to pre-dosing values (207±8 sec). Control experiments without any anticoagulation were also performed, and resulted in high platelet accumulation rates of 1.72x109 min-1 and total deposition of 72±9x109 platelets (n=2), with one device completely occluding during the 60 min study period. Heparinization resulted in a total deposition of 52±11x109 platelets and an accumulation rate of 1.20x109 platelets min-1 over 60 min (n=4). AB052 alone (n=3) or in combination with UFH (n=3) significantly reduced total platelet deposition (28±3x109 and 24±5x109, respectively) as well as the platelet accumulation rates (0.60x109 min-1 and 0.56x109 min-1, respectively) compared to heparinization alone. Fibrin content within the ECMO device was also significantly lower in the AB052 treated groups when compared with either UFH alone or non-anticoagulated controls (45±22 mg (UFH), 15±3 mg (AB052), 15±9 mg (AB052 plus UFH), and 60±11 mg (non-anticoagulated). Furthermore, measurement of systemic thrombin anti-thrombin (TAT) complexes at the end of the experiments showed a marked 63% decrease in TAT levels in the AB052 treatment group and a 67% decrease in the combination treatment group compared to heparinization alone. Importantly, template bleeding time tests (Surgicutt) that were used to assess hemostasis impairment during the experiments showed no prolongation associated with AB052 administration. In summary, our data suggest that selective inhibition of FXII activation may be effective and safe when used alone or in combination with UFH for pharmacological thromboprophylaxis during contact initiated thrombosis. DisclosuresWallisch:Aronora, Inc: Employment, Equity Ownership. Tucker:Aronora, Inc: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees. Lorentz:Aronora, Inc: Employment, Equity Ownership. Carris:Aronora, Inc: Employment, Equity Ownership. Gruber:Aronora, Inc: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees.

P1402USE OF ROMOSOZUMAB FOR OSTEOPOROSIS IN HEMODIALYSIS PATIENTS

Abstract Background and Aims Recently, the importance of osteoporosis and chronic kidney disease-mineral bone disorder (CKD-MBD) has been highlighted as a cause of increased fracture risk in dialysis patients. However, many osteoporosis drugs require careful administration or are contraindicated in dialysis patients, and evidence for their use in dialysis patients is scarce. Here, we administered the new human anti-sclerostin antibody preparation romosozumab to hemodialysis patients with osteoporosis, and examined changes in bone density, bone metabolism markers and other values. Method Romosozumab was administered once per month to 12 hemodialysis patients (mean age: 72.6 ± 9.4 years old; 4 males, 8 females) who were diagnosed with osteoporosis while visiting our hospital and who had no history of cardiovascular complications in the preceding year. Bone density YAM value was measured every 3 months by dual energy X-ray absorptiometry (DXA). TRACP-5b, BAP, P1NP and whole-PTH values were measured every month as bone metabolic markers. Values at the start of observation were regarded as pre-dosing values and the mean values during the 6 months after administration were measured as post-treatment values. The results were then compared. Results At 6 months after the start of treatment, no change was seen in bone density of the proximal femur compared to the pre-dosing value. In contrast, lumbar spine bone density YAM value (%) was significantly improved, from 63.8% ± 9.4% to 70.9% ± 5.2% (P = 0.012). Regarding bone metabolism markers, mean value of bone resorption marker TRACP-5b after the start of administration did not significantly differ from that at the start of observation. BAP value increased significantly from 16.2 ± 8.7 to 25.2 ± 11.0 (p < 0.01) while whole-PTH significantly increased from 103.5 ± 52.8 to 236.1 ± 100.8 (p < 0.01). In addition, serial time changes for each bone metabolic marker, whole-PTH value, and corrected Ca value during the observation period are also reported. Conclusion Romosozumab may improve bone density in dialysis patients with osteoporosis.

Outer retinal involvement in N-methyl-D-aspartate-induced inner retinal injury in rabbits assessed by optical coherence tomography.

