The safety of high-dose buprenorphine administered subcutaneously in cats.

The safety of a proprietary formulation of buprenorphine hydrochloride administered subcutaneously (SC) to young cats was investigated in a blinded, randomized study. Four cohorts of eight cats aged approximately 4 months were administered saline, 0.24, 0.72 or 1.20 mg/kg/day buprenorphine SC for nine consecutive days, representing 0×, 1×, 3× and 5× of the intended dose. Cats were monitored daily for evidence of clinical reactions, food and water intake and adverse events (AEs). Physical examinations, clinical pathology, vital signs and electrocardiograms (ECGs) were evaluated at protocol-specified time points. Complete necropsy and histopathologic examinations were performed following humane euthanasia. Four buprenorphine-treated cats experienced AEs during the study, two unrelated and two related to study drug administration. The two cats with AEs considered related to drug administration had clinical signs of hyperactivity, difficulty in handling, disorientation, agitation and dilated pupils in one 0.24 mg/kg/day cat and one 0.72 mg/kg/day cat. All of these clinical signs were observed simultaneously. There were no drug-related effects on survival, injection response, injection site inspections, body weight, food or water consumption, bleeding time, urinalysis, respiration rate, heart rate, ECGs, blood pressures, body temperatures, macroscopic examinations or organ weights. Once daily buprenorphine s.c. injections at doses of 0.24, 0.72 and 1.20 mg/kg/day for 9 consecutive days were well tolerated in young domestic cats.

The 2008 ACVP role delineation survey and initial data analysis: from the Role Delineation Task Force.

The American College of Veterinary Pathologists commissioned a role delineation survey to define the specialized tasks, knowledge, and tools that define the current practice of veterinary clinical pathology and veterinary anatomic pathology. The survey also identified when competence was acquired for each task (i.e., before certification or after certification). The response rate by diplomates was high, with approximately 50% of practicing pathologists within each specialty responding to each survey. Using the survey results, all tasks for each specialty were classified as either appropriate or unsuitable for testing in the certifying examinations. The role delineation survey data will facilitate the creation of test plans that objectively define the content in each certifying examination, the evaluation and enhancement of training curricula, and the optimization of continuing education opportunities for practicing veterinary pathologists.

Ecotypic and genetic variation in poplar bark storage protein gene expression and accumulation.

Bark storage proteins (BSP) store nitrogen (N) translocated from senescing leaves in autumn, and supply reduced N for spring growth. Expression of bsp and BSP accumulation are associated with short day photoperiod. To determine if photoperiod-associated bsp expression varies among poplars native to different latitudes, Populus deltoides Bartr. clones originating from six latitudes were grown under natural conditions at a common location. Relative amounts of BSP mRNA in these clones were measured at 2-week intervals from August 7 to October 16. The date of maximum BSP mRNA accumulation was correlated with latitude of origin, and maximum accumulation of BSP mRNA occurred earlier in clones native to northern latitudes than in clones native to southern latitudes. This pattern of variation is consistent with photoperiodic responses of plants native to temperate climates. Genotypic variations in BSP accumulation, bark protein concentration and bark N concentration were compared among clones of six hybrid poplar (Populus trichocarpa Torr. and Gray x P. deltoides) full-sib families (three F(2) families, two F(1) families and one BC(1) family) after 6 weeks in a short day photoperiod and at midwinter. Significant differences in BSP accumulation occurred among clones within four of the six full-sib families after 6 weeks in a short day photoperiod and also at midwinter for outdoor-grown plants. Bark protein and bark N concentrations also varied significantly among clones within certain families. In general, the greatest variation was found in F(2) and BC(1) families. Within several families, relative BSP amounts were positively correlated with bark protein concentration and total bark N concentration. These results indicate a role of photoperiod in regulating bsp expression and demonstrate a genetic component underlying seasonal BSP accumulation. The results could have significance in selecting for clones with improved N storage capacity and N-use efficiency.

Increased macrophage chemoattractant protein-1 in cerebrospinal fluid precedes and predicts simian immunodeficiency virus encephalitis.

