Pharmacokinetics of chlortetracycline in maternal plasma and in fetal tissues following oral administration to pregnant ewes.

The purpose of this study was to determine if concentrations of chlortetracycline could be detected in fetal plasma or tissues after administering an oral dose of chlortetracycline (CTC; 500 mg/head/day) reported to be effective in controlling Campylobacter spp. abortions. Five pregnant ewes were administered 250 mg/head twice a day (total dose 500 mg/hd/d) for 7 days. On the beginning of day 7, intravenous catheters were surgically implanted or inserted into the fetus and dam. Plasma samples were collected from the ewe and fetus at various time points before and up to 36 hr after the last dose of CTC. All ewes were then sacrificed, and tissues were harvested from the fetus for drug analysis. Concentrations of CTC in maternal plasma were consistent with our previous study and below the minimum inhibitory concentration of Campylobacter abortion isolates. Concentrations of CTC were below the limit of detection in three of five fetal plasma samples and all of the placenta, amniotic fluid, and fetal stomach contents. Low concentrations were detectable in fetal kidney and liver, suggesting that CTC reaches the fetus, although at a variable and low ratio when compared to maternal concentrations.

Pharmacokinetics of tulathromycin in nonpregnant adult ewes.

The objectives of this study were to determine plasma concentrations and pharmacokinetic parameters of tulathromycin after a single subcutaneous administration in the cervical region in sheep using the cattle labeled dose of 2.5 mg/kg. Six adult healthy ewes were administered tulathromycin on day 0. Blood samples were collected just prior to dosing and at selected time points for 360 h. Plasma samples were analyzed to determine tulathromycin concentrations, and noncompartmental analysis was performed for pharmacokinetic parameters. The mean maximum plasma concentration was 3598 ng/mL, the mean time to maximum concentration was 1.6 h, and the apparent elimination half-life ranged from 68.1 to 233.1 h (mean 118 h). When comparing our results to goats and cattle, it appears sheep are more similar to cattle in regard to the concentrations observed and pharmacokinetic parameters. In summary, the pharmacokinetics of tulathromycin in sheep appear to be similar enough to those in goats and cattle to recommend similar dosing (2.5 mg/kg SC), assuming that the target pathogens have similar inhibitory concentrations.

Pharmacokinetics of oral chlortetracycline in nonpregnant adult ewes.

The objectives of this study were to determine plasma concentrations and pharmacokinetic parameters of feed-grade chlortetracycline (CTC) in sheep after oral administration of 80 or 500 mg/head daily, divided into two equal doses given at 12-h intervals for 8 days. These are the approved, and commonly used but unapproved, feed additive doses, respectively, in the United States for the prevention of ovine infectious abortion. Blood samples were collected just prior to dosing at 0, 12, 24, 72, 96, and 192 h, as well as 4, 8, 12, 24, and 36 h after the last dose, and noncompartmental pharmacokinetic analysis was performed to estimate elimination half-life and area under the plasma concentration-time curve (AUC). Mean observed maximum CTC concentrations (Cmax ) were 20.0 ng/mL (80 mg dose) and 101 ng/mL (500 mg dose). Mean apparent elimination half-life was 18 h (80 mg dose) and 20 h (500 mg dose). Although published data do not exist to estimate plasma CTC concentrations necessary for the prevention of ovine infectious abortion, concentrations reached in our study suggest that either the FDA-approved and FDA-unapproved dosages are not high enough or that the pharmacodynamic parameter relating preventive dose to pathogen minimum inhibitory concentrations is yet to be determined.

Caprine abscess model of tulathromycin concentrations in interstitial fluid from tissue chambers inoculated with Corynebacterium pseudotuberculosis following subcutaneous or intrachamber administration.

