Relationship between testicular transferrin and plasma estradiol-17beta concentrations of dogs with azoospermia and dogs with sertoli cell tumors.

Testicular Transferrin (Tf) and peripheral plasma estradiol-17beta (E2) concentrations were measured in 3 dogs with azoospermia (AZ dogs), 3 dogs with Sertoli cell tumors (SC dogs), and 5 normal male Beagles. The mean Tf concentrations in the testes of the AZ dogs and the affected testes of the SC dogs, and the plasma E2 concentrations in both these groups of dogs were significantly higher than the values in normal dogs (P<0.05, 0.01 and 0.01, respectively). Therefore, excessive E2 secretion by hyperfunctioning Sertoli cells is thought to have caused the azoospermia in the 3 dogs.     

Intracranial germ cell tumours in two dogs

Intracranial germ cell tumours were identified in two middle-aged, male dogs. The predominant clinical signs were progressive visual and pupillary abnormalities; one dog was blind. At post mortem examination a discrete extramedullary tumour was found at the base of the brain in each dog. Expansion of the mass had produced severe destruction of the diencephalon and brain stem. Histopathological examination revealed a mixed cellular pattern, consistent with a neoplasm derived from germ cells.     

Effects of stage and number of tumours on prognosis of dogs with cutaneous mast cell tumours

Between 1997 and 1999, 280 dogs with mast cell tumours were identified, of which 59 (21 per cent) had multiple tumours. Follow-up data for survival analysis were available for 145 dogs with single tumours and 50 dogs with multiple tumours. There was no significant difference between the survival times of the two groups; the survival rates after 12 and 24 months were 88 per cent and 83 per cent, respectively, for the dogs with single tumours, and 86 per cent at both intervals for the dogs with multiple tumours. Eight of the dogs with single tumours had lymph node metastases (stage II disease) and these dogs had a median survival time of 431 days, whereas the 50 dogs with multiple tumours (classified as stage III disease) and the dogs with single tumours (classified as stage I disease) had not reached their median survival times. Golden retrievers appeared to be predisposed to developing multiple tumours in the population studied, with an odds ratio of 3.8. This study found no evidence that dogs with multiple tumours had different survival times than those with single tumours, although there was evidence that the presence of lymph node metastasis generally carried a poorer prognosis.     

Successful long term culture of immature porcine sertoli cells in the reconstructed testicular cell cord

The ultimate goal of this study was to establish an in vitro system to produce sperms. To pursue this goal, immature porcine testicular cells were cultured in stereostructural form and cultured testicular cord was investigated morphologically. At 4 weeks of age, the seminiferous tubules of the porcine testes consisted of undifferentiated germ cells (gonocytes and undifferentiated spermatogonia) and immature Sertoli cells. The interstitial tissue was largely occupied by Leydig cells. The testes were enzymatically digested, and the dispersed cells were encapsulated with alginate either immediately or after freeze-thawing. The resulting testicular cell cords were cultured for up to 10 weeks. After 2 weeks of culture, Sertoli cells, which were identified by their inhibin-positive reaction in immunohistochemistry, and Leydig cells, which were identified by their morphological characteristics, were observed in the cords. Neither undifferentiated nor differentiated types of germ cells were detected. The number of cells in the cords progressively decreased during the culture period. In order to discover the fate of the Sertoli cells, the level of inhibin in the spent media was determined. Inhibin in the media was at a detectable level after 2 days of culture. The levels increased and peaked at 2 weeks. When frozen-thawed testicular cells were applied to the culture, the peak level was maintained for over 8 weeks, in contrast to the gradual decrease of inhibin level when fresh cells were cultured. These results indicate that the culture conditions can sustain the survival of Sertoli cells. Further improvement is required for proliferation and differentiation of germ cells.     

Recurrence rates and clinical outcome for dogs with grade II mast cell tumours with a low AgNOR count and Ki67 index treated with surgery alone

Grade II mast cell tumours (MCT) are tumours with variable biologic behaviour. Multiple factors have been associated with outcome, including proliferation markers. The purpose of this study was to determine if extent of surgical excision affects recurrence rate in dogs with grade II MCT with low proliferation activity, determined by Ki67 and argyrophilic nucleolar organising regions (AgNOR). Eighty‐six dogs with cutaneous MCT were evaluated. All dogs had surgical excision of their MCT with a low Ki67 index and combined AgNORxKi67 (Ag67) values. Twenty‐three (27%) dogs developed local or distant recurrence during the median follow‐up time. Of these dogs, six (7%) had local recurrence, one had complete and five had incomplete histologic margins. This difference in recurrence rates between dogs with complete and incomplete histologic margins was not significant. On the basis of this study, ancillary therapy may not be necessary for patients with incompletely excised grade II MCT with low proliferation activity.     