Macrophage chemoattractant protein-1 (MCP-1) may be a key trigger for the influx of macrophages into the brain in human immunodeficiency virus (HIV) encephalitis. In this study, simian immunodeficiency virus-infected macaques that developed moderate-to-severe encephalitis had significantly higher MCP-1 levels in cerebrospinal fluid (CSF) than in plasma as early as 28 days after inoculation, which was before the development of brain lesions. In contrast, CSF:plasma MCP-1 ratios remained constant at preinoculation levels in macaques that developed minimal or no encephalitis. Abundant MCP-1 protein and mRNA were detected in both macrophages and astrocytes in the brain. Macaques with increased MCP-1 in CSF had significantly greater expression of markers of macrophage and microglia activation and infiltration (CD68; P= .003) and astrocyte activation (glial fibrillary acidic protein; P= .019 and P= .031 in white and gray matter, respectively). The results suggest that the CSF:plasma MCP-1 ratio may be a valuable prognostic marker for the development of HIV-induced central nervous system disease.

The poplar bark storage protein gene (Bspa) promoter is responsive to photoperiod and nitrogen in transgenic poplar and active in floral tissues, immature seeds and germinating seeds of transgenic tobacco.

In Populus, seasonal nitrogen storage involves the accumulation of a 32 kDa bark storage protein (BSP) in the inner bark parenchyma and xylem rays. Poplar BSPs are encoded by a multigene family and one member, bspA, has been cloned and sequenced. The regulation of bspA was investigated by transforming either hybrid poplar or tobacco with a chimeric gene consisting of the 2.8 kb bspA promoter fused to the coding region of beta-glucuronidase (uidA). In transformed poplar, the bspA 2.8 kb promoter conferred both short-day (SD) and nitrogen (N) inducibility to GUS and activity was localized to the bark (primary and secondary phloem, and cortex) and xylem rays. Night-break treatments inhibited SD induction of GUS. Deletion of the 1.6 kb distal DNA sequences from the bspA promoter eliminated SD induction of GUS while some N induction was retained. These results indicate that although poplar BSP is encoded by a multigene family, transcriptional activation of bspA per se can account for bsp expression in bark and xylem rays in response to either SD or N treatment. These results also show that the elements responsible for SD or N induction are separable. Because of the long generation intervals associated with trees, the developmental regulation of bspA in flowers, developing seeds, and germinating seeds was investigated by transforming the 2.8 kb bspA-promoter::uidA chimeric gene into tobacco. The bspA promoter was active in developing tobacco floral tissues and in seeds during early stages of embryogenesis, decreased progressively during seed maturation and regained activity upon seed germination. Although seed storage proteins of poplar share some similarities to poplar BSP, the observed developmental expression patterns in tobacco are consistent with a role for bspA in vegetative rather than seed storage protein storage.

Phytochrome-mediated photoperiod perception, shoot growth, glutamine, calcium, and protein phosphorylation influence the activity of the poplar bark storage protein gene promoter (bspA).

In poplars (Populus), bspA encodes a 32-kD bark storage protein that accumulates in the inner bark of plants exposed to either short-day (SD) photoperiods or elevated levels of nitrogen. In this study, poplars transformed with a chimeric gene consisting of the bspA promoter fused to beta-glucuronidase (uidA) were used to investigate the transcriptional regulation of the bspA promoter. Photoperiodic activation of the bspA promoter was shown to involve perception by phytochrome and likely involves both a low fluence response and a parallel very low fluence response pathway. Activity of the bspA promoter was also influenced by shoot growth. High levels of bspA expression usually occur in the bark of plants during SD but not long day or SD with a night break. When growth was inhibited under growth permissive photoperiods (SD with night break) levels of bark beta-glucuronidase (GUS) activity increased. Stimulating shoot growth in plants treated with SD inhibited SD-induced increases in bark GUS activity. Because changes in photoperiod and growth also alter carbon and nitrogen partitioning, the role of carbon and nitrogen metabolites in modulating the activity of the bspA promoter were investigated by treating excised stems with amino acids or NH4NO3 with or without sucrose. Treatment with either glutamine or NH4NO3 resulted in increased stem GUS activity. The addition of sucrose with either glutamine or NH4NO3 resulted in synergistic induction of GUS, whereas sucrose alone had no effect. Glutamine plus sucrose induction of GUS activity was inhibited by EGTA, okadaic acid, or K-252A. Inhibition by EGTA was partially relieved by the addition of Ca2+. The Ca2+ ionophore, ionomycin, also induced GUS activity in excised shoots. These results indicate that transcriptional activation of bspA is complex. It is likely that SD activation of bspA involves perception by phytochrome coupled to changes in growth. These growth changes may then alter carbon and nitrogen partitioning that somehow signals bspA induction by a yet undefined mechanism that involves carbon and nitrogen metabolites, Ca2+, and protein phosphorylation/dephosphorylation.