This study was aimed to investigate morphological alteration of the retina with N-methyl-D-aspartate (NMDA)-induced injury in rabbits by optical coherence tomography (OCT). The right and left eyes of a total of 12 rabbits received single-intravitreal injection of vehicle and NMDA, respectively. Four out of the 12 animals underwent OCT and quantification of plasma microRNA repeatedly (4, 48, and 168 hr after dosing), followed by ocular histopathology at the end of the study. Ocular histopathology was also conducted in the eyes collected 4 or 48 hr after dosing from 4 animals at each time period. OCT revealed hyper-reflective ganglion cell complex and thickened inner retina in NMDA-treated eyes 4 hr after dosing; the inner retina shifted to thinning at later time points. The eyes given NMDA also exhibited greater thickness of the outer retina, which contains photoreceptors, after treatment, and thickened and obscured ellipsoid zone 168 hr after dosing. The plasma levels of miR-182 and miR-183, which are known to be highly expressed in photoreceptors, were higher 4 hr after dosing than pre-dosing values. Histopathologically, NMDA-induced inner retinal damage was confirmed: single-cell necrosis was observed in the ganglion cell layer and the inner nuclear layer 4 hr after dosing, the incidence of which decreased thereafter. At 168 hr after dosing, reduced number of ganglion cells was noted. No change was histopathologically observed in the outer retina. In conclusion, our results suggest involvement of photoreceptors in NMDA-induced inner retinal injury. Additionally, OCT revealed acute inner retinal findings suggestive of temporary edema.

Repeat Dose Study of the Cancer Chemopreventive Agent Resveratrol in Healthy Volunteers: Safety, Pharmacokinetics, and Effect on the Insulin-like Growth Factor Axis

Resveratrol, a naturally occurring polyphenol, has cancer chemopreventive properties in preclinical models. It has been shown to downregulate the levels of insulin-like growth factor-1 (IGF-I) in rodents. The purpose of the study was to assess its safety, pharmacokinetics, and effects on circulating levels of IGF-I and IGF-binding protein-3 (IGFBP-3) after repeated dosing. Forty healthy volunteers ingested resveratrol at 0.5, 1.0, 2.5, or 5.0 g daily for 29 days. Levels of resveratrol and its metabolites were measured by high performance liquid chromatography-UV in plasma obtained before and up to 24 hours after a dose between days 21 and 28. IGF-I and IGFBP-3 were measured by ELISA in plasma taken predosing and on day 29. Resveratrol was safe, but the 2.5 and 5 g doses caused mild to moderate gastrointestinal symptoms. Resveratrol-3-O-sulfate, resveratrol-4'-O-glucuronide, and resveratrol-3-O-glucuronide were major plasma metabolites. Maximal plasma levels and areas under the concentration versus time curve for the metabolites dramatically exceeded those for resveratrol, in the case of areas under the concentration versus time curve, by up to 20.3-fold. Compared with predosing values, the ingestion of resveratrol caused a decrease in circulating IGF-I and IGFBP-3 (P<0.04 for both), respectively, in all volunteers. The decrease was most marked at the 2.5 g dose level. The results suggest that repeated administration of high doses of resveratrol generates micromolar concentrations of parent and much higher levels of glucuronide and sulfate conjugates in the plasma. The observed decrease in circulating IGF-I and IGFBP-3 might contribute to cancer chemopreventive activity.

Effects of tetracycline on water quality, soil and gases in aerated and unaerated leachfield mesocosms

