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Biomedical subjects

E W Howerth

Publications and source records attributed to E W Howerth.

At least 19 recordsLinked to original sources
PubMed2004

Survivin: a bifunctional inhibitor of apoptosis protein.

Survivin is a recently discovered protein belonging to the inhibitor of apoptosis (IAP) gene family. IAP molecules are characterized by both the presence of a zinc-binding fold termed the baculoviral IAP repeat and the ability to suppress apoptosis. In addition to inhibiting apoptosis, survivin is essential for proper cell division. Survivin is expressed during embryonal development but is absent in most normal, terminally differentiated tissues. Survivin is also upregulated in a variety of human cancers, and its expression in tumors is associated with a more aggressive phenotype, shorter survival times, and a decreased response to chemotherapy. The exact mechanism behind the ability of survivin to inhibit apoptosis is still unclear. Furthermore, it is not known why this protein is upregulated in cancer. The purpose of this article is to provide an overview of the current knowledge of survivin, including its role in cell division and its expression in normal and neoplastic tissues. Although much of the current research in this field is focused on human medicine, this area also has potential significance for veterinary species.

Animals↗
PubMed2002

Failure of porcine reproductive and respiratory syndrome virus to replicate in porcine endothelial cell cultures.

Clinical, gross, and microscopic pathologic and immunohistochemical findings in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV) suggest that PRRSV may replicate in endothelial cells. Endothelial cell cultures from porcine aorta and pulmonary artery were tested for susceptibility to various strains of PRRSV. Cultures were identified as endothelium by light microscopy and immunohistochemical staining for P-selectin and von Willebrand factor. Five strains of PRRSV, i.e., the National Veterinary Services Laboratories, Ames, IA PRRSV strain 130-PDV and 4 field strains isolated from pneumonic lungs, failed to replicate in these porcine large-vessel endothelial cell cultures.

Animals↗
PubMed2001

Photosensitive chemical and laser light treatments decrease epizootic hemorrhagic disease virus associated with in vitro produced bovine embryos.

Photoinactivation was employed to eliminate EHDV-2 from in vitro produced bovine embryos experimentally exposed to this virus. Immature oocytes were matured, fertilized, and cultured in chemically defined conditions. All treatments were performed on zygotes. Developmental potential of zygotes and cell numbers of resulting hatched blastocysts were assessed after exposure to a 1 mW helium neon laser (633 nm, red) for 1, 5, 10, and 15 min; the photosensitive chemicals hematoporphyrin (15 microM) and hypericin (1 and 10 microM) for 15 min; a combination of 10 microM hypericin and laser light for 1, 3, or 5 min; and a combination of 15 microM hematoporphyrin and laser light for 1, 2, or 3 min. There were no significant differences among proportions of embryos developing or cell numbers after treatment with or without exposure to laser light alone for up to 10 min. No differences were observed after exposure of zygotes to photosensitive chemicals alone. Exposure to 10 microM hypericin and 5 min of laser light or 15 microM hematoporphyrin and 2 min of laser light compromised zygote developmental potential. After exposure to 10(6) TCID50/mL EHDV-2 for 90 min groups of 10 zygotes were exposed to 10 microM hypericin or 15 microM hematoporphyrin and laser light to inactivate the virus. Hematoporphyrin was effective with 3 min light exposure at reducing the percentage of EHDV-2 contaminated zygote pools (16.7%) as compared to EHDV-2 exposed pools without treatment (88.9%) but hematoporphyrin + 1 min light was ineffective. Hypericin + 3 min light provided an intermediate effect (55.6%).

Animals↗
PubMed2001

Pathogenesis of experimental vesicular stomatitis virus (New Jersey serotype) infection in the deer mouse (Peromyscus maniculatus).

