A novel MCT1 and MCT4 dual inhibitor reduces mitochondrial metabolism and inhibits tumour growth of feline oral squamous cell carcinoma

Monocarboxylate transporters (MCTs) support tumour growth by regulating the transport of metabolites in the tumour microenvironment. High MCT1 or MCT4 expression is correlated with poor outcomes in human patients with head and neck squamous cell carcinoma (HNSCC). Recently, drugs targeting these transporters have been developed and may prove to be an effective treatment strategy for HNSCC. Feline oral squamous cell carcinoma (OSCC) is an aggressive and treatment‐resistant malignancy resembling advanced or recurrent HNSCC. The goals of this study were to investigate the effects of a previously characterized dual MCT1 and MCT4 inhibitor, MD‐1, in OSCC as a novel treatment approach for feline oral cancer. We also sought to determine the potential of feline OSCC as a large animal model for the further development of MCT inhibitors to treat human HNSCC. In vitro, MD‐1 reduced the viability of feline OSCC and human HNSCC cell lines, altered glycolytic and mitochondrial metabolism and synergized with platinum‐based chemotherapies. While MD‐1 treatment increased lactate concentrations in an HNSCC cell line, the inhibitor failed to alter lactate levels in feline OSCC cells, suggesting an MCT‐independent activity. In vivo, MD‐1 significantly inhibited tumour growth in a subcutaneous xenograft model and prolonged overall survival in an orthotopic model of feline OSCC. Our results show that MD‐1 may be an effective therapy for the treatment of feline oral cancer. Our findings also support the further investigation of feline OSCC as a large animal model to inform the development of MCT inhibitors and future clinical studies in human HNSCC.     

Characterization of the cDNA Encoding alphaIIb and beta3 in normal horses and two horses with Glanzmann thrombasthenia

Glanzmann thrombasthenia (GT) is an inherited, intrinsic platelet defect characterized by a quantitative or qualitative change in the platelet glycoprotein complex IIb-IIIa (integrin alpha(IIb)beta3). The subunits are encoded by separate genes and both subunits must be expressed for a stable complex to form on the platelet surface; therefore, a defect in either gene can result in GT.     

Development of an indirect enzyme-linked immunosorbent assay for detecting equine serum antibodies to the lipopolysaccharide of Salmonella abortusequi

An indirect enzyme-linked immunosorbent assay (IELISA) was developed for the detection of equine serum antibodies to lipopolysaccharide of Salmonella enterica subsp. enterica serovar Abortusequi (LPS), a causative organism of Equine Paratyphoid. The data presented demonstrates that horses immunized with S. abortusequi LPS developed antibodies detectable by the IELISA. By comparison, the tube agglutination test (TAT) did not detect antibody to S. abortusequi LPS as consistently as the IELISA. The data suggests that the IELISA may be a more suitable test for the detection of serum antibodies to S. abortusequi than the TAT.     

Effects of level of feed intake and Fusarium toxin-contaminated wheat on rumen fermentation as well as on blood and milk parameters in cows

The aims of this study were to examine the effects of and possible interactions between dry matter (DM) intake and feeding Fusarium toxin-contaminated wheat on ruminal fermentation, serum chemical parameters and milk yield of dairy cows. Fourteen dairy cows equipped with ruminal and duodenal cannulas were analysed. All animals were fed the same ration, the daily feed amounts being adjusted to current performance. On DM basis, the ration consisted of 60% concentrate including 55% wheat [Fusarium-contaminated wheat (mycotoxin period) or control wheat (control period)] and was completed with 40% maize and grass silage. Each cow was fed the contaminated wheat [deoxynivalenol (DON), 8.21 mg/kg DM and zearalenone (ZON), 0.09 mg/kg DM] and the control wheat (0.25 mg DON/kg DM and 51 microg ZON/kg DM). As expected, a higher organic matter (OM) intake decreased the amounts of fermented crude nutrients related to the respective intakes. An increased amount of crude protein degraded (p < 0.05) and a lower molar percentage of propionate in the rumen fluid were observed when feeding the Fusarium toxin-contaminated wheat at increased OM intakes in comparison with the control wheat. The activities of serum aspartate aminotransferase (ASAT; p < 0.001), glutamate dehydrogenase (GLDH; p < 0.01) and gamma glutamyl transferase (gamma-GT; p < 0.01) increased with increasing OM intake and were not related to the mycotoxin contamination of the wheat.     

