Genetic aspects of canine mixed leukocyte cultures.

Recent data stress the importance of matching donor and recipient of an organ graft for both the serologically defined (SD) and lymphocyte-defined (LD) determinants. To allow experimental evaluation of the effect of these SD and LD structures in a noninbred experimental animal, mixed leukocyte culture tests were performed between SD identical and nonidentical dogs to clarify the LD system in these animals. The results of these experiments can be summarized as follows: (a) In the dog there is a LD locus distinct from the known SD loci, which in all probability is localized outside the first (SD-1) series locus on the chromosome. (b) The crossing-over frequency between the SD and LD loci on the chromosome is low. (c) Studies in SD identical unrelated dogs and random unrelated dogs show an apparent high linkage disequilibrium between SD and LD loci. (d) The LD system in dogs is polymorphic.

[Hodgkin and the classification of malignant lymphomas: how the British physician also became associated with later discovered lymphomas]. / Hodgkin en de classificatie van maligne lymfomen.

Thomas Hodgkin was the first to describe a malignant lymphoma, which would later carry his name. Over the course of time, other lymphoid malignancies were recognised that showed no similarity with Hodgkin's disease. They were subsequently named after the - at that time - applicable morphological nomenclature of the associated cells. Later, nomenclature also took immunological features into consideration. However, we still describe the group of lymphomas recognised after Hodgkin's discovery as 'not being Hodgkin's disease', i.e. non-Hodgkin lymphoma. We feel it is unjust that not many people know about the man behind this prominent disease. In this article, an historic overview is given of Thomas Hodgkin, 'his' lymphoma and the other malignant lymphomas.

[Indications for bone marrow biopsy in adults]. / Indicaties voor beenmergbiopsie bij volwassenen.

Bone marrow biopsies are more and more often part of the work-up of patients with haematological disorders. The most important reason for this is the fact that a biopsy supplies important additional information compared to an aspirate alone. Biopsies are superior for the assessment of the bone marrow architecture, the vascularisation, the cellularity, the localisation and the extent of infiltrates and the degree of fibrosis. In addition, biopsy is a good way to evaluate the effects of therapy in the course of the disease. As is the case with aspirates, examination of a biopsy alone is usually sufficient for a correct diagnosis. However, a combination of both techniques makes possible an optimal assessment of the nature and extent of the disease process in the often very serious haematological conditions that we are dealing with here.

The effects of long term testosterone administration on pulsatile luteinizing hormone secretion and on ovarian histology in eugonadal female to male transsexual subjects.

Polycystic ovarian disease (PCOD) is associated with elevated serum LH and (sub)normal FSH levels, while serum androgen levels are often elevated. To clarify the role of androgens in this abnormal pattern of gonadotropin secretion, LH secretion was studied in 1) 9 eugonadal female to male transsexual subjects before and during long term (6 months) testosterone (T) administration (250 mg/2 weeks, im), and 2) in a woman with an androgen-secreting ovarian tumor both before and after surgical removal of the tumor. Finally, we studied the effects of high serum androgen levels on ovarian histology in 3) 26 transsexual subjects after long term (9-36 months) T administration (250 mg/2 weeks, im) to assess whether T-induced ovarian abnormalities are similar to those that occur in women with PCOD. Long term T treatment in the nine female to male transsexual subjects resulted in increases in the mean serum T level from 1.7 +/- 0.8 (+/- SD) to 40.8 +/- 31.9 nmol/L (P less than 0.01), the mean serum dihydrotestosterone level from 0.6 +/- 0.2 to 3.3 +/- 1.5 nmol/L (P less than 0.02), and the mean serum free T level from 9.5 +/- 5.2 to 149 +/- 46 pmol/L (P less than 0.02). Mean serum estrone and estradiol levels were similar before and during T treatment. The mean serum LH level decreased from 6.3 +/- 2.0 to 2.9 +/- 1.1 U/L (P less than 0.01), and the mean FSH levels decreased from 6.6 +/- 2.0 to 3.7 +/- 2.2 U/L (P less than 0.02). Pulsatile LH secretion before and during T treatment was studied in five subjects. Neither the mean nadir LH interval nor the LH pulse amplitude changed significantly in these five subjects. The serum T level in the woman with the androgen-secreting ovarian tumor was 9.6 nmol/L, and it declined to normal after removal of the tumor. Her mean serum LH and FSH levels, the mean nadir LH interval, and LH pulse amplitude were in the normal range before and after removal of the tumor. Studies of ovarian histopathology in 26 transsexual subjects after long term androgen treatment revealed multiple cystic follicles in 18 subjects (69.2%), diffuse ovarian stromal hyperplasia in 21 subjects (80.8%), collagenization of the tunica albuginea in 25 subjects (96.2%), and luteinization of stromal cells in 7 subjects (26.9%). Findings consistent with criteria for the pathological diagnosis of polycystic ovaries, that is 3 of the 4 findings listed above, were present in 18 of the 26 subjects (69.2%).(ABSTRACT TRUNCATED AT 400 WORDS)

