Disseminated histiocytic sarcoma with hemophagocytosis in a rabbit.

A 7-year-old female domestic rabbit suffered from labored respiration, poor appetite, mild anemia and thrombocytopenia. Radioscopic examination revealed masses in multiple locations including the intrapleural cavity and spleen. Forty-three days after the first visit to a private veterinary clinic, the rabbit died of severe respiratory distress. Microscopically, all of the masses were composed of round to polygonal neoplastic cells with distinct cell borders that were arranged in a sheet pattern. Multinucleated giant neoplastic cells were often observed. Some neoplastic cells had phagocytozed one or more erythrocytes. Immunohistochemical staining revealed that the neoplastic cells expressed vimentin, CD204, Iba-1 and lysozyme, but not CD163. Based on the morphological and immunohistochemical findings, this case was diagnosed as disseminated histiocytic sarcoma with hemophagocytosis.

Murine leukemia virus Gag localizes to the uropod of migrating primary lymphocytes.

UNLABELLED: B and CD4(+) T lymphocytes are natural targets of murine leukemia virus (MLV). Migrating lymphocytes adopt a polarized morphology with a trailing edge designated the uropod. Here, we demonstrate that MLV Gag localizes to the uropod in polarized B cells and CD4(+) T cells. The uropod localization of MLV Gag was dependent on plasma membrane (PM) association and multimerization of Gag but independent of the viral glycoprotein Env. Basic residues in MA that are required for MLV Gag recruitment to virological synapses between HEK293 and XC cells were dispensable for uropod localization in migrating B cells. Ultrastructural studies indicated that both wild-type and basic-residue mutant Gag localized to the outer surface of the PM at the uropod. Late-domain mutant virus particles were seen at the uropod in form of budding-arrested intermediates. Finally, uropods mediated contact between MLV-infected B cells and uninfected T cells to form virological synapses. Our results suggest that MLV, not unlike HIV, accumulates at the uropod of primary lymphocytes to facilitate viral spreading through the formation of uropod-mediated cell-cell contacts. IMPORTANCE: Viruses have evolved mechanisms to coordinate their assembly and budding with cell polarity to facilitate their spreading. In this study, we demonstrated that the viral determinants for MLV Gag to localize to the uropod in polarized B cells are distinct from the requirements to localize to virological synapses in transformed cell lines. Basic residues in MA that are required for the Gag localization to virological synapses between HEK293 and XC cells are dispensable for Gag localization to the uropod in primary B cells. Rather, plasma membrane association and capsid-driven multimerization of Gag are sufficient to drive MLV Gag to the uropod. MLV-laden uropods also mediate contacts between MLV-infected B cells and uninfected T cells to form virological synapses. Our results indicate that MLV accumulates at the uropod of primary lymphocytes to facilitate viral spreading through the formation of uropod-mediated cell-cell contacts.

Underlying mechanisms involved in the decrease of milk secretion during Escherichia coli endotoxin induced mastitis in lactating mice.

Mastitis, the inflammation of mammary glands resulting from bacterial infection, disrupts milk production in lactating mammary glands. In this study, we injected lipopolysaccharide (LPS), one of the endotoxins from Escherichia coli into mouse mammary glands to disrupt milk production, and we investigated the influence of LPS on nutrient uptake, synthesis, and secretion processes for milk component production in alveolar epithelial cells (AEC). The expression of genes relevant to the three-staged milk component production process (nutrient uptake, synthesis, and secretion of milk components) were down-regulated within 12 h after LPS injection in AEC. The internalization of glucose transporter 1 (GLUT-1) from the basolateral membrane to the cytoplasm occurred in accordance with the down-regulation of gene expression 3 h after LPS injection. The abnormal localization of adipophilin and beta-casein was also observed in the LPS-injected mammary glands. SLC7A1, an amino acid transporter, was up-regulated 3 and 6 h after LPS injection. Furthermore, the inactivation of signal transducer and activator of transcription 5 (STAT5) and the activation of STAT3 and nuclear factor-kappa B (NFkappaB) occurred 3 h after LPS injection. These results indicate that the nutrient uptake, synthesis, and secretion of milk components in AEC are rapidly shut down in the lactating mammary glands after LPS injection.

