RESUMO
Scrapie is a transmissible spongiform encephalopathy that can be transmitted amongst susceptible sheep. The prion protein gene (PRNP) profoundly influences the susceptibility of sheep to the scrapie agent. This study reports the failure to detect PrP(Sc) in nervous or lymphoid tissues of Suffolk sheep of the PRNP ARQ/ARR genotype after oral inoculation with a U.S. scrapie isolate. Lambs were inoculated within the first 24 h of birth with 1 ml of a 10% (wt./vol.) brain homogenate derived from a clinically affected ARQ/ARQ sheep. The inoculated sheep were observed daily throughout the experiment for clinical signs suggestive of scrapie until they were necropsied at 86 months post inoculation. Tissues were collected for examination by immunohistochemistry and enzyme immunoassay, but all failed to demonstrate evidence of scrapie infection. Neonatal sheep of the ARQ/ARQ genotype receiving the same inoculum developed scrapie within 24 months. Lambs of the ARQ/ARR genotype that received the same inoculum by intracranial inoculation develop scrapie with a prolonged incubation period and with abnormal prion present within the central nervous system, but not peripheral lymphoid tissues. Results of this study suggest that ARQ/ARR sheep are resistant to oral infection with the scrapie isolate used even during the neonatal period.
Assuntos
Sistema Nervoso Central/fisiopatologia , Genótipo , Tecido Linfoide/fisiopatologia , Proteínas PrPSc/metabolismo , Scrapie/genética , Animais , Sistema Nervoso Central/metabolismo , Técnicas Imunoenzimáticas/veterinária , Imuno-Histoquímica/veterinária , Tecido Linfoide/metabolismo , Ovinos , Vacinação/veterináriaRESUMO
Sheep scrapie is a transmissible spongiform encephalopathy that can be transmitted horizontally. The prion protein gene (PRNP) profoundly influences the susceptibility of sheep to the scrapie agent and the tissue levels and distribution of PrPSc in affected sheep. The purpose of this study was to compare the survival time and PrPSc tissue distribution in sheep with highly resistant and highly susceptible PRNP genotypes after intracranial inoculation of the agent of scrapie. Five sheep each of genotype VRQ/VRQ, VRQ/ARR or ARQ/ARR were inoculated. Sheep were euthanized when clinical signs of scrapie became severe. Clinical signs, microscopic lesions, and western blot profiles were uniform across genotypes and consistent with manifestations of classical scrapie. Mean survival time differences were associated with the 171 polymorphic site with VRQ/VRQ sheep surviving 18 months, whereas VRQ/ARR and ARQ/ARR sheep survived 60 and 56 months, respectively. Labeling of PrPSc by immunohistochemistry revealed similar accumulations in central nervous system tissues regardless of host genotype. Immunoreactivity for PrPSc in lymphoid tissue was consistently abundant in VRQ/VRQ, present but confined to tonsil or retropharyngeal lymph node in 4/5 VRQ/ARR, and totally absent in ARQ/ARR sheep. The results of this study demonstrate the susceptibility of sheep with the ARQ/ARR genotype to scrapie by the intracranial inoculation route with PrPSc accumulation in CNS tissues, but prolonged incubation times and lack of PrPSc in lymphoid tissue.