Multifocal rhabdomyoma of the neck. Report of a case studied by fine-needle aspiration, light and electron microscopy, histochemistry, and immunohistochemistry.

A 32-year-old Caucasian man presented with a tumor in the right side of the neck. Preoperative fine-needle aspiration showed large multinucleated cells with abundant granular cytoplasm that were consonant with the features of a granular cell tumor. At surgery, two separate tumors were found. Histologically, the tumor proved to be an adult rhabdomyoma, the 11th such multifocal case reported. The striated muscle origin of this benign tumor was confirmed by immunohistochemical and ultrastructural studies. The tumor cells were desmin-, myoglobin-, and actin-positive. They showed variable numbers of thick and thin filaments, as well as hypertrophic Z-band material. Histochemical studies showed the presence of basophilic muscle cells, vesicular nuclei, ragged red fibers, and diffuse acid phosphatase positivity. These features, together with the absence of actual muscle cell proliferation and the assumption that the mass of the tumor could be explained by the enormous swelling of the muscle cells, lead us to conclude that an adult rhabodomyoma is merely the result of a process of disorderly degeneration and regeneration rather than a real neoplasm or a hamartomatous lesion.

A generic sequencing based typing approach for the identification of HLA-A diversity.

Sequencing Based Typing (SBT) is a generic approach for the identification of HLA-A polymorphism. This approach includes the high resolution typing of the HLA-A broad reacting groups, HLA-A subtypes and will identify new alleles directly. The SBT approach described here uses a locus specific amplification of DNA from exon 1 to exon 5. The resulting 2,022 bp PCR product serves as a template for the subsequent sequencing reactions. Amplification is followed by direct sequencing of exons 2, 3 and 4 in both orientations with fluorescently labeled primers to define all polymorphic positions leading to a high resolution typing result. In this study the sequence of exons 2 and 3 of a panel of 49 cell lines was determined. In addition, the exon 4 region of 35 cell lines was also sequenced to evaluate the exon 4 polymorphism. The HLA-A type of most of the cells could be identified by sequencing only exons 2 and 3. However, the sequence of exon 4 was required to discriminate A*0201 from A*0209 and A*0207 from A*0215N. In this panel, an identical new "HLA-A*0103" was identified in two Caucasian samples.

neu overexpression correlates with extent of disease in large cell ductal carcinoma in situ of the breast.

In a retrospective study of ductal carcinoma in situ (DCIS) of the breast, the expression of the neu oncogene was determined immunohistochemically in 76 women treated by local excision or mastectomy. The histopathological features, including the extent of the lesion, histological subtype, cell type, and number of mitoses, were related to neu overexpression. Immunopositivity was found only in DCIS of large cell type, where it correlated with extent of disease but not with mitotic rate. Our findings, together with previous experimental evidence, suggest that this relationship is a consequence of the effect of the neu protein on cell motility.

The biology of the HL-A system and the association with malignant lymphomas.

Present knowledge of the HL-A system as it relates to lymphoproliferative disease is reviewed briefly. Preliminary results from a current study demonstrate an elevated frequency of Aw33 and depressed frequency of B17 associated with non-Hodgkin's lymphoma-leukemia. In somewhat weaker association, Aw24 and Aw33 are elevated in follicular-center-cell lymphomas, while Bw44 is depressed. It is concluded that more specific subclassification of the lymphomas according to immunologic concepts may aid in correlating the disease with HL-A type as it has for Hodgkin's disease.

Immunoglobulin inclusions in non-Hodgkin's lymphomas.