Mutation of the Theiler's virus leader protein zinc-finger domain impairs apoptotic activity in murine macrophages.

The Theiler's murine encephalomyelitis virus (TMEV) leader (L) protein zinc-finger domain was mutated to study its role in cell death in infection of the murine macrophage cell line M1-D, revealing that an intact zinc-finger domain is required for full apoptotic activity. A functional L zinc-finger domain was also required for activation of p38 MAPK that results in phosphorylation and activation of p53, and in turn, alteration of the conformation of the anti-apoptotic proteins Puma and Mcl-1, leading to the release of pro-apoptotic Bax and apoptosis through the intrinsic pathway. TMEV infection also inhibits host protein synthesis, a stress shown by others to induce apoptosis. Since inhibition of host protein synthesis follows rather than precedes activation of MKK3/6 and p38, it seems less likely that it triggers apoptosis in infected cells. Finally, we showed that the levels of reactive oxygen species following infection were consistent with apoptotic rather than necrotic cell death. Thus, these experiments support an important role for the TMEV L protein zinc-finger domain in apoptosis in an infected murine macrophage line.

Virion endocytosis is a major target for murid herpesvirus-4 neutralization.

Herpesviruses consistently transmit from immunocompetent carriers, implying that their neutralization is hard to achieve. Murid herpesvirus-4 (MuHV-4) exploits host IgG Fc receptors to bypass blocks to cell binding, and pH-dependent protein conformation changes to unveil its fusion machinery only after endocytosis. Nevertheless, neutralization remains possible by targeting the virion glycoprotein H (gH)-gL heterodimer, and the neutralizing antibody responses of MuHV-4 carriers are improved by boosting with recombinant gH-gL. We analysed here how gH-gL-directed neutralization works. The MuHV-4 gH-gL binds to heparan sulfate. However, most gH-gL-specific neutralizing antibodies did not block this interaction; neither did they act directly on fusion. Instead, they blocked virion endocytosis and transport to the late endosomes, where membrane fusion normally occurs. The poor endocytosis of gH-gL-neutralized virions was recapitulated precisely by virions genetically lacking gL. Therefore, driving virion uptake appears to be an important function of gH-gL that provides a major target for antibody-mediated neutralization.

Overexpression of PDGFA and its receptor during carcinogenesis of Opisthorchis viverrini-associated cholangiocarcinoma.

Cholangiocarcinoma (CCA) is a crucial health problem in northeastern part of Thailand, which is caused by a combination of Opisthorchis viverrini infection and nitrosamine. A better understanding of its molecular mechanism is an important step to discover and develop the new diagnostics and therapies for CCA. To reveal the involvement of potential genes in the development of CCA, the present study investigated the expression kinetics of platelet-derived growth factor alpha (Pdgfa) and its receptor (Pdgfra) during the tumorigenesis of CCA induced by O. viverrini infection with quantitative RT-PCR, and confirmed the expression with immunohistological staining. The results showed that in the hamster model of opisthorchiasis-associated CCA, the expression of Pdgfa was increased after infection plus N-nitrosodimethylamine (NDMA) administration, reached its peak at 2 months post infection, and remained at the high level until 6 months. Similarly, the expression of Pdgfra was increased time-dependently. The positive immunostaining for PDGFA proteins was observed in the cytoplasm of epithelial tumor cells of hamster CCA. Moreover, the analysis of the expression of these genes in 10 cases of human opisthorchiasis-associated CCA showed that Pdgfa was overexpressed in 80%, and Pdgfra was overexpressed in 40% cases (>3.0 folds, compared with the expressions of adjacent normal tissues). This result suggests that PDGFA is likely involved in the tumorigenesis of opisthorchiasis-associated CCA, and may be a promising candidate biomarker for diagnosis and treatment strategies of CCA.

The antiapoptotic protein Mcl-1 controls the type of cell death in Theiler's virus-infected BHK-21 cells.

