Serum Chemistry Changes Research Articles

Intrathecal enzyme replacement therapy in a patient with mucopolysaccharidosis type I and symptomatic spinal cord compression

In mucopolysaccharidosis I, deficiency of alpha-L-iduronidase can cause spinal cord compression (SCC) due to storage of glycosaminoglycans (GAGs) within the cervical meninges. As intravenous enzyme replacement therapy (ERT) is not likely to provide enzyme across the blood-brain barrier, standard treatment for this complication is usually surgical, which has a high morbidity and mortality risk. We report on the use of intrathecal (IT) laronidase in a MPS I patient with SCC who refused the surgical treatment. Assessments were performed at baseline, with clinical and biochemical evaluations, 4-extremity somatosensory evoked potentials, 12 min walk test and MRI studies of the CNS. Changes on these parameters were evaluated after 4 IT infusions of laronidase administered monthly via lumbar puncture. To our knowledge, this was the first MPS patient who received IT ERT. No major adverse events were observed. There were no clinically significant changes in serum chemistries. CSF GAG results revealed pretreatment values slightly above normal standards: 13.3 mg/L (NV < 12 mg/L) which after IT laronidase infusions were within normal levels (10.3 mg/L). 12MWT presented a 14% improvement, with better performance on stability and gait control. Maximum voluntary ventilation showed 55.6% improvement considering the percentage of predicted (26.7% at baseline compared to 41.9%); Maximum Inspiration Pressure improved 36.6% of predicted (26.8% at baseline to 36.7%); Pulmonary diffusion improved 17.6% of predicted %. In conclusion, although the improvement observed in this case with IT laronidase should be confirmed in further patients, this procedure seems to be a safe treatment for SCC in MPS I.

The effects of changes in salinity on osmoregulation and chloride cell morphology of juvenile Australian snapper, Pagrus auratus

The effect of rapid transfer of juvenile Australian snapper, Pagrus auratus from ambient seawater (30‰) to concentrated hyperosmotic (45‰) and diluted hyperosmotic (15‰) environments on serum osmolality, serum [Na+], [K+], [Cl−], blood haematocrit and branchial chloride cell morphology was assessed during 168 h after transfer. Serum osmolality, [Na+], [K+] and [Cl−] increased after 24 h in 45‰. In contrast, after 24 h in 15‰, [K+] did not change but serum osmolality, [Na+] and [Cl−] decreased. The serum chemistry changes were transient and had returned to near initial levels after 168 h in 45‰ and 15‰. Transfer from 30‰ to 45‰ and 15‰ did not affect blood haematocrit. Branchial chloride cells were identified in both filament and lamellar epithelia of snapper held in all salinity treatments by an immunocytochemical staining technique using an antiserum specific for Na+, K+–ATPase. In 45‰, the number of filament and lamellar chloride cells did not change, but filament chloride cells were more abundant than lamellar chloride cells. In contrast, filament chloride cells had increased in size after 72 h and by 168 h after transfer from 30‰ were 1.4-fold larger than the initial size. In 15‰, the number of filament chloride cells and the size of both filament and lamellar chloride cells had decreased after 72 h. Our results demonstrate that snapper can osmoregulate in a wide range of salinity and provide indirect evidence that both filament and lamellar chloride cells are responsible for excretion of excess salt from snapper in hyperosmotic environments. The ability for snapper to adapt rapidly and maintain homeostasis in a wide range of salinities supports the fact that snapper are a suitable species for land-based aquaculture in ponds, where rapid fluctuation in salinity can occur.