The pathogenesis of vesicular stomatitis virus (VSV) infection has not been investigated previously in native New World rodents that may have a role in the epidemiology of the disease. In the present study, 45 juvenile and 80 adult deer mice (Peromyscus maniculatus) were inoculated intranasally with VSV New Jersey serotype (VSV-NJ) and examined sequentially over a 7-day period. Virus was detected by means of immunohistochemistry and in situ hybridization in all tissues containing histologic lesions. Viral antigen and mRNA were observed initially in olfactory epithelium neurons, followed by olfactory bulbs and more caudal olfactory pathways in the brain. Virus also was detected throughout the ventricular system in the brain and central canal of the spinal cord. These results support both viral retrograde transneuronal transport and viral spread within the ventricular system. Other tissues containing viral antigen included airway epithelium and macrophages in the lungs, cardiac myocytes, and macrophages in cervical lymph nodes. In a second experiment, 15 adult, 20 juvenile, and 16 nestling deer mice were inoculated intradermally with VSV-NJ. Adults were refractory to infection by this route; however, nestlings and juveniles developed disseminated central nervous system infections. Viral antigen also was detected in cardiac myocytes and lymph node macrophages in these animals. Viremia was detected by virus isolation in 35/72 (49%) intranasally inoculated juvenile and adult mice and in 17/36 (47%) intradermally inoculated nestlings and juveniles from day 1 to day 3 postinoculation. The documentation of viremia in these animals suggests that they may have a role in the epidemiology of vector-borne vesicular stomatitis.

Animals↗
PubMed2001

Contact transmission of vesicular stomatitis virus New Jersey in pigs.

OBJECTIVE: To determine how viral shedding and development or lack of clinical disease relate to contact transmission of vesicular stomatitis virus New Jersey (VSV-NJ) in pigs and determine whether pigs infected by contact could infect other pigs by contact. ANIMALS: 63 pigs. PROCEDURE: Serologically naive pigs were housed in direct contact with pigs that were experimentally inoculated with VSV-NJ via ID inoculation of the apex of the snout, application to a scarified area of the oral mucosa, application to intact oral mucosa, or ID inoculation of the ear. In a second experiment, pigs infected with VSV-NJ by contact were moved and housed with additional naive pigs. Pigs were monitored and sampled daily for clinical disease and virus isolation and were serologically tested before and after infection or contact. RESULTS: Contact transmission developed only when vesicular lesions were evident. Transmission developed rapidly; contact pigs shed virus as early as 1 day after contact. In pens in which contact transmission was detected, 2 of 3 or 3 of 3 contact pigs were infected. CONCLUSIONS AND CLINICAL RELEVANCE: Transmission was lesion-dependent; however, vesicular lesions often were subtle with few or no clinical signs of infection. Contact transmission was efficient, with resulting infections ranging from subclinical (detected only by seroconversion) to clinical (development of vesicular lesions). Long-term maintenance of VSV-NJ via contact transmission alone appears unlikely. Pigs represent an efficient large-animal system for further study of VSV-NJ pathogenesis and transmission.

Animals↗
PubMed2001

A rapid and strong laparoscopic-assisted gastropexy in dogs.

OBJECTIVE: To develop a technique for laparoscopic gastropexy in dogs and evaluate effects on stomach position and strength of the adhesion between the stomach and abdominal wall. ANIMALS: 8 healthy dogs. PROCEDURE: Dogs were anesthetized, and the abdomen was insufflated with carbon dioxide. A laparoscope was placed through a cannula inserted on the abdominal midline caudal to the umbilicus. Babcock forceps placed through a cannula inserted lateral to the right margin of the rectus abdominus muscle were used to exteriorize the pyloric antrum, a longitudinal incision was made through the serosa and muscular layer of the pyloric antrum, and the seromuscular layer of the pyloric antrum was sutured to the transversus abdominus muscle. After surgery, positive-contrast gastrography was used to evaluate stomach position and the onset of gastric emptying, and ultrasonography was used to assess stomach wall activity and mobility. Dogs were euthanatized 1 month after surgery, and tensile strength of the adhesion was tested. RESULTS: In all dogs, stomach position and the onset of gastric emptying were normal 25 days after surgery, and the pyloric antrum was firmly attached to the abdominal wall 30 days after surgery. Mean +/- SD ultimate load of the adhesion in tension was 106.5 +/- 45.6 N. CONCLUSIONS AND CLINICAL RELEVANCE: The laparoscopic gastropexy technique described in the present study could be performed quickly and easily by an experienced surgeon, resulted in a strong fibrous adhesion between the stomach and abdominal wall, and appeared to cause minimal stress to the dogs.