We examined the effects of tetracycline (TET) addition on the function of mesocosms representing aerated and unaerated septic system leachfields. Replicate mesocosms (n = 3) were filled with soil and either vented to a leachfield (LEACH) or aerated intermittently to maintain an O2 level of ∼ 0.21 mol mol− 1 (AIR). All mesocosms were dosed every 6 h for 10 d with 3 cm of domestic wastewater amended with 5 mg TET L− 1. Water quality parameters, headspace gas composition, and soil properties were measured prior to and during the dosing period, and for 42 days after the last antibiotic dose. No significant effect of TET was observed on the pH, level of dissolved O2 or dissolved organic carbon (DOC) in drainage water from either treatment. In contrast, levels of Fe2 + and SO4 in drainage water from LEACH mesocosms decreased in response to TET dosing, with lower levels persisting until Day 52. Persistent increases were observed in the level of NO3 in drainage water from AIR lysimeters and in NH4 in LEACH mesocosms in response to TET additions. Removal of total P and DOC were unaffected by TET dosing in either treatment. Nitrogen removal in AIR mesocosms decreased during the TET dosing period, returning to pre-dosing values by Day 52. In contrast, TN removal in LEACH mesocosms increased during TET dosing, returning to pre-dosing values by Day 52. The composition of headspace gases in AIR mesocosms was not affected by tetracycline dosing. TET dosing resulted in significant increases in soil NH4 concentration in LEACH mesocosms, whereas significant decreases were apparent in AIR mesocosms. Elevated levels of H2S and CH4 in the headspace of LEACH mesocosms coincided with TET dosing and returned to pre-dosing levels when antibiotic dosing ceased. The effects of tetracycline on leachfield mesocosms differed as a function of aeration. Although most effects were transient, with values returning to pre-dosing levels after a 6-week recovery period in both treatments, persistent effects were observed in LEACH mesocosms.

Acute phase response in toxicity studies. I. Survey of beagle dogs subjected to single-dose toxicity studies.

In the field of routine single-dose toxicity studies, we occasionally meet with transient leukocytosis associated with an increase in fibrinogen in beagle dogs within a few days after treatment with the test article. Only a little is known, however, about the toxicological significance of these changes. However, these changes were thought to belong to the category of "Acute Phase Response, APR," which has been known for a long time in connection with injury, trauma or infection. Aiming at proper understanding of these experiences, we surveyed 25 single-dose toxicity studies (7 intravenous bolus, 5 intravenous infusion, 12 oral and 1 subcutaneous treatment, hereafter referred to simply as i.v. bolus, i.v. infusion, oral and s.c.) in beagle dogs, provided with data from hematological examinations. We set the following criteria as a positive response in the present survey: increases of 50% or more in either or both WBC or fibrinogen compared to the predosing value, transiently from Day 1 to Day 3 of the study. Among 25 studies surveyed, about 1/2 of the studies exhibited increases of 50% or more in either or both fibrinogen or WBC counts compared to the predosing values showing dose-dependency transiently on Day 1 or Day 2. These changes were remarkable after intravenous application. Oral application produced similar effects, although the incidence and severity were low compared to the i.v. routes. Regarding blood chemical and hematological changes other than changes in fibrinogen and WBC counts, there were no essential differences between the groups of studies with and without the changes in fibrinogen and WBC counts. These changes were thought to be characteristic and to have occurred as incidents unrelated to other changes. The reported changes seen in single-dose toxicity studies may belong to the category of APR as the non-specific mechanism of living bodies as stated by Burns et al. (1996).

Ovarian Response to Exogenous Gonadotropins in Infant Cynomolgus Monkeys Hidenobu Sameshima

The response of the ovaries to exogenous gonadotropins was examined by determining serum estradiol (E2) and progesterone (P) levels and by observing the development of follicles by using 5-to 7-month-old cynomolgus females. The serum E2 levels began to increase 2 h after a single injection of 15 IU of human menopausal gonadotropin (h MG) and reached peak levels at 6 h after dosing, but the E2levels decreased to near predosing values at 24 h. The serum E2 levels increased markedly due to the repeated injections of 15 IU h MG for 10 days (days 0-9), but the P levels were low and equal to the levels observed at the follicular stage in adult females. The E2 levels decreased after an injection of 1,000 IU h CG on day 10, and the levels on day 21 were almost equal to the predosing values on day 0. On the other hand, the P levels increased transiently on day 16, which suggested ovulation. In the histopathological examination of the ovaries obtained on day 10, large antrum follicles were observed. These results confirmed that the ovary of infant cynomolgus monkeys reacted in the same way as the ovary of adult monkeys to exogenous gonadotropins and suggested that this method could become a useful means of evaluating the ovarian function of infant females.

