Synthetic vascular grafts seeded with genetically modified endothelium in the dog: evaluation of the effect of seeding technique and retroviral vector on cell persistence in vivo.

Unique characteristics of endothelium make it an attractive target cell for gene transfer. Genetically modified endothelial cells (ECs) seeded on synthetic vascular grafts offer the potential to control neointimal hyperplasia, decrease graft thrombogenicity and improve small diameter graft patency. This study addresses the issue of synthetic vascular graft colonization with endothelial cells transduced with noninducible retroviral marker genes in the dog. Autologous endothelial cells were enzymatically harvested and transduced with either the bacterial NeoR gene or human growth hormone gene using retroviral vectors. All transduced cells were positive by polymerase chain reaction (PCR) amplification for the transduced gene sequence prior to graft seeding. Transduced ECs were seeded on Dacron grafts (n = 3) preclotted with autologous blood. These grafts exhibited complete endothelialization at times from 250 to 360 days. Recovered DNA, however, was negative for the transduced gene sequence when analyzed by PCR and Southern blotting. Expanded polytetrafluoroethylene (ePTFE) was evaluated (n = 8) using several different cell seeding protocols. Grafts were seeded at 3 densities (ranging from 6 x 10(3) to 1.5 x 10(5) cells/cm2) and 2 different adherence times. Seeding substrate was also evaluated. Grafts were either preclotted with whole blood or incubated with 20 or 120 micrograms/ml fibronectin for 60 min. Graft biopsies were evaluated from 2 to 52 wk. Limited endothelialization was present in 4 dogs as early as 2 wk, but never progressed to full luminal coverage. The remaining dogs failed to ever exhibit any luminal EC adherence. Two dogs with limited EC coverage had positive DNA by PCR for the NeoR gene sequence at 2 and 3 wk. In contrast to transduced EC's, nontransduced EC colonization of ePTFE was complete at 2 wk when seeded under conditions that transduced cells had failed to persist. Neither seeding density, adherence time, seeding substrate or retroviral vector used influenced the uniformly poor graft coverage seen with transduced cells. Results of this study indicate that despite successful gene transfer using 4 different retroviral vectors, transduced endothelial cells seeded under varying conditions appear altered in their ability to stably adhere and colonize synthetic vascular grafts in vivo.

Social support and anxiety in pregnant drug abusers and nonusers: unexpected findings of few differences.

Drug-abusing (n = 25) and nonusing (n = 55) pregnant women from a publicly supported prenatal clinic were tested for level of social support and of pregnancy anxiety during the last half of pregnancy. Differences found between the groups were fewer than expected. Drug abusers did not differ from nonusers in overall level of social support or in Appraisal, Belonging, or Tangible subscales. Abusers were found to report lower levels of self esteem; lower self esteem was predicted by drug abuse, having more children and lower socioeconomic status. Drug abusers did not differ from nonusers in their overall feelings of pregnancy anxiety, but they did indicate higher fears for themselves and for the baby, and there was a tendency for higher depression and withdrawal.

Treatment of murine cutaneous melanoma with near infrared light.

Treatment of cutaneous melanoma (M-3 and B16-F10 implanted in mice) with rapidly-scanned, tightly-focused near infrared light elicits selective destruction of tumor tissue. A single laser treatment yielded complete eradication in >90% of B16-F10 tumors with thicknesses of approximately 3 mm; amelanotic M-3 tumors proved less responsive (ca 25% clearance rate). In addition to local tumor destruction, laser treatment of B16-F10 tumors in immunocompetent mice stimulated enhanced cytokine levels (interleukin-2 and interleukin-10) within treated tumor tissues and rejection of tumor cells upon a subsequent challenge dose. Such an antitumor immune response may lead to improved outcomes at both the treatment site and at sites of distant metastasis.

Metastatic pheochromocytoma in a parakeet.

Pheochromocytoma, a tumor of the adrenal medulla, was diagnosed in a budgerigar based on gross, histopathologic, and electron microscopic findings.

Radiographic and scintigraphic evidence of focal pulmonary neoplasia in three cats with hyperthyroidism: diagnostic and therapeutic considerations.

