Role of mastitis in on-farm deaths of Finnish dairy cows.

According to our recent necropsy-based study, mastitis is the most common underlying diagnosis of on-farm deaths in Finnish dairy cows. However, it remained unanswered to what extent mastitis has contributed to death of all necropsied cows. In the present study, based on histopathology, we detected one-third of the necropsied dairy cows having active inflammatory udder lesions (n = 110). The role of mastitis varied and was interpreted by causes of death (underlying, intermediate, immediate, other significant). Mastitis was most commonly the underlying (28%) or both immediate and underlying cause of death (48%), and only seldom was it the immediate (4%) or intermediate (4%) cause of death. Mastitis occurred either as the only cause leading to death (mastitis only, MO; 39%), or with many other contributing diseases (multiple diseases, MD; 61%), which were interacting with mastitis and together leading to death. Between these groups (MO vs. MD), time of mastitis occurrence during lactation, producer-reported duration of illness, clinical signs, and medication differed, as well the histopathologic severity of mastitis. The cases where mastitis was the only initial insult occurred evenly throughout the entire lactation, but the cases with many interacting diseases were clustered in early lactation. In MD cows, cases of mastitis occurred concurrently with metritis (31%), aspiration pneumonia (24%), acute trauma or dystocia (15%), or with other diseases, such as ketosis, hepatic lipidosis, rumenitis, and abomasal diseases. For a pathologist, the gross mastitis diagnosis was most challenging at the beginning of the lactation, especially if inflammation was mild to moderate, suggesting the value of histopathologic examination being highest at that time. Also, producers reported mastitis signs less frequently if cow had many simultaneously occurring diseases. Therefore, even if clinical signs of other diseases are present, the udder should be considered a potential cause of illness and should be examined, especially in dry and transition-period cows.

Necropsy-based study on dairy cow mortality-Underlying causes of death.

The worldwide problem of increasing dairy cow mortality is widespread in modern production systems, it causes economic losses, and indicates problems with herd health and welfare. Most studies on causes of dairy cow mortality are limited as they are based on secondary register data, or questionnaires for producers or veterinarians, and neither necropsies nor histopathologic analyses are usually performed. For this reason, no definite causes for dairy cow deaths have been determined making it difficult or impossible to implement effective preventive measures. The objectives of this study were to (1) determine the causes of on-farm mortality of Finnish dairy cows, (2) determine the usefulness of routine histopathologic analysis in bovine necropsies, and (3) assess how reliable producers' perception about the cause of death is. Underlying diagnoses of on-farm deaths were determined through necropsy of 319 dairy cows at an incineration plant. The necropsy data were combined with background information obtained from online questionnaires covering cow and herd records. Mastitis was the most common underlying diagnosis of death (26.6%), followed by digestive disorders (15.4%), other known disorders (13.8%), calving-associated disorders (12.2%), and locomotion disorders (11.9%). The underlying diagnoses of death varied during different stages of lactation and with parity. A large proportion of the study cows (46.7%) died during the first 30 d after calving, and of those, 63.6% died during the first 5 d. A routine histopathologic analysis was performed in every necropsy, and it changed the preliminary gross diagnosis in 18.2% of the cases. Producers' perception about the cause of death agreed with the necropsy-based underlying diagnosis of death in 42.8% of the cases. It was most consistent for mastitis, calving disorders, locomotion diseases, and accidents. In cases where producers had no insights about the cause of death, necropsy revealed the final underlying diagnosis in 88.2% of cases, demonstrating the usefulness of necropsy. Based on our findings, necropsies provide useful and reliable information to develop control programs for cow mortality. Including routine histopathologic analysis in necropsies, more accurate information may be obtained. Furthermore, targeting preventive measures on transitional cows may be most effective, as the number of deaths was highest at this time.

Mechanisms behind the varying severity of Aleutian mink disease virus: Comparison of three farms with a different disease status.

Aleutian mink disease virus (AMDV) is distributed widely among mink farms and wild mustelids despite ongoing attempts to stop the spread. The severity of Aleutian disease (AD) varies from subclinical to fatal but the reasons for its varying severity are complex and unclear. Recently, breeding of tolerant mink has drawn attention as the possible solution to reduce the effects of AD in farms. The aim of this study was to gather information on the effects of breeding based on overall health, production traits, and antibody titer on AD severity by comparing a positive farm (farm 1) that has been breeding for tolerance in mink to an infected farm without tolerance selection, and an AMDV-free farm. During the 2.5-year follow-up, the mink in farm 1 remained mostly free of clinical AD, had normal pelt quality and litter size, and had low virus copy numbers in tissues and low antibody titers in ELISA. In histopathological studies, most of the farm 1 mink had no/mild lesions in their kidneys. 29-43% of the mink were ELISA negative but PCR positive throughout the follow-up and frequent changes in virus strains and coinfections were observed. Several differences in gene expression between animals from different farms were also detected. These results indicate that the disease burden of AMDV can be reduced, with seemingly normal health and production rates, despite continual circulation of ADMV in cases where eradication attempts are unsuccessful.