Theiler's murine encephalomyelitis virus (TMEV) results in a persistent central nervous system infection (CNS) and immune-mediated demyelination in mice. TMEV largely persists in macrophages (Ms) in the CNS, and infected Ms in vitro undergo apoptosis, whereas the infection of other rodent cells produces necrosis. We have found that necrosis is the dominant form of cell death in BeAn virus-infected BHK-21 cells but that ~20% of cells undergo apoptosis. Mcl-1 was highly expressed in BHK-21 cells, and protein levels decreased upon infection, consistent with onset of apoptosis. In infected BHK-21 cells in which Mcl-1 expression was knocked down using silencing RNAs there was a 3-fold increase in apoptotic cell death compared to parental cells. The apoptotic program switched on by BeAn virus is similar to that in mouse Ms, with hallmarks of activation of the intrinsic apoptotic pathway in a tumor suppressor protein p53-dependent manner. Infection of stable Mcl-1-knockdown cells led to restricted virus titers and increased physical to infectious particle (PFU) ratios, with additional data suggesting that a late step in the viral life cycle after viral RNA replication, protein synthesis, and polyprotein processing is affected by apoptosis. Together, these results indicate that Mcl-1 acts as a critical prosurvival factor that protects against apoptosis and allows high yields of infectious virus in BHK-21 cells.

Capillaria hepatica in man--an overview of hepatic capillariosis and spurious infections.

Capillaria hepatica (syn. for Calodium hepaticum) is a zoonotic nematode parasitizing in the livers of rodents as main hosts and in numerous other mammals including humans. It is the causative agent of the rare conditions of hepatic capillariosis and spurious C. hepatica infections in humans. In this review, 163 reported cases of infestations with this parasite (72 reports of hepatic capillariosis, 13 serologically confirmed infestations and 78 observations of spurious infections) are summarized with an overview on the distribution, symptoms, pathology, diagnosis, serology and therapy of this rare human pathogen.

Spontaneous dwarf rat: a novel model for aging research.

AIM: Dwarf animal models can provide new models for aging research. For the spontaneous dwarf rat (SDR), a dwarf strain derived from the Sprague-Dawley (SD) rat, no data relevant to aging research are available. The present study aimed to examine its growth, hormonal background, lifespan and age-related diseases. METHODS: Male SDR and SD rats were used for growth comparison and for immunohistochemistry and plasma hormonal analysis. SDR of each sex were maintained until natural death and then inspected pathologically. RESULTS: SDR showed an apparent dwarfism in their youth. Immunohistochemistry indicated that the development of growth hormone (GH)-positive cells in the pituitary was insufficient in SDR. In SDR, plasma GH levels were lower than in SD rats. Moreover, both insulin-like growth factor-1 (IGF-1) and insulin levels were decreased compared to levels in SD rats. Male and female SDR showed a mean lifespan of 29.3 +/- 3.3 and 26.8 +/- 5.3 months, respectively. The main neoplastic lesions in SDR were pituitary and mammary tumors. Major non-neoplastic lesions were incisor malocclusion, heart disease, chronic nephropathy (male) and cerebral hemorrhage (female). Most cases of chronic nephropathy were mild. CONCLUSION: Compared with longevity data and pathological data reported for SD rats, the lifespan in SDR was increased by 20-40% in males and 10-20% in females, and SDR had characteristic decreases in pituitary and mammary tumors as well as in severe chronic nephropathy. The SDR, differing in endocrinology, longevity and pathology from the SD rat, is potentially a new animal model for aging research.

Reciprocal affiliation among adolescent rats during a mild group stressor predicts mammary tumors and lifespan.