Corynebacterium pseudotuberculosis causes chronic, suppurative, abscessing conditions in livestock and humans. We used an in vivo model to evaluate antimicrobial efficacy for focal abscesses caused by C. pseudotuberculosis. Tissue chambers were surgically implanted in the subcutaneous tissues of the right and left paralumbar fossa of 12 goats to serve as a model for isolated, focal abscesses. For each goat, one tissue chamber was inoculated with C. pseudotuberculosis, while the contralateral chamber served as an uninoculated control. Six goats were administered a single dose of tulathromycin at 2.5 mg/kg of body weight subcutaneously, while the other six received the same dose by injection directly into the inoculated chambers. Our objective was to compare the effects and tulathromycin concentrations in interstitial fluid (IF) samples collected from C. pseudotuberculosis-infected and control chambers following subcutaneous or intrachamber injection of tulathromycin. In addition, the effects of tulathromycin on the quantity of C. pseudotuberculosis reisolated from inoculated chambers were assessed over time. Tulathromycin IF concentrations from C. pseudotuberculosis-infected and control tissue chambers were similar to those in plasma following subcutaneous administration. Following intrachamber administration, tulathromycin IF concentrations in infected chambers were continuously above the MIC for the C. pseudotuberculosis isolate for 15 days. There were no significant differences for plasma area under the curve and elimination half-lives between subcutaneous and intrachamber administration. Six of the 12 infected chambers had no growth of C. pseudotuberculosis 15 days postadministration. Results of this study indicate that tulathromycin may be beneficial in the treatment of focal infections such as those caused by C. pseudotuberculosis.

Neurological disease in lambs associated with exposure to high environmental temperature and humidity.

BACKGROUND: We detected a pattern of lambs presenting with hyperthermia and neurological signs during the summer. OBJECTIVES: The main objectives of this study were to compare clinical findings and results of diagnostic testing and to identify a potential etiology. ANIMALS: Fifteen clinical cases of lambs less than 12 months of age presenting with neurological signs, tachypnea, and hyperthermia over 4 summers. METHODS: Retrospective case series. Medical records were searched for lambs less than 12 months of age that presented with neurological signs including the following: kyphosis, pelvic limb hyperextension, treading of feet, muscle tremors and recumbency, and hyperthermia of greater than 104°F. A grading system was established to describe severity of presenting neurological signs. Weather data were collected from weather stations near the farm of origin for 3 days prior to presentation. RESULTS: The lambs were from 7 flocks in central Texas. All cases occurred between July and September, with a median heat index of 90.5 for the 3 days before presentation. Complete blood count, serum chemistry, necropsy examination, rumen content, virology, brain MRI, liver copper, selenium, and vitamin E failed to identify a consistent etiology for the signs presented. The only common factor was high heat and humidity. Histopathological examination identified axonal degeneration and skeletal muscle necrosis in some lambs. CONCLUSIONS AND CLINICAL IMPORTANCE: These clinical cases appeared similar to the Australian disease humpyback and indicate that lambs exposed to high environmental temperatures and humidity might be at risk of developing the described clinical presentation.

A critical body residue approach for predicting persistent bioaccumulative toxicant effects on reproduction and population dynamics of meiobenthic copepods.

Critical body residues (CBRs) are the measured tissue toxicant concentrations yielding a median dose-response on a dry-weight or lipid-normalized basis. They facilitate management decisions for species protection using tissue analysis. Population CBR is the mean dose yielding 50% population suppression and was predicted here in Amphiascus tenuiremis for fipronil sulfide (FS) using lifetables and the Leslie matrix. Microplate bioassays (ASTM E-2317-14) produced biomass sufficient for dry mass and lipid-normalized CBR estimates of reproduction (fertility) and population growth suppression. Significant FS toxic effects were delayed naupliar development (at ≥0.10 µg L(-1)), delayed copepodite development (at 0.85 µg L(-1)), decreased reproductive success (at ≥ 0.39 µg L(-1)), and decreased offspring production (at 0.85 µg L(-1)). A reproductive median effective concentration (EC50) of 0.16 µg L(-1) (95% CI: 0.12-0.21 µg L(-1)) corresponded to an adult all-sex CBR and lipid-normalized CBR of 0.38 pg FS · µg(-1) dry weight (95% CI: 0.27-0.52 pg FS · µg(-1)) or 2.8 pg FS · µg(-1) lipid (95% CI: 2.2-3.6 pg FS · µg(-1)), respectively. Copepod log bioconcentration factor (BCF) = 4.11 ± 0.2. Leslie matrix projections regressed against internal dose predicted fewer than five gravid females in a population by the third generation at 0.39 and 0.85 µg FS · L(-1) (i.e., 9.6-10.2 µg FS · µg(-1) lipid), and 50% population suppression at a CBR of 1.6 pg FS · µg(-1) lipid. This more integrative population CBR as a management tool would fall 1.75 times below the CBR for the single most sensitive endpoint-fertility rate.

