Clinical utility of liver biopsies in dogs undergoing splenectomy.

OBJECTIVE: To determine the prevalence of neoplasia detected on liver biopsy obtained at the time of splenectomy in dogs with splenic masses. MATERIALS AND METHODS: Retrospective study of medical records of dogs with splenic masses from which liver biopsies were taken following splenectomy. RESULTS: Malignant splenic neoplasia was detected in 50 of 113 (44.2%) of the dogs undergoing splenectomy. Neoplastic liver disease was detected on biopsy from 1 of 40 (2.5%) dogs with a grossly normal liver and from 20 of 69 (28.9%) dogs with a grossly abnormal liver. Dogs with a grossly abnormal liver had a ~ 16 times (95% CI: 2.5-170) higher chance of being diagnosed with liver neoplasia on biopsy. Haemoabdomen was also associated with an increased likelihood of liver neoplasia on biopsy at the time of splenectomy. CLINICAL SIGNIFICANCE: A liver biopsy taken from grossly normal liver is a low-yield diagnostic test but liver biopsy is recommended following splenectomy if the liver appears abnormal at surgery.

The use of megavoltage radiation therapy in the treatment of thymomas in rabbits: 19 cases.

An overall median survival time (MST) and prognostic factors in rabbits with thymomas treated with megavoltage radiation therapy (RT) were determined in this multi-institutional retrospective case analysis. Medical records for 19 rabbits with suspected or confirmed thymomas treated with RT were evaluated for data including signalment, haematological and serum biochemistry abnormalities, presence of pleural effusion, radiation plan, body weight, total radiation dose and institution administering RT. Statistical significance of these factors related to overall survival was assessed. An overall MST for all 19 rabbits was 313 days; exclusion of 3 rabbits that died acutely during the first 14 days of RT yielded a MST of 727 days. The only factor associated with a significantly decreased survival time was having a body weight lower than mean body weight of 1.57 kg. Radiation treatment-associated complications were infrequent and included radiation-induced myocardial failure, radiation pneumonitis and alopecia.

Direct evidence for the role of centrosomally localized p53 in the regulation of centrosome duplication.

Abnormal amplification of centrosomes is the major cause of mitotic defects and chromosome instability in cancer cells. Centrosomes duplicate once in each cell cycle, and abrogation of the regulatory mechanism underlying centrosome duplication leads to centrosome amplification. p53 tumor suppressor protein is involved in the regulation of centrosome duplication: loss of p53 as well as expression of certain p53 mutants result in deregulated centrosome duplication and centrosome amplification. p53 at least in part depends on its transactivation function to control centrosome duplication, primarily via upregulation of p21 cyclin-dependent kinase (CDK) inhibitor, which prevents untimely activation of CDK2/cyclin E, a key initiator of centrosome duplication. However, numerous studies have shown the presence of p53 at centrosomes, yet the role of the centrosomally localized p53 in the regulation of centrosome duplication had been enigmatic. Here, we comparatively examined wild-type p53 and p53 mutants that are transactivation(+)/centrosome-binding(-), transactivation(-)/centrosome-binding(+) and transactivation(-)/centrosome-binding(-) for their abilities to control centrosome duplication. We found that the transactivation(+)/centrosome-binding(-) and transactivation(-)/centrosome-binding(+) mutants suppress centrosome duplication only partially compared with wild-type p53. Moreover, the transactivation(-)/centrosome-binding(-) mutant almost completely lost the ability to suppress centrosome duplication. These observations provide direct evidence for the centrosomally localized p53 to participate in the regulation of centrosome duplication in a manner independent of its transactivation function in addition to its transactivation-dependent regulation of centrosome duplication.

Spontaneous conception in the presence of stage IIIC endometrioid ovarian cancer.