The Utility of a Rodent Model in Detecting Pediatric Drug-Induced Nephrotoxicity

A multi-age rat model was used to identify potential age-related differences in renal injury following exposure to gentamicin (GM). In this study, 10-, 25-, 40-, and 80-day-old Sprague-Dawley rats were dosed with GM at 0, 50, or 100 mg kg(-1) body weight per day (mkd) sc for 6 or 14 days. Urine samples were collected up to 72 h after initial dosing. The maximum tolerated dose was lower in 10-day-old rats than for other ages (none survived 11 days of treatment). Eighty-day-old rats given the highest dose showed a diminished rate of growth and an increase in serum creatinine, blood urea nitrogen (BUN), urinary kidney injury molecule-1 (Kim-1), and renal pathology. Ten- and 40-day-old rats given 100 mkd of GM for 6- or 14 days also had increased levels of serum BUN and Cr and renal pathology, whereas only mild renal alterations were found in 25-day-old rats. After 6 days of treatment with 100 mkd GM, significant increases in Havcr-1 (Kim-1) gene expression were detected only in 10- and 80-day-old rats. In urine samples, nuclear magnetic resonance and ultra performance liquid chromatography/mass spectrometry analysis detected changes related to GM efficacy (e.g., hippurate) and increases in metabolites related to antioxidant activity, which was greatest in the 80-day-old rats. The magnitude of the genomic, metabonomic, and serum chemistry changes appeared to correlate with the degree of nephropathy. These findings indicate that an experimental animal model that includes several developmental stages can detect age-related differences in drug-induced organ toxicities and may be a useful predictor of pediatric drug safety in preclinical studies.

Serum chemistry changes in weanling kittens fed high‐ or moderate‐protein diets

Serum chemistry changes in weanling kittens fed high‐ or moderate‐protein diets

Limitations of cyclosporine C2 monitoring in pediatric heart transplant recipients

Monitoring CSA levels at two h after dosing (C2) has been shown effective in providing adequate CSA-based immunosuppression in clinical trials in adult transplant recipients, but there is limited data regarding C2 monitoring in pediatric transplant recipients. Given the differences in CSA pharmacokinetics between children and adults, a cohort of stable pediatric transplant recipients was converted from monitoring CSA trough (C0) to C2 levels, to establish the clinical utility and safety of C2 monitoring. After an abbreviated AUC(0-5) to establish baseline exposure, subsequent CSA dosing was adjusted based on C2 levels. Additional evaluation included monitoring for rejection, changes in CSA dose, toxicity, serum chemistries, and infection. Twelve heart transplant recipients were enrolled, with mean age 4.8 yr (range: 0.6-14.0). All patients received microemulsified CSA (Neoral((R)); Novartis Pharmaceuticals, East Hanover, NJ, USA) twice daily. Baseline CSA dose was 5.39 +/- 2.05 mg/kg/day (mean +/- s.d.), with mean C0 = 267 +/- 112, C2 = 1065 +/- 565, and AUC(0-5) = 3817 +/- 1435. Only seven participants showed clear CSA peak levels at two h, with five exhibiting delayed peaks at three to five h post-dose. These seven participants completed 48 wk of study, with mean CSA dose decreasing to 4.55 +/- 3.61 mg/kg/day, maintaining mean C2 599 +/- 211 (vs. target C2 = 800). No significant change in serum creatinine was observed, although GFR increased from 76.9 to 107.6 mL/min/1.73 m(2) (p = 0.11). Five patients failed to achieve target C2 levels (>800) during the first four wk, despite comparable AUC values, and were maintained on trough monitoring (C0). Mean systolic and diastolic blood pressures fell slightly, three minor infections were noted during the study period, and one episode of acute rejection occurred, despite stable CSA dosing. Nearly 50% of stable pediatric transplant recipients failed to achieve adequate peak C2 CSA levels during conversion from C0 to C2 monitoring. Age-dependent differences in CSA absorption and/or clearance pharmacokinetics may explain these findings.

Relationship of Thermal Status to Productivity in Heat-Stressed Dairy Cows Given Recombinant Bovine Somatotropin