Assuntos
Tecido Linfoide/metabolismo , Proteínas PrPSc/metabolismo , Scrapie/metabolismo , Animais , Tronco Encefálico/metabolismo , Tronco Encefálico/patologia , Estudos de Associação Genética , Predisposição Genética para Doença , Tecido Linfoide/patologia , Especificidade de Órgãos , Proteínas PrPSc/genética , Scrapie/genética , Scrapie/patologia , Carneiro DomésticoRESUMO
Cattle could be exposed to the agent of chronic wasting disease (CWD) through contact with infected farmed or free-ranging cervids or exposure to contaminated premises. The purpose of the current study was to assess the potential for CWD derived from elk to transmit to cattle after intracranial inoculation. Calves (n = 14) were inoculated with brain homogenate derived from elk with CWD to determine the potential for transmission and to define the clinicopathologic features of disease. Cattle were necropsied if clinical signs occurred or at the end of the study (49 months postinoculation; MPI). Clinical signs of poor appetite, weight loss, circling, and bruxism occurred in 2 cattle (14%) at 16 and 17 MPI, respectively. Accumulation of abnormal prion protein (PrP(Sc)) occurred in only the 2 clinically affected cattle and was confined to the central nervous system, with the most prominent immunoreactivity in midbrain, brainstem, and hippocampus with lesser immunoreactivity in the cervical spinal cord. The rate of transmission was lower than in cattle inoculated with CWD derived from mule deer (38%) or white-tailed deer (86%). Additional studies are required to fully assess the potential for cattle to develop CWD through a more natural route of exposure, but a low rate of transmission after intracranial inoculation suggests that risk of transmission through other routes is low. A critical finding is that if CWD did transmit to exposed cattle, currently used diagnostic techniques would detect and differentiate it from other prion diseases in cattle based on absence of spongiform change, distinct pattern of PrP(Sc) deposition, and unique molecular profile.
Assuntos
Doenças dos Bovinos/etiologia , Cervos , Suscetibilidade a Doenças/veterinária , Doença de Emaciação Crônica/transmissão , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/patologia , Transmissão de Doença Infecciosa/veterinária , Doença de Emaciação Crônica/diagnóstico , Doença de Emaciação Crônica/patologiaRESUMO
Sheep scrapie susceptibility or resistance is a function of genotype, with polymorphisms at codon 171 in the sheep prion gene playing a major role. Glutamine (Q) at codon 171 contributes to scrapie susceptibility, while arginine (R) is associated with resistance. In some breeds, lysine (K) occurs at codon 171, but its effect on scrapie resistance has not been determined. Charge and structural similarities between K and R suggest that they may contribute to prion disease susceptibility in a similar way, but studies have not been performed to confirm this. The purpose of the current study was to compare susceptibility and incubation times of AA(136)RR(154)QQ(171) (where the letter denotes the amino acid and the number the position) with AA(136)RR(154)QK(171) sheep after inoculation with scrapie. Barbado AA(136)RR(154)QQ(171) and AA(136)RR(154)QK(171) sheep were inoculated with scrapie intracerebrally to assess their susceptibility to scrapie. After inoculation, sheep were observed daily for clinical signs and were euthanized and necropsied after clinical signs were unequivocal. Tissues were collected at necropsy for immunohistochemistry and Western blot analyses. The QQ(171) sheep had clinical signs approximately 12 months after inoculation, whereas QK(171) animals had an average incubation time of 30 months to onset of clinical signs. The distribution of abnormal prion protein was similar in QQ(171) and QK(171) sheep. Results of the study indicate that sheep with a single K allele at codon 171 are susceptible to scrapie but with a prolonged incubation time. Work is currently underway to examine relative scrapie susceptibility or resistance of KK(171) sheep.
Assuntos
Lisina/genética , Príons/genética , Scrapie/genética , Animais , Western Blotting , Encéfalo/patologia , Códon , Predisposição Genética para Doença , Genótipo , Imuno-Histoquímica/veterinária , Polimorfismo Genético , Scrapie/patologia , OvinosRESUMO
Scrapie of sheep and chronic wasting disease (CWD) of cervids are transmissible prion diseases. Milk and placenta have been identified as sources of scrapie prions but do not explain horizontal transmission. In contrast, CWD prions have been reported in saliva, urine and feces, which are thought to be responsible for horizontal transmission. While the titers of CWD prions have been measured in feces, levels in saliva or urine are unknown. Because sheep produce ~17 L/day of saliva, and scrapie prions are present in tongue and salivary glands of infected sheep, we asked if scrapie prions are shed in saliva. We inoculated transgenic (Tg) mice expressing ovine prion protein, Tg(OvPrP) mice, with saliva from seven Cheviot sheep with scrapie. Six of seven samples transmitted prions to Tg(OvPrP) mice with titers of -0.5 to 1.7 log ID50 U/ml. Similarly, inoculation of saliva samples from two mule deer with CWD transmitted prions to Tg(ElkPrP) mice with titers of -1.1 to -0.4 log ID50 U/ml. Assuming similar shedding kinetics for salivary prions as those for fecal prions of deer, we estimated the secreted salivary prion dose over a 10-mo period to be as high as 8.4 log ID50 units for sheep and 7.0 log ID50 units for deer. These estimates are similar to 7.9 log ID50 units of fecal CWD prions for deer. Because saliva is mostly swallowed, salivary prions may reinfect tissues of the gastrointestinal tract and contribute to fecal prion shedding. Salivary prions shed into the environment provide an additional mechanism for horizontal prion transmission.
