Toxins of concern to animals and people.

Historically veterinarians have diagnosed accidental poisonings and identified possible terrorist events before they have come to the attention of public health authorities. There are many toxins that pose a threat to both humans and animals and the authors examine several of them here, namely, anthrax, tricothecenes, staphylococcal enterotoxin B, botulinum toxins, ricin, saxitoxin and dinoflagellate toxins. By discussing exposure routes, clinical signs and differential diagnoses the authors demonstrate how veterinarians are in a unique position to recognise zoonotic diseases, toxin exposure, and acts of bioterrorism. The work of veterinarians protects the food supply and contributes to human health and this article highlights the importance of coordination and communication between veterinarians and physicians. Sharing information is critical in confirming diagnoses and, in the case of intentional toxin attacks, could also be beneficial in identifying the perpetrators of the crime.

The molecular basis of canine pyruvate kinase deficiency.

Inherited hemolytic anemia due to pyruvate kinase (PK) deficiency is an autosomal recessive disease of the Basenji dog that closely resembles human PK deficiency. Characterization of transcriptional and translational expression of PK isozymes and sequencing of DNA from normal and mutant dogs were performed to identify the genetic defect in Basenji dogs. Measurement of erythrocytic PK activity by ion exchange chromatography, substrate kinetics, immunologic reactivity, and electrophoretic mobility suggests that M2-type PK is the major form of PK activity in erythrocytes of PK-deficient dogs, in contrast to normal dogs having only R-type PK activity. Both R-type and M2-type PK mRNA are detectable in reticulocytes of PK-deficient dogs, suggesting that the aberrant isozyme expression is not due to a failure in the erythroid maturational switch from M2- to R-type isozymes. Nucleotide sequence data from wild-type and mutant R-type PK cDNA identified a single nucleotide deletion, delta C433, in the mutant cDNA. The deduced amino acid sequence predicts a truncated mutant protein devoid of all residues contributing to the catalytic site of the wild-type protein. In the absence of R-type PK activity, there is anomalous compensatory expression of M2-type PK in erythroid cells of PK-deficient Basenjis. The PK-deficient Basenji dog may be valuable in somatic cell gene therapy trials involving manipulation of hematopoietic stem cells.

Cutaneous lymphoid hyperplasia and cutaneous marginal zone lymphoma: comparison of morphologic and immunophenotypic features.

Cutaneous marginal zone lymphoma (MZL) is a recently described low-grade B-cell lymphoma that usually follows an indolent course. This tumor shares many histologic and clinical features with cutaneous lymphoid hyperplasia (CLH), a benign reactive lymphoid proliferation. Sixteen biopsy specimens from 14 patients with CLH were studied, and compared with 16 cases of cutaneous MZL (9 primary cutaneous, 7 with secondary involvement of the skin) to determine whether there were features that would permit their distinction on routinely fixed, paraffin-embedded tissue sections. Both disorders showed a female preponderance (CLH: 9 F, 5 M; MZL: 11 F, 5 M). The median age was also similar (CLH: 54 years; cutaneous MZL: 55 years). CLH was most common on the arm (8) and the head and neck (7) but also involved the trunk (1); primary cutaneous MZL most often involved the limbs (3), trunk (3), and head and neck (3). Lymphoma did not develop in any of the 14 CLH patients (follow-up ranging from 9 to 246 months, mean 62 months). Six of 9 patients with primary cutaneous MZL and all 7 patients with secondary cutaneous MZL experienced relapses, most commonly isolated to skin or a subcutaneous site. On hematoxylin-eosin stained sections, a diffuse proliferation of marginal zone cells (p < 0.0001), zones of plasma cells (p = 0.01), the absence of epidermal change (p = 0.01), reactive germinal centers (p = 0.03), and a diffuse pattern of dermal or subcutaneous infiltration (p = 0.03) were more often seen in cutaneous MZL. A dense lymphocytic infiltrate, bottom-heavy or top-heavy growth pattern, eosinophils, and a grenz zone were seen equally often in both disorders. Dutcher bodies were observed only in cutaneous MZL. Immunoperoxidase stains on formalin-fixed paraffin-embedded tissue sections showed monotypic expression of immunoglobulin light chains by plasma cells in 11 of 16 MZL cases. By definition, no case with monotypic plasma cells was diagnosed as CLH. In CLH, T cells usually outnumbered B cells, and a B:T cell ratio > or = 3:1 was not observed in any case. By contrast, 40% of the MZL cases showed a B:T cell ratio > or = 3:1. No coexpression of CD20 and CD43 was seen in any case of either MZL or CLH. In summary, the clinical presentations of CLH and MZL are similar. In contrast to historical criteria for diagnosing cutaneous lymphoid infiltrates, the presence of reactive follicles favors a diagnosis of cutaneous B-cell lymphoma (CBCL). In addition, a bottom-heavy or top-heavy growth pattern is not a distinctive finding. Marginal zone cells and zones or sheets of plasma cells are strong morphologic indicators of marginal zone lymphoma. The diagnosis of CBCL can be supported in 40% of the cases by demonstrating a B:T cell ratio of > or = 3:1, and confirmed in 70% of the cases by demonstrating monotypic light chain expression of plasma cells on paraffin sections.