Abdominal Muscles↗
PubMed2001

Persistent Ehrlichia chaffeensis infection in white-tailed deer.

Four white-tailed deer (Odocoileus virginianus) were inoculated intravenously with a deer-origin isolate (15B-WTD-GA) of Ehrlichia chaffeensis. The course of infection was monitored using indirect fluorescent antibody (IFA), polymerase chain reaction (PCR), and culture over a 9 m period. All deer became rickettsemic within 24 days post inoculation (DPI), and all developed antibody titers >1:64 to E. chaffeensis by 17 DPI. Titers in all deer fell below 1:64 during 87 to 143 DPI. One deer exhibited a second period of seropositivity (peak titer of 1:256) from 207 to 271 DPI but was culture and PCR negative during this period. Rickettsemia was confirmed by reisolation of E. chaffeensis as late as 73 to 108 DPI in three deer. Positive PCR results were obtained from femur bone marrow of one deer and from rumenal lymph node of another (leer at 278 DPI. None of the deer developed clinical signs, hematologic abnormalities, or gross or microscopic lesions attributable to E. chaffeensis. Two uninoculated control deer were negative on all tests through 90 DPI at which time they were removed from the study. Herein we confirm that white-tailed deer become persistently infected with E. chaffeensis, have initial rickettsemias of several weeks duration and may experience recrudescence of rickettsemia, which reaffirm the importance of deer in the epidemiology of E. chaffeensis.

Animals↗
PubMed1999

Ehrlichia chaffeensis in archived tissues of a white-tailed deer.

White-tailed deer (Odocoileus virginianus) play an integral role in the natural history of Ehrlichia chaffeensis, the causative agent of human monocytic ehrlichiosis (HME). Paraffinized tissues from a white-tailed deer submitted as a diagnostic case to the Southeastern Cooperative Wildlife Disease Study (Athens, Georgia, USA) in October of 198.5 and originally described as infected with an unidentified rickettsial organisim were re-examined by specific nested polymerase chain reaction (PCR) for evidence of infection with Ehrlichia spp. Ehrlichia chaffeensis was identified from the bone marrow and inguinal lymph node of this deer based on amplification of a characteristic sequence-confirmed 16S rDNA fragment from these tissues. Parallel PCR tests on the same samples were negative for 16S rDNA fragments of the agent of human granulocytic ehrlichiosis (HGE) and for an Ehrlichia-like organism widely distributed in white-tailed deer populations. This report describes detection of E. chaffeensis in archived tissue from a deer collected before the index case of human monocytic ehrlichiosis was established.

Animals↗
PubMed1999

Susceptibility of red and gray foxes to infection by Ehrlichia chaffeensis.

Red foxes (Vulpes vulpes) and gray foxes (Urocyon cinereoargenteus) were evaluated for their susceptibility to experimental infection with Ehrlichia chaffeensis, the causative agent of human monocytotropic ehrlichiosis. Two red foxes and three gray foxes were inoculated intravenously with E. chaffeensis (15B-WTD-GA strain) and were monitored at 7, 14, 21, and 28 days post inoculation (DPI) for evidence of infection using an indirect fluorescent antibody (IFA) assay, light microscopy, polymerase chain reaction (PCR), and cell culture methods. One red fox and one gray fox served as negative controls. Red foxes were susceptible to infection based on reisolation of E. chaffeensis from blood at 7 and 14 DPI, seroconversion by 7 DPI, and positive PCR assays on spleen and lymph nodes at 28 DPI. Morulae were not found in circulating leukocytes and clinical signs or lesions of ehrlichiosis were not observed. In contrast, gray foxes were refractory to infection based on negative results on all culture, PCR, serologic, and microscopic examinations. These findings imply that red foxes, but not gray foxes, are potential vertebrate reservoirs for E. chaffeensis. These findings also illustrate the need to verify serologic evidence of E. chaffeensis infection among wild animals.