OBJECTIVE: To describe a rare case of spontaneous conception in a patient with a preexisting metastatic ovarian cancer. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 39-year-old Asian woman who conceived while undergoing an evaluation for primary infertility and newly detected bilateral adnexal masses. INTERVENTION(S): Staging laparotomy and total abdominal hysterectomy and bilateral salpingo-oophorectomy. MAIN OUTCOME MEASURE(S): Anatomic pathology diagnosis. RESULT(S): Blighted ovum and stage IIIC endometrioid adenocarcinoma of ovary. CONCLUSION(S): Metastatic ovarian cancer does not prevent either spontaneous ovulation or spontaneous conception.

Individual variability in the zinc inducibility of metallothionein-IIA mRNA in human lymphocytes.

The metallothionein-III gene (MT-IIA) is a major member of the human MT gene family. Metallothioneins (MTs) are low-molecular-weight, cysteine-rich proteins that bind and detoxify heavy metals. At least two different MT-IIA polymorphisms have been identified in humans, one or both of which may affect susceptibility to metal toxicity. The purpose of this study was to investigate whether these different genotypes affect the inducibility of MT-IIA mRNA in human lymphocytes treated with zinc (Zn), the major known inducer of MT-IIA in vitro. Fresh lymphocytes obtained from 16 healthy volunteers, aged 23-38 yr, were genotyped for the MT-IIA gene and tested for expression. A 43.5-bp HindIII-Taql fragment of the MT-IIA promoter was used to probe for the two known polymorphisms (a 7.8-kb vs. a 5.3-kb fragmnent, and a 1.7-kb vs. a 1.6-kb fragment). The allele frequencies of the 16 subjects were 14%, for 5.3-kb allele and 19% for 1.6-kb allele. In Northern blotting experiments, MT-II mRNA levels were induced over a wide range of Zn concentrations during 2-h exposures; specifcally, levels increased by 9- to 115-fold with exposure to 100 microM ZnCl, and by 16- to 311-fold with exposure to 200 microM ZnCl2. However, no significant differences in MT-IIA inducibility were found between the 7.8/5.3-kb allele pair (n = 4) and the 7.8/7.8-kb allele pair (n = 12) or between the 1.7/1.6-kb allele pair (n = 5) and the 1.7/1.7-kb allele pair (n = 11). Thus. MT-IIA is strongly inducible by Zn in human lymphocytes, but individual variations exceed those that can be attributed to the known promoter-region polymorphisms.

Ureterotomy for removal of two ureteroliths in a parrot.

A 21-year-old male double-yellowheaded Amazon parrot was referred because of possible urolithiasis. The bird had strained to void since it was young, and recently, signs of depression and inappetence had appeared. Radiography revealed 2 mineralized opacities in the left caudal portion of the celomic cavity. A left lateral celiotomy revealed that the left ureter was dilated and contained a calculus, which was later determined to be composed of monosodium uric acid crystals and proteinaceous material. The second mineralized mass could not be located at that time, but was detected in the caudal aspect of the celomic cavity after surgery by use of radiography. Manual attempts to maneuver the uterolith from the ureter through the cloaca were unsuccessful. Five days after the initial surgery, the uterolith was removed by cloacotomy and ventral midline laparotomy. Excretory urography performed 10 and 31 days after surgery revealed that the left ureter was homogeneously opacified and gradually decreased to 3 to 6 mm in diameter. The bird remained healthy and seemed to strain less severely during voiding. Nonspecific signs associated with ureteroliths may result in delay in diagnosis. Surgical removal of ureteroliths may be an effective treatment for this uncommon condition, but it is complicated by certain anatomic features of birds and may result in ureteral stricture.

In vitro elution of gentamicin, amikacin, and ceftiofur from polymethylmethacrylate and hydroxyapatite cement.