The occurrence and characteristics of intracellular immunoglobulin inclusions in non-Hodgkin's lymphomas are described. Lymphoma cells from three patients with immunoblastic sarcoma contained classic cytoplasmic, periodic acid-Schiff (PAS)-positive Russell bodies. Immunoperoxidase staining revealed monoclonal IgG (k) in two cases and a polyclonal pattern in the third case, where the tumor evolved from a reactive lesion. Unusual cytoplasmic inclusions were observed in the neoplastic cells of two patients with follicular center cell lymphoma. In one case large globular structures lacking a distinct limiting membrane were seen, while the cells of the other patient contained "signet-ring-like" vacuoles filled with microvesicles. Both were PAS-negative and showed monoclonal immunglobulin staining. In two other cases PAS-positive intranuclear inclusions consisting of monoclonal immunoglobulin IgM(k) could be demonstrated. The possible significance of these findings in B-lymphocyte derived neoplasms is discussed.

Encoded latent membrane protein 1 of Epstein-Barr virus on follicular dendritic cells in residual germinal centres in Hodgkin's disease.

AIMS: To determine if there is an association between Epstein-Barr virus (EBV) infection and Hodgkin's disease. METHODS: Fifty cases of Hodgkin's disease and 25 reactive lymph nodes were screened for the presence of EBV-RNA (EBER) using in situ hybridisation, and for the expression of EBV encoded latent membrane protein 1 (LMP-1) by immunohistochemistry. RESULTS: In 42% of the cases of Hodgkin's disease, EBER was detected in the nuclei of the malignant cells, and in LMP-1 expression was found 36%. Both EBER and LMP-1 positivity were seen in 34% of the cases. An additional finding was the presence of LMP-1 on follicular dendritic cells in residual germinal centres in two cases of Hodgkin's disease. EBER was not detected in these germinal centres. In reactive lymph nodes only occasional EBER positive, small, lymphoid cells were found, without LMP-1 expression. CONCLUSIONS: These results show a strong correlation between the presence of EBER and the LMP-1 expression in the Reed-Sternberg cells. They corroborate a role for EBV in at least some cases of Hodgkin's disease. LMP-1 is probably presented as an immune complex in the germinal centres, as part of an immune response against EBV.

Expression of Epstein-Barr virus encoded latent genes in nasal T cell lymphomas.

AIMS: To determine the expression of Epstein-Barr (EB) virus encoded latent genes in nasal T-cell lymphomas in The Netherlands. METHODS: Seven europid (Dutch) cases of nasal T cell lymphoma were investigated for the presence of EB virus by RNA in situ hybridisation (EBER). The expression of the EB virus encoded genes BARF0, EBNA1, EBNA2, LMP1, LMP2A, LMP2B, and ZEBRA was studied at the mRNA level using reverse transcriptase polymerase chain reaction. At the protein level the expression was investigated of EBNA2 and LMP1 by immunohistochemistry. RESULTS: In all seven nasal T cell lymphomas EBER was detected in the nuclei of virtually all tumour cells. BARF0 mRNA was detected in all samples. EBNA1 mRNA was found in six cases, LMP1 mRNA in five, LMP2A mRNA in three, LMP2B mRNA in one, and ZEBRA mRNA in one. EBNA2 mRNA was not found in any case. At the protein level occasional LMP1 positive tumour cells were seen in only one case. The EBNA2 protein was not detected. CONCLUSIONS: Nasal T cell lymphomas in The Netherlands are strongly associated with EB virus. The virus shows a type II latency pattern (EBNA1+, LMP1+, EBNA2-) that seems to be similar to the EB virus associated nasal T cell lymphomas in oriental countries.

Is preoperative transrectal ultrasonography of value in localised prostatic carcinoma? A blind comparative study between preoperative transrectal ultrasonography and the histopathological radical prostatectomy specimen.

Digital rectal examination (DRE) for staging is subjective and unreliable. Understaging has been reported in 25-72% and clinical overstaging of T3 tumors varies from 24-50%. In the present study of 15 patients, transrectal ultrasound (TRUS) staging was compared, in a blind comparative fashion, with pathological staging of the surgical specimen. Multifocal lesions were present in 8/15 patients (53%). A distinction was made between capsular involvement and 'clear' capsular penetration. TRUS was more sensitive in predicting capsular involvement than DRE (83% vs 17%), but the specificity was low (67% vs 100%). If capsular perforation was considered, the sensitivity and specificity of TRUS are 43% and 91%, respectively. Sensitivity and specificity for seminal vesicle tumor involvement by TRUS was 63% and 86%. Using TRUS the overall staging was improved by 33% compared with DRE and therefore TRUS is considered to be a valuable acquisition in localising and staging prostate cancer.

The diagnostic accuracy of core biopsy in palpable and non-palpable breast lesions.