Immunohistochemical localization of distinct angiotensin II AT1 receptor isoforms in the kidneys of the Sprague-Dawley rat and the desert rodent Meriones crassus

Employing a purified lgG fraction of a polyclonal anti-AT1 receptor anti-body, raised against a synthetic octapeptide encompassing residues 14-21 of the first extracellular domain of the AT1 polypeptide, selective AT1 receptor expression was immunohistochemically demonstrable within renal structures in Sprague-Dawley (SD) rats and the desert rodent Meriones crassus. In both animal models, prominent AT1 receptor labelling was evident in renal vascular elements, particularly cortical inter-lobular arteries (IA) as well as vasa recta bundles in the inner stripe of the outer medulla. Less intense labelling was observed among peritubular capillary endothelia within the deep cortex, and at both the outer stripe and the inter-bundle regions of the inner stripe of the outer medulla. The binding of the anti-peptide anti-body was, however, lacking among glomeruli and, except for the intense labelling confined to basement membranes of Bowman's capsule of deep nephrons, was virtually absent in all renal tubular structures of both animal models. Structural assessment of the expressed AT1 receptors by two-dimensional Western blotting revealed that a spectrum of structurally distinct AT1 receptor isoforms is expressed in the renal tissues of both animal models. This spectrum was constituted by isoforms of equal size (70 kDa) but distinct pls in SD rats, and of both different sizes (67-73 kDa) and isoelectric points in M. crassus. In either species, the charge and/or size heterogeneity of AT1 receptor isoforms may be attributed in part to differential post-translational glycosylation mechanisms of the AT1 receptor polypeptide backbone. The potential for the differential glycosylation state of AT1 receptors to alter recognition properties may add another level of complexity to tissue-specific and/or species-specific mechanisms underlying angiotensin II interactions in the kidney.     

Renal tubular-cell neoplasms in black-footed ferrets (Mustela nigripes)--38 cases

Thirty-eight cases of renal tubular cell neoplasms were diagnosed in 184 captive, adult (>1-year-old), black-footed ferrets (Mustela nigripes) examined from 1985 to 1996. This prevalence (20.7%) is one of the highest reported for this neoplasm in a population of animals. These tumors rarely metastasized (1/38), and usually were incidental postmortem findings, associated clinical disease being present in only 3 (8%) of the 38 cases. The prevalence of renal tubular cell neoplasms found at postmortem examination increased linearly with age, up to 67% in ferrets >8 years old. Both males (prevalence = 19%) and females (prevalence = 24%) were affected. Multiple renal tumors were common, and seven ferrets (18.4% of affected animals) had bilateral tumors. The cause of this neoplastic syndrome could not be determined. Since most of the animals affected by this condition were in their postreproductive years of life, the impact of this neoplastic syndrome on the captive propagation of this species is negligible.     

