Quantitative expression analysis of genes regulated by both obesity and leptin reveals a regulatory loop between leptin and pituitary-derived ACTH.

Absence of the hormone leptin leads to dramatic increases in appetite, food intake, and adiposity. The primary site of action, at least with respect to appetite, is the hypothalamus. Leptin also has significant effects on the function(s) of peripheral organs involved in maintaining body composition. Some of these effects are mediated through direct interaction of leptin with its receptor on the target tissue, and some effects are indirectly mediated through secondary hormonal and neural pathways. Few of the genes that are responsible for regulating body composition and the peripheral effects of leptin are known. We have used a new gene profiling technology to characterize gene expression changes that occur in the pituitary, hypothalamus, fat, muscle, and liver in response to both obesity and treatment with exogenous leptin. These differences were then overlaid to allow the identification of genes that are regulated by obesity and at least partially normalized by leptin treatment. By using this process we have identified five genes (POMC, PC2, prolactin, HSGP25L2G, and one novel) that are both abnormally expressed in the pituitaries of obese mice and are sensitive to the effects of leptin. We also show that adrenocorticotropic hormone appears to be involved in a regulatory loop involving leptin.

IFN-alpha induces autoimmune T cells through the induction of intracellular adhesion molecule-1 and B7.2.

Insulin-dependent diabetes is an autoimmune disease characterized by the loss of the insulin-producing beta cells and the appearance of autoreactive (anti-islet) T cells. The mechanism by which these autoimmune T cells become activated has not been resolved. We demonstrate that the expression of IFN-alpha by the pancreatic beta cells leads to the development of CD4+ T cells that proliferate in the presence of islet Ags. These autoreactive T cells have a Th1 phenotype and are able to lyse islet cells, possibly through an indirect, cytokine-mediated mechanism. We also demonstrate that the pancreatic infiltrating leukocytes in the transgenic mice express increased levels of 87.2 and ICAM-1 and that IFN-alpha can directly induce these two costimulatory molecules on nontransgenic splenic APCs. Treatment of the transgenic mice with Abs against these costimulatory molecules demonstrated that B7.2 is essential for the induction of autoreactive T cells, whereas ICAM-1 contributes to but is not essential for the formation of these autoreactive cells. As B7.2 and ICAM-1 are able to synergize to provide costimulatory signals to naive T cells under conditions of limiting Ag, we propose that IFN-alpha expression normally contributes to the development of an anti-viral cellular immune response, but that uncontrolled expression of this cytokine will induce autoimmunity.

Genetic absence of gamma-interferon delays but does not prevent diabetes in NOD mice.

Cytokines, particularly interferons, may participate in the development of type I diabetes. This involvement could be from direct cytotoxic actions of the interferons on the pancreatic beta-cells or from an indirect influence on the number, activity, or type of inflammatory cells that invade the islets in type I diabetes. To examine directly the role of interferon (IFN)-gamma in a mouse model of type I diabetes, we have introduced an inactivating mutation in the IFN-gamma gene (ifg) into NOD mice. The genetic absence of IFN-gamma does not prevent either insulitis or diabetes in the NOD mice, but it does increase the time to onset. Although it might have been predicted that the absence of IFN-gamma in these mice would lead to an increase in expression of Th2 T-helper cell-related cytokines, we found instead a profound decrease in the expression of two of the characteristic Th2 cytokines, interleukin (IL)-4 and IL-10. We also demonstrate that the splenocytes taken from IFN-gamma-deficient diabetic mice are fully capable of transferring diabetes to naive recipients.

Transgenic knockouts reveal a critical requirement for pancreatic beta cell glucokinase in maintaining glucose homeostasis.

The secretion of insulin is controlled by the rate of glucose metabolism in the pancreatic beta cells. As phosphorylation by glucokinase (GLK) appears to be the rate-limiting step for glucose catabolism in beta cells, this enzyme may be the glucose sensor. To test this possibility and to resolve the relative roles of liver and beta cell GLK in maintaining glucose levels, we have generated mice completely deficient in GLK and transgenic mice in which GLK is expressed only in beta cells. In mice with only one GLK allele, blood glucose levels are elevated and insulin secretion is reduced. GLK-deficient mice die perinatally with severe hyperglycemia. Expression of GLK in beta cells in the absence of expression in the liver is sufficient for survival. These mice demonstrate the critical need for beta cell GLK in maintaining normal glucose levels and provide a novel model for one form of noninsulin-dependent diabetes.

Islet expression of interferon-alpha precedes diabetes in both the BB rat and streptozotocin-treated mice.