Three cats were diagnosed as hyperthyroid based on clinical signs, historical findings, laboratory abnormalities, and basal serum thyroxine (T4) concentrations, and/or nuclear thyroid scans. Additionally, a presumptive diagnosis of thyroid carcinoma with pulmonary metastasis was made in each cat based on radiographic or scintigraphic evaluation. All three cats had solitary pulmonary nodules 1.5 to 2 cm in diameter on survey thoracic radiographs; one cat also had chylous pleural effusion and pulmonary lobar consolidation. Focal pulmonary accumulation of sodium pertechnetate (99mTcO4-) and/or radioiodine (131I) corresponding to radiographic lesions were seen in all cats. Two cats were treated with single ablative doses (1111 to 1480 MBq) of 131I; the remaining cat was euthanatized. One of the treated cats died 8 days later; the other cat was euthanatized 22 weeks following treatment. Histopathologic examination of tissue obtained at necropsy confirmed metastatic thyroid carcinoma in one cat and bronchogenic adenocarcinoma in two cats. Our findings indicate that increased radionuclide uptake in focal pulmonary lesions and cytologic evaluation of tissue obtained by fine-needle aspiration are not specific for thyroid tissue.

Acquired hydrocephalus and hydromyelia in a cat with feline infectious peritonitis: A case report and brief review.

A one-year-old domestic long-haired cat was referred to the New York State College of Veterinary Medicine because of acute onset of paraparesis and hyperesthesia associated with trauma. Myelography and cerebrospinal fluid analysis revealed severe hydromyelia and myelitis, respectively. The definitive diagnosis of feline infectious peritonitis was made by histological examination at necropsy. Lesions were confined exclusively to the brain and spinal cord. Partial occlusion of the third and fourth ventricles with pyogranulomatous debris caused hydrocephalus and subsequent hydromyelia. The hydromyelia may have been the primary means of compensation for the hydrocephalus, thus masking subclinical disease.

Intestinal mast cell tumor in a cat: presentation as eosinophilic enteritis.

A 10-year-old, neutered male domestic shorthair was diagnosed as having eosinophilic enteritis following two separate surgical biopsies. Although treatment was aggressive and a partial response was seen, the cat's condition deteriorated. Postmortem histopathology revealed an intestinal mast cell tumor.

A centering balloon for photodynamic therapy of esophageal cancer tested in a canine model.

Delivery of uniform circumferential light is desirable during photodynamic therapy of early or advanced esophageal cancer in human beings. Studies were performed in the canine esophagus to investigate whether use of a centering balloon would improve circumferential illumination of esophageal mucosa for photodynamic therapy of esophageal cancer. When the centering balloon was used, photodynamic therapy produced uniform and circumferential injury. With the cylindrical diffuser used in human studies, non-uniform and focal esophageal injury occurred. Placement of isotropic probes on the balloon wall allowed measurement and verification of relatively uniform light doses delivered to esophageal mucosa during balloon photodynamic therapy. The centering balloon has a potential role in improving light dosimetry during esophageal photodynamic therapy.

Photodynamic therapy for esophageal cancer using a 180 degrees windowed esophageal balloon.

Although delivery of uniform circumferential light is desirable during photodynamic therapy of advanced esophageal cancer in humans, early esophageal cancer may need only targeted treatment. Studies were performed in the canine esophagus of eight animals to investigate whether use of a "windowed" (shaded) centering balloon would improve targeted illumination of esophageal mucosa for photodynamic therapy. Shaded balloons were developed with a 2-cm-long, 360 degrees or 180 degrees clear "window." Photofrin 4 mg/Kg was used as the photosensitizer. Light at 630 nm was delivered at 300 J/cm or 600 J/cm. Isotropic probes placed on the balloon wall allowed real-time measurement and verification of relatively uniform light doses delivered to esophageal mucosa during balloon photodynamic therapy. With the windowed balloon, targeted delivery of photodynamic therapy was possible. Using the 180 degrees balloon, mucosa exposed to illumination was destroyed, whereas mucosa protected from light by the balloon shading was undamaged. Healing was complete and strictures did not occur. The shading of the balloon protected normal mucosa and prevents the formation of esophageal strictures. The "windowed" centering balloon provides a technology and technique that allows targeted delivery of uniform light during esophageal PDT.

Clinical and pathological characterization of an osteoma in a barred owl.

Osteomas are rare in birds. An osteoma of the left proximal radius was diagnosed in an adult barred owl based on gross, radiographic, and pathologic findings.

Comparative study between pulsed and continuous wave lasers for Photofrin photodynamic therapy.