Histopathological risk factors for ipsilateral breast events after breast conserving treatment for ductal carcinoma in situ of the breast--results from the Swedish randomised trial.

AIM: The primary aims were to study risk factors for an ipsilateral breast event (IBE) after sector resection for ductal carcinoma in situ of the breast (DCIS) in a trial comparing adjuvant radiotherapy to no therapy and to assess predictive factors for response to radiotherapy. Secondary aims were to analyse reproducibility of the histopathological evaluation and to estimate correctness of diagnosis in the trial. SETTING: A randomised trial in Sweden (the SweDCIS trial), including 1046 women with a median of 5.2 years of follow-up in a population, offered routine mammographic screening. METHODS: A case-cohort design with a total of 161 cases of IBE (42 of those being members of the subcohort) and 284 sampled for the sub-cohort. Ninety five percent of the participants' slides could be retrieved and were re-evaluated by three experienced pathologists. RESULTS: Low nuclear grade (NG 1-2) and absence of necrosis halves the risk of IBE in both irradiated and non-irradiated patients. Lesion size, margins of excision and age at diagnosis did not modify these associations. The presence of necrosis modified the effect of radiotherapy: relative risk was 0.40 with necrosis present and 0.07 with necrosis absent (p-value for interaction 0.068). In all subsets of prognostic factors, radiotherapy conferred a substantial benefit. The risk factors for in situ and invasive IBE were similar. The agreement between pathologists was moderate (kappa=0.486). Correctness of diagnosis in the subcohort of SweDCIS was 84.8%. CONCLUSION: Although nuclear grade and necrosis carry prognostic information, we could not define a group with very low risk after sector resection alone. Radiotherapy has a protective effect in all substrata of risk factors studied. The interaction between the presence of necrosis and radiotherapy is a clinically and biologically relevant research area.

The p53 gene in breast cancer: prognostic value of complementary DNA sequencing versus immunohistochemistry.

BACKGROUND: Mutations in the p53 tumor suppressor gene (also known as TP53) have been detected in a wide variety of human cancers. In breast cancer, the presence of p53 gene alterations has been associated with worse prognosis. PURPOSE: We compared a complementary DNA (cDNA)-based sequencing method and an immunohistochemical (IHC) method for their abilities to detect p53 mutations in breast cancer specimens. In addition, we determined the prognostic value of information obtained when these two methods were used. METHODS: Specimens from 316 primary breast tumors were evaluated for the presence of mutant p53 protein by use of the mouse monoclonal antibody Pab 1801 (that recognizes both wild-type and mutant forms of p53) and standard IHC methods. In addition, the entire coding region of p53 genes expressed in these tumors was screened for mutations by combining reverse transcription, the polymerase chain reaction, and DNA sequencing. Probabilities for overall survival (OS), breast cancer-corrected survival (BCCS; death from breast cancer is the considered event), and relapse-free survival (RFS) were estimated by use of the Kaplan-Meier method, and survival curves for different patient subgroups were compared by use of the logrank method. All reported P values are from two-sided tests. RESULTS: Sixty-nine (22%) of 316 tumors had p53 gene mutations detected by the cDNA-based sequencing method; only 31 (45%) of these mutations were located in evolutionarily conserved portions of the p53 coding region. Sixty-four tumors (20% of the total) had elevated levels of p53 protein as detected by IHC, suggesting the presence of mutations. Of the sequencing-positive tumors (i.e., p53 mutant), 23 exhibited negative IHC reactions, indicating that IHC failed to detect 33% of the mutations. Furthermore, 19 of the IHC-positive tumors were sequencing negative (i.e., p53 wild-type), suggesting a 30% false-positive frequency with IHC. Four tumors (1.3% of the total) could not be analyzed by the cDNA-based sequencing method, and three tumors (1% of the total) could not be analyzed by IHC. The 5-year estimates for RFS, BCCS, and OS were significantly shorter for patients with p53 sequencing-positive tumors than for patients with sequencing-negative tumors (P = .001, P = .01, and P = .0003, respectively). Patients with IHC-positive tumors showed reduced survival in all three categories when compared with those with IHC-negative tumors, but the differences were not statistically significant. CONCLUSIONS: Use of a cDNA-based sequencing method to determine the status of the p53 gene in primary breast cancers yielded better prognostic information than IHC performed with the Pab 1801 monoclonal antibody.