The responses of lactating Holstein cows to daily administration of bovine somatotropin (bST) were measured at thermoneutrality (Tn) and under both constant and cycled heat-stress conditions to determine the relationship between thermal status and bST-induced shifts in milk production. All tests included a 5-d acclimation period at Tn (18°C), followed by a 2-d increase in ambient temperature to 28.5°C. After d 3, ambient temperature was cycled between 28.5 (day) and 25.5°C (night) for 4 d. Daily injections with either 31mg of bST or saline began on d 1 of the experiment. Milk production, feed intake, and respiratory rate (RR) were measured daily. Intraperitoneal, telemetric temperature transmitters were used for a continuous measure of core body temperature (Tcore). Blood samples were collected during each phase to evaluate the changes in serum chemistry in response to bST and heat stress. Following a 15-d recovery, cows were switched across injection treatments and the study was repeated. Milk production decreased by ∼18.4% below the initial yield at Tn by the end of 7 d of heat challenge. Although a reduction in milk production occurred during heat stress in both groups, milk production was higher in bST-treated cows compared with control cows during periods of constant and cyclic heat. Likewise, bST treatment during the entire period increased the milk-to-feed ratio over the control level by ∼11.3%. Plasma insulin-like growth factor 1 and serum nonesterified fatty acids accompanied the increased growth hormone level with bST treatment (∼122.0 and 88.8%, respectively), whereas plasma urea nitrogen was reduced by ∼13.3% to reflect the shift to lipid metabolism. There was no difference in Tcore of the treatment and control groups at Tn. Both bST and control cows increased RR and Tcore above the Tn level by ∼94.8 and 2.9%, respectively, during constant heat, with a greater increase in Tcore of bST-treated compared with control cows (∼0.6%). The increase in RR during heat stress preceded Tcore by 1 d for both groups. During cyclic heat, Tcore decreased by ∼0.4% compared with constant heat in both the control and bST-treated groups. Bovine somatotropin treatment increased milk production similarly during the Tn and heat-stress periods, ∼8.3% over the control; however, the bST-induced increase in milk-to-feed ratio was greatest during the continuous and cyclic heat-stress phases, ∼16.2%. This increase occurred together with the elevation in Tcore.

Hypercalcaemia resulting from the use of tigecycline in the treatment of multidrug-resistant Acinetobacter in patients with multiorgan failure

Tigecycline (Wyeth) is a new glycylcycline antimicrobial that has been used in the treatment of deep-seated multidrug-resistant Acinetobacter (MDRA) infections. Unexpected changes in routine hematology or serum chemistry have not been reported.

Oral V2 receptor antagonist (RWJ‐351647) in patients with cirrhosis and ascites: a randomized, double‐blind, placebo‐controlled, single ascending dose study

RWJ-351647 is a selective V2 receptor antagonist that inhibits vasopressin-induced water reabsorption in the kidney. To investigate the safety and tolerability of RWJ-351647 compared with placebo after single oral dose administration to patients with cirrhosis and ascites, on a stable treatment with furosemide and spironolactone. Single oral doses of 1, 2 and 5 mg of RWJ-351647 were administered to 24 patients with ascites on stable concomitant diuretic treatment. RWJ-351647 had a tmax of 1 to 1.1 h and mean half-life of 10.4-17.4 h. There was no affect on the pharmacokinetics of concomitant diuretics. Increases in cumulative urine volume and free water excretion, and a decrease in urine osmolality were noted in a dose-dependent manner reaching the statistical significance at the 5-mg dose. Four patients exhibited a decrease of > 2 kg in weight in the 24 h after dosing. RWJ-351647 was well tolerated, with no evidence of a dose-related increase in adverse events when compared with placebo. No changes in either serum chemistry or plasma AVP (arginine vasopressin) and renin levels were observed despite the observed aquaresis. RWJ-351647 is an effective aquaretic causing dose-dependent increases in urine output and free water clearance, when co-administered with conventional diuretics in patients with cirrhosis and ascites.

(45) Safety and pharmacokinetics of human serum butyrylcholinesterase in guinea pigs

Human serum butyrylcholinesterase (Hu BChE) has been demonstrated to be a highly effective detoxifying enzyme for counteracting the acute toxicity of organophosphorus (OP) nerve agents. In order to initiate an investigational new drug (IND) application for human use, the safety and pharmacokinetic properties of the enzyme were assessed in guinea pigs. Sixty milligrams per kilogram of Hu BChE was administered to guinea pigs by either i.p. or i.m. injection. Blood was drawn at various time points for up to 2 weeks following enzyme injection for the measurement of blood BChE activity. Hu BChE displayed a mean residence time of 110 h, regardless of the route of administration and the enzyme activity remained almost 10-fold above baseline level even after 2 weeks post enzyme injection. Fourteen days post Hu BChE administration, all animals were subjected to 20 panel serum chemistry, hematology, and complete gross/histopathology examination. Results showed no toxic effects as measured by general observation, serum chemistry, hematology, and gross and histological tissue changes. In conclusion, Hu BChE displays a long-lasting stability in the circulation of guinea pigs, and is devoid of any toxic side effects. These results provide convincing data for the safe and effective use of Hu BChE as a bioscavenger to protect humans against all OP nerve agents.