Morphine-3-glucuronide: evidence to support its putative role in the development of tolerance to the antinociceptive effects of morphine in the rat

Antinociceptive tolerance to morphine (MOR) was induced in groups of Sprague-Dawley rats receiving continuous intravenous infusions of morphine sulphate administered by 3 different MOR dosing regimes. At appropriate intervals throughout each infusion period, antinociceptive testing was performed using the tail-flick latency test and blood samples were collected. Groups of saline (SAL)-infused control rats also underwent antinociceptive testing and blood sample collection. Complete antinociceptive tolerance developed during each MOR infusion period and was characterized by a marked decline in the degree of antinociception from values greater than 90% of the maximum possible effect (%MPE) to pre-dosing baseline values. By contrast, %MPE values in SAL-infused control animals and in sham-operated rats were not significantly different from pre-dosing values throughout the infusion period, indicating that the experimental procedures themselves did not contribute to the development of antinociceptive tolerance to MOR. In addition, the rate of MOR tolerance development was inversely proportional to the MOR infusion rate. A very significant inverse relationship was observed between the mean degree of antinociception (%MPE) and the mean plasma molar concentration ratio, [morphine-3-glucuronide]/[MOR], for each of the 3 MOR dosing regimes and for the cumulated data. This relationship showed that near-maximum antinociception was attainable at ratio values less than approximately 0.50, whilst at ratio values above approximately 1.5, little or no antinociception was observed. Although %MPE was highly inversely correlated with the mean plasma morphine-3-glucuronide (M3G) concentrations for rats receiving regimes A and B, this was not the case for rats receiving regime C where antinociceptive tolerance was partially reversed by an increase in the morphine infusion rate part-way through the infusion period. In addition, a poor relationship was observed between %MPE and the mean plasma MOR concentration, possibly due to the confounding presence of M3G in all samples. Thus, we may conclude from this study in Sprague-Dawley rats that irrespective of the rate of antinociceptive tolerance development, the level of antinociception achievable appears to be highly inversely correlated with the mean [M3G]/[MOR] plasma molar concentration ratio and poorly correlated with the plasma MOR concentration, consistent with the notion that it is perhaps the balance between the excitatory effects of M3G and the inhibitory effects of MOR at the functional level which is the important determinant. Further research is required in carefully conducted studies in cancer patients to evaluate the possible contribution of the MOR metabolites. M3G and morphine-6-glucuronide (MbG), to increasing dosing requirements of MOR. Other factors such as disease progression and increasing psychological distress must also be borne in mind, as these will have a major influence on increasing dosing requirements of MOR in cancer patients (Portenoy 1994).

In vivo inhibition of serum dopamine-beta-hydroxylase by CaNa2 EDTA injection.

1. In humans CaNa2EDTA increases urinary excretion of lead as well as that of essential metals such as zinc, ferrum and manganese but not that of copper. 2. We studied the effect of CaNa2EDTA injection on serum dopamine-beta-hydroxylase, a copper-dependent enzyme, in three male lead welders hospitalized for suspected lead poisoning. 3. Injection of CaNa2EDTA (1000 mg i.v.) resulted in rapid reduction of serum dopamine-beta-hydroxylase activity which returned to normal level within 24 hours. 4. Copper sulphate (8 microns) but not ethylmaleimide (10 mM) restored in vitro serum dopamine-beta-hydroxylase activity to pre-dosing values which indicates that CaNa2EDTA inhibits dopamine-beta-hydroxylase by removing copper from the enzyme. 5. An inverse dose effect relationship between injected CaNa2EDTA and residual serum dopamine-beta-hydroxylase activity was observed. 6. In spite of the reported lack of changes in copper serum concentration, CaNa2EDTA significantly affects the activity of a copper-dependent enzyme in the serum. 7. As CaNa2EDTA is currently the drug of choice for the treatment of asymptomatic lead poisoning in children, the effect of CaNa2EDTA on copper-enzymes requires further investigation.