Animals↗
PubMed1999

Potential for contact and mechanical vector transmission of vesicular stomatitis virus New Jersey in pigs.

OBJECTIVE: To determine whether swine could be infected with vesicular stomatitis virus New Jersey (VSV-NJ) by routes simulating mechanical vector and contact transmission, and to document clinical response and measure source, duration, and extent of associated viral shedding. ANIMALS: 48 approximately 10-kg male pigs. PROCEDURE: 7 inoculation routes were tested at 3 virus concentrations (10(6), 10(4), and 10(2) median tissue culture-infective doses [TCID50]). Inoculation routes included intradermal (snout), topical application of virus to a scarified area (ear or mucous membrane of the lip), mucosal (oral or conjunctiva) application, and nasal instillation (drop or aerosol). Swine were monitored daily for clinical disease, seroconversion, and virus isolation. RESULTS: Virus was isolated from swab specimens of nasal planum, nasal cavity, saliva, tonsil, and feces from 15 pigs infected by intradermal snout inoculation (10(6) and 10(4) TCID50), scarification of the lip (10(6) and 10(4) TCID50), and oral route (10(6) TCID50). Virus was not isolated from plasma. Vesicular lesions were restricted to pigs inoculated via intradermal snout and lip scarification routes. Seroconversion was detected in 23 pigs inoculated by all routes except intranasal aerosol and conjunctival. Frequency of virus isolation from these pigs was dependent on serologic status and presence of vesicular lesions. CONCLUSIONS: Viral titers obtained from swab specimens and infective doses indicated that sufficient virus is shed from infected swine for contact or mechanical transmission to occur. Amount of virus is highest when lesions are present and before seroconversion. Pigs may represent an effective large-animal model for studying pathogenesis and transmission of VSV-NJ.

Animals↗
PubMed1998

Diseases diagnosed in red foxes from the southeastern United States.

Diagnostic findings on 51 red foxes (Vulpes vulpes) from the southeastern United States examined from 1967 to 1995 were reviewed. Etiologic diagnoses included sarcoptic manage (n = 33), traumatic injury and associated complications (n = 5), suspected canine distemper (n = 2), capture myopathy (n = 1), congenital absence of guard hairs (n = 1), intradermal tick infestation (n = 1), otodectic manage (n = 1), and toxicosis (n = 1). The cause of morbidity was not determined for three of the foxes, and three others were classified as normal animals. Sarcoptic manage was diagnosed in 65% of the red foxes, was found in foxes submitted from four of the eight southeastern states represented, and was seen in 19 of 29 yr covered by this study.

Alopecia↗
PubMed1997

Cell-mediated immune response and IL-2 production in white-tailed deer experimentally infected with hemorrhagic disease viruses.