Marginal zone lymphoma (low-grade B-cell lymphoma of mucosa-associated lymphoid tissue type) of skin and subcutaneous tissue: a study of 15 patients.

Extranodal low-grade B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) type occurs in the gastrointestinal tract, salivary gland, thyroid, orbit, lung, and breast. We report 15 patients with MALT-type lymphomas involving skin and subcutaneous tissue. All patients had tumors with histologic features of low-grade B-cell lymphoma of MALT type, including marginal zone cells (15 of 15 cases), plasmacytic differentiation (10 of 15 cases), Dutcher bodies (three of 15 cases), and reactive germinal centers (10 of 15 cases). All expressed pan B-cell antigens and monotypic immunoglobulin. Seven patients (five women, two men) aged 29 to 86 years (median, 53 years) had primary MALT-type lymphoma of skin (6) or subcutaneous tissue (1). One patient had persistent disease, and four patients had relapses involving skin, subcutaneous tissue, breast, orbit, and lymph node. At last follow-up (11-121 months; median, 36 months), one patient was alive with disease, and six patients had no evidence of disease. Three patients (two women, one man) aged 36 to 67 years (median, 57 years) had concurrent MALT-type lymphoma involving both subcutaneous tissue and extracutaneous sites at primary diagnosis, including lung, breast, orbit, lymph node, and bone marrow. One patient responded to treatment but relapsed with lymphoma of the skin and breast. The other two patients had persistent disease despite treatment. One patient died of disease at 25 months, and, at last follow-up (7 and 46 months), two patients were alive with disease. Five patients (four women and one man) aged 29 to 72 years (median, 63 years) had secondary skin or subcutaneous involvement by MALT-type lymphoma with primary tumors of ocular adnexa (3) or parotid gland (2). All five patients had relapses, which involved skin or subcutaneous tissue, parotid gland, lacrimal gland, breast, and lymph node. At last follow-up (61-137 months), two patients were alive with disease and three were alive with no evidence of disease. Low-grade B-cell lymphomas of MALT type may arise in or secondarily involve the skin and subcutaneous tissue and have a tendency to affect middle-aged to older women. These tumors are characterized by multiple extranodal relapses and are associated with long patient survival. Patients with primary MALT-type lymphoma of skin or subcutaneous tissue without extracutaneous involvement at diagnosis were more likely to experience prolonged disease-free survival than patients with extracutaneous spread at presentation (p < 0.03).

Ofloxacin clinical pharmacokinetics.

Ofloxacin is a new fluoroquinolone with a spectrum of activity similar to other fluoroquinolones with activity which includes Chlamydia trachomatis, Mycobacterium spp., Mycoplasma spp. and Legionella pneumophila. Through its additional mechanisms of action, ofloxacin may be less susceptible to the development of resistance from Staphylococcus aureus commonly seen with currently available fluoroquinolones. The impact of these findings cannot be evaluated without further clinical experience. The pharmacokinetics of ofloxacin are characterised by almost complete bioavailability (95 to 100%), peak serum concentrations in the range of 2 to 3 mg/L after a 400mg oral dose and an average half-life of 5 to 8h. In comparison with other available quinolones, elimination is more highly dependent on renal clearance, which may lead to more frequent dosage adjustments in patients with impaired renal function. Ofloxacin appears less likely to affect the pharmacokinetics of drugs (e.g. theophylline) which commonly interact with fluoroquinolones such as ciprofloxacin and enoxacin. The properties of ofloxacin make it a therapeutic alternative to currently available fluoroquinolones.