Rhinosporidiosis in a domestic cat.

Rhinosporidiosis was diagnosed in a domestic shorthair cat from a suburb of Washington DC, USA. The clinical presentation of protracted sneezing and epistaxis was associated with a polypoid lesion in the right nostril. Light microscopic examination revealed a polypoid lesion with numerous sporangia containing maturing endospores. Free endospores were present in the stroma of the polyp and lumen of the nasal cavity. Transmission electron microscopy revealed ultrastructural features typical of Rhinosporidium seeberi. The case was followed clinically for a total of 70 months and there were five attempts at surgical excision. This is the first reported case of rhinosporidiosis in a domestic cat.

Cystic prostatic disease associated with adrenocortical lesions in the ferret (Mustela putorius furo).

A review of case materials at the Armed Forces Institute of Pathology identified six cases of prostatitis in domestic ferrets (Mustela putorius furo). Five of these ferrets (83%) had one or more cysts and the remaining ferret had a paraprostatic cyst. Three of the six ferrets (50%) exhibited various degrees of prostatic squamous metaplasia. Inflammation ranged from subacute to chronic-active or pyogranulomatous. In addition to the prostatic lesions, hyperplastic and/or neoplastic adrenocortical lesions were present in 4/6 (66%) ferrets; 1/6 (17%) ferrets had previously been clinically diagnosed with adrenal gland-associated endocrinopathy. The remaining ferret had previously had the right adrenal gland removed, but the reason for the removal is unknown. Based upon the histologic findings in these six ferrets, there appears to be an association between proliferative adrenal lesions and cystic prostatitis in domestic ferrets.

T-cell-rich B-cell lymphoma in a cat.

T-cell-rich B-cell lymphoma is a neoplasm recognized in humans in which a neoplastic proliferation of large B lymphocytes is present amid a background of reactive T lymphocytes. A 13-year-old Domestic Shorthair cat developed a mass in the region of the left parotid gland. Histologically, the mass was composed of scattered large atypical cells within a dense background of uniform small lymphoid cells. Immunohistochemically, the large cells were uniformly labeled using antiserum directed against the B-lymphocyte marker BLA.36, whereas labeling of nearly all of the small cells was limited to the T-lymphocyte marker CD3. The histomorphologic and immunohistochemical features of this unique feline neoplasm are characteristic of T-cell-rich B-cell lymphoma of humans.

Does the genome of Corylus avellana L. contain sequences homologous to the self-incompatibility gene of Brassica?

Self-incompatibility is a genetic mechanism enforcing cross-pollination in plants. Hazelnut (Corylus avellana L.) expresses the sporophytic type of self-incompatibility, for which the molecular genetic basis is characterized only in Brassica. The hypothesis that the hazelnut genome contains homologs of Brassica self-incompatibility genes was tested. The S-locus glycoprotein gene (SLG) and the kinase-encoding domain of the S-receptor kinase (SRK) gene of B. oleracea L. were used to probe blots of genomic DNA from six genotypes of hazelnut. Weak hybridization with the SLG probe was detected for all hazelnut genotypes tested; however, no hybridization was detected with PCR-generated probes corresponding to two conserved regions of the SLG gene. One of these PCR probes included the region of SLG encoding the 11 invariant cysteine residues that are an important structural feature of all S-family genes. The present evidence suggests that hazelnut DNA hybridizing to SLG differs significantly from the Brassica gene, and that the S-genes cloned from Brassica will not be useful for exploring self-incompatibility in hazelnut.

Poplar Bark Storage Protein and a Related Wound-Induced Gene Are Differentially Induced by Nitrogen.