OBJECTIVE: Although the detrimental physical health effects of social isolation have been known for three decades, the answers to how and why social relationships generally improve health remain elusive. Social relationships are not always beneficial, and we examined a structural dimension that may bring about their salubrious effects: affiliative reciprocity during a stressor. METHODS: In a lifespan study, female rats lived with their sisters and were tested for temperament, affiliative reciprocity during an everyday stressor at puberty, corticosterone response to a stressor, mammary tumor development and diagnosis, and death. RESULTS: Rats that affiliated more reciprocally during a mild group stressor survived longer (p = .0005), having exhibited a lower corticosterone peak in response to an acute novel stressor in late adulthood (p = .0015), and longer time to the development of spontaneous mammary tumors (p = .02). These effects could not be explained solely by the number of affiliative interactions or individual temperament. Indeed, affiliative reciprocity and neophobia were independent and predicted mortality additively (p = .0002). CONCLUSIONS: Affiliative reciprocity during a stressor, a structural quality of social interactions, protected females from early mammary tumor development (the primary pathology in Sprague-Dawley rats) and early all-cause mortality. Conversely, lack of reciprocity (whether disproportionately seeking or receiving attempted affiliation) was as potent a risk factor as neophobia. Thus a social role increased risk additively with individual temperament. Our data indicate that affiliative reciprocity functions as a buffer for everyday stressors and are likely mediated by attenuated reactivity of the hypothalamic-pituitary-adrenal axis.

Balb/Cj male mice do not feminize after infection with larval Taenia crassiceps.

Balb/cJ mice fail to mount an immune response capable of clearing infection with larval Taenia crassiceps. Additionally, male Balb/cJ mice display a lag in larval growth of approximately 3 wk as compared to growth in female mice. It has been reported that male Balb/ cAnN mice generate a protective immune response early in infection, and become permissive to larval growth after they feminize (200-fold increase in serum estradiol and 90% decrease in serum testosterone). To determine if a different strain of Balb/c mice (Balb/cJ) also feminize, serum was collected from infected male mice for 16 wk and levels of 17-beta-estradiol and testosterone were measured via ELISA. In addition, the mounting responses of 12- and 16-wk infected male mice, as well as uninfected control mice, were determined after isolation with a female mouse. The results of these experiments show that male Balb/cJ mice do not feminize during infection with larval T. crassiceps. There was no significant change in serum levels of either 17-beta-estradiol or testosterone during the course of infection (> 16 wk). Moreover, there was no significant decrease in the number of times infected male mice mounted the female mouse as compared to uninfected controls. These results suggest that there may be variances between the substrains of Balb/c mice that lead to the phenotypic differences reported for male Balb/cJ and Balb/cAnN mice.

Implications of increased susceptibility to predation for managing the sylvatic cycle of Echinococcus multilocularis.

The ability to increase the chances that infectious prey are taken by predators is an observed feature of many parasites that rely on one or more predator-prey relationships to complete their life-cycle. In the sylvatic life-cycle of Echinococcus multilocularis - the causative agent of human alveolar echinococcosis-- foxes are the final host, with voles acting as intermediate hosts. Here we review the evidence that E. multilocularis causes increased susceptibility to predation and present a general mathematical model for the sylvatic life-cycle. The ability to increase susceptibility to predation in the intermediate host reduces the sensitivity of the parasite population to adverse conditions. For example, there is no critical density of foxes below which the parasite is expected to die out, even if the effect of the parasite on infected prey is very small. We suggest that increased susceptibility to predation is a plausible explanation for the observed resilience of E. multilocularis during and following field trials of praziquantel baiting.

Clinical impact of persistent Bartonella bacteremia in humans and animals.

Bartonella spp. are emerging vector-borne pathogens that cause persistent, often asymptomatic bacteremia in their natural hosts. As our knowledge progresses, it appears that chronic infection may actually predispose the host to mild, insidious nonspecific manifestations or induce, in selected instances, severe diseases. Persistent asymptomatic bacteremia is most common in animals that serve as the main reservoir for the specific Bartonella. In humans, these organisms are B. bacilliformis and B. quintana. Other Bartonella species, for which humans are not the natural reservoir, tend to cause persistent bacteremia only in immunodeficient individuals. In some of these individuals, endothelial cell proliferation may create lesions such as bacillary angiomatosis or bacillary peliosis. In cats, bacteremia of variable level and continuity may last for years. Some strains of B. henselae may induce clinical manifestations, including fever, mild neurological signs, reproductive disorders, whereas others do not induce clinically obvious disease. Reproductive disorders have also been reported in mice experimentally infected with B. birtlesii. Finally, canids constitute the most interesting naturally occurring animal model for the human disease. Like immunocompetent people, healthy dogs only occasionally demonstrate long-term bacteremia when infected with Bartonella spp. However, some dogs develop severe clinical manifestations, such as endocarditis, and the pathologic spectrum associated with Bartonella spp. infection in domestic dogs is rapidly expanding and resembles the infrequently reported clinical entities observed in humans. In coyotes, persistent bacteremia is more common than in domestic dogs. It will be of interest to determine if coyotes develop clinical or pathological indications of infection.