Linezolid for treatment of vancomycin-resistant enterococcal peritonitis.

A 42-year-old woman on continuous ambulatory peritoneal dialysis (CAPD) developed peritonitis secondary to vancomycin-resistant Enterococcus faecium and Candida albicans while hospitalized for pneumonia. She was treated successfully with intravenous linezolid, fluconazole given by nasogastric tube, and removal of the peritoneal catheter. A concentration of linezolid above the minimum inhibitory concentration for most gram-positive pathogens, including vancomycin-resistant E faecium, was achieved in the dialysate fluid after an oral loading dose of 1200 mg. Additional data are needed to establish the role of linezolid in treating CAPD-associated peritonitis.

Exaggerated systemic antibody response to mucosal Helicobacter pylori infection in IgA nephropathy.

Numerous studies in the literature report aberrant immune responsiveness in immunoglobulin A (IgA) nephropathy. However, all these studies investigate immune responses invoked by an artificially engineered antigen challenge. For the first time in IgA nephropathy, we report the systemic humoral responses generated as part of an active mucosal immune response against a common environmental pathogen, Helicobacter pylori (Hp). We studied 22 patients with IgA nephropathy and 9 controls without renal disease who were shown to be infected with Hp, using a 13C-urea breath test. Hp antigen-specific enzyme-linked immunosorbent assays were established to measure the anti-Hp IgA, IgG, and IgA and IgG subclass antibody levels. In addition, anti-Hp responses in the monomeric and polymeric (pIgA) fractions of serum IgA were measured after separation by gel filtration high-performance liquid chromatography. IgA nephropathy was associated with both a greater rate of IgA anti-Hp seropositivity (P < 0.05) and a more pronounced IgA anti-Hp antibody response (P < 0.01). In almost all cases, IgA anti-Hp was IgA1, and more than 90% was polymeric. There was no difference in the frequency of IgG anti-Hp seropositivity, but patients produced a much greater IgG anti-Hp response (P < 0.01). In addition, the IgG subclass responses were markedly different, with IgG1 predominant in controls and IgG2 and IgG3 the major subclasses produced in IgA nephropathy. We have shown an exaggerated systemic antibody response to mucosal infection caused by Hp in patients with IgA nephropathy, predominantly consisting of pIgA1, IgG2, and IgG3. This suggests that in IgA nephropathy, not only is pIgA1 production poorly controlled, but regulation of IgG isotype switching in response to mucosal pathogens is also deranged.

Thermal injury in rats alters glucose utilization by skin, wound, and small intestine, but not by skeletal muscle.

The effects of thermal injury in rats on glucose utilization (Rg) by skin, wound, small intestine, and muscle in vivo has been determined using 2-[18F]-fluoro-2-deoxy-D-glucose (18FDG) 6 hours, 24 hours, and 3 weeks after injury. These results were compared with serum glucose and insulin levels at the same time points and with hexokinase activity in the tissues. Thermal injury had no significant effects on serum glucose levels; however, serum insulin levels were lower than sham values 6 hours after injury, the same as sham values 24 hours after injury, and significantly higher than sham values 3 weeks after injury. Rg of unburned skin was not changed at 6 or 24 hours after injury, but was increased at 3 weeks after injury. The Rg of the wound was almost zero at 6 and 24 hours after injury; however, at 3 weeks after injury, the wound area had a Rg two to three times higher than that of the normal skin of sham animals. Small intestine Rg was decreased 6 and 24 hours after injury, but was essentially normal by 3 weeks after injury. The Rg of muscle was unchanged at all of the time points tested. Total Rg was significantly higher in the animals 3 weeks after injury, with the increase primarily due to the increases in skin and wound. The increases in Rg were associated with changes in tissue hexokinase activity. The present data suggest that thermal injury to rats results in dramatic alterations in glucose utilization by the skin and wound, changes that may contribute to the overall alterations in carbohydrate metabolism during burn trauma.

Carbohydrate and the cytokine response to 2.5 h of running.