OBJECTIVE: To compare the elution characteristics of ceftiofur and liquid and powdered gentamicin and amikacin from polymethylmethacrylate (PMMA) and from hydroxyapatite cement (HAC). METHODS: PMMA and HAC beads in triplicate were impregnated with various amounts and formulations of antibiotics. Beads were immersed in 5 mL of phosphate buffered saline that was replaced at 1, 3, 6, and 12 hours, and 1, 2, 3, 5, 7, 10, 14, 18, 22, 26, and 30 days. The eluent was stored at -70 degrees C until assayed within 2 weeks by microbiological assay (gentamicin and amikacin) or capillary electrophoresis (ceftiofur). RESULTS: Rate of elution for all beads was greatest within the first 24 hours. Cumulative release of total antibiotic dose from beads over 30 days was significantly greater from HAC than PMMA. Antibiotic elution was directly related to the amount of antibiotic incorporated into the cement. Powdered and liquid forms of gentamicin had similar elution rates from PMMA. Elution of amikacin from PMMA beads was greater when the powdered form was used compared with liquid amikacin. Eluent concentrations of ceftiofur were similar to those of the aminoglycosides during the first 3 to 7 days but then decreased precipitously by comparison. CONCLUSIONS: Elution of antibiotics from HAC was greater than from PMMA. Gentamicin- and amikacin-impregnated PMMA and HAC released bactericidal concentrations of antibiotic for at least 30 days. Ceftiofur-impregnated PMMA or HAC is unlikely to provide long-term bactericidal concentrations. CLINICAL RELEVANCE: Gentamicin and amikacin elute effectively from PMMA and HAC.

Intestinal volvulus and stricture associated with a leiomyoma in a green turtle (Chelonia mydas).

A previously stranded 30-kg female green turtle (Chelonia mydas) was referred to the Veterinary Medical Teaching Hospital at the University of Florida following a 2-mo history of anorexia, intermittent regurgitation, decreased fecal production, and positive buoyancy of the right side. Radiographs confirmed gaseous distension of bowel loops suggestive of intestinal obstruction. The coelom was surgically approached through a plastron osteotomy, and a 540 degrees volvulus of the small intestine was identified and derotated. An intestinal stricture was present at the site of the volvulus, and segments of bowel orad to the stricture were greatly distended. Following resection of abnormal bowel, the osteotomy was stabilized using self-tapping screws and figure-eight wire, and the defect was sealed with fiberglass cloth and fast-drying epoxy resin. A leiomyoma associated with the focal stricture was identified by histology. Appetite and defecation returned to normal. Six months after surgery, the turtle had regained normal buoyancy and showed no further clinical signs of gastrointestinal obstruction.

Thoracic vertebral chordoma in a domestic ferret (Mustela putorius furo).

A castrated adult male 1.0-kg domestic ferret (Mustela putorius furo) was referred for evaluation of a mass of >5 mo duration that was associated with the thoracic spine. The ferret had motor dysfunction and no conscious proprioception or pain perception in either hind limb. Segmental reflexes were present. Survey radiography, myelography, and magnetic resonance imaging showed a mass involving the ninth and 10 thoracic vertebrae and compressing the spinal cord. Decompressive surgery was performed, but clinical signs persisted. A diagnosis of chordoma was made using histopathology and immunohistochemistry. This is the first reported case of a chordoma involving the thoracic vertebrae in a domestic ferret.

Preparation and equipment useful for surgery in small exotic pets.

Because many exotic pets are so small, the instrumentation and equipment used for surgery are different from in larger animals. Magnification greatly improves the surgeon's ability to accomplish surgeries in small patients and helps to provide hemostasis because small amounts of blood are significant in these tiny patients. Microsurgical instrumentation is important to allow for proper tissue handling and suture placement. Various other considerations, instrumentation, and equipment are useful when performing surgery on small exotic pets.

Nonreproductive surgery in reptiles.

As reptiles have gained popularity as pets, veterinarians are more frequently called on to perform surgical procedures on these pets. Veterinarians must have a basic understanding of the anatomic and physiologic differences between reptilian and mammalian companion animals. This article reviews some basic points about reptilian anatomy and physiology that are important to consider when performing surgery on reptilian patients. This article covers approaches to the coelomic cavity in the different groups of reptiles and some basic concepts regarding surgery on different, nonreproductive systems in these reptiles.