Mixed pituitary adenoma-gangliocytoma in a female albino rat.

A mixed intrasellar pituitary adenoma-gangliocytoma was found incidentally in an aged female Sprague-Dawley-derived rat. The animal was killed at the end of a 104-week carcinogenicity study. At necropsy, the pituitary fossa was occupied by a large, hemorrhagic nodule compressing and displacing the base of the brain. The lesion consisted of large areas of a prolactin-secreting adenoma surrounding a central island of gangliocytoma. In the latter, ganglion-like cells of varying size exhibited, occasionally, beta-tubulin and neurofilament protein immunoreactivity in their perikarya, while their cell processes expressed intense neurofilament immunoreactivity. Accompanying satellite cells in the neuropil immunostained for glial acidic and S-100 proteins. To the best of our knowledge, the presence of mixed pituitary adenoma-gangliocytoma has not been previously reported in rats.

Occurrence of the parathyroid cyst in golden hamsters.

Parathyroid cyst is a rare lesion, but has clinical significance because of it's ability to mimic a thyroid mass and it's association with hyperparathyroidism. The occurrence and morphology of parathyroid cysts in golden hamsters from neonatal to senile periods were investigated using light and electron microscopy. The results demonstrate the presence of chief cell cysts in the parathyroid glands of 5-day-old hamsters. Some chief cells lining the cyst wall showed mitosis and apoptosis. The existence of chief cell cysts may represent the rapid proliferation of the parathyroid chief cells in 5-day-old hamsters. Ciliated cysts were observed in the parathyroid glands of 5-day-, 1- and 3-month-old hamsters. Three cell types were distinguished in the wall of the ciliated cyst: Ciliated, mucous and basal cells. Ciliated cysts possessed the features of the pharyngeal epithelia without endocrine cells and may arise from embryological remnants of pharyngeal pouches in the neck undergoing cystic degeneration and entrapping portions of parathyroid tissue. The frequency of parathyroid cysts decreased with age.

Inhibitory neurotransmission in lethal spotted mutant mice: a model for Hirschsprung's disease.

BACKGROUND & AIMS: The pathogenesis of Hirschsprung's disease is not well understood. The suitability of the animal model for the unknown pathogenesis of inhibitory neurotransmission in Hirschsprung's disease was investigated. METHODS: Circular smooth muscle strips from the internal anal sphincter (IAS) and distal colon (2, 6, 8, 16, and 24 mm from the anal verge) from normal and Ls/Ls mice (mice homozygous for the lethal spotting mutation that develop fetal megacolon after aganglionosis of the terminal colon) were prepared to record changes in isometric tensions in response to different agents and nonadrenergic, noncholinergic nerve stimulation by electrical field stimulation. RESULTS: Bethanechol was used to produce contraction of the smooth muscle strips of distal colon to record a decrease in the tension. Conversely, the IAS smooth muscle strips developed spontaneous tone. In the normal homozygous mice, electrical field stimulation caused a biphasic response, an initial decrease followed by an after-contraction, whereas in Ls/Ls mice, the predominant response was contraction. All smooth muscle strips from normal and Ls/Ls mice produced relaxation in response to sodium nitroprusside and vasoactive intestinal polypeptide. CONCLUSIONS: Ls/Ls mice may serve as an appropriate animal model to investigate the pathogenesis of the inhibitory neurotransmission in Hirschsprung's disease in the distal colon and IAS.