Normal chromosomal aberration frequencies in peripheral lymphocytes of healthy human volunteers exposed to a maximum daily dose of paracetamol in a double blind trial

Paracetamol (acetaminophen) has been examined for mutagenic potential in numerous studies: gene mutation tests consistently gave negative results while in vitro chromosomal aberration tests showed equally consistently positive effects. In vivo studies for chromosome breaking activity gave clearly negative, equivocal or weakly positive results. In particular two reports have indicated that human volunteers taking a maximum daily dose of paracetamol (3 × 1000 mg over 8 h) exhibited significantly elevated frequencies of chromatid breaks in their peripheral lymphocytes 24 h later. In the one study evaluating the time course, levels returned to normal between 3 and 7 days later. We performed a carefully controlled double-blind study in which volunteers were pre-screened for normal liver function, they all were non-smoking and their diet and environmental exposures were controlled during the study. Cell-cycle kinetics were monitored and paralleled and a placebo group was included. Although a larger number of cells than in the other studies was analysed we were unable to reproduce their findings. No significant increases in structural chromosome aberrations (CA) were found either when the paracetamol group (male, female or both) post-dosing values were compared with pre-dosing values, or when treated groups at any sampling time were compared with the placebo groups. There was not even any evidence that individuals responded to the clastogenic potential of paracetamol or that a group response may have been masked by non-responders. In conjunction with the recently published results of the NTP bioassay, showing no carcinogenic activity in mice and no carcinogenic activity in rats except an increase of mononuclear cell leukaemia in female rats which is of doubtful relevance, the study presented here argues that paracetamol does not pose an unacceptable (if any) genotoxic/carcinogenic risk to man.

Effect of onion ingestion on anti-oxidizing agents in dog erythrocytes.

The effect of onion ingestion on red cell anti-oxidizing agents such as superoxide dismutase (SOD), catalase, glucose-6-phosphate dehydrogenase (G-6-PD), glutathione peroxidase (GPx), NADH-methemoglobin reductase and reduced glutathione (GSH) was studied in relation to Heinz body hemolytic anemia in dogs. To dogs was given the onion soup equivalent to 30g of raw onion/kg body weight once a day for three days. A few days after the initiation of feeding, there was a transient decrease of SOD, catalase and G-6-PD activities to 60%, and GSH value to 70% of their predosing values with a sudden and marked Heinz body formation. These values then began to increase when Heinz bodies were observed in almost every red cells. On day 10, when red cells carrying Heinz bodies were almost cleared from blood circulation, catalase and G-6-PD reached peak values of nearly 250% of their predosing levels. Changes of GPx and NADH-methemoglobin reductase were not correlated with Heinz body counts. Methemoglobin concentration was significantly elevated and maintained in the level of 2-3% during the presence of Heinz bodies in the red cells.

The effect of xylazine upon hepatic glucose production and blood flow rate in the lactating dairy cow.

The effect of xylazine (Rompun) upon hepatic glucose production and blood flow rate was measured in two cows. Doses of 0-16 or 0-18 mg/kg bodyweight increased blood glucose concentrations by 200 per cent and hepatic glucose production by 400 per cent. Maximum blood glucose concentrations were reached approximately 40 minutes after dosing and did not start to fall until 185 minutes. Concentrations were near normal 24 hours after dosing. The increase in hepatic glucose production was greatest 20 minutes after dosing and production had returned to control rates 150 minutes after dosing. Visceral glucose utilisation was also increased. Blood flow rates in the hepatic and portal veins were reduced to 50 to 60 per cent of their predosing values. It is concluded that the prolonged hyperglycaemia which persists beyond 150 minutes is produced either by continued glucose production from sites other than liver and viscera or by reduced utilisation of the blood glucose by peripheral tissue.