Assuntos
Cervos/metabolismo , Príons/metabolismo , Saliva/metabolismo , Proteínas e Peptídeos Salivares/metabolismo , Ovinos/metabolismo , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Determinação de Ponto Final , Estimativa de Kaplan-Meier , Camundongos , Camundongos Transgênicos , Scrapie/patologia , Scrapie/transmissão , Fatores de TempoRESUMO
Here we describe gross and microscopic sweat gland tumors found in a transgenic mouse model of breast cancer, which had transforming growth factor α under the control of mouse mammary tumor virus promoter (MMTV-TGFα). Initially, 20% of the mice in the colony were affected. Cystic lesions formed on the phalanges, palmar surfaces of the metacarpals, and plantar surfaces of the metatarsals. The lesions were multifocal and nonulcerated with straw-colored fluid, ranging in size from 1 to 30 mm at the largest dimension. The colony was monitored for 6 mo; during that time, the prevalence of lesions increased to 52% of the mice. Histologically, in most cases the cyst walls were lined by 1 or 2 layers of normal-appearing epithelial cells that resembled basal cells, indicating adenoma. However, 2 cysts from 2 different mice had papillary proliferative projections and extensive disorganized glandular structures that protruded into the cyst cavities, indicating adenocarcinoma. In these 2 cases, the neoplastic cells revealed architectural and cytologic atypia with rare mitoses. Similar findings have previously been observed in sweat gland tumors; however, multiple sweat-gland tumors have not been reported in mice.
Assuntos
Adenocarcinoma/patologia , Neoplasias da Mama/genética , Cistos/patologia , Extremidades/patologia , Neoplasias das Glândulas Sudoríparas/patologia , Adenocarcinoma/epidemiologia , Animais , Cistos/epidemiologia , Feminino , Vírus do Tumor Mamário do Camundongo/genética , Camundongos , Camundongos Transgênicos , Prevalência , Regiões Promotoras Genéticas/genética , Neoplasias das Glândulas Sudoríparas/epidemiologia , Fator de Crescimento Transformador alfa/genética , Fator de Crescimento Transformador alfa/metabolismoRESUMO
BACKGROUND: Three distinct forms of bovine spongiform encephalopathy (BSE), defined as classical (C-), low (L-) or high (H-) type, have been detected through ongoing active and passive surveillance systems for the disease.The aim of the present study was to compare the ability of two sets of immunohistochemical (IHC) and Western blot (WB) BSE confirmatory protocols to detect C- and atypical (L- and H-type) BSE forms.Obex samples from cases of United States and Italian C-type BSE, a U.S. H-type and an Italian L-type BSE case were tested in parallel using the two IHC sets and WB methods. RESULTS: The two IHC techniques proved equivalent in identifying and differentiating between C-type, L-type and H-type BSE. The IHC protocols appeared consistent in the identification of PrPSc distribution and deposition patterns in relation to the BSE type examined. Both IHC methods evidenced three distinct PrPSc phenotypes for each type of BSE: prevailing granular and linear tracts pattern in the C-type; intraglial and intraneuronal deposits in the H-type; plaques in the L-type.Also, the two techniques gave comparable results for PrPSc staining intensity on the C- and L-type BSE samples, whereas a higher amount of intraglial and intraneuronal PrPSc deposition on the H-type BSE case was revealed by the method based on a stronger demasking step.Both WB methods were consistent in identifying classical and atypical BSE forms and in differentiating the specific PrPSc molecular weight and glycoform ratios of each form. CONCLUSIONS: The study showed that the IHC and WB BSE confirmatory methods were equally able to recognize C-, L- and H-type BSE forms and to discriminate between their different immunohistochemical and molecular phenotypes. Of note is that for the first time one of the two sets of BSE confirmatory protocols proved effective in identifying the L-type BSE form. This finding helps to validate the suitability of the BSE confirmatory tests for BSE surveillance currently in place.