T-cell-rich B-cell lymphoma in a ring-tailed lemur (Lemur catta).

A 13-yr-old ring-tailed lemur (Lemur catta) was evaluated for depression, anorexia, polyuria, and polydipsia. The lemur was in poor body condition and was anemic, hypoalbuminemic, and hyponatremic. Cytologic examination of aspirates of the spleen, liver, and bone marrow and histopathologic examination of liver and bone marrow biopsies revealed a disseminated round cell tumor. After euthanasia, necropsy revealed hepatomegaly, splenomegaly, and mesenteric lymphadenomegaly. Neoplastic cells were present within the spleen, liver, kidneys, multiple lymph nodes, bone marrow, lung, small intestine, pancreas, and testicle and were composed of large anaplastic round cells in a background of small well-differentiated lymphocytes. Immunohistochemical analysis revealed that the small well-differentiated lymphocytes labeled for the anti-human T-cell marker, CD3, and the large anaplastic round cells labeled with the anti-human B-cell marker, CD79a. On the basis of the immunohistochemical staining results and morphologic appearance, a diagnosis of a T-cell-rich B-cell lymphoma was made.

Surgical management of bifid sternum in two African grey parrots.

Transposition of the pectoral muscles for treatment of caudal bifid (cleft) sternum in 2 unrelated African Grey Parrots is described. The birds did not have clinical signs of ventilatory compromise prior to surgery; however, both had cutaneous ulcers over the defects. The pectoral muscles provided a thick pad over the heart, minimizing the risk of trauma to the heart.

Removal of a thymoma via median sternotomy in a rabbit with recurrent appendicular neurofibrosarcoma.

A 9-year-old rabbit was evaluated for a mass on the right elbow. As a result of preoperative radiography, another mass was identified in the cranial portion of the mediastinum. Ultrasound guided fine needle aspiration was used for cytologic evaluation of the mass in the mediastinum and resulted in a presumptive diagnosis of thymoma. A median sternotomy approach was chosen to allow wide exposure for meticulous dissection in the cranial portion of the thoracic region. Histologic examination and immunoperoxidase labeling of lymphocytes with CD3 (a T cell marker) confirmed the diagnosis. The mass on the right elbow was subsequently removed and identified as a neurofibrosarcoma. Multiple appendicular masses that developed at separate locations and distinct times were excised and classified as locally invasive primary neurofibrosarcomas. Postmortem examination 9 months after initial evaluation confirmed recurrence of the neurofibrosarcoma at previous surgical sites. Gross or histologic evidence of thymoma was not evident. Prognosis is dismal for incompletely excised soft tissue sarcomas, because they are locally invasive and often recur.

Incidence of contralateral versus ipsilateral neurological signs associated with lateralised Hansen type I disc extrusion.

Asymmetrical neurological signs were noted in 50 dogs presenting with Hansen type I thoracolumbar disc extrusion. Thoracolumbar myelograms and surgical decompression were performed in all cases. Dogs were divided into two groups (acute and chronic) based on the duration of clinical signs prior to presentation to the University of Georgia. Lateralising extradural cord compressive lesions were noted on all myelograms. In the acute group, 35 per cent of the dogs had asymmetrical neurological signs contralateral to the myelographic and surgical lesion, while in the chronic group only 11 per cent had neurological signs contralateral to the lesion. There was found to be no significant difference in frequency of contralateral asymmetrical clinical signs between the two groups (Fischer's exact test; P = 0.095). The high frequency of contralateral signs documents the importance of thoracolumbar myelography for accurate localisation of the disc material before decompressive surgery.

Regulation of eukaryotic abasic endonucleases and their role in genetic stability.