Poplars (Populus deltoides Bartr. ex Marsh) accumulate a 32-kD bark storage protein (BSP) in phloem parenchyma and xylem ray cells during autumn and winter. Accumulation of poplar BSP is associated with short-day (SD) photoperiods. Poplar BSP shares sequence similarity with the product of the wound-inducible poplar gene win4. The influence of nitrogen availability and photoperiod on the levels of BSP, BSP mRNA, and win4 mRNA was investigated. In long-day (LD) plants BSP, BSP mRNA, and win4 mRNA levels were correlated with the amount of NH4NO3 provided to the plant. BSP mRNA and BSP were detected only in bark, whereas win4 mRNA was detected only in leaves. In LD plants treated with NH4NO3, BSP mRNA levels were significantly greater than those of win4. In nitrogen-deficient plants exposed to SD conditions, the accumulation of BSP mRNA and BSP was delayed for 2 weeks. This delay was eliminated by further SD exposure, and after 6 weeks of SD treatment similar levels of BSP and BSP mRNA were detected in the bark of SD plants regardless of the level of NH4NO3 treatment. win4 mRNA levels declined to undetectable levels in young leaves of SD plants but increased in mature leaves. These results indicate that BSP accumulation in both LD and SD plants is influenced by nitrogen availability. Although both BSP and win4 appear to be involved in nitrogen storage, our data suggest that BSP is probably the primary protein involved in both seasonal and short-term nitrogen storage in poplar. These results also suggest that nitrogen cycling and storage in poplar could involve a two-component system. In this system the win4 gene product may modulate accumulation and mobilization of leaf nitrogen, whereas BSP is involved in seasonal and short-term nitrogen storage during periods of excess nitrogen availability.

A family of wound-induced genes in Populus shares common features with genes encoding vegetative storage proteins.

Two wound-inducible cDNAs from poplar leaves show sequence identity to vegetative storage proteins (VSP) that accumulate seasonally in poplar bark tissues. We have compared the genomic organization, cDNA sequences and expression of the genes encoding the wound-inducible cDNAs (win4) with that of a bark VSP (called bark storage protein, or BSP). There appear to be several win4 genes in the poplar genome which segregate as a single locus and are therefore likely to be clustered. The same is true of the BSP genes. The win4 locus is linked (map distance of 5 cM) to the BSP locus, consistent with a common evolutionary origin of the genes. A near full-length win4 cDNA shows 75% sequence identity to BSP cDNAs. Both win4 and BSP are systemically wound-inducible; win4 transcripts accumulate in leaves and stems, whereas BSP transcripts accumulate almost exclusively in stems. A phloem transport-dependent signaling mechanism appears to be involved in systemic win4 expression after wounding. In contrast to BSP gene expression, win4 genes are not expressed in response to short day conditions. The data indicate win4 and BSP genes are differentially regulated, and their products may play important roles in the storage and reallocation of nitrogen in perennial plants.

Physiological and Environmental Requirements for Poplar (Populus deltoides) Bark Storage Protein Degradation.

In poplar (Populus deltoides Bartr. ex Marsh), a 32-kD bark storage protein (BSP) accumulates in the bark during autumn and winter and declines during spring shoot growth. We investigated the physiological and environmental factors necessary for the degradation of poplar BSP. Poplar plants were exposed to short-day (SD) photoperiods for either 28 or 49 d. Plants exposed to short days for 28 d formed a terminal bud but were not dormant, whereas exposure to short days for 49 d induced bud dormancy. BSP accumulated in bark of plants exposed to both SD treatments. The level of BSP declined rapidly when nondormant plants were returned to long days. BSP levels did not decline in dormant plants that were exposed to long-day (LD) conditions. If dormant plants were first treated with either low temperatures (0[deg]C for 28 d) or with 0.5 M H2CN2 to overcome dormancy and then returned to long days, the level of BSP declined. Removal of buds from non-dormant or dormant plants in which dormancy had been overcome inhibited the degradation of BSP in LD conditions. BSP mRNA levels rapidly declined in plants exposed to long days, irrespective of the dormancy status of the plants or the presence or absence of buds. These results indicate that the buds of poplars are somehow able to communicate with bark storage sites and regulate poplar BSP degradation. These results further support an association of BSP mRNA levels with photoperiod because short days stimulate BSP mRNA accumulation, whereas long days result in a decline of BSP mRNA abundance.

Complementary DNA cloning of poplar bark storage protein and control of its expression by photoperiod.