This randomized, double-blind, placebo-controlled study was designed to determine the influence of 6% carbohydrate (C) vs. placebo (P) beverage ingestion on cytokine responses (5 total samples over 9 h) to 2.5 h of high-intensity running (76.7 +/- 0.4% maximal O2 uptake) by 30 experienced marathon runners. For interleukin-6 (IL-6), a difference in the pattern of change between groups was found, highlighted by a greater increase in P vs. C immediately postrun (753 vs. 421%) and 1.5 h postrun (193 vs. 86%) [F(4,112) = 3.77, P = 0.006]. For interleukin-1-receptor antagonist (IL-1ra), a difference in the pattern of change between groups was found, highlighted by a greater increase in P vs. C 1.5 h postrun (231 vs. 72%) [F(2,50) = 6.38, P = 0.003]. No significant interaction effects were seen for bioactive IL-6 or IL-1 beta. The immediate postrun plasma glucose concentrations correlated negatively with those of plasma cortisol (r = -0.67, P < 0.001); postrun plasma cortisol (r = 0.70, P < 0.001) and IL-6 levels (r = 0.54, P = 0.003) correlated positively with levels of IL-1ra. Taken together, the data indicate that carbohydrate ingestion attenuates cytokine levels in the inflammatory cascade in response to heavy exertion.

Economic evaluation of three methods of treating urogenital chlamydial infections in the emergency department.

We attempted to determine the economic impact of three alternatives for the treatment of chlamydial infections in the emergency department: a written prescription for 7 days of doxycycline therapy (D-RX); a prepacked 7-day supply of doxycycline (D-ED); or a single 1-g dose of azithromycin (AZI). Data inputs for the model were obtained from both patient experience and literature sources. Primary health outcomes of the model were number of infection relapses. Economic outcomes were costs for initial treatment, treatment of relapses, and treatment of complications of relapse. For every 1000 patients, D-ED and AZI resulted in 21.6 (-10 to -41) and 36.2 (-25 to -63) fewer relapses than D-RX, respectively; AZI resulted in 14.6 (-35 to -4) fewer relapses than D-ED. Total costs were decreased for D-ED and AZI versus D-RX by $18,879 (-$39,000 to -$8000) and $24,039 (-$59,000 to -$10,000), respectively, and AZI resulted in a total cost decrease of $5160 (-$35,000 to +$6000) versus D-ED. Both D-ER and AZI decreased infection relapses and overall health care costs compared with D-RX. Also, AZI resulted in additional decreases in relapses versus D-ED, although the incremental impact on cost was inconclusive.

The triazole antifungal agents: a review of itraconazole and fluconazole.

Fluconazole and itraconazole are two investigational triazole antifungal agents that are currently undergoing clinical trials in the United States. Both are active orally, have favorable pharmacokinetics (i.e., good bioavailability, long half-life, low plasma protein binding), and possess activity against several systemic fungal pathogens. In addition, preliminary information suggests that these agents are substantially less toxic than currently available azole compounds. Fluconazole and, to a lesser degree, itraconazole have been shown to be highly effective for the treatment of cryptococcal meningitis. The potential for drug interactions is much lower with these agents compared to drugs such as ketoconazole.

Combination antifungal therapy for invasive aspergillosis infection in liver transplant recipients: report of two patients.

Invasive aspergillosis in solid organ transplant recipients is associated with mortality of approximately 100%. The search for optimal therapy has led clinicians to administer antifungal combinations. Two orthotopic liver transplant recipients developed invasive aspergillosis (pulmonary and perivertebral) after transplantation and were treated with combination antifungal therapy consisting of liposomal amphotericin B and itraconazole. Although both patients were initially stabilized, they died after 94 and 138 days of antifungal therapy, respectively. Presumably, aspergillosis was the principal cause of death. Antifungal serum concentrations and fungicidal titers in both patients indicated that the drugs may have been antagonistic and thus detrimental.

The developmental toxicity of an isolate from the plant Solanum dimidiatum (potato-weed) in Syrian golden hamsters.

The developmental toxicity of an ethanol extract from Solanum dimidiatum was investigated in date-bred Syrian golden hamsters. Neural tube defects were seen in three Syrian golden hamster fetuses, from one litter out of 12, at a maternal dose of 1.52 mg of plant isolate per g of body weight. Other malformations observed were syndactyly in a single fetus, from one litter out of ten, in the 0.45 mg/g dose group and gastroschisis in a single fetus, from one litter out of nine, in the 1.01 mg/g dose group. These findings suggest minimal developmental toxicity for this plant isolate.