Acute phase protein levels in dogs with mast cell tumours and sarcomas

The acute phase proteins (APP) form part of a non-specific host response to inflammation. They may be induced by a range of different causes, including infection, inflammation, cancer and trauma. As they form part of the earliest response to such insults, they have potential for early identification of disease. In people, APP levels have been shown to correlate both with the extent of disease and also the prognosis in several forms of neoplasia, including prostate, oesophageal and colorectal cancer. As such, they can be used as prognostic and monitoring tools. To date, similar studies in veterinary patients have been limited, largely retrospective in nature and many are non-specific for tumour type. The purpose of this study was to evaluate a panel of four APPs in dogs with naturally occurring mast cell tumours (MCTs) and sarcomas to identify in the first instance whether increased levels of individual APPs, or identifiable combinations of APPs, was linked with the presence of disease. In the patients with MCTs, C-reactive protein (CRP) and α-1 acid glycoprotein levels increased, with a concurrent drop in serum amyloid A levels. In the sarcoma patients, CRP, α-1 acid glycoprotein and haptoglobin were increased. These findings suggest that specific solid tumour types in dogs may be associated with specific changes in APP profiles.     

Ultrasonographic diagnosis of non-palpable Sertoli cell tumours in infertile dogs

Three previously fertile dogs were examined with a history of recent infertility. Semen evaluation revealed marked spermatozoal abnormalities, including lesions of the mid-piece region, poor spermatozoal motility and a low total spermatozoal output. The mean number of live normal spermatozoa was 21·1 ± 17·4 × 10⁶ spermatozoa. Examination of the testes with diagnostic B-mode ultrasound demonstrated solitary mass lesions which in each case were not palpable. Plasma oestrogen concentrations were elevated (mean, 43·0 ± 8·s 5 pg/ml), and histologically the masses were shown to be Sertoli cell tumours. Several months after unilateral orchidectomy there was an increase in the percentage of normal spermatozoa and an increase in the spermatozoal motility and total spermatozoal output. The mean number of live normal spermatozoa was 149·8 ± 22·9 × 10⁶ spermatozoa and all the dogs subsequently returned to fertility. Ultrasonographic examination of the testes should be considered part of the routine investigation of the male reproductive tract.     

Chlorambucil and prednisolone chemotherapy for dogs with inoperable mast cell tumours: 21 cases

O bjectives : To evaluate the response of measurable canine mast cell tumours unsuitable for other treatment modalities to a chemotherapy protocol comprising chlorambucil and prednisolone.
M ethods : Dogs bearing measurable mast cell tumours, unsuitable for treatment by surgery or radiotherapy, were treated with orally administered prednisolone and chlorambucil, and their responses assessed.
R esults : Twenty-one dogs were enrolled in the study; 13 had intermediate-grade mast cell tumour, six were high grade and two were diagnosed by cytology alone. Eight dogs had multiple tumours and 13 dogs had single tumours, and six dogs had lymph node metastases and no dogs had visceral metastases detected. Three dogs achieved complete remission, five achieved partial remission (overall response rate 38 per cent), nine had static disease and four dogs had progressive disease. Median progression-free interval for the eight responders was 533 days, and median survival time for all dogs in the study was 140 days. Progression-free interval and median survival time were not influenced by the age, sex, weight or neutering status of the patient, by the grade or stage of the tumour or whether the patient had single or multiple tumours. No toxicity was detected.
C linical S ignificance : Response and survival rates of inoperable canine MCT to chlorambucil and prednisolone are comparable to previously described protocols, with no apparent toxicity.     

European consensus document on mast cell tumours in dogs and cats

In preparing this document the Authors aimed to pool current information on canine and feline mast cell disease. The information was gathered from international studies and a emphasis was placed on material and opinion with a strong evidence base. We intend it to form the basis of our understanding in this disease at the current time and we anticipate that it will be particularly useful for the general practitioner. It should be emphasized that the authors are presenting this work from a European perspective.     

Cytologic features of testicular tumours in dog

In this paper, we report on our experience of cytology of fine needle biopsies performed on 92 dogs with testicular tumours during the period from 1998 to 2002. Cytological diagnosis was consistent with seminoma in 20 cases, sertolioma in 16 cases, Leydig cell tumours in 50 cases and mastocytoma in one case. Five cases could not be diagnosed by cytology. Cytological observations were confirmed after surgery by histopathological examination in 87 cases. Cytology provided a sensitivity of 95% for seminoma, 88% for sertolioma and 96% for Leydig cell tumours. The specificity was 100% for all three tumour types. In our experience cytology of fine needle aspirations of testicular tumours is a very reliable technique.     