A study was conducted in the normal canine esophagus to compare continuous wave (CW) and pulsed laser light for photodynamic therapy with Photofrin (4 mg/kg). Forty-eight hours post-injection, 630 nm laser light (CW light from an argon-pumped dye-laser and pulsed light from a KTP/532-pumped dye-laser) was delivered using a 24 mm diameter cylindrical esophageal PDT balloon positioned at either distal or proximal esophagus. A 1.0 cm cylindrical diffuser placed in the center of the balloon delivered 300 J/cm of light at an intensity of 400 mW/cm. Three dogs received CW light proximally and pulsed light distally. Four dogs received CW light distally and pulsed light proximally. The light dose delivered to the esophageal mucosa was measured using three isotropic probes placed on the balloon wall. Laser-induced fluorescence technique was used to compare photosensitizer fluorescence intensities at distal and proximal locations. Similar mucosal light doses and drug fluorescence intensities were verified for sites receiving pulsed or CW laser light. Two days after light delivery, the dogs were endoscoped to evaluate the severity of the lesions. While some response variability was observed among different animals, endoscopic examination of the lesions revealed comparable injury from CW and pulsed light in each subject. The animals were then euthanized and necropsies were performed. Based on the gross and histological examination of the lesions, the CW and pulsed laser-induced injuries could not be distinguished.

Quantitative 99mTc-pertechnetate thyroid scintigraphy in normal beagles.

This study was done to investigate the validity of published canine thyroid/salivary (T/S) ratios of approximately 1 in normal dogs and to determine thyroid uptake of 99mTc-pertechnetate (pertechnetate) measured as percent uptake of injected dose. These parameters were evaluated in 13 Beagle dogs over a 4 hour period. Mean +/- standard deviation (SD) and median T/S ratios of 1.2 +/- 0.3 and 1.1 were essentially the same at twenty minutes and 1 hour. T/S values ranged from 0.9 to 2.2 at 20 minutes and from 0.8 to 2.4 at 1 hour. T/S ratio values progressively declined over the subsequent time intervals with mean +/- SD and median values of 0.6 +/- 0.2 and 0.6, respectively, measured at 4 hours. The mean +/- SD maximum percent dose uptake of pertechnetate within the thyroid gland was 0.55% +/- 0.15% with a range of 0.28% to 0.90%. The mean +/- SD time interval from injection of pertechnetate to maximum uptake within the thyroid gland was 160 +/- 55 minutes with a range 31-240 minutes. The data derived from this study of normal dogs may be useful in the evaluation of dogs with thyroidal diseases including hypothyroidism.

Centering balloon to improve esophageal photodynamic therapy.

A cylindrical balloon was developed to improve delivery of circumferential light for photodynamic therapy (PDT) of esophageal carcinoma. The balloon consisted of a 36-mm-long clear cylindrical membrane and a central tube to hold a cylindrical diffuser in the center of the lumen. Three isotropic probes were placed on the outside of the balloon to allow measurement of delivered light dose to the esophageal mucosa. The balloon was tested in the normal esophagus of 8 dogs that were injected with 4.0 mg/kg of PHOTOFRINR. Endoscopy was performed 48 hours following the injection, and under endoscopic observation the balloon assembly was passed, fixed in place, and inflated. A 1-cm cylindrical diffuser was passed into the central tube and 150, 300, and 600 Joules/cm of 630 nm laser light was delivered at 25 cm, 15 cm, and 5 cm proximal to the gastroesophageal junction. One control dog was illuminated using the cylindrical diffuser alone at doses of 300 and 600 Joules/cm of diffuser. Complete circumferential tissue response was obtained when the balloon was used. Relatively uniform light intensities were measured around the lumen. In contrast, noncircumferential and unpredictable PDT responses were generated when the cylindrical diffuser was used without the balloon.

Unusual presentation of an Amazon parrot (Amazona species) with hepatocellular carcinoma.

Non-haematopoietic hepatic malignancies are uncommon in birds. The clinical presentation (i.e. chronic buphthalmos)and non-specific radiographic findings observed in this adult Amazon parrot (Amazona spp.) were not consistent with previous reports describing the natural behaviour of metastatic hepatocellular carcinoma in birds.

Drug-using and nonusing women: potential for child abuse, child-rearing attitudes, social support, and affection for expected baby.

Eighty pregnant women (25 substance using, 55 nonusing) from an American prenatal clinic serving lower-income to working-class women responded to questionnaire measures of child-rearing attitudes. The drug users' primary substance of misuse was cocaine (68%), alcohol (16%), amphetamines (12%), or sedatives (4%); polydrug use was documented for 80% of the women. The two (user and nonuser) groups were not different on demographic (age, race, marital status, education, SES, source of income) or obstetrical factors (number of pregnancies, number of children). Drug-using women scored significantly higher on a measure of child abuse potential; more than half scored in the range of clinical criterion for extreme risk. As their babies were not yet born, no actual physical abuse was documented, only a higher potential for abuse. The subgroup who were both drug users and had lower social support scored higher on child abuse potential than all other subgroups. The drug users also had lower self-esteem scores than the nonusers. The two groups did not differ on measures of overall social support, authoritarian/democratic child-rearing beliefs, or affection for the expected baby.