Hemorrhagic disease, caused by various serotypes of two closely related orbivirus serogroups, the epizootic hemorrhagic disease viruses (EHDV) and the bluetongue viruses (BTV), is a major cause of morbidity and mortality in white-tailed deer (WTD) in the United States. Despite the importance of hemorrhagic disease in WTD, little is known about host defense mechanisms triggered by infection with either causative virus or how that immune response is modulated by challenge with closely related orbiviruses, as can occur under natural conditions. Initial experimental data from our laboratory showed WTD infected with EHDV serotype 2 (EHDV-2) had responded serologically but often became lymphopenic and had a reduced lymphocyte proliferative response in vitro to T-cell mitogens, suggesting possible suppression of cell-mediated immunity. The primary objective of this study was to more closely examine cell-mediated immunity of WTD when experimentally infected with EHDV-2 and subsequently challenged with BTV serotype 10 (BTV-10). The cell-mediated response was evaluated via in vitro lymphocyte proliferation and interleukin-2 (IL-2) production assays, and in vivo delayed type hypersensitivity tests. Deer infected with either EHDV-2 or BTV-10 responded similarly in all assays. Infected deer had decreased lymphocyte counts between post-infection days (PID) 6 and 10, with concurrent diminished lymphocytic response to concanavalin A in lymphocyte proliferation assays and phytohemagglutinin in delayed, type hypersensitivity tests. However, IL-2 production by peripheral blood lymphocytes of infected deer was comparable with that of non-infected control deer as measured using a IL-2-dependent bovine cell line (BT2). This suppression of T-cell proliferation, but not IL-2 production suggests selective inhibition of T-cells probably via altered signal transduction for either expression of the IL-2 receptor or for IL-2 receptor signal-induced T-cell proliferation.

Animals↗
PubMed1997

Purified outer membranes of Serpulina hyodysenteriae contain cholesterol.

We have isolated outer and inner membranes of Serpulina hyodysenteriae by using discontinuous sucrose density gradients. The outer and inner membrane fractions contained less than 1 and 2%, respectively, of the total NADH oxidase activity (soluble marker) in the cell lysate. Various membrane markers including lipooligosaccharide (LOS), the 16-kDa outer membrane lipoprotein (SmpA), and the C subunit of the F1F0 ATPase indicated that the lowest-density membrane fraction contained outer membranes while the high-density membrane fraction contained inner membranes and that both are essentially free of contamination by the periplasmic flagella, a major contaminant of membranes isolated by other techniques. The outer membrane fractions (rho = 1.10 g/cm3) contained 0.25 mg of protein/mg (dry weight), while the inner membrane samples (rho = 1.16 g/cm3) contained significantly more protein (0.55 mg of protein/mg [dry weight]). Lipid analysis revealed that the purified outer membranes contained cholesterol as a major component of the membrane lipids. Treatment of intact S. hyodysenteriae with different concentrations of digitonin, a steroid glycoside that interacts with cholesterol, indicated that the outer membrane could be selectively removed at concentrations as low as 0.125%.

Bacterial Outer Membrane Proteins↗
PubMed1997

Isolation of Ehrlichia chaffeensis from wild white-tailed deer (Odocoileus virginianus) confirms their role as natural reservoir hosts.

Field and experimental studies have implicated white-tailed deer (Odocoileus virginianus) as probable reservoir hosts for Ehrlichia chaffeensis, the causative agent of human monocytic ehrlichiosis, but natural infection in deer has not been confirmed through isolation of E. chaffeensis. Thirty-five white-tailed deer collected from three Amblyomma americanum-infested populations in Georgia were examined for evidence of E. chaffeensis infection by serologic, molecular, cell culture, and xenodiagnostic methods. Twenty-seven deer (77%) had E. chaffeensis-reactive indirect fluorescent-antibody assay titers of > or = 1:64; and the blood, spleens, or lymph nodes of seven (20%) deer were positive in a nested PCR assay with E. chaffeensis-specific primers. E. chaffeensis was isolated in DH82 cell cultures from the blood of five (14%) deer, including two deer that were PCR negative. Combination of culture and PCR results indicated that six (17%) deer were probably rickettsemic and that nine (26%) were probably infected. Restriction digestion of PCR products amplified from deer tissues and cell culture isolates resulted in a banding pattern consistent with the E. chaffeensis 16S rRNA gene sequence. The sequences of all PCR products from deer tissues or cell culture isolates were identical to the sequence of the Arkansas type strain of E. chaffeensis. Xenodiagnosis with C3H mice inoculated intraperitoneally with deer blood, spleen, or lymph node suspensions was unsuccessful. When viewed in the context of previous studies, these findings provide strong evidence that E. chaffeensis is maintained in nature primarily by a tick vector-vertebrate reservoir system consisting of lone star ticks and white-tailed deer.