Distribution of ceftiofur into Mannheimia haemolytica-infected tissue chambers and lung after subcutaneous administration of ceftiofur crystalline free acid sterile suspension.

The objective of this study was to evaluate the penetration of ceftiofur- and desfuroylceftiofur-related metabolites (DCA) into sterile and infected tissue chambers, lung tissue and disposition of DCA in plasma across four different sacrifice days postdosing. Twelve healthy calves were utilized following implantation with tissue chambers in the paralumbar fossa. Tissue chambers in each calf were randomly inoculated with either Mannheimia haemolytica or sterile PBS. All calves were dosed with ceftiofur crystalline free acid sterile suspension (CCFA-SS) subcutaneously in the ear pinna. Calves were randomly assigned to 4 groups of 3 to be sacrificed on days 3, 5, 7 and 9 postdosing. Prior to euthanasia, plasma and tissue chamber fluid were collected, and immediately following euthanasia, lung tissue samples were obtained from four different anatomical sites DCA concentration analysis. Results of our study found that, in general, DCA concentrations followed a rank order of plasma > infected tissue chamber fluid > noninfected tissue chamber fluid > lung tissue. Data also indicated DCA concentrations remained above the therapeutic threshold of 0.2 microg/mL for plasma and chamber fluid and 0.2 microg/g for lung tissue for at least 7 days post-treatment.

Hepatocellular carcinoma patients are advantaged in the current liver transplant allocation system.

Patients with hepatocellular carcinoma (HCC) within Milan criteria receive priority on the liver transplant waiting list (WL) and compete with non-HCC patients. Dropout from the WL is an indirect measure of transplant access. Competing risks (CR) evaluation of dropout for HCC and non-HCC patients has not previously been reported. Patients listed between 16 March 2005 and 30 June 2008 were included. Probability of dropout was estimated using a CR technique as well as a Cox model for time to dropout. Overall, non-HCC patients had a higher dropout rate from the WL than HCC patients (p < 0.0001). This was reproducible throughout all regions. In Cox regression, tumor size, model for end-stage liver disease (MELD) score and alpha fetoprotein (AFP) were associated with increased dropout risk. Multivariable analysis with CR showed that MELD score and AFP, were most influential in predicting dropout for HCC patients. The index of concordance for predicting dropout with the CR was 0.70. HCC patients appear to be advantaged in the current allocation scheme based on lower dropout rates without regard to geography. A continuous score incorporating MELD, AFP and tumor size may help to prioritize HCC patients to better equate dropout rates with non-HCC patients and equalize access.

Pore characterization through propagator-resolved transverse relaxation exchange.

We use the propagator-resolved transverse relaxation exchange experiment to characterize the pore space and fluid behavior of water saturated, tight-packed quartz sand. The experiment uses T2 exchange plots to observe the number of molecules that shift their environment for a range of mixing times. The propagator dimension allows us to determine how far the molecules have moved. The peak intensities are integrated and then plotted as a function of displacement and mixing time. We also model our system using both a probabilistic pore-hopping simulation and a spreading Gaussian model. We use the results of these simulations to interpret the peak intensity plots. From this, we can estimate pore features such as characteristic time, pore radii, and interpore spacing. The tortuosity of the different pore sizes can then be calculated from these values.

The dependence on magnetic field strength of correlated internal gradient relaxation time distributions in heterogeneous materials.

Magnetic susceptibility differences in porous media produce local gradients within the pore space. At high magnetic fields, these inhomogeneities have the potential to greatly affect nuclear magnetic resonance measurements. We undertake a study using a new NMR technique to measure the internal gradients present in highly heterogeneous samples over a wide range of magnetic field strengths. Our results show that even at ultra-high fields there can exist signal at internal gradient strengths sufficiently small that techniques for suppressing unwanted side effects have the possibility to be used. Our findings encourage the use of these high and ultra-high field strengths for a broader range of samples. Our results also give experimental evidence to support the theory of internal gradient scaling as a function of field strength within pores.

Propagator resolved transverse relaxation exchange spectroscopy.