Human serum butyrylcholinesterase: In vitro and in vivo stability, pharmacokinetics, and safety in mice

The use of exogenously administered cholinesterases (ChEs) as bioscavengers of highly toxic organophosphate (OP) nerve agents is now sufficiently well documented to make them a highly viable prophylactic treatment against this potential threat. Of the ChEs evaluated so far, human serum butyrylcholinesterase (HuBChE) is most suitable for human use. A dose of 200 mg (3 mg/kg) of HuBChE is envisioned as a prophylactic treatment in humans that can protect from an exposure of up to 2 × LD 50 of soman. In addition to its use as a prophylactic for a variety of wartime scenarios, including covert actions, it also has potential use for first responders (civilians) reacting to terrorist nerve gas release. We recently, developed a procedure for the large-scale purification of HuBChE, which yielded approximately 6 g of highly purified enzyme from 120 kg of Cohn fraction IV-4. The enzyme had a specific activity of 700–750 U/mg and migrated as a single band on SDS–PAGE. To provide data for initiating an investigational new drug (IND) application for the use of this enzyme as a bioscavenger in humans, we established its pharmacokinetic properties, examined its safety in mice, and evaluated its shelf life at various temperatures. In mice administered various doses up to 90 mg/kg, enzyme activity reached peak levels in circulation at 10 and 24 h following i.p. and i.m. injections, respectively. The enzyme displayed a mean residence time (MRT) of 40–50 h, regardless of the route of administration or dose of injected enzyme. Mice were euthanized 2 weeks following enzyme administration and tissues were examined grossly or microscopically for possible toxic effects. Results suggest that HuBChE does not exhibit any toxicity in mice as measured by general observation, serum chemistry, hematology, gross or histologic tissue changes. The shelf life of this enzyme stored at 4, 25, 37, and 45 °C was determined in lyophilized form. The enzyme was found to be stable when stored in lyophilized form at −20, 4, 25, or 37 °C to date (2 years), as measured by specific activity and SDS polyacrylamide gel electrophoresis. The effect of storage on circulatory stability was determined by measuring MRT in mice; there was no change in the MRT of lyophilized enzyme stored at −20 °C to date (2 years). These results provide convincing data that HuBChE is a safe bioscavenger that can provide protection against all OP nerve agents. Efforts are now underway to prepare the required documentation for submission of an IND application to the United States Food and Drug Administration (USFDA).

Efficacy and Safety of a Continuous-Use Regimen of Levonorgestrel/Ethinyl Estradiol: North American Phase 3 Study Results

LNG/EE Continuous, which contains the well-studied hormones levonorgestrel (LNG) and ethinyl estradiol (EE) at lower daily doses than currently available, is the first low-dose oral contraceptive (OC) intended to inhibit menses and alleviate cycle-related symptoms. Primary objectives were to evaluate safety and efficacy; other objectives included evaluation of menses inhibition and cycle-related symptoms. This phase 3, single-treatment, open-label, 1-year trial was conducted at 92 sites in the United States and Canada Healthy sexually active women aged 18 to 49 years were enrolled. Subjects had to have had regular (21- to 35-day) menstrual cycles in the 3-month period before study entry. There were no restrictions on weight, history of vaginal bleeding, or smoking, however, women older than 34 years who smoked >15 cigarettes per day were excluded. Subjects took 1 pill daily of LNG 90 μg/EE 20 μg starting on the first day of their first menstrual cycle and continuing for 12 months (13 pill packs) with no pill-free intervals. The primary efficacy measure was the number of unintended pregnancies. Safety was based on physical exams, clinical laboratory tests, and adverse event records. Subjects kept a written daily diary for vaginal bleeding and spotting. Amenorrhea was defined as the absence of bleeding or spotting; no bleeding was defined as the absence of bleeding, with or without spotting. A total of 2,134 subjects took at least 1 dose of study drug. Nineteen (19) subjects became pregnant during use of this regimen. The Pearl Index for the continuous-use regimen will be calculated from pooled results of all phase 3 studies, but for this single study the Pearl Index was1.60 (95% CI, 0.96, 2.49) based on 15,461 pill packs and 13 pill packs per year. Additionally, LNG/EE Continuous was effective in inhibiting menses by inducing amenorrhea and no bleeding. There was a steady increase from baseline in the percentage of subjects who reported amenorrhea and no bleeding: 44.8% and 70.8%, respectively, by pill pack 7. LNG/EE Continuous was also effective in reducing the number of bleeding and spotting days. The median number of bleeding days decreased to 0 (range 0 to 28) by pill pack 3. Six (6) serious adverse events (AEs) occurred that were at least possibly related to LNG/EE: cholecystitis (2), deep vein thrombosis/pulmonary embolism (1), ectopic pregnancy (1), prolonged vaginal bleeding (1) and enlarged uterine fibroids (1). There was 1 unrelated death from an ovarian teratoma. Discontinuations due to AEs occurred in 17% of subjects. Except for treatment-emergent AEs related to vaginal bleeding, the overall incidence of AEs was consistent with that commonly seen with a 21-day cyclic regimen that includes a 7-day pill-free interval. There were no clinically important changes in serum chemistries, hematology, or hemostasis factors, and no apparent differences from a 21-day cyclic regimen. Lipid profile changes were typical of OCs containing LNG/EE. LNG/EE Continuous was safe, efficacious, and well tolerated, and demonstrated the ability to inhibit menses by incrementally inducing amenorrhea and no bleeding (with or without spotting) with increased duration of use. The regimen offers women a safe and effective contraceptive option that reduces the number of bleeding days and symptoms commonly associated with the menses.