The urinary light-chain ladder pattern. A product of improved methodology that may complicate the recognition of Bence Jones proteinuria.

OBJECTIVE: --To describe the recently reported urinary light-chain "ladder" pattern and to indicate that this phenomenon, which may give rise to confusion with Bence Jones protein (BJP), may be observed during routine examination of 50-fold concentrated urine samples tested by high-resolution agarose gel electrophoresis and immunofixation. METHODS: --Urine samples that were usually submitted for examination for the presence of BJP were concentrated 50-fold. Concentrated urine samples were subjected to immunoelectrophoresis and agarose gel electrophoresis. Samples that failed to show a BJP on immunoelectrophoresis but which did show a faint banding pattern in the stained agarose gel were subjected to immunofixation. RESULTS--Samples of urine from 23 patients failed to show a distinct BJP. Nevertheless, these samples did show a kappa, with or without a lambda, light-chain banding pattern. The urine samples came from patients with serum M components associated with neoplasms of either plasma cells (n = 2) or lymphocytes (n = 2) or with M components of undetermined significance (n = 6). The remainder came from patients with infectious (n = 8), inflammatory (n = 4), or neoplastic (n = 1) processes. Some of these patients had no apparent renal disease, while others had variably altered renal function. CONCLUSIONS--The urinary light-chain ladder pattern was found by routine examination of 50-fold concentrated urine samples subjected to agarose gel electrophoresis and immunofixation. The pattern probably reflects the limited heterogeneity of normal human light chains. Detection in the urine samples of some patients may reflect increased synthesis, failure of resorption/degradation in the kidney, or the interference in proximal tubular function by substances producing transient tubular proteinuria. The presence of the light-chain ladder pattern in urine may prevent the detection of small amounts of BJP sharing the electrophoretic mobility of one of the normal light-chain bands.

Steroid hormone concentration profiles in healthy intact and neutered dogs before and after cosyntropin administration.

The purpose of this study was to determine steroid hormone concentration profiles in healthy intact and neutered male and female dogs. Seventeen intact female dogs, 20 intact male dogs, 30 spayed female dogs, and 30 castrated male dogs were used in this study. Serum samples were collected before and 1h after cosyntropin administration, and serum concentrations were determined for cortisol, progesterone, 17-OH progesterone (17-OHP), dehydroepiandrosterone sulfate (DHEAS), androstenedione, testosterone, and estradiol. Intact male dogs had greater concentrations of DHEAS, androstenedione, and testosterone. Intact female dogs had greater concentrations of progesterone. There was no significant difference in estradiol concentration among the four groups. Intact male dogs had lower concentrations of cortisol post-stimulation. DHEAS and testosterone did not increase in response to ACTH in intact males, and estradiol concentrations did not increase in response to ACTH in any group. Results from this study will enhance interpretation of suspected adrenal and/or gonadal disorders of dogs. Because estradiol concentrations were similar in all groups of dogs, measuring estradiol may not be a useful diagnostic test. Cortisol concentrations for intact male dogs with hyperadrenocorticism may be lower than those of female or neutered dogs.

The role of lysosomes in the pathogenesis of copper-induced hepatotoxicity: morphological studies.

Copper-induced hepatotoxicity was evaluated in beige and conventional mice which were injected intraperitoneally with aqueous copper chloride (8 mg per kg body weight per day). Hepatic lesions developed more quickly and were more severe in beige mice and consisted of karyomegaly, progressive accumulation of lipofuscin within hepatocytes and Kupffer cells, acute necrosis of random hepatocytes, and aggregation of leukocytes within the sinusoids. In both groups of mice the hepatic lesion had a centrilobular distribution and closely paralleled the appearance and distribution of copper within hepatocytes. Overall, beige mice had more severe copper-mediated hepatotoxicity, suggesting that normal lysosomes have protective effects for hepatocytes by sequestering copper from other cytoplasmic constituents.

Assessment of the mitogenic potential of the alkaloids produced by endophyte (Acremonium coenophialum)-infected tall fescue (Festuca arundinacea) on bovine vascular smooth muscle in vitro.