RESUMO
BACKGROUND: Formalin-fixed paraffin-embedded tissue is regularly employed in the diagnosis of transmissible spongiform encephalopathies (TSE) by immunohistochemistry (IHC), the standard by which all other TSE diagnostic protocols are judged. While IHC affords advantages over diagnostic approaches that typically utilize fresh or frozen tissue, such as Western blot and ELISA, the process of fixing, staining, and analyzing individual sections by hand does not allow for rapid or high throughput screening. However, preservation of tissues in formalin is not dependent upon the availability of refrigeration. FINDINGS: Formalin-fixed paraffin-embedded tissues from TSE transmission studies of scrapie in sheep, chronic wasting disease in white-tailed deer or transmissible mink encephalopathy in cattle were cut at 5 µm thickness. Samples containing the tissue equivalent of as little as one 5 µm section can be used to readily discriminate positive from negative samples. CONCLUSIONS: This approach cannot replace IHC but may be used along with IHC as both a more rapid and readily high throughput screen where fresh or frozen tissues are not available or impractical.
RESUMO
Prion diseases or transmissible spongiform encephalopathies (TSEs) of animals include scrapie of sheep and goats; transmissible mink encephalopathy (TME); chronic wasting disease (CWD) of deer, elk and moose; and bovine spongiform encephalopathy (BSE) of cattle. The emergence of BSE and its spread to human beings in the form of variant Creutzfeldt-Jakob disease (vCJD) resulted in interest in susceptibility of cattle to CWD, TME and scrapie. Experimental cross-species transmission of TSE agents provides valuable information for potential host ranges of known TSEs. Some interspecies transmission studies have been conducted by inoculating disease-causing prions intracerebrally (IC) rather than orally; the latter is generally effective in intraspecies transmission studies and is considered a natural route by which animals acquire TSEs. The "species barrier" concept for TSEs resulted from unsuccessful interspecies oral transmission attempts. Oral inoculation of prions mimics the natural disease pathogenesis route whereas IC inoculation is rather artificial; however, it is very efficient since it requires smaller dosage of inoculum, and typically results in higher attack rates and reduces incubation time compared to oral transmission. A species resistant to a TSE by IC inoculation would have negligible potential for successful oral transmission. To date, results indicate that cattle are susceptible to IC inoculation of scrapie, TME, and CWD but it is only when inoculated with TME do they develop spongiform lesions or clinical disease similar to BSE. Importantly, cattle are resistant to oral transmission of scrapie or CWD; susceptibility of cattle to oral transmission of TME is not yet determined.
Assuntos
Encefalopatia Espongiforme Bovina/transmissão , Doenças Priônicas/transmissão , Animais , Bovinos , Doenças dos Bovinos/transmissão , Síndrome de Creutzfeldt-Jakob/transmissão , Síndrome de Creutzfeldt-Jakob/veterinária , Cervos , Suscetibilidade a Doenças/veterinária , Doenças das Cabras/transmissão , Cabras , Humanos , Príons/patogenicidade , Scrapie/transmissão , Ovinos , Doenças dos Ovinos/transmissão , Doença de Emaciação Crônica/transmissãoRESUMO
The raccoon (Procyon lotor) is almost ubiquitous in North America. In recent times, it was introduced in many parts of the world where it has now become largely feral. Since the outbreak of raccoon rabies epizootic in eastern United States and Canada, most diagnostic laboratories have had increased numbers of raccoon carcasses or raccoon brain submissions for diagnosis of rabies. However, since a number of other diseases that affect the central nervous system and have similar clinical signs as rabies have been documented in this species, the current review attempts to bring together the published information on neurologic disorders of raccoons.