Safety and immunological efficacy of a prostate cancer plasmid DNA vaccine encoding prostatic acid phosphatase (PAP)

Prostatic acid phosphatase (PAP) is a prostate tumor antigen currently being investigated as a target antigen in several human vaccine trials, some with evidence of clinical benefit. We have previously demonstrated that plasmid DNA vaccines encoding either human or rat PAP can elicit antigen-specific cellular and humoral immunity in rat models. The current study was performed to determine the safety and potential immunological efficacy in rodents of large and repetitive doses of a GMP-grade plasmid DNA vaccine encoding human PAP, pTVG-HP. Fifty-four male Lewis rats were immunized intradermally at 2-week intervals with 100, 500, or 1500 μg pTVG-HP with 5 μg recombinant rat GM-CSF protein given as a vaccine adjuvant. An additional 12 male Lewis rats served as controls with groups immunized with 1500 μg of a parental DNA vector not encoding human PAP, and a group that received GM-CSF protein only without plasmid DNA. Groups of animals ( n = 3–6) were euthanized after two, four, or six immunizations with collections of tissues and blood for toxicity assessment and immunological analysis. No significant toxicities were observed in terms of animal weights, histopathology, hematological changes, or changes in serum chemistries. Six of fifty-four were found to have subtle evidence of possible renal toxicity, however these findings were not statistically different from control animals. The vaccine was found to be effective in eliciting PAP-specific CD4 and CD8 T cells, predominantly Th1 in type, in all immunized animals at all doses and numbers of immunizations. PAP-specific IgG were detected in a dose-dependent fashion, with titers increasing after multiple immunizations. These studies demonstrate that, in rats, immunization with the pTVG-HP vaccine is safe and effective in eliciting PAP-specific cellular and humoral immune responses. These findings support the further clinical evaluation of pTVG-HP in patients with prostate cancer.

A chronic dietary toxicity study of DAG (diacylglycerol) in Beagle dogs

The potential chronic toxic effects of DAG (diacylglycerol) when administered orally for 12 months were evaluated in this dietary study in Beagle dogs. DAG is a cooking oil which contains >80% diglycerides, <20% triglycerides and ⩽5% monoglycerides. For this study, a special diet was prepared with no dietary fat so that all of the dietary fat could be provided by DAG, at various concentrations together with a control oil. The control oil, TG (triacylglycerol), was prepared to contain >85% triglycerides, <10% diglycerides and ⩽5% monoglycerides. The fatty acid composition for DAG and TG was closely matched. Dietary concentrations of 0% DAG/9.5% TG (TG control), 1.5% DAG/8.0% TG, 5.5% DAG/4.0% TG, and 9.5% DAG/0% TG were presented daily, seven days per week, for 52 weeks. A second concurrent control group received the standard basal diet (Certified Canine LabDiet ® 5007, which has a fat content of 9.5%). The basal diet, control article-treated and DAG-treated groups each consisted of four male and four female dogs. Treatment was initiated in prejuvenile (2.5-month-old) dogs. Statistical evaluations compared the DAG-treated groups both to the basal diet and 9.5% TG control groups. The clinical condition of the animals, body weights, body weight gains and food consumption were unaffected by DAG. Hematology and urinalysis parameters were unaffected. No serum chemistry changes indicative of a toxic effect were observed. There were no effects noted on ECG data. No test article-related gross or histopathologic findings or changes in organ weights were observed. While there were no identifiable differences between the effects of TG and DAG, both caused some differences relative to the basal diet (lower food consumption, higher alkaline phosphatase, cholesterol and triglycerides). These differences were not toxicologically significant and were attributed to the differences in the diet rather than the fat source. Thus, DAG at dietary concentrations up to 9.5% for one year had no effect on normal canine growth and development, in comparison to TG.