We present in this communication a novel propagator-resolved transverse relaxation exchange experiment. This experiment enhances the previous technique of transverse relaxation exchange by enabling spatial resolution. Hence, we are able to obtain separate, and remarkably different, T2-T2 exchange plots, corresponding to different spatial displacement of the spin bearing water molecules in a porous sand matrix. This experiment is the first to combine two inverse Laplace dimensions with a Fourier dimension, opening the door to a host of new experiments combining Fourier and inverse Laplace spectroscopy.

Modified RIFLE criteria in critically ill children with acute kidney injury.

A classification system has been proposed to standardize the definition of acute kidney injury in adults. These criteria of risk, injury, failure, loss, and end-stage renal disease were given the acronym of RIFLE. We have modified the criteria based on 150 critically ill pediatric RIFLE (pRIFLE) patients to assess acute kidney injury incidence and course along with renal and/or non-renal comorbidities. Of these children, 11 required dialysis and 24 died. Patients without acute kidney injury in the first week of intensive care admission were less likely to subsequently develop renal Injury or Failure; however, 82% of acute kidney injury occurred in this initial week. Within this group of 123 children, 60 reached pRIFLEmax for Risk, 32 reached Injury, and 31 reached Failure. Acute kidney injury during admission was an independent predictor of intensive care; hospital length of stay and an increased risk of death independent of the Pediatric Risk of Mortality (PRISM II) score (odds ratio 3.0). Our results show that a majority of critically ill children develop acute kidney injury by pRIFLE criteria and do so early in the course of intensive care. Acute kidney injury is associated with mortality and may lead to increased hospital costs. We suggest that the pRIFLE criteria serves to characterize the pattern of acute kidney injury in critically ill children.

Clinical Trial of the Pan-Caspase Inhibitor, IDN-6556, in Human Liver Preservation Injury.

Cold ischemia/warm reperfusion (CI/WR) injury remains a problem in liver transplantation. The aim of the current study was to assess the utility of the pan-caspase inhibitor IDN-6556 on CI/WR injury during human liver transplantation. This report is a post hoc analysis of a Phase II, multi-center, randomized, placebo-controlled, double-blinded, parallel group study. Subjects were assigned to four treatment groups: Group 1 (Organ storage/flush: Placebo-Recipient: Placebo); Group 2 (Organ storage/flush: 15 microg/mL-Recipient: Placebo); Group 3 (Organ storage/flush: 5 microg/mL-Recipient: 0.5 mg/kg); and Group 4 (Organ storage/flush: 15 microg/mL-Recipient: 0.5 mg/kg). Liver cell apoptosis was assessed by serum concentrations of the apoptosis-associated CK18Asp396 ('M30') neo-epitope, TUNEL assay and caspase 3/7 immunohistochemistry. Liver injury was assessed by serum AST/ALT determinations. Serum markers of liver cell apoptosis were reduced in all groups receiving drug as compared to placebo. However, TUNEL, caspase 3/7 positive cells and serum AST/ALT levels were only consistently reduced in Group 2 (drug exposed to organ only). This reduction in serum transaminases was significant and observed across the study. In conclusion, IDN-6556 when administered in cold storage and flush solutions during liver transplantation offers local therapeutic protection against CI/WR-mediated apoptosis and injury. However, larger studies are required to confirm these observations.

Tracking pore to pore exchange using relaxation exchange spectroscopy.

We observe the movement of water over time between pores of differing sizes in Castlegate sandstone. To achieve this, we perform an NMR transverse relaxation exchange experiment for several mixing times. The resulting data are converted to 2D T2 distributions using a 2D inverse Laplace transform (ILT). We show for the first time that quantitative analysis of ILT distributions enables one to extract characteristic times for different pores sizes. This information is potentially useful for permeability determination as well as better understanding of exchange between specific pore subpopulations.

P2 purinergic receptors signal to STAT3 in astrocytes: Difference in STAT3 responses to P2Y and P2X receptor activation.