The mechanism(s) leading to beta cell dysfunction in type I diabetes has not been defined. We have investigated whether islet expression of IFN alpha could be a cause of the lesions that are hallmarks of type I diabetes. Streptozotocin induces the expression of interferon-alpha by pancreatic islets prior to the diabetes induced by streptozotocin. Increased IFN alpha, induced by poly I/C or expressed from a transgene will exacerbate the diabetogenic effects of streptozotocin. In another rodent model of type I diabetes (the BB rat), islet expression of IFN alpha precedes lymphocytic infiltration and diabetes. As in the streptozotocin model, in the BB rats poly I/C will induce islet expression of IFN alpha and accelerate the onset of diabetes. These results are consistent with the hypothesis that islet expression of IFN alpha participates in causing type I diabetes.

Neutrophil and B cell expansion in mice that lack the murine IL-8 receptor homolog.

Interleukin-8 (IL-8) is a proinflammatory cytokine that specifically attracts and activates human neutrophils. A murine gene with a high degree of homology to the two known human IL-8 receptors was cloned and then deleted from the mouse genome by homologous recombination in embryonic stem (ES) cells. These mice, although outwardly healthy, had lymphadenopathy, resulting from an increase in B cells, and splenomegaly, resulting from an increase in metamyelocytes, band, and mature neutrophils. Thus, this receptor may participate in the expansion and development of neutrophils and B cells. This receptor was the major mediator of neutrophil migration to sites of inflammation and may provide a potential therapeutic target in inflammatory disease.

Induction of type I diabetes by interferon-alpha in transgenic mice.

Type I diabetes is an autoimmune disease involving an interaction between an epigenetic event (possibly a viral infection), the pancreatic beta cells, and the immune system in a genetically susceptible host. The possibility that the type I interferons could mediate this interaction was tested with transgenic mice in which the insulin-producing beta cells expressed an interferon-alpha. These mice developed a hypoinsulinemic diabetes associated with a mixed inflammation centered on the islets. The inflammation and the diabetes were prevented with a neutralizing antibody to the interferon-alpha. Thus, the expression of interferon-alpha by the beta cells could be causal in the development of type I diabetes, which suggests a therapeutic approach to this disease.

Dioxin intoxication from chronic exposure of horses to pentachlorophenol-contaminated wood shavings.

Investigations into the cause of health problems on a horse-breeding farm led to the discovery of high concentrations (630 to 9,810 mg/kg of bedding) of pentachlorophenol in wood shavings used as bedding for horses over a period of 2 to 4 years. Toxicologic signs in the horses were characteristic of toxic effects associated with exposure of polychlorinated dibenzo-p-dioxins and dibenzofurans. Tissue residue analysis confirmed presence of toxic polychlorinated dibenzo-p-dioxin and dibenzofuran isomers known to be in pentachlorophenol, substantiating the bioavailability of polychlorinated dibenzo-p-dioxins and dibenzofurans in the wood shavings. The findings provide evidence that residue concentrations in the range of 2 ng/g of toxic polychlorinated dibenzo-p-dioxin isomers in liver or fat correlate with toxicologic effects in horses.

Osteochondrosis and juvenile spavin in equids.

Thirty-six of 50 young equids examined at necropsy for gross pathologic and histopathologic evidence of osteochondrosis were determined to have lesions characteristic of this disorder in the distal joints of the tarsus. Abnormalities ranged from retained endochondral cores underlying undisturbed articular cartilage surfaces to clefts, subchondral osseous cyst-like lesions, and cartilage ulceration. Our findings supported the conclusion that osteochondrosis may cause spavin in the juvenile equid.

Clinical, viral, and genetic evaluation of equine degenerative myeloencephalopathy in a family of Appaloosas.

A clinical, viral, hematologic , and genetic study was conducted over a 4-year period on a family of Appaloosas with high incidence of clinical ataxia and pathologic features of equine degenerative myeloencephalopathy. Marginal to deficient serum vitamin E (alpha-tocopherol) and blood selenium values were the only other consistent antemortem abnormalities in the affected horses. Members of this family were all descendants of a clinically normal mare and were raised in 3 separate environments with variable quality of feed. All horses had access to pasture grasses. Normal chromosomal karyotypes were found in 11 affected and/or related horses examined. Equine herpesvirus type 2 was isolated from 4 of the horses, but evidence for a role of this virus in the pathogenesis of the disease was not found. The role of antioxidant deficiency in the pathogenesis of neurologic dysfunction in this equine family and in others reported to be affected with equine degenerative myeloencephalopathy remains speculative.

Chemical involution of the equine parotid salivary gland.

The efficacy of eliminating parotid salivary secretions by retrograde infusion of 10% formalin, 2% chlorhexidine, or 2% or 3% silver nitrate solutions was evaluated in 10 horses. Solutions were kept within the parotid salivary gland for 90 seconds after infusion through parotid duct cannulae and then allowed to drain freely. Severed parotid ducts and surgical incisions were left to heal by second intention. All agents eliminated glandular secretions. There was less necrosis and suppurative inflammation after formalin infusion than after chlorhexidine and silver nitrate. Silver nitrate (2% and 3%) caused the most necrosis and inflammation.

Gastrointestinal cancer among cement workers. A case-referent study.