OBJECTIVE: To assess the accuracy of large-core needle biopsy in evaluating palpable and nonpalpable breast lesions. METHODS AND PATIENTS: Stereotaxic and ultrasound (US) guided core biopsies were performed in 103 breast lesions in 97 patients. Subsequently, all patients underwent surgery. All specimens (core biopsy and surgical) underwent radiography for evaluation of microcalcifications. The histopathologic findings of the core biopsies and the surgical specimens were correlated. RESULTS: Core biopsies and surgery findings were concordant in 100% of the 27 palpable lesions and in 89% of the 76 nonpalpable lesions. One case of malignancy, ductal carcinoma in situ (DCIS), was not diagnosed by core biopsy. In 102 (99%) of the 103 breast lesions, a correct choice for additional diagnostic procedure or definitive treatment could have been made upon histopathologic findings of core biopsy. CONCLUSION: Stereotaxic and ultrasound-guided core biopsy are safe, reliable and less traumatic than excisional biopsy. Special attention is necessary when atypical ductal hyperplasia (ADH) or DCIS without invasive carcinoma is found. Radiography of the biopsy specimens for detection of microcalcifications is essential.

Neural network-based screening (NNS) in cervical cytology: no need for the light microscope?

Neural network-based screening (NNS) of cervical smears can be performed as a so-called "hybrid screening method," in which parts of the cases are additionally studied by light microscope, and it can also be used as "pure" NNS, in which the cytological diagnosis is based only on the digital images, generated by the NNS system. A random enriched sample of 985 cases, in a previous study diagnosed by hybrid NNS, was drawn to be screened by pure NNS. This study population comprised 192 women with (pre)neoplasia of the cervix, and 793 negative cases. With pure NNS, more cases were recognized as severely abnormal; with hybrid NNS, more cases were cytologically diagnosed as low-grade. For a threshold value > or = HSIL (high-grade squamous intraepithelial lesions), the areas under the receiver operating characteristic (ROC) curves (AUC) were 81% (95% CI, 75-88%) for pure NNS vs. 78% (95% CI, 75-81%) for hybrid NNS. For low-grade squamous intraepithelial lesions (LSIL), the AUC was significantly higher for hybrid NNS (81%; 95% CI, 77-85%) than for pure NNS (75%; 95% CI, 70-80%). Pure NNS provides optimized prediction of HSIL cases or negative outcome. For the detection of LSIL, light microscopy has additional value.

Negative cervical smears before CIN 3/carcinoma. Reevaluation with the PAPNET Testing System.

OBJECTIVE: To test the effectiveness of the PAPNET testing system in identifying false negative smears, using archival cervical cytologic smears from women with histologically proven diagnoses of high grade lesions and carcinoma of the uterine cervix. STUDY DESIGN: Forty-six negative smears from women who developed a high grade cervical intraepithelial lesion (CIN 3) or carcinoma of the uterine cervix within three years were retrieved from the archives, plus 20 consecutive control smears for each case. The smears were analyzed with the PAPNET testing system, and the selected cells were reviewed by a cytotechnologist using a strict protocol. RESULTS: With the PAPNET testing system, 9 of 46 (20%) smears were positive. Seven were reclassified as low grade and two reclassified as high grade squamous intraepithelial lesion (SIL). One of the 31 initially positive smears in the control group of 920 smears was not recognized as such. In the control group of 889 negative smears, 14 newly identified positive cases (1.6%) were detected, all low grade SIL. CONCLUSION: The PAPNET testing system is a good tool for detecting false negative smears and, when used as an adjunct to conventional screening, can reduce the false negative rate.

Cost analysis of PAPNET-assisted vs. conventional Pap smear evaluation in primary screening of cervical smears.

OBJECTIVE: To assess the difference in costs between PAPNET-assisted and conventional microscopy of cervical smears when used as a primary screening tool. STUDY DESIGN: We performed time measurements of the initial screening of smears by four cytotechnologists in one laboratory. Time was measured in 816 conventionally screened smears and in 614 smears with PAPNET-assisted screening. Data were collected on the components of initial screening, clerical activities and other activities in the total work time of cytotechnologists in the routine situation and on resource requirements for both techniques. RESULTS: PAPNET saved an average of 22% on initial screening time per smear. Due to costs of processing and additional equipment, the costs of PAPNET-assisted screening were estimated to be $2.85 (and at least $1.79) higher per smear than conventional microscopy. The difference in costs is sensitive to the rate of time saving, the possibility of saving on quality control procedures and the component of the initial screening time in the total work time of cytotechnologists. CONCLUSION: Although PAPNET is time saving as compared with conventional microscopy, the associated reduction in personnel costs is outweighed by the costs of scanning the slides and additional equipment. This conclusion holds under a variety of assumptions. Using PAPNET instead of conventional microscopy as a primary screening tool will make cervical cancer screening less cost-effective unless the costs of PAPNET are considerably reduced and its sensitivity and/or specificity are considerably improved.