Extracellular ATP, released upon tissue damage to the CNS, can evoke reactive astrogliosis. The released ATP activates P2 purinergic receptors associated with the proliferation of normally quiescent astrocytes, although the underlying mechanisms remain to be fully elucidated. Signal transducer and activator of transcription 3 (STAT3) has been implicated in reactive astrogliosis and plays an important role in cell cycle regulation. Therefore, we investigated whether extracellular ATP and purinergic receptors regulate STAT3 signaling. Using immunoblot analysis, we found that addition of ATP to primary cultures of rat cortical astrocytes increased Ser-727 phosphorylation of STAT3 in a time-sensitive and concentration-dependent manner. ATP-stimulated Ser-727 STAT3 phosphorylation was mediated through P2 receptor activation since suramin, an antagonist of P2 receptors, diminished this response, whereas 8-(para-sulfo-phenyl)-theophylline (8PSTP), an antagonist of P1 receptors, did not. We found that UTP, an agonist of P2Y(2/4/6) receptors, stimulated rapid and robust phosphorylation of Ser727-STAT3, whereas BzATP, an agonist for P2X receptors, exhibited a delayed and weaker response. In contrast, both P2Y and P2X agonists stimulated phosphorylation of Tyr705-STAT3 to a similar extent. P2 receptors can couple to extracellular signal-regulated protein kinases (ERK) and we found that inhibition of ERK signaling blocked phosphorylation of Ser727-STAT3. Further characterization of the Ser727-STAT3 phosphorylation response to P2Y receptor activation supported a role for P2Y2 and P2Y4, but not P2Y6, receptors as well as a partial role for P2Y1 receptors. Because phosphorylation of Ser727-STAT3 can promote DNA transcriptional activity of cell cycle regulatory genes, the differences in phosphorylation of Ser727-STAT3 may contribute to the mechanism by which P2Y receptors promote, whereas P2X receptors inhibit, astrocyte proliferation. In support of this hypothesis, inhibition of STAT3 activation by cucurbitacin I prevented ATP-stimulated mitogenesis. We conclude that P2 receptors stimulate STAT3 activation and suggest that P2 receptor/STAT3 signaling may play an important role in astrocyte proliferation and reactive astrogliosis.

Penetration of ceftiofur into sterile vs. Mannheimia haemolytica-infected tissue chambers in beef calves after subcutaneous administration of ceftiofur crystalline free acid sterile suspension in the ear pinna.

The effect of Mannheimia haemolytica infection on the penetration of ceftiofur and desfuroylceftiofur metabolites into tissue chambers was studied in cattle after subcutaneous administration of ceftiofur crystalline free acid sterile suspension (CCFA-SS). Four tissue chambers were implanted subcutaneously in each of 12 calves. Approximately 45 days after implantation, two chambers were inoculated with M. haemolytica (10(6) colony-forming units per chamber) while the remaining two chambers were inoculated with sterile phosphate-buffered saline. Twenty-four hours after inoculation, CCFA-SS was administered subcutaneously in the middle third of the caudal ear pinna of each calf. Chamber fluid and blood samples were collected at predetermined times for 10 days following dosing and analyzed for ceftiofur and desfuroylceftiofur metabolites by high-performance liquid chromatography. Concentrations of ceftiofur and desfuroylceftiofur metabolites in plasma and tissue chamber fluid remained above a threshold of 0.2 microg/mL for at least 8 days. Infected tissue chamber fluid concentrations of ceftiofur and desfuroylceftiofur metabolites were significantly higher than those in non-infected tissue chamber fluid, which correlated with significantly higher total protein concentration in infected tissue chambers. These results indicate that single subcutaneous administration of CCFA-SS at 6.6 mg/kg can be expected to provide effective therapy of susceptible bacterial infections for a period of at least 1 week.

A familial peripheral neuropathy and glomerulopathy in Gelbvieh calves.

Nine Gelbvieh calves originating in four herds and clinically presenting with rear limb ataxia/paresis had histopathologically confirmed peripheral neuropathy and a proliferative glomerulopathy. Degenerative lesions were severe in peripheral nerves, dorsal and ventral spinal nerve roots, and less marked in dorsal fasciculi of the spinal cord. Cell bodies of spinal ganglia were minimally diseased; ventral horn neurons occasionally had central chromatolysis and nuclear displacement. Glomerular lesions ranged from mild mesangial hypercellularity to glomerulosclerosis. Pedigree analysis of affected animals from one herd indicated a strong familial relationship and probable hereditary basis for the syndrome.