Detection of chlamydiae in boar semen and genital tracts

Chlamydiae cause abortion and reproductive disorders in sows. Although organisms can infect the male genital tract, little is known about the disease situation in boars. Hence, we examined the prevalence of chlamydial infection in semen and genital tracts of boars. Samples collected from Swiss boars (group A: n=42), and boars from Germany (group B: n=39) were examined by bacteriology, LPS-ELISA, immunohistochemistry (IHC) and polymerase chain reaction (PCR). The latter methodology involved use of three PCR assays including 16Sig rDNA, IGS-S (intergenic spacer 16S/23S-Short) and IGS-L (intergenic spacer 16S/23S-Long) PCR for comparison methods. PCR sensitivity and the presence of potential PCR inhibitors were determined by spiking semen with Chlamydophila (Cp.) abortus DNA. Detection limits of the 16Sig and IGS-S PCR were 10 templates, while the IGS-L PCR was less sensitive (100 templates). Of 25 semen samples that were collected from group A, one semen sample was positive for Cp. psittaci and two were positive for Chlamydia-like organisms by 16Sig PCR. Screening of sera from Swiss boars revealed three animals with positive reactions in the LPS-ELISA, although we failed to detect chlamydiae within organs of these or sera-negative animals by IHC or IGS-S PCR. In group B, 10 ejaculates were positive for Chlamydia (C.) suis and two were positive for Chlamydia-like organisms by 16S PCR. The identification of DNA from Chlamydia-like organisms in semen from both groups of boars was surprising and a role for these bacteria in reproductive diseases requires further assessment. In conclusion, the prevalence of chlamydial infection was low in group A animals indicating that venereal transmission may not be significant for Chlamydia-associated reproductive diseases in pigs, although rare cases may occur.     

Residues of fluoroquinolone drugs in the cloacal gland and other tissues of Japanese quail

1. In this study we investigated the residues of fluoroquinolone drugs (ciprofloxacin and pefloxacin) in the cloacal gland (a site of foam synthesis) and other tissues such as breast muscle, testes, brain, kidney and plasma. 2. Fifty-four healthy male Japanese quail were selected at random from a flock, maintained under uniform husbandry conditions and divided into three groups, each of 18 birds. Group I (control) received 1 ml vehicle (normal saline 0.9% (w/v) NaCl) daily for 12 d through the intraperitoneal route. Birds of groups II and III received ciprofloxacin and pefloxacin by the same route at the rate of 10 and 12 mg/kg body weight, respectively, every day for a similar period. 3. Birds from each group were killed, at 1, 5 and 10 d after the cessation of treatment, to collect the cloacal gland together with other tissues that were analysed for residual drugs. 4. Cloacal gland retained the maximum drug residues of ciprofloxacin (60%) and pefloxacin (80%) on d 10 compared with that on d 1 after drug withdrawal. The drug residues were found 60 and 80% in ciprofloxacin and pefloxacin groups, respectively, in the cloacal gland tissue even on d 10 after withdrawal of the treatment. 5. In the ciprofloxacin-treated group, all tissues except cloacal gland contained very small amounts of the drug residues on d 10 after treatment ended. In the pefloxacin group the cloacal gland, breast muscle and kidney retained a fairly high amount of drug even on d 10 after treatment ceased. No residues of pefloxacin were detectable in testes and brain throughout. 6. In conclusion, the cloacal gland in Japanese quail acted as the largest sink for the fluoroquinolone drugs. Ciprofloxacin was more widely distributed in different tissues and persisted for a shorter period than pefloxacin.     

Concentration of conjugated linoleic acid in grazing sheep and goat milk fat

The conjugated linoleic acid (CLA) content of grazing sheep and goat milk fat, throughout their lactation period, was examined. Six sheep and six goat representative farms were selected at random and milk samples were taken at monthly intervals for fatty acids profile determination. Sheep and goat nutrition was based on natural grazing and on supplementary feeding during the winter months. From April onwards, grazing native pastures was the only source of feed for sheep and goats. The University farm whose sheep are kept indoors all year round without any grazing, was also used as reference. Fifteen individual milk samples were also taken in April from a sheep and goat farm respectively, in order to see the variability of CLA inside the farm. The results showed that: a. the CLA content of grazing sheep and goat milk fat increased significantly in April–May (early growth stage of grass) and then declined while that of indoors kept sheep was more or less constant during the same period, b. the isomers cis-9, trans-11 and trans-10, cis-12 of CLA were found in grazing sheep milk fat, while in indoors kept sheep and goats' milk fat only the cis-9, trans-11 isomer was found, c. the CLA content of sheep milk fat was much higher than that of goats, d. a negative correlation between sheep milk fat and CLA content was found and e. there was considerable variation in milk fat CLA content between sheep and goat farms and inside the farms.