Animals↗
PubMed1997

Experimental vesicular stomatitis in swine: effects of route of inoculation and steroid treatment.

An enzootic focus of vesicular stomatitis virus New Jersey serotype (VSV-NJ) exists on Ossabaw Island, Georgia. Many questions regarding the epizootiology of this virus at this focus still exist, but evidence suggests that the vector for this virus is a phlebotomine sand fly (Lutzomyia shannoni), with feral swine serving as a potential source of virus for the sand fly and for other swine via contact transmission. We conducted 2 experimental trials in domestic swine using VSV-NJ isolated from a sand fly from Ossabaw Island to determine if route of inoculation or immunosuppression via steroid administration affected the development of disease, viremia, viral shedding, or the neutralizing antibody response. In a third trial, we studied the potential for contact transmission among swine using this isolate. Virus isolations were made from nasal cavity or palatine tonsil of the soft palate, and VSV-NJ neutralizing antibodies developed when pigs were inoculated intradermally in the apex of the snout, ear, or coronary band, intravenously, intranasally, or via scarification of the apex of the snout or coronary band. Vesicles developed only in pigs inoculated in the apex of the snout or coronary band, and these vesicles were at the site of inoculation. Steroid treatment did not potentiate the development of secondary vesicles and did not prolong the period of virus shedding from VSV-NJ-infected swine. Contact transmission, as determined by shedding of virus from the tonsil of the soft palate and the development of VSV-NJ neutralizing antibodies, occurred in pigs in contact with animals inoculated in the apex of the snout but not in contact animals exposed to pigs inoculated intradermally in the coronary band or intranasally. These trials show that contact transmission can occur and VSV-NJ can be shed without the development of clinical disease (i.e., vesicle formation). Viremia was never detected in any of the experimental pigs, suggesting that swine may not be a good amplifying host for VSV-NJ.

Animals↗
PubMed1997

Evaluation of low-level aflatoxin in the diet of white-tailed deer.

We evaluated the response of white-tailed deer (WTD) (Odocoileus virginianus) to dietary aflatoxin. Fourteen 4-to-5-mo-old WTD were used in this 8-wk study, conducted between November 1993 and January 1994. Seven animals received a ration containing 800 parts per billion (ppb) total aflatoxin (AF). Seven control animals received the same ration without AF. At 0, 1, 3, 6 and 8 wk, feed consumption, feed conversion, liver enzymes, bile acid levels, and immune function via lymphocyte proliferation assays and delayed type hypersensitivity reactions were determined. At the conclusion of the 8-wk feeding trial, deer were euthanized and necropsied. Clinical illness was not evident in any of the animals, but by the end of the study, AF-fed deer had reduced feed consumption and body weight as compared to control deer; the differences were not statistically significant. The AF-exposed group had a significant increase (P = 0.03) in serum bile acid concentration as compared to control deer. Two AF-exposed deer had gross and histologic hepatic lesions indicative of a mild degenerative hepatopathy. Residues of an aflatoxin metabolite, aflatoxin M1, were found in the livers of all treated animals. No differences in immune function were detected between the two groups. We conclude that consumption of 800 ppb AF in the diet of young WTD over an 8-wk period can produce subclinical hepatic injury.

Administration, Oral↗
PubMed1997

Shell disease in river cooters (Pseudemys concinna) and yellow-bellied turtles (Trachemys scripta) in a Georgia (USA) lake.

A disfiguring shell disease was detected in river cooters (Pseudemys concinna) and yellow-bellied turtles (Trachemys scripta) from Lake Blackshear, Georgia (USA). The turtles used were part of a mark-recapture study conducted from September 1991 to June 1993. Histologic changes on four turtles included acute segmental necrosis of the epidermis, followed by ulceration, necrosis of the underlying dermis and dermal bone, and exaggerated remodeling of bone. Additional findings included visceral inflammatory lesions and bacterial infection, sepsis and marked trematode ova granulomatosis. The cause of the shell lesions was not determined.

Animals↗