The objective of these experiments was to test the hypothesis that the major alkaloid classes found in endophyte-infected tall fescue could act as growth promoters for vascular smooth muscle. Bovine vascular smooth muscle cells (VSMC) from the dorsal metatarsal artery were grown in vitro and exposed to five concentrations (10(-6), 10(-8), 10(-9), 10(-11) and 0 M) of ergonovine, alpha-ergocryptine, ergovaline, and N-acetyl loline for 48 h. The mitogenic potential of the alkaloids was tested on both actively growing cells (serum fed, 10% fetal bovine serum) and quiescent cells (serum starved, 0.1% serum) to assess the need for serum constituents for alkaloid induced growth or toxicity. Ergonovine stimulated (P < .07) VSMC growth at 10(-6) M concentration in growing and quiescent culturs and at 10(-8) M concentration in quiescent cultures. alpha-Ergocryptine stimulated (P < .01) growth at 10(-6) M concentration in growing cultures and at 10(-8) and 10(-9) M concentrations in quiescent cultures. Ergovaline exhibited a dual activity on the growth of VSMC in culture, stimulating (P = .06) growth of quiescent cells at 10(-9) M concentration but inhibiting (P < .05) growth of growing cultures at concentrations of 10(-6) and 10(-9) M. This duality of activity was also noted for N-acetyl loline: N-acetyl loline stimulated (P < .05) growth of quiescent cultures at concentrations of 10(-8), 10(-9), and 10(-11) M but inhibited (P < .05) growth of growing cultures at concentrations of 10(-8) and 10(-9) M. The growth effects of the alkaloids in vitro on VSMC support in part the hypothesis that the alkaloids may contribute to the vascular complications noted in cattle grazing endophyte-infected tall fescue through hyperplasia of the intima. This would result in a decreased luminal diameter of the blood vessels and a resultant decrease in blood flow to the afflicted tissues. The diminished blood flow to tissues would result in tissue death and reduced ability to dissipate heat.

Succinylcholine chloride as a euthanatizing agent in dogs.

In order to determine the efficacy of succinylcholine chloride as a euthanatizing agent, the possible situation encountered in mass euthanasia was recreated. While anesthetized or awake, sixteen 2-year-old Beagles of both sexes were subcutaneously injected with either 1.1 g or 11 mg of succinylcholine chloride/kg of body weight. Positive pressure respiration was given to 2 dogs, otherwise the treatment was allowed to run its course. Electrocardiogram, electroencephalogram, and impedance pneumogram recordings were obtained for time comparisons and for determination of the ultimate cause of death.

Phototoxic effect of Thamnosma texana (Dutchman's breeches) in sheep.

Oral exposure of sheep to the airdried, aerial portions of Thamnosma texana (Dutchman's breeches) resulted in severe photosensitization. Sheep fed the plant at 9 or 12 g/kg of body weight/day and held in direct sunlight exhibited signs of phototoxicosis within 24 to 48 hours. The clinical signs manifested were increased body temperature; photophobia; edema of the muzzle, ears, and vulva; keratoconjunctivitis with edema of the cornea; and exudative dermatitis of the skin of the ears, muzzle, and vulva. Lesser dosages of the plant produced similar effects after several days, except that corneal edema and opacity were not seen. Histopathologic studies indicated no hepatic lesions, consistent with primary photosensitization. The photosensitizing effects of T texana can be attributed to the presence of photosensitizing linear furocoumarins (psoralens) in the plant.

Photosensitization of cattle in southeast Texas: identification of phototoxic activity associated with Cooperia pedunculata.

A microbiological assay (Candida albicans) was used to screen plants in southeast Texas where bovine photosensitization (PS) of unknown cause was a recurring problem. Phototoxic activity was identified associated with dead leaf tips of Cooperia pedunculata, a native, perennial forb of the Amaryllis family (Amaryllidaceae) from central, southeast, and south Texas and parts of Mexico. A syndrome compatible with naturally occurring PS in cattle was induced in laboratory mice after oral administration of dead leaf material from C pedunculata. Availability and phototoxic activity of dead leaf material of C pedunculata corresponded with occurrence of PS in cattle. Seemingly, C pedunculata was involved in recurring PS.