Assuntos
Doenças do Sistema Nervoso/veterinária , Guaxinins , Animais , Doenças do Sistema Nervoso/diagnóstico , Doenças do Sistema Nervoso/epidemiologia , Doenças do Sistema Nervoso/patologia , América do Norte/epidemiologiaRESUMO
Final observations on experimental transmission of chronic wasting disease (CWD) from elk (Cervus elaphus nelsoni) and white-tailed deer (Odocoileus virginianus) to fallow deer (Dama dama) are reported herein. During the 5-year study, 13 fawns were inoculated intracerebrally with CWD-infected brain material from white-tailed deer (n = 7; Group A) or elk (n = 6; Group B), and 3 other fawns were kept as uninoculated controls (Group C). As described previously, 3 CWD-inoculated deer were euthanized at 7.6 mo post-inoculation (MPI). None revealed presence of abnormal prion protein (PrP(d)) in their tissues. At 24 (Group A) and 26 (Group B) MPI, 2 deer were necropsied. Both animals had a small focal accumulation of PrP(d) in their midbrains. Between 29 and 37 MPI, 3 other deer (all from Group A) were euthanized. The 5 remaining deer became sick and were euthanized between 51 and 60 MPI (1 from Group A and 4 from Group B). Microscopic lesions of spongiform encephalopathy (SE) were observed in only these 5 animals; however, PrP(d) was detected in tissues of the central nervous system by immunohistochemistry, Western blot, and by commercial rapid test in all animals that survived beyond 24 MPI. This study demonstrates that intracerebrally inoculated fallow deer not only amplify CWD prions, but also develop lesions of spongiform encephalopathy.
Assuntos
Transplante de Tecido Encefálico/veterinária , Cervos , Príons/isolamento & purificação , Doença de Emaciação Crônica/transmissão , Animais , Príons/administração & dosagem , Doença de Emaciação Crônica/patologiaRESUMO
Hepatocellular carcinoma (HCC) and pancreatic carcinoma (PC) cells often have inherent urea cycle defects rendering them auxotrophic for the amino acid l-arginine (l-arg). Most HCC and PC require extracellular sources of l-arg and undergo cell cycle arrest and apoptosis when l-arg is restricted. Systemic, enzyme-mediated depletion of l-arg has been investigated in mouse models and human trials. Non-human enzymes elicit neutralizing antibodies, whereas human arginases display poor pharmacological properties in serum. Co(2+) substitution of the Mn(2+) metal cofactor in human arginase I (Co-hArgI) was shown to confer more than 10-fold higher catalytic activity (k(cat)/K(m)) and 5-fold greater stability. We hypothesized that the Co-hArgI enzyme would decrease tumor burden by systemic elimination of l-arg in a murine model. Co-hArgI was conjugated to 5-kDa PEG (Co-hArgI-PEG) to enhance circulation persistence. It was used as monotherapy for HCC and PC in vitro and in vivo murine xenografts. The mechanism of cell death was also investigated. Weekly treatment of 8 mg/kg Co-hArgI-PEG effectively controlled human HepG2 (HCC) and Panc-1 (PC) tumor xenografts (P = .001 and P = .03, respectively). Both cell lines underwent apoptosis in vitro with significant increased expression of activated caspase-3 (P < .001). Furthermore, there was evidence of autophagy in vitro and in vivo. We have demonstrated that Co-hArgI-PEG is effective at controlling two types of l-arg-dependent carcinomas. Being a nonessential amino acid, arginine deprivation therapy through Co-hArgI-PEG holds promise as a new therapy in the treatment of HCC and PC.