Safety and pharmacokinetics of recombinant factor XIII in healthy volunteers

Factor XIII (FXIII) is a plasma transglutaminase that covalently cross-links fibrin proteins to one another and to other proteins, increasing the mechanical strength of blood clots. Endogenous FXIII is the final enzyme in the clotting cascade and circulates as a heterotetramer comprising 2 FXIII-A subunits and 2 FXIII-B subunits. Recombinant human FXIII A2 (rFXIII) homodimer is produced in Saccharomyces cerevisiae. Upon injection, rFXIII combines with the free FXIIIB subunit that circulates in excess to form the rA2B2 tetramer. In this placebo-controlled, double-blind, multi-dose study, the safety, pharmacokinetics, and pharmacodynamics of rFXIII were studied in 24 healthy volunteers, who were randomized in 2 cohorts of 12 subjects each. In each cohort, 8 subjects received 5 daily intravenous doses of rFXIII (10 or 25 U/kg), and 4 subjects received placebo. Recombinant FXIII was well tolerated. No deaths or serious adverse events occurred. The type and frequency of adverse events showed no pattern or dose response. No clinically significant changes in haematology, serum chemistry, or urinalysis laboratory values were observed. No clinical coagulopathy or thrombosis was observed. Recombinant FXIII did not produce any anti-yeast or anti-FXIII antibodies. After 5 daily doses of rFXIII, accumulation indices indicated a 3 to 4fold accumulation of rFXIII in plasma. The elimination half-life, estimated for rFXIII as the heterotetramer, ranged from 228-346 hours for the 10U/kg dose group and 167-197 hours for the 25U/kg dose group. The safety, pharmacokinetic, and immunogenic profile of rFXIII suggests it may have potential use in patients with congenital or acquired FXIII deficiency.

FC‐9 Coat funk in Alaskan malamutes

The objective of this study was to determine if a condition called coat funk by Alaskan malamute breeders has clinical, microscopic, haematologic, serum chemistry, urine and/or hormonal changes that allow affected dogs to be distinguished from normal dogs. Alaskan malamute breeders provided five dogs that met the breed standard for a normal pelage and five dogs considered to have coat funk. All animals were given complete physical examinations, and skin biopsies were collected. In addition, clipped hair was examined by optical‐based fibre diameter analysis. The skin biopsy samples were analysed by descriptive and morphometric analyses. Complete blood counts, serum chemistry profiles, urinalyses, and endocrine evaluations (thyroid profiles, urine cortisol:creatinine ratios, somatomedin C levels, and pre‐ and post‐ACTH stimulation concentrations of cortisol and adrenal sex hormones) were performed on all dogs. Clinically, affected dogs had varying degrees of involvement of the dorsal neck, tail and trunk. This was the only consistent difference noted between normal and affected dogs. In spite of the detailed analyses used in this investigation, the procedures failed to always separate normal from affected dogs and precluded separating coat funk from other hair loss diseases of Alaskan malamutes. This could be due to the lack of real differences between the two groups, failure of the procedures to allow separation of the two groups, or small sample size. Larger numbers of dogs followed over time are needed to better define the clinical and microscopic features of this disease and its related hair loss. Funding: The Alaskan Malamute Club of America.