Increased p53 mutation frequency during tumor progression--results from a breast cancer cohort.

The mutational patterns of the p53 gene for exons 4-9 were analyzed in 30 recurring tumors compared with the p53 status of the corresponding 30 primary breast cancers. The prevalence of p53 mutations was higher, although not statistically significant (P = 0.07), in the evaluable recurring tumors compared with the corresponding primaries, 12 of 29 (41%) versus 7 of 30 (23%). Twenty-one of the patients had unchanged p53 mutation status in the recurring compared with the primary tumors, whereas 8 had an altered mutational status or pattern in the sequential tumor. These findings indicate that p53 mutations may be an important factor for tumor progression in human breast cancer.

Lack of the latent transforming growth factor beta binding protein in malignant, but not benign prostatic tissue.

Transforming growth factor beta (TGF-beta) is a family of proteins which act as a potent growth inhibitor for most cell types including epithelial cells. TGF-beta is synthesized as latent high molecular weight complexes, composed of TGF-beta, the NH2-terminal part of the TGF-beta precursor and the third molecule, the latent TGF-beta binding protein (LTBP). We here ascertain that TGF-beta is expressed in human prostatic cancer tissue as well as in cystectomized prostatic tissue and in materials from transurethral resections with benign prostatic hyperplasia, analyzed by immunohistochemistry. TGF-beta is observed in both epithelial cells and stromal cells. No significant correlation was obtained between TGF-beta expression in tumor cells and their degree of differentiation. However, analysis by immunohistochemistry using antibodies against LTBP revealed that specimens from histopathologically verified human prostatic cancer are mostly negative for this molecule, although it is expressed in cystectomized prostatic and benign prostatic hyperplasia tissues. These results indicate that in cystectomized prostatic and benign prostatic hyperplasia tissues, TGF-beta may be produced in a complex associated with LTBP; whereas in prostatic carcinoma, TGF-beta is produced without associating with LTBP. The biological significance of the production of TGF-beta in relation to LTBP and the possible association with prognosis are discussed.

10-Year results after sector resection with or without postoperative radiotherapy for stage I breast cancer: a randomized trial.

PURPOSE: To study the long-term effectiveness of postoperative radiotherapy after sector resection for breast cancer in a randomized trial in which mammography is a major pathway to diagnosis. PATIENTS AND METHODS: Three hundred eighty-one women with a unifocal breast cancer < or = 20 mm in diameter on the preoperative mammogram and without histopathologic signs of axillary metastases were treated by sector resection plus axillary dissection. Of these patients, 184 women were randomized to receive postoperative radiotherapy to the breast (XRT group), and 197 women received no further treatment (non-XRT group). RESULTS: The local recurrence rate was 8.5% (95% confidence interval [CI], 3.9% to 13.1%) in the XRT group and 24.0% (95% CI, 17.6% to 30.4%) in the non-XRT group (P =.0001). Survival free from regional and distant recurrence was 83. 3% in the XRT group (95% CI, 77.5% to 89.1%) and 80.0% in the non-XRT group (95% CI, 73.9% to 86.1%) (P =.23). Overall survival was 77.5% in the XRT group (95% CI, 70.9% to 84.1%) and 78% in the non-XRT group (95% CI, 71.7% to 84.3%) (P =.99). A subgroup analysis suggested that women older than 55 years of age without comedo or lobular carcinomas had a low risk of local recurrence of 6.1% (95% CI, 0.1% to 9.1%) in the XRT-group and 11.0% (4.0% to 18.0%) in the non-XRT group (P =.16). CONCLUSION: Sector resection plus radiotherapy resulted in an absolute reduction in local recurrence of 16% at 10 years compared with surgery alone. Women older than 55 years of age without comedo or lobular carcinomas may have a low risk of local recurrence. Postoperative radiotherapy was not shown to reduce distant recurrences or improve overall survival.

Expression of EGFR, HER2, HER3, and HER4 in metastatic squamous cell carcinomas of the oral cavity and base of tongue.