Abasic (AP) sites in DNA arise from spontaneous reactions or the action of DNA glycosylases and represent a loss of genetic information. The AP sites can be mutagenic or cytotoxic, and their repair is initiated by class II AP endonucleases, which incise immediately 5' to AP sites. The main enzyme of S. cerevisiae. Apn1, provides cellular resistance to oxidants (e.g., H2O2) or alkylating agents, and limits the spontaneous mutation rate. AP endonucleases from other species can replace Apn1 function in yeast to different extents. We studied the main human enzyme, Ape, with respect to its incision specificity in vitro and the expression of the APE gene in vivo. The results suggest that Ape evolved to act preferentially on AP sites compared to deoxyribose fragments located at oxidative strand breaks and that the incision modes of Ape and Apn1 may be fundamentally different. We also defined the functional APE promoter, and showed that APE expression is transiently downregulated during the regeneration of epidermis after wounding. This latter effect may lead to a window of vulnerability for DNA damage and perhaps mutagenesis during the healing of epidermal and other wounds. Such unexpected effects on the expression of DNA repair enzymes need to be taken into account in analyzing the susceptibility of different tissues to carcinogens.

Construction of a vector containing a site-specific DNA double-strand break with 3'-phosphoglycolate termini and analysis of the products of end-joining in CV-1 cells.

Previous studies have shown that linearized SV40-based shuttle vectors transfected into mammalian cells are efficiently recircularized by an error-prone end-joining pathway. To determine whether and with what specificity free radical-mediated double-strand breaks are rejoined by this pathway, a structural mimic of such a break was introduced at a specific site in an SV40-based shuttle vector, by ligating purified 3'-phosphoglycolate-terminated oligonucleotides into 3' recessed ends generated in the linearized vector. These terminally blocked linear vectors were efficiently repaired and replicated when transfected into simian CV-1 cells. Sequencing across the repair joints in progeny plasmid indicated that, for a blunt-ended vector, the most frequent mechanism of rejoining was splicing at a terminal 4-base homology; however, a significant fraction of the joints retained all bases from both ends of the break, consistent with a mechanism involving simple 3'-phosphoglycolate removal, followed by blunt-end ligation. For the analogous 3'-hydroxyl terminated break, the fraction of simple blunt-end ligations was considerably higher. For a phosphoglycolate-terminated vector with cohesive ends the most frequent repair mechanism was simple ligation of the annealed cohesive ends, presumably preceded by phosphoglycolate removal. For all these substrates, the remaining repair joints showed small or large deletions from one or both of the ends, usually with apparent annealing at short (1-4-base) homologies. The results suggest that while breaks with 3'-phosphoglycolates can be repaired, these blocked termini represent a significant barrier to DNA end-joining, and can significantly alter its specificity. The presence of cohesive ends appears to improve markedly the fidelity of rejoining for terminally blocked double-strand breaks.

Immunohistochemical localization of transforming growth factor-alpha, epidermal growth factor (EGF), and EGF receptor in the human fetal ovary.

Peptide growth factors are thought to be involved in adult ovarian regulatory functions. However, little is known about the role of growth factors in human fetal ovarian development. This study is an attempt to identify and localize transforming growth factor-alpha (TGF alpha), epidermal growth factor (EGF), and EGF receptor (EGF-R) in human fetal ovaries. Ovaries were obtained from first and second trimester elective abortuses. Immunohistochemistry was performed on paraffin sections of these specimens after fixation. We examined the sections microscopically using the specific antibodies against TGF alpha, EGF, and EGF-R. Phosphate-buffered saline and preimmune IgG were used as negative controls. First and second trimester ovaries stained positively for all three proteins. Staining was significantly more intense in the oocytes than in the stroma. Negative controls did not stain. These results combined with our previous demonstration of messenger ribonucleic acid for these growth factors suggest roles for TGF alpha, EGF, and EGF-R in human fetal ovarian development. The strong staining in the oocytes suggests a possible autocrine or paracrine role of these growth factors in human oocyte growth in utero.