Bark storage proteins accumulate in the bark of many woody plants during autumn and winter. In poplar (Populus deltoides Bartr. ex Marsh), the accumulation of the 32-kilodalton bark storage protein is controlled by photoperiod. We have isolated a full-length cDNA encoding for the poplar 32-kilodalton bark storage protein and determined its nucleotide sequence. The derived amino acid sequence shows that poplar bark storage protein is rich in serine, leucine, phenylalanine, and lysine. Poplar bark storage protein is similar to the poplar wound-induced cDNA clone 4 and clone 16 (TJ Parsons, HD Bradshaw, MP Gordon [1989] Proc Natl Acad Sci USA 86: 7895-7899). DNA gel blot analysis suggests that poplar bark storage protein is encoded by a multigene family of about five genes. Poplar plants grown in long days contained low levels of mRNA for the bark storage protein. Exposure to short days resulted in an increase in bark storage protein mRNA within 7 days. After 21 days of short day exposure, high levels of mRNA were detected. The accumulation of bark storage protein mRNA in response to short days was also observed in plants exposed to natural shortening daylengths. Our results indicate that the accumulation of poplar bark storage protein mRNA is controlled by photoperiod. This finding will provide a useful system for investigating photoperiodism in woody plants.

Photoperiod control of poplar bark storage protein accumulation.

Bark storage proteins (BSPs) accumulate in the inner bark parenchyma of many woody plants during autumn and winter. We investigated the effect of a short-day (SD) photoperiod on the accumulation of the 32-kilodalton bark storage protein of poplar (Populus deltoides Bart. ex Marsh.) under controlled environmental and natural growing conditions. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and protein gel blot analysis revealed that 10 days of SD exposure (8 hours of light) resulted in a 20% increase in the relative abundance of the 32-kilodalton bark storage protein of poplar. After 17 days of SD exposure, the 32-kilodalton bark storage protein accounted for nearly one-half of the soluble bark proteins. In natural field conditions, accumulation of the 32-kilodalton bark storage protein was observed to start by August 18 (daylength 14.1 hours). Immunoprecipitation of in vitro translation products with anti-BSP serum revealed that the SD protein accumulation was correlated with changes in the pool of translatable mRNA. A survey of poplar clones from different geographic origins revealed the presence of the 32-kilodalton BSP in the dormant bark of all the clones tested. These results demonstrate that a SD photoperiod induces, whether directly or indirectly, rapid changes in woody plant gene expression, leading to the accumulation of BSP.

Vaxstrate: an anti-reproductive vaccine for cattle.

This paper describes the development of a vaccine for the prevention of pregnancy in female cattle. The vaccine is based on the established principle that antibodies to the hypothalamic releasing hormone, gonadotrophin releasing hormone (GnRH) block the action of GnRH on pituitary secretion of luteinizing hormone and follicle stimulating hormone, leading to gonadal atrophy in mammals. The vaccine comprises an immunogenic GnRH:ovalbumin conjugate formulated in a novel double adjuvant system and is administered in a two-dose treatment regimen. Field trials have confirmed efficacy and the product, Vaxstrate, has now been registered and commercialized.

Axillary shoot proliferation and growth of Populus deltoides shoot cultures.

Shoot cultures of four genotypes of Populus deltoides Bartr. ex Marsh. were established from adventitious shoots regenerated from internodal stem explants. Stable shoot cultures for all four genotypes were maintained in a continuous culture regime for over one year. The stable shoot cultures were used as explants to investigate the effects of zeatin concentration and genotype on axillary shoot production and growth. The concentration of zeatin significantly affected the production of axillary shoots, with 1.0 mgL(-1) zeatin producing the greatest number of shoots (31.0 shoots per culture vessel) while 0.25 mgL(-1) zeatin produced the greatest growth (5.9 mg per axillary shoot) when measured by dry weight accumulation per shoot. Genotypic differences were significant in the production and growth of axillary shoots.

In vitro shoot regeneration of Populus deltoides: effect of cytokinin and genotype.

Treatment differences were observed in the in vitro adventitious shoot regeneration response from internodal explants of three genotypes of Populus deltoides cultured on media supplemented with five concentrations each of the cytokinins 6-benzyladenine, 2-isopentyladenine, and zeatin. For each of the three genotypes, the greatest number of shoots were consistently regenerated on media containing the cytokinin zeatin. Tissue necrosis resulted when explants from any of the three genotypes were cultured on media supplemented with 6-benzyladenine. A zeatin concentration by genotype interaction was also observed. Genotypic differences in shoot regeneration were observed for 16 genotypes of Populus deltoides when cultured on medium supplemented with 0.5 mgL(-1) zeatin. Six genotypes were highly recalcitrant and failed to regenerate shoots. The percent of explants regenerating was greater than 50% for four genotypes.