The expressions of all four receptors in the epidermal growth factor receptor family, EGFR. HER2, HER3, and HER4 were evaluated by immunohistochemistry in 19 cases of metastatic squamous cell carcinoma of the oral cavity and base of tongue. EGFR had a similar and high expression in both primary tumours and the corresponding metastases, while the expression in normal epithelium was lower in most cases. HER2 was not expressed to the same extent as EGFR. However, when HER2 was well expressed, it was in most cases expressed to the same extent and intensity in the primary tumours, metastases, and normal epithelium. The expression of HER3 and HER4 varied and was mainly cytoplasmic in all cases studied. No overexpression of HER3 and HER4 in tumours was seen as compared to normal epithelium. In order to further investigate the distribution of HER3, two HER3 expressing cell lines originating from tongue cancer were analysed in vitro, using radiolabelled anti-HER3 antibodies directed to the extracellular domains of the receptor. The results indicated that HER3 was not present in measurable amounts in the cellular membrane. There is a need for improved diagnostics and therapy for the studied type of tumours, e.g. using radiolabelled antibodies or ligands, and EGFR seemed suitable as target since the expression was high, membrane associated and similar in the primary tumours and the corresponding metastases.

Ductal carcinoma in situ of the breast from a population-defined cohort: an evaluation of new histopathological classification systems.

The increased incidence of ductal carcinoma in situ of the breast (DCIS) in the era of mammography screening requires a deeper knowledge of the biology of the disease and calls for a suitable classification system to optimise therapy. Our aim was to evaluate the correlation to prognosis for two new classification systems of DCIS. The histopathological specimens from 195 women consecutively diagnosed between 1986 and 1994 with a primary DCIS were re-classified by two separate observers using the system proposed by an European Organization for Research and Treatment of Cancer (EORTC) working group and the Van Nuys system. The relapse-free survival (RFS) by histopathological subgroup and by nuclear grade only was estimated for women treated with breast conserving surgery (n = 149). Thirty-two local recurrences occurred among 149 women (mean follow-up time 59 months). No distant recurrences or breast cancer deaths were reported. The women in the group with the highest differentiation according to the EORTC classification had no recurrences. RFS did not differ appreciably between the two other groups. This was true also after stratification for radiotherapy. We found no statistically significant difference in RFS between the three groups in the Van Nuys classification. There was an overall agreement between the observers in 79% and 64% of the cases, according to the EORTC and Van Nuys systems, respectively. We were able to define one group with highly differentiated lesions and an excellent prognosis with the EORTC classification. Further classification into intermediate and low differentiated lesions did not help predict RFS.

Histopathological and immunohistochemical analysis of lung tumors: description of a convenient technique for use with fine needle biopsy.

In this paper we describe a convenient procedure for use with fine-needle biopsy in the diagnosis of lung and medistinal tumors. Cells obtained from a fine-needle aspiration are suspended in a formaldehyde solution and allowed to sediment onto a polycarbonate filter, using a filtering device specially designed for this purpose. The filter with the aspirated cells is then embedded in paraffin and thereafter sectioned and stained in a routine manner. The main advantages with this technique, instead of cytology, are: 1. the cells are kept mainly intact, mostly in undisrupted clusters, and this increases the possibility of obtaining a specific diagnosis, a prerequisite for the diversified clinical management of malignant tumors: 2. possibilities exist, when necessary, of further immunohistochemical characterization of tumor cells in order to obtain a specific diagnosis without thoractomy. This could be of major importance when the management of certain tumors and infiltrates (i.e. those preferably treated by radio- and chemotherapy) does not require surgical intervention.

Guidelines for prophylactic radical lymph node dissection in cases of carcinoma of the external ear.

All cases of squamous-cell carcinoma of the external ear that were treated at the ENT Clinic, University Hospital, Lund, Sweden, between 1970 and 1977 were analyzed retrospectively to determine parameters for predicting the development of metastases. These cases included 62 males and 3 females, with a mean age of 77 years. The frequency of metastases for these patients was 16.4%. The carcinomas were graded according to four parameters: depth of growth, mode of invasion, cellular differentiation, and cellular plasmolymphocytic response. We concluded that depth of growth and mode of invasion were the most useful of these parameters and that prophylactic lymph node dissection should be performed in cases of tumors larger than 4 cm in diameter, of tumors infiltrating the cartilage, and of smaller tumors with maximum scores for depth of growth and mode of invasion.

Human milk fat globule antigen 1 (HMFG1) expression in prostatic carcinoma and immunotargeting with a radiolabelled monoclonal anti-HMFG1 antibody.

BACKGROUND: Prostatic cancer is the leading cause of death in Swedish men. Approximately 50% have disseminated disease at diagnosis. Radiolabelled antibodies could possibly be a treatment modality for disseminated prostatic cancer, so that in this study the expression of the human milk fat globulin 1 (HMFG1) antigen in prostate cancer was examined. MATERIALS AND METHODS: An immunohistochemistry technique with a murine monoclonal antibody was used, as well as the human prostate cancer cell line DU-145, which expresses this cell surface antigen. TUR specimens from patients with prostate cancer were also examined. RESULTS: Eighteen out of 22 (82%) patients exhibited an HMFG1-positive tumour. An inhomogenity in the immunostaining could occasionally be seen, with smaller apparently negative areas. The immunolocalisation properties of the antibody were investigated using a radiolabelled antibody injection into nude mice bearing heterotransplants of the DU-145 cell line. The highest accumulation of the antibody was seen in the tumour tissue and the liver. CONCLUSION: The results obtained form a basis for further investigations with the goal of using the antibodies for staging and therapy for prostate cancer.