Immunohistochemical expression of dogTERT in canine testicular tumours in relation to PCNA,ki67 and p53 expression.

The objective of the study was to determine the immunohistochemical expression of canine TERT in canine testicular tumours comparing two different antibodies for TERT, and to correlate them with well established markers specific to dividing cells such as PCNA and ki67, and with expression of the p53 tumour suppressor gene. The study included 36 cases of canine testicular tumours, which were categorized as 12 Sertoli Cell Tumours (SCT), 20 seminomas, 3 interstitial cell tumours and 1 mixed germ cell-sex cord stromal tumour (MT). Two antibodies for hTERT were examined; a highly specific TERT antibody, RCK-hTERT, was evaluated for the first time. Immunodetection of RCK-hTERT was observed in 31% of tumours examined (6/20 Seminomas, 4/12 SCT, 1/3 interstitial cell tumour and 0/1 mixed germ cell-sex cord stromal tumour), while the NCL-hTERT in 67% of them (15/20 Seminomas, 6/12 SCT, 3/3 interstitial cell tumour and 0/1 ΜΤ). PCNA immunoreactivity was detected in all cases. Regarding ki67, 3 SCT, 12 seminomas and all interstitial cell tumours showed clear immunoreaction. p53 immunoreactivity was detected in 6 SCT, 15 seminomas and all interstitial cell tumours. The immunohistochemical expression of both TERT antibodies are discussed and compared in order to clarify their potential usefulness in canine testicular malignancies in relation to the expression of well known cell cycle markers. Our results indicate that TERT and PCNA are useful proliferation markers but not helpful to evaluate prognosis. Instead of that ki67 and p53 could be used for predicting aggressiveness in this group of tumours     

Association of Ki67 index with prognosis for intermediate‐grade canine cutaneous mast cell tumours*

Intermediate‐grade mast cell tumours (MCT) represent a heterogeneous population of tumours. The prognosis for the majority of dogs is excellent following surgical excision, but a minority die because of their disease. A previous study identified Ki67 expression as a predictor of prognosis in all three grades of MCT. The purpose of this study was to validate those results in a new group of dogs, with intermediate‐grade MCT only. Ki67 immunohistochemistry was performed on intermediate‐grade MCT from 163 dogs with known outcome. Digital microscopy images were taken from each tumour, and an index calculated of Ki67‐positive cells. Ki67 index as a binary variable with a cut‐off value of 1.8% was confirmed to be associated with prognosis (hazard ratio = 19.1, P < 0.0001) for this cohort of dogs. The 1‐year, 2‐year and 3‐year survival probabilities (with standard errors) of 127 dogs with a Ki67 index ≤1.8% were [0.95 (0.024), similar for all] and for 36 dogs with a Ki67 index >1.8% were 0.54 (0.100), 0.45 (0.101) and 0.33 (0.104), respectively.     

Treatment of spontaneous malignant tumours in dogs with PTMC

The chemical analysis of the caffeine salt of a phosphomolybdic-tungstic acid compound (PTMC) is described. Twenty-seven dogs with advanced malignant tumours have been treated with PTMC by intramuscular or intravenous injection. Five dogs - one case of metastatic osteosarcoma, two cases of metastatic mammary carcinoma, one case of metastatic squamous cell carcinoma and one case of primary fibrosarcoma showed objective tumour regression for periods varying from four weeks to 20 months. Only two of the 27 dogs treated showed toxic effects.     

Cytological lymph node evaluation in dogs with mast cell tumours: association with grade and survival*

The purpose of this retrospective cohort study is to describe the association of cytological assessment of lymph node metastasis with survival and tumour grade in dogs with mast cell tumours. Regional lymph node aspirates of 152 dogs diagnosed with a mast cell tumour were reviewed and classified according to specific cytological criteria for staging. 97 dogs (63.8%) had stage I tumours, and 55 (36.2%) had stage II tumours. Stage II dogs had a significantly shorter survival time than dogs with stage I disease (0.8 and 6.2 years, respectively; P < 0.0001). Dogs with grade III mast cell tumours were more likely to have stage II disease (P = 0.004). These results suggest that cytological evaluation of lymph nodes in dogs with mast cell tumours provides useful and valuable clinical information, and the results correlate with tumour grade and outcome thus providing a practical and non‐invasive method for staging.     