Epidemiological characterization of newly recognized rat parvovirus, "rat orphan parvovirus".

Newly recognized rat parvovirus (rat orphan parvovirus: ROPV) was examined for viral excretion and persistence in infected rats, and also for infectivity to mice and hamsters. The virus appeared to replicate mainly in lymphoid or hematopoietic tissues, and was detected in feces, urine and oropharynx of the infected rats at 1 to 4 weeks postinfection. The infective virus was also detected in peripheral leukocytes and various tissues at an acute phase of infection, and decreased in every tissue at 8 weeks postinfection. Viral DNA, however, was persistent in lymphoid tissues at least up to 24 weeks postinfection. When the virus was inoculated to mice and hamsters, no evidence of viral production and antibody response was demonstrated. ROPV is assumed to be a variant of the known rat parvovirus which resulted to alter cell tropism and persist in lymphoid or hematopoietic tissues, in order to escape from host immune system.

Inherited prolonged bleeding time and platelet storage pool deficiency in the subtle gray (sut) mouse.

A high proportion of mouse mutants with diluted pigmentation have severely prolonged bleeding times due to platelet storage pool deficiency. The deficiency is associated with concomitant abnormalities in platelet dense granules and coat pigment granules. The coat color of the subtle gray (sut) mouse is diluted to a relatively minor degree. Analysis of platelet serotonin concentration established that this dense granule component similarly is reduced a relatively small amount in this mutant. The subtle gray mouse thus allowed a test of the hypothesis that relatively small changes in platelet dense granule contents may cause discernible increases in bleeding times. Bleeding times of mutant mice were significantly prolonged (3.4-fold) in comparison with those in normal sut/+ controls. These bleeding times were significantly reduced in comparison with other mouse pigment dilution mutants with more severe storage pool deficiency. These results establish the subtle gray mouse as an appropriate animal model for mild storage pool deficiency and human Hermansky-Pudlak syndrome. They indicate, together with related experiments, that bleeding times are highly sensitive to concentrations of platelet dense granule components such as serotonin.

Murine models of polycystic kidney disease.

The current knowledge of human polycystic kidney disease (PKD)--its morphology as well as the current biochemical and molecular understanding of the disease- has been enormously aided by the existence of a variety of animal models. In mice, several spontaneous mutations have been identified that give rise to PKD. Furthermore, it has been possible to create experimental models of renal cystic disease by genetic manipulation. All these different models have been very informative in studying the role of growth hormones, cell differentiation and hyperplasia, ionic transport, oncogene expression and changes in extracellular matrix (ECM) composition during the development of PKD. Furthermore, they have allowed investigators to test different therapeutic approaches in vivo. This article will review the characteristics of the most common murine models of PKD, some of their current uses and the future role of these animal models in the understanding of human renal cystic disease.

Behavioral and physiologic effects of inapparent wound infection in rats.

There is a common notion that rats are resistant to postoperative wound infection because many recover from surgery performed under nonsterile conditions. As a result, nonaseptic surgical techniques are used commonly in rat surgery. Our aim was to determine if these techniques cause wound infection and, if so, whether or not the infection, inapparent to casual observation, creates measurable changes in rat physiology and behavior. Rats subjected to craniotomies or laparotomies and inoculated with 10(8) Staphylococcus aureus or Pseudomonas aeruginosa or sterile saline were tested for open-field activity, freezing behavior, home-cage behavior score, and wheel-running activity. Physiologic indices included lactate dehydrogenase, blood glucose, plasma fibrinogen, complete blood counts, wound bacterial counts and histology scores, body temperature, and body weight. Although no clinical signs were detected by postoperative observation, rats inoculated with bacteria were significantly less active in the open field and the duration of freezing behavior was shorter. Plasma fibrinogen, serum glucose, total white blood cell counts, and wound histology scores were significantly altered in the bacteria-inoculated rats. These findings underscore the need for sterile techniques in rat surgery to avoid confounding experimental data.