Structural and Ultrastructural Alterations in BALB/c Mice: Effects of Penicillium CitrinumMetabolites

The aims of this work were to determine the effect of feeding BALB/c mice a diet containing culture materials of a citrinin producing strain of Penicillium citrinum (Thom). Changes in hematological parameters, serum chemistry and histological changes in liver, kidney and heart were determined. After 60 days, control treated (CT) mice appeared normal in all respects, whereas, the mice fed the feeds supplemented with Penicillium (CMT) showed decreased weight gain, lower hematocrits, increased serum alanine aminotransferase (ALT) and clear signs of renal and hepatotoxicity based on histological changes. Changes observed in the liver of CMT mice included portal and lobular infiltration of polymorphonuclear cells, with concomitant hepatocellular necrosis, hepatic steatosis, prominent Kupffer's cells, hemosiderin granules in the cytoplasm of periportal hepatocytes and other lipid inclusions in the surrounding mitochondria were also observed. Our findings suggest that in vivo, P. citrinum Thom metabolites, which contain citrinin, could cause illnesses such as toxic hepatitis or intravascular hemolysis.

Adenovirus Encoding Human Platelet-Derived Growth Factor-B Delivered in Collagen Exhibits Safety, Biodistribution, and Immunogenicity Profiles Favorable for Clinical Use

We have developed a therapeutic approach to wound repair involving immobilization of gene transfer vectors within biocompatible matrices (gene-activated matrix, or GAM). The matrix also serves as a scaffold for cellular in-growth and subsequent gene uptake and expression. An adenoviral vector encoding human platelet-derived growth factor-B delivered in collagen (AdPDGF-B/GAM) has demonstrated efficacy in models of wound repair. The safety, biodistribution, and immunogenicity profiles of AdPDGF-B/GAM were examined using a rabbit dermal wound model. Four weekly doses at 1 × 1010 and 1 × 1011 viral particles/cm2 of wound surface stimulated dose-related increases in granulation tissue formation and cell proliferation. In situ hybridization and immunostaining demonstrated concordant expression of human PDGF-B mRNA and protein. No treatment-related changes in hematology, serum chemistry, or histopathology were observed. Although AdPDGF-B DNA and PDGF-B mRNA were detected in wounds and axillary lymph nodes of treated animals, no AdPDGF-B was detected in blood or other organs. No immunologic responses against collagen were observed; however, as expected, IgG responses to AdPDGF-B and human PDGF-BB protein were detected. In adenovirus-preimmunized rats, attenuation of the wound healing response was modest (∼16%). Collectively, these observations indicate that repeat doses of AdPDGF-B/GAM are well tolerated and lead to robust, localized tissue repair.

Phase I Trial of Intramuscular Injection of a Recombinant Adeno-Associated Virus Alpha 1-Antitrypsin (rAAV2-CB-hAAT) Gene Vector to AAT-Deficient Adults

A recombinant virus vector constructed from adeno-associated virus (AAV) that has been altered to carry the human alpha1-antitrypsin (hAAT) gene expressed from a hybrid chicken beta-actin promoter with a cytomegalovirus enhancer has been developed. The construct has been shown to initiate the production of hAAT in animal models closely matching the proposed human trial. The proposed clinical trial is an open-label, phase I study administering recombinant adeno-associated virus alpha1-antitrypsin (rAAV2-CB-hAAT) gene vector intramuscularly to AAT-deficient human subjects where gene expression can be measured directly in blood samples to assess safety. Safety parameters will be measurement of changes in serum chemistries and hematology, urinalysis, pulmonary function testing, semen assay for vector genomes, immunologic response to AAT, and AAV, as well as reported subject history of any symptoms.

Cardiopulmonary, blood and peritoneal fluid alterations associated with abdominal insufflation of carbon dioxide in standing horses.