Immunohistochemical detection of proliferating cell nuclear antigen and Ki-67 in mast cell tumors from dogs

OBJECTIVE: To determine whether immunohistochemical detection of proliferating cell nuclear antigen (PCNA) and Ki-67 correlated with prognosis for dogs with cutaneous mast cell tumors (MCT). DESIGN: Case series. ANIMALS: 120 dogs with solitary cutaneous MCT that were excised. PROCEDURE: Information on signalment, history, and outcome was obtained by sending a questionnaire to referring veterinarians. Tumors were graded histologically, and immunohistochemical staining for Ki-67 and PCNA was performed. RESULTS: Survival rates 12, 18, and 24 months after surgery were significantly different among groups when dogs were grouped on the basis of histologic grade. Although mean number of PCNA-positive nuclei/1,000 tumor nuclei was significantly higher for dogs that died of MCT than for those that survived, there was great overlap in values. Mean number of Ki-67-positive nuclei/1,000 tumor nuclei was significantly higher for dogs that died of MCT than for those that survived, without any overlap in values between groups, and number of Ki-67-positive nuclei/1,000 tumor nuclei was significantly different among groups when tumors were grouped on the basis of histologic grades. For dogs with grade-II tumors, number of Ki-67-positive nuclei/1,000 tumor nuclei (< 93 vs > or = 93) was significantly associated with outcome (survived vs died). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that for dogs with solitary cutaneous MCT, determining number of Ki-67-positive nuclei may be useful in predicting prognosis, particularly for dogs with grade-II tumors.     

Incorporation of sentinel lymph node mapping in dogs with mast cell tumours: 20 consecutive procedures

The study hypothesis is that incorporation of sentinel lymph node (SLN) mapping in dogs presenting for mast cell tumour (MCT) removal would impact the recommended adjuvant therapy offered. Nineteen dogs were enrolled having either spontaneously occurring or incompletely excised MCTs. Staging included regional lymph node aspiration. SLN mapping was done with regional lymphoscintigraphy combined with intra‐operative lymphoscintigraphy and blue dye. Twenty MCTs in 19 dogs were excised with SLN mapping. Eight dogs had SLNs different from the closest node. Twelve dogs had metastasis in extirpated SLNs, seven occurred in MCTs with a MI ≤ 5. No correlation was noted between patient stage and the c‐KIT proto‐oncogene. Because of SLN staging, 8 of 19 dogs were offered additional therapy that would have otherwise been excluded. Anatomic sampling of lymph nodes in dogs with MCTs does not accurately reflect which lymph nodes are most likely to be receiving the draining tumour lymph.     

Pretreatment leukocyte ratios and concentrations as predictors of outcome in dogs with cutaneous mast cell tumours

Leukocyte ratios correlate with outcome in several human cancers. Little is known about their prognostic significance in mast cell tumour (MCT). The aim of the study was to evaluate the prognostic significance of pretreatment leukocyte concentrations and their ratios in dogs with MCT for survival. Medical records of 92 dogs with MCT were retrospectively reviewed. Tumour diagnosis was made by tumour biopsy or fine‐needle aspirate. Only dogs without prior treatment were included. Eosinophil, lymphocyte, monocyte and neutrophil concentration were obtained by ADVIA 2120? (Siemens Healthcare, Vienna, Austria). Neutrophil‐to‐eosinophil ratio (NER), lymphocyte‐to‐monocyte ratio (LMR) and neutrophil‐to‐lymphocyte ratio (NLR) were calculated from collected leukocyte concentrations. Relative eosinophil concentration (REC), NER (P < 0.001), NLR (P = 0.001) and LMR (P < 0.001) were significant prognostic factors for outcome in univariate analysis. REC (P = 0.008) and NER (P = 0.001) remained independent predictors of survival in multiple analyses. Leukocyte concentrations and ratios, especially REC and NER may serve as prognostic indicators in MCT.     

Long-term follow-up of dogs with patent ductus arteriosus

Postocclusion survival data from dogs with left-to-right shunting patent ductus arteriosus (PDA) was available from 80 dogs, diagnosed from 1990 to 2000. Of these, 37 had undergone a procedure to close the ductus and were re-evaluated at the time of this study; clinical data from the follow-up examination was compared with that from the original examination. Radiographically, the right ventricle remained apparently enlarged, and the aortic bulge associated with dilation of the descending aorta did not disappear after closure. On M-mode echocardiography, left ventricular chamber diameter in diastole and systole and left ventricular posterior wall in systole decreased significantly. Mitral endocardiosis was a common feature. Residual flow was evident in 46 per cent of the animals. Late closure occurred in 8 per cent of the dogs, and trivial recanalisation in 19 per cent. The maximum survival time postclosure was 168 months and, after non-occlusion, 114 months, suggesting that dogs with PDA follow an unpredictable course. However, there was a significant difference in survival times between the corrected and non-corrected group.