RESUMO
In nature, free-ranging raccoons typically do not live longer than 2 y; most raccoons in the wild die young due to accidents and diseases. Therefore, few data are available regarding lesions associated with advancing age in raccoons. This communication documents the lesions present in raccoons (7 male; 3 female) that were older than 7 y and had been used as breeders at a commercial facility in central Iowa. The most frequent microscopic lesions in these raccoons included accumulation of iron pigment in livers and spleens (10 of 10 animals evaluated), neuroaxonal degeneration in caudal medulla (10 of 10), vascular mineralization (psammoma body) in choroid plexus (9 of 10), myocardial inclusions (7 of 8), and cystic endometrial hyperplasia (2 of 3). Other conditions were seen with less prevalence. Except for the detection of gastritis with bacteria in the gastric mucosa of 1 raccoon, the presence of inflammatory cells in 3 choroid plexuses, and the presence of Lafora bodies in the brain of 1 animal, all conditions observed had previously been reported in raccoons. Surprisingly, islet-cell amyloidosis, previously observed as common incidental finding in older captive raccoons, was not seen in any of the raccoons we examined. Because free-ranging raccoons are distributed over wide geographic areas, their local environment may have considerable influence on the range of spontaneous lesions that would occur in raccoons obtained from a specific location. Therefore, the lesions found in these raccoons from central Iowa may differ from those of other raccoon populations.
Assuntos
Envelhecimento/patologia , Plexo Corióideo/patologia , Miocárdio/patologia , Sistema Nervoso/patologia , Guaxinins/fisiologia , Envelhecimento/fisiologia , Animais , Cruzamento , Plexo Corióideo/fisiopatologia , Feminino , Coração/fisiopatologia , Iowa , Ferro/metabolismo , Fígado/metabolismo , Fígado/patologia , Masculino , Sistema Nervoso/fisiopatologia , Baço/metabolismo , Baço/patologiaRESUMO
The primary objective of this study was to determine whether or not Spiroplasma mirum would be capable of producing lesions of transmissible spongiform encephalopathy (TSE) when inoculated in raccoons (Procyon lotor) and, if that was possible, to compare the clinicopathological findings with those of transmissible mink encephalopathy (TME) in the same experimental model. For this purpose, 5 groups (n = 5) of raccoon kits were inoculated intracerebrally with either S. mirum and/or TME. Two other groups (n = 5) of raccoon kits served as sham-inoculated controls. All animals inoculated with TME, either alone or in combination, showed clinical signs of neurologic disorder and were euthanized within 6 mo post-inoculation (MPI). None of the carcasses revealed gross lesions. Spongiform encephalopathy was observed by light microscopy and the presence of abnormal disease-causing prion protein (PrP(d)) was detected by immunohistochemistry (IHC) and Western blot (WB) techniques in only the raccoons administered TME. Raccoons inoculated with Spiroplasma, but not administered TME agent, were euthanized at 30 MPI. They did not show clinical neurologic signs, their brains did not have lesions of spongiform encephalopathy, and their tissues were negative for S. mirum by polymerase chain reaction (PCR) and for PrP(d) by IHC and WB techniques. The results of this study indicate that Spiroplasma mirum does not induce TSE-like disease in raccoons.
Assuntos
Doenças Priônicas/veterinária , Guaxinins/microbiologia , Spiroplasma/patogenicidade , Animais , Encéfalo/metabolismo , Encéfalo/microbiologia , Encéfalo/patologia , Doenças Priônicas/microbiologia , Doenças Priônicas/patologia , Doenças Priônicas/fisiopatologia , Príons/análise , Distribuição AleatóriaRESUMO
Here we document the case of a domestic ferret (Mustela putorius) that survived experimental inoculation with rabies virus of skunk origin. The ferret showed initial clinical signs of rabies (hindlimb paralysis) on day 81 after inoculation. The animal survived with paraplegia but otherwise was in an adequate nutritional state until the end of the observation period (PI day 181). At necropsy, no gross lesions were observed. Microscopic lesions were found in sections of cerebrum and spinal cord. In both tissues, the lesions were similar but were more severe with loss of neuronal parenchyma in the spinal cord. The lesions consisted of locally extensive areas with proliferation of astrocytes and moderate numbers of glial cells. Severely affected areas also contained clearly defined vacuoles in the neuropil. Multifocal areas of involvement showed mononuclear cuffing of blood vessels. In a few areas, the cuffing extended to the meninges. Rabies virus antigen was not detected by immunohistochemistry of tissue sections.