A matched case-referent study was conducted to investigate the risk of gastrointestinal cancer in cement workers. All male cases of cancer in esophagus, stomach, colon and rectum in the period 1922-1988 from two parishes surrounding a cement plant were identified. For each case, four referents were chosen, matched to age and year of death. Employment in the cement plant was confirmed as from plant records or job title noted in the parish books. Of 25 cases with a definite colorectal tumour, 40% had been blue-collar cement workers for 25 years or more, compared with 20% of their referents [odds ratio (OR) 3.2; 95% confidence limits (CI) 1.1-9.4]. There were indications of a dose (time of employment)-response relationship. For 18 cases with an esophageal or stomach tumour, the OR for those ever employed as cement workers was 3.2 (CI 1.0-11), but short-term employment predominated, thus making a causal relationship less likely.

Pharmacokinetics of recombinant human tumor necrosis factor-alpha in rats. Effects of size and number of doses and nephrectomy.

The serum pharmacokinetics of recombinant human tumor necrosis factor-alpha (rHuTNF) were determined in male and female rats under various conditions. The clearance of rHuTNF after iv administration was determined to be saturable over the dose range of 10-63 micrograms/kg. Multiple iv administration did not significantly change the clearance of rHuTNF. Nephrectomy significantly reduced but did not eliminate the clearance of rHuTNF. The data suggest that sites other than the kidney also contribute to the elimination of rHuTNF. Female rats showed significantly reduced clearance of rHuTNF compared to male rats at all doses after single and multiple iv administration in control and nephrectomized animals.

Pericardial haemangiosarcoma in a horse.

A primary pericardial haemangiosarcoma with metastasis to the parietal pleura and left adrenal gland is described in a 22-year-old Clydesdale x Percheron gelding. Although haemangiosarcomas have been reported in horses, the pericardium has not hitherto been described as the primary site of origin.

Coccidioidomycosis in a Sonoran gopher snake, Pituophis melanoleucus affinis.

Coccidioides immitis was isolated from lesions in the lung of a Sonoran Gopher snake, Pituophis melanoleucus affinis. A lactophenol cotton blue mount of a fungal colony grown on Sabouraud's agar revealed coarse, branching, septate hyphae with alternating thick-walled arthroconidia. Histologically, the lung lesions contained spherical, non-budding, double-walled spherules in various stages of maturity. Some spherules contained endospores. Spherules were also present in granulomas associated with the thyroid and pancreas. A diagnosis was made of disseminated coccidioidomycosis, a condition not previously reported in a reptile.

Cervical dorsal spondylosis with spinal cord compression in a black swan (Cygnus atratus).

Cervical dorsal spondylosis with spinal cord compression in a black swan (Cygnus atratus) was confirmed radiographically and morphologically. Clinically ataxia was associated. Noninflammatory, degenerative changes of the synovial joint cartilage were associated with the spondylosis.

Pathology of Campylobacter jejuni abortion in sheep.

Campylobacter jejuni was inoculated intravenously into pregnant ewes on gestation days 114 and 123 to reproduce ovine abortion. All ewes aborted 7-12 days post-inoculation. High numbers of C. jejuni were isolated from ewe tissues (caruncle, bile, cecal feces), fetal tissues, and placenta. C. jejuni colonies were identified in caruncles and placenta by light microscopy and immunoperoxidase techniques. Histologically, inoculated ewes had a severe purulent endometritis with vasculitis. Placentas from inoculated ewes and field cases showed necrosis and purulent inflammation; however, placentas from inoculated ewes had large numbers of bacterial colonies compared to few bacteria found in field cases. Histologically, only one fetus from the inoculated ewes showed lesions (purulent bronchopneumonia), whereas all fetuses from field cases had a distinct bronchopneumonia, and one fetus showed multifocal hepatic necrosis. These results suggest that C. jejuni (serotypes Penner 1 and Lior 2) is an important abortifacient organism for sheep.

Nasomaxillary fibrosarcomas in three young horses.

Nasomaxillary fibrosarcoma was diagnosed in 3 young horses. Clinical signs included epiphora, facial swelling, dyspnea, unilateral serosanguineous nasal discharge, or an external mass. Physical examination, radiography, and cytology were important diagnostic adjuncts. The definitive diagnosis in each case was based on biopsy. In one case, surgical removal of the tumor and extensive curettage of the affected sinus resulted in extended remission from neoplastic disease.

Juvenile granulosa cell tumour in an equine weanling.

A juvenile granulosa cell tumour is described in a 3-month-old Arabian filly. The 32 X 27 X 27 cm tumour had replaced the right ovary. The neoplasm was composed of multiple nodules of neoplastic granulosa cells which formed microfollicles, macrofillicles and cysts, as well as solid nests and trabeculae. The supporting stroma contained theca cells. Mixing of the neoplastic granulosa cells and theca cells in disorderly fashion was noted. The diagnosis was based on age of occurrence and histological appearance.