Rett syndrome: spinal cord neuropathology.

The morphologic changes of the spinal cord in Rett syndrome are described in 2 young women who died at 20 and 30 years of age. Both patients had been in a severely disabled state for many years with tetraparesis and extreme muscle wasting. Degeneration and loss of spinal ganglion nerve cells, in addition to gliosis of both the white and gray matter of the spinal cord, were evident. The number of motor neurons appeared to be reduced and axonal changes suggestive of degeneration were observed in both the ascending and descending tracts.

Electron microscopy of fine needle aspiration biopsy from soft tissue tumors.

The value of electron microscopy as an adjunct to fine needle aspiration cytology in the preoperative diagnosis of soft tissue tumors was studied in 22 cases. In 19 of these, the aspirated material obtained was adequate for electron microscopy analysis. The aspiration procedure did not damage the cells, and the aspirates were well suited to electron microscopic examination. In many cases this examination enhanced the precision of diagnosis and subclassification. This was the case in two intramuscular myxomas diagnosed as myxoid liposarcomas with light microscopy and in two sarcomalike squamous cell carcinomas diagnosed as polymorphous sarcomas in the ordinary smears.

Electron microscopy of fine needle aspiration biopsy from a malignant fibrous histiocytoma. Report of a case.

A case is reported in which electron microscopy was utilized to make a preoperative differential diagnosis of a soft tissue sarcoma on material obtained by fine-needle aspiration biopsy. The value of electron microscopy as an adjunct to cytologic study of such material is pointed out, as is the suitability of aspirated material for electron microscopic examination.

Expression of estramustine-binding protein (EMBP) and the proliferation-associated antigen Ki-67 in prostatic carcinomas.

The expression of EMBP and the proliferation-associated antigen Ki-67 was studied in in vitro cultured prostatic carcinoma cells and in tumor tissues removed by transurethral electroresection (TUR). EMBP was found to be expressed predominantly in the moderately differentiated carcinomas. A technique based on the immunohistochemical analysis of fine needle specimens was also evaluated. This technique is of potential interest in prospective studies and in monitoring the effect of therapy. Ki-67 was found to be expressed in the prostatic carcinoma cell line (DU-145) studied, as well as in TUR specimens. This antigen reflects the proliferative characteristics of the tumor and may prove useful with respect to prognostic information and choice of appropriate therapy.

EGFR, HER2 and HER3 expression in primary colorectal carcinomas and corresponding metastases: Implications for targeted radionuclide therapy.

Members of the epidermal growth factor receptor, EGFR, family are interesting as targets for radionuclide therapy using targeting agents labeled with α- or ß-emitting radionuclides, especially when EGFR-positive colorectal carcinomas, CRC, are resistant to EGFR inhibiting agents like cetuximab and various tyrosine kinase inhibitors. The expression of EGFR, HER2 and HER3 was therefore analyzed in CRC samples from primary tumors, corresponding lymph node metastases and, in a few cases, liver metastases. The expression of HER2 and EGFR was scored from immunohistochemical preparations using the HercepTest criteria 0, 1+, 2+ or 3+ for cellular membrane staining while HER3 expression was scored as no, weak or strong cytoplasm staining. Material from 60 patients was analyzed. The number of EGFR 2+ or 3+ positive primary tumors was 16 out of 56 (29%) and for lymph node metastases 8 out of 56 (14%) whereas only one out of nine (11%) liver metastases were positive. Thus, there was lower EGFR positivity in the metastases. Only one among 53 patients was strongly HER2 positive and this in both the primary tumor and the metastasis. Eight out of 49 primary tumors (16%) were strongly HER3 positive and the corresponding numbers for lymph node metastases were 9 out of 49 (18%) and for liver metastases 2 out of 9 (22%). The observed number of strongly EGFR positive cases was somewhat low but EGFR might be, for the cases with high EGFR expression in metastases, a target for radionuclide therapy. HER2 seems not to be of such interest due to rare expression, neither HER3 due to mainly expression in the cytoplasm. The requirements for successful EGFR targeted radionuclide therapy are discussed, as well as patient inclusion criteria related to radionuclide therapy.