Assuntos
Furões/virologia , Vírus da Raiva/fisiologia , Raiva/veterinária , Animais , Antígenos Virais/análise , Cérebro/patologia , Cérebro/virologia , Paraplegia/veterinária , Paraplegia/virologia , Raiva/patologia , Raiva/virologia , Recuperação de Função Fisiológica , Medula Espinal/patologia , Medula Espinal/virologiaRESUMO
There is a paucity of data regarding the safety of administering solid gold nanoparticles (AuNPs) in large animal tumor models. We assessed the acute toxicity and biodistribution of 5 nm and 25 nm solid AuNPs in New Zealand White rabbits (n = 6 in each) with implanted liver Vx2 tumors 24 h after intravenous injection. Gold concentration was determined by inductively coupled plasma atomic emission spectrometry (ICP) and imaged with transmission electron microscopy (TEM). There was no clinico-pathologic evidence of renal, hepatic, pulmonary, or other organ dysfunction. After 25 nm AuNP administration, the concentration of white blood cells increased after treatment (p = 0.001). Most other blood studies were unchanged. AuNPs were distributed to the spleen, liver, and Vx2 tumors, but not to other tissues. The urinary excretion of AuNPs was bimodal as measured by ICP. 25 nm AuNPs were more evenly distributed throughout tissues and may be better tools for medical therapy.
Assuntos
Ouro/farmacocinética , Ouro/toxicidade , Neoplasias Hepáticas Experimentais/metabolismo , Nanopartículas Metálicas/toxicidade , Animais , Contagem de Células Sanguíneas , Ouro/administração & dosagem , Ouro/urina , Histocitoquímica , Injeções Intravenosas , Neoplasias Hepáticas Experimentais/química , Neoplasias Hepáticas Experimentais/patologia , Espectrometria de Massas , Nanopartículas Metálicas/administração & dosagem , Microscopia Eletrônica de Transmissão , Tamanho da Partícula , Coelhos , Distribuição TecidualRESUMO
PURPOSE: Pancreatic carcinoma is one of the deadliest cancers with few effective treatments. Gold nanoparticles (AuNP) are potentially therapeutic because of the safety demonstrated thus far and their physiochemical characteristics. We used the astounding heating rates of AuNPs in nonionizing radiofrequency (RF) radiation to investigate human pancreatic xenograft destruction in a murine model. EXPERIMENTAL DESIGN: Weekly, Panc-1 and Capan-1 human pancreatic carcinoma xenografts in immunocompromised mice were exposed to an RF field 36 hours after treatment (intraperitoneal) with cetuximab- or PAM4 antibody-conjugated AuNPs, respectively. Tumor sizes were measured weekly, whereas necrosis and cleaved caspase-3 were investigated with hematoxylin-eosin staining and immunofluorescence, respectively. In addition, AuNP internalization and cytotoxicity were investigated in vitro with confocal microscopy and flow cytometry, respectively. RESULTS: Panc-1 cells demonstrated increased apoptosis with decreased viability after treatment with cetuximab-conjugated AuNPs and RF field exposure (P = 0.00005). Differences in xenograft volumes were observed within 2 weeks of initiating therapy. Cetuximab- and PAM4-conjugated AuNPs demonstrated RF field-induced destruction of Panc-1 and Capan-1 pancreatic carcinoma xenografts after 6 weeks of weekly treatment (P = 0.004 and P = 0.035, respectively). There was no evidence of injury to murine organs. Cleaved caspase-3 and necrosis were both increased in treated tumors. CONCLUSIONS: This study demonstrates a potentially novel cancer therapy by noninvasively inducing intracellular hyperthermia with targeted AuNPs in an RF field. While the therapy is dependent on the specificity of the targeting antibody, normal tissues were without toxicity despite systemic therapy and whole-body RF field exposure.