Abdominal insufflation is performed routinely during laparoscopy in horses to improve visualisation and facilitate instrument and visceral manipulations during surgery. It has been shown that high-pressure pneumoperitoneum with carbon dioxide (CO2) has deleterious cardiopulmonary effects in dorsally recumbent, mechanically ventilated, halothane-anaesthetised horses. There is no information on the effects of CO2 pneumoperitoneum on cardiopulmonary function and haematology, plasma chemistry and peritoneal fluid (PF) variables in standing sedated horses during laparoscopic surgery. To determine the effects of high pressure CO2 pneumoperitoneum in standing sedated horses on cardiopulmonary function, blood gas, haematology, plasma chemistry and PF variables. Six healthy, mature horses were sedated with an i.v. bolus of detomidine (0.02 mg/kg bwt) and butorphanol (0.02 mg/kg bwt) and instrumented to determine the changes in cardiopulmonary function, haematology, serum chemistry and PF values during and after pneumoperitoneum with CO2 to 15 mmHg pressure for standing laparoscopy. Each horse was assigned at random to either a standing left flank exploratory laparoscopy (LFL) with CO2 pneumoperitoneum or sham procedure (SLFL) without insufflation, and instrumented for measurement of cardiopulmonary variables. Each horse underwent a second procedure in crossover fashion one month later so that all 6 horses had both an LFL and SLFL performed. Cardiopulmonary variables and blood gas analyses were obtained 5 mins after sedation and every 15 mins during 60 mins baseline (BL), insufflation (15 mmHg) and desufflation. Haematology, serum chemistry analysis and PF analysis were performed at BL, insufflation and desufflation, and 24 h after the conclusion of each procedure. Significant decreases in heart rate, cardiac output and cardiac index and significant increases in mean right atrial pressure, systemic vascular resistance and pulmonary vascular resistance were recorded immediately after and during sedation in both groups of horses. Pneumoperitoneum with CO2 at 15 mmHg had no significant effect on cardiopulmonary function during surgery. There were no significant differences in blood gas, haematology or plasma chemistry values within or between groups at any time interval during the study. There was a significant increase in the PF total nucleated cell count 24 h following LFL compared to baseline values for LFL or SLFL at 24 h. There were no differences in PF protein concentrations within or between groups at any time interval. Pneumoperitoneum with CO2 during standing laparoscopy in healthy horses does not cause adverse alterations in cardiopulmonary, haematology or plasma chemistry variables, but does induce a mild inflammatory response within the peritoneal cavity. High pressure (15 mmHg) pneumoperitoneum in standing sedated mature horses for laparoscopic surgery can be performed safely without any short-term or cumulative adverse effects on haemodynamic or cardiopulmonary function.

Effects of Intraperitoneal Hyaluronan on Peritoneal Fluid and Solute Transport in Peritoneal Dialysis Patients

Hyaluronan (HA) is a glycosaminoglycan found in connective tissues and tissue spaces, including the peritoneal cavity. In vivo studies in a rat model of peritoneal dialysis (PD) have shown that addition of HA to PD solution during an intraperitoneal dwell can alter peritoneal fluid transport and protect the peritoneal membrane from the effects of inflammation and repeated infusions of dialysis solution. The current study sought to evaluate the safety of intraperitoneal HA and its effect on peritoneal fluid and solute transport when administered during a dialysis dwell in humans. 13 PD patients were enrolled in a prospective, randomized crossover study involving three dialysis treatments using the following PD solutions: (1) a commercially available PD solution (Dianeal PD-4, 1.36% glucose; Baxter Healthcare Corporation, Alliston, Ontario, Canada); (2) Dianeal PD-4 containing 0.1 g/L HA, and (3) Dianeal PD-4 containing 0.5 g/L HA. Each 6-hour dialysis exchange was separated from the other exchanges by a 2-week washout period. Radioiodinated human serum albumin (RISA) was administered with the dialysis solution to evaluate intraperitoneal volume, net ultrafiltration (UF), and fluid reabsorption. Peritoneal clearances, dialysate/plasma ratios (D/P), and mass transfer area coefficients (MTACs) were determined for sodium, urea, creatinine, albumin, and glucose. Safety was evaluated by monitoring adverse events and changes in serum chemistries. Ten patients completed all three dialysis exchanges and two additional patients completed at least one treatment exchange. There were no reported adverse events related to HA administration and no significant changes in serum chemistries. There were no significant differences in net UF or peritoneal volume profiles among the three treatments. Mean net UF calculated using residual volumes, estimated by RISA dilution, tended to be slightly higher during treatment with solution containing 0.1 g/L HA and 0.5 g/L HA [74 +/- 86 (SE) and 41 +/- 99 mL, respectively] compared to control treatment (-58 +/- 129 mL). Although not statistically significant, there was a trend toward decreased fluid reabsorption during treatment with HA. Solute clearances, D/P ratios, and MTACs were similar for the three treatments. Serum levels of HA were also unaffected by the two treatment solutions. These data support the acute safety of HA when administered intraperitoneally with the dialysis solution to PD patients. Due to the small sample size and variability in net UF and fluid reabsorption, statistically significant differences were not demonstrated for these parameters. However, a trend toward decreased fluid reabsorption was observed, suggesting that HA may act by a mechanism similar to that observed in animal studies. Further studies are necessary to evaluate whether the beneficial effects of HA observed in animal studies can be shown in humans.