Hypertrophic osteopathy in a dog with a chronic lung abscess.

A 2-year-old dog was examined because of gradual lameness of all 4 limbs and weight loss. Hypertrophic osteopathy was diagnosed. Radiography revealed a mass in the caudal lobe of the right lung. At necropsy, the mass was determined to be a chronic abscess. Corynebacterium pyogenes was cultured from the lesion. Although hypertrophic osteopathy in dogs is commonly associated with a thoracic lesion, most often neoplastic, the chronic lung abscess in this dog might have been formed as a result of a paralaryngeal abscess that was treated 3 months before the onset of the lameness.

[Autopsies as an important indicator for quality control]. / Obducties als kwaliteitsinstrument serieus nemen.

The decreasing number of autopsies, in the Netherlands as well, is deplorable because with it an important instrument of medical quality control is likely to disappear. For this not only the relatives, but also the attending physicians and the pathologists are to blame. To turn the tide we need some drastic changes in our attitude towards autopsies. The families should known that an autopsy is a right they have in order to check the quality of diagnosis and treatment of their beloved, it is not a favour towards the physician. A physician who does not see a reason for autopsy, should explain that to the family. Pathologists should think about and realize a subspecialty of autopsy pathology with a thorough training in pathophysiology and intensive care medicine. Autopsy reports should be of the highest quality and reach the physician within a few weeks. A required autopsy percentage should be introduced into the certification process of medical specialists and hospitals and the possibility of Continuous Medical Education credit points for physicians with a certain autopsy percentage should be considered.

[Truth after death]. / De waarheid na de dood.

Diagnosis is central to medicine. In spite of tremendous diagnostic technological advances, no infallible test exists and in the complex diagnostic process the physician may well get lost. The ultimate feedback on the accuracy of diagnosis is the autopsy. Five patients illustrate that the autopsy may disclose unexpected results. The first patient was a 9-year-old girl who suffered from daily abdominal spasmodic pain but each time recovered. She died suddenly; autopsy revealed intestinal intussusception. A 46-year-old man who was treated for hypertension developed pain in the chest and the lower back, but there were no other signs of myocardial infarction. He died suddenly; autopsy revealed a dissecting aortic aneurysm with rupture in the left pleural cavity. A 21-year-old woman, an excellent swimmer, drowned during a swim in the sea. Autopsy revealed severe widespread coronary disease with multiple myocardial infarction. A 32-year-old Surinam woman developed acute coma and died from cardiorespiratory arrest. At autopsy she had massive pulmonary embolism and generalized lymphadenopathy due to sarcoidosis. The last patient, a 32-year-old woman suffered from fatigue after her fourth child was born. She was admitted with severe dyspnoea and her chest X-ray showed interstitial fibrosis. She died presently and autopsy revealed metastatic colon carcinoma with pulmonary lymphangitis carcinomatosa. Systematic reviews of the results of autopsies show no decline in the percentage of false diagnoses and/or unexpected findings in spite of the enormous growth of the diagnostic armamentarium. Although we may radiologically 'slice' the body in incredible detail or investigate human cells at the molecular level, the autopsy has by no means become obsolete and is an invaluable tool for quality control and teaching.

[Digital roentgen-guided large-needle biopsy as an alternative to surgical biopsy in patients with nonpalpable mammographic abnormalities]. / Digitale röntgengestuurde dikkenaaldbiopsie als alternatief voor chirurgische biopsie bij patiënten met niet-palpabele mammografische afwijkingen.

OBJECTIVE: To compare histopathological findings from stereotaxic needle core biopsies and from excision biopsies performed on patients with impalpable breast lesions suggestive of carcinoma. DESIGN: Prospective and descriptive, with a description of the stereotaxic needle core biopsy procedure. SETTING: Academic Hospital Utrecht, the Netherlands. METHODS: In 36 patients core biopsies were done with 14-gauge biopsy needles, followed by a localisation procedure with surgical biopsy. The pathological features of the core and excision specimens were compared. RESULTS: Results of 35 (97%) core biopsies corresponded to those of excision biopsies. Stereotaxic core biopsy was not possible due to technical problems in 2 cases. CONCLUSION: Stereotaxic core biopsy appears to be an acceptable alternative to excision biopsy.