Assuntos
Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/radioterapia , Anticorpos Monoclonais/administração & dosagem , Ouro/uso terapêutico , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/radioterapia , Terapia por Radiofrequência , Adenocarcinoma/patologia , Animais , Anticorpos Monoclonais Humanizados , Antineoplásicos/administração & dosagem , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Linhagem Celular Tumoral , Cetuximab , Terapia Combinada , Sistemas de Liberação de Medicamentos/métodos , Ouro/administração & dosagem , Ouro/efeitos adversos , Humanos , Nanopartículas Metálicas/administração & dosagem , Nanopartículas Metálicas/efeitos adversos , Nanopartículas Metálicas/uso terapêutico , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Neoplasias Pancreáticas/patologia , Ondas de Rádio/efeitos adversos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Here we describe gross and histopathologic findings in a laboratory-confined adult male raccoon (Procyon lotor) with microscopic ossified areas in pulmonary alveoli. At the time of necropsy, gross lesions were present in the kidneys and in one thyroid gland. Noteworthy microscopic findings included multifocal foci of osseous tissue within the alveoli of the lungs, bilateral thyroid adenomas, pancreatic islet cell amyloidosis, cortical kidney infarcts, cystic adenomatous hyperplasia of urinary bladder, and mineralizations (psommama bodies) of small blood vessels of meninges and choroid plexus. Pulmonary ossification in raccoons has not been reported previously. The other histopathologic lesions have been documented to occur as incidental findings in raccoons and do not appear to have any apparent association with the formation of osseous foci in the lungs of the animal described.
Assuntos
Pneumopatias/patologia , Pneumopatias/veterinária , Ossificação Heterotópica/patologia , Ossificação Heterotópica/veterinária , Alvéolos Pulmonares/patologia , Guaxinins , Animais , Autopsia/veterinária , Pneumopatias/complicações , MasculinoRESUMO
Gross and histopathologic findings are described in an adult male raccoon (Procyon lotor) with Staphylococcus aureus infection affecting several organs. At the time of necropsy the carcass was jaundiced and in poor nutritional state. A large, raised, irregular necrotic mass was present in the spleen. The liver had small multifocal, pale yellow, randomly distributed foci throughout the parenchyma. Coagulase-positive Staphylococcus aureus was cultured from both the liver and the spleen. Microscopic lesions were present in spleen, liver, kidney, and heart, which suggested systemic infection. Although S. aureus has been cultured from raccoons previously, the bacterium did not appear to have been associated with pathology. In this case, S. aureus was obtained as a pure culture, but microscopic examination did not show large numbers of bacterial colonies as seen in other species with such lesions.
Assuntos
Animais Selvagens/microbiologia , Guaxinins/microbiologia , Infecções Estafilocócicas/veterinária , Animais , Evolução Fatal , Masculino , Especificidade da Espécie , Infecções Estafilocócicas/diagnóstico , Infecções Estafilocócicas/epidemiologia , Staphylococcus aureus/isolamento & purificaçãoRESUMO
Chronic wasting disease (CWD), a class of neurodegenerative transmissible spongiform encephalopathies (TSE) occurring in cervids, is found in a number of states and provinces across North America. Misfolded prions, the infectious agents of CWD, are deposited in the environment via carcass remains and excreta, and pose a threat of cross-species transmission. In this study tissues were tested from 812 representative mammalian scavengers, collected in the CWD-affected area of Wisconsin, for TSE infection using the IDEXX HerdChek enzyme-linked immunosorbent assay (ELISA). Only four of the collected mammals tested positive using the ELISA, but these were negative when tested by Western blot. While our sample sizes permitted high probabilities of detecting TSE assuming 1% population prevalence in several common scavengers (93%, 87%, and 87% for raccoons, opossums, and coyotes, respectively), insufficient sample sizes for other species precluded similar conclusions. One cannot rule out successful cross-species TSE transmission to scavengers, but the results suggest that such transmission is not frequent in the CWD-affected area of Wisconsin. The need for further surveillance of scavenger species, especially those known to be susceptible to TSE (e.g., cat, American mink, raccoon), is highlighted in both a field and laboratory setting.