[Suicidal adolescents can benefit from Family Group Conferences]. / Eigen Kracht-conferenties bij suïcidale adolescenten.

A risk factor among suicidal adolescents is generally their inability to form and maintain relationships with other people. In the case that we investigated, the technique known as Family Group Conference (FGC) was used successfully to break through the adolescent's passivity and social isolation. The FGC also helped to alter the adolescent's conviction of being a burden to relatives, friends and other people. The results are in line with additional studies that used FGC with other target groups. This case study suggests that FGC is a promising type of intervention that can reduce the passivity and isolation of suicidal adolescents, strengthening their relationships and boosting a feeling of belonging. These factors as well as the results of the case investigated are currently stimulating further research into the use of FGCs to foster feelings of belonging and togetherness among suicidal adolescents.

Hyperthermic isolated limb perfusion with tumor necrosis factor-alpha and melphalan in patients with locally advanced soft tissue sarcomas: treatment response and clinical outcome related to changes in proliferation and apoptosis.

Hyperthermic isolated limb perfusion with tumor necrosis factor-alpha and melphalan (HILP-TM) with or without IFN-gamma is a promising local treatment in patients with locally advanced extremity soft tissue sarcomas (STSs), with response rates of up to 84%. The mechanisms of the treatment response are poorly understood. Here, we determined the HILP-TM-induced changes in mitotic activity, proliferation, and apoptosis in 37 STSs; the additional effect of IFN-gamma; and the association of HILP-TM with treatment response and clinical outcome. On archival material, obtained before and 6-8 weeks after HILP-TM with (n = 15) or without (n = 22) IFN-gamma, the number of mitoses was counted, and the proliferation fraction was determined by immunohistological staining for the proliferation associated Ki-67 antigen (MIB1). Apoptosis was visualized by enzymatic detection of DNA fragmentation (terminal deoxynucleotidyl transferase-mediated nick end labeling method). Clinical and histological response, follow-up status, and survival were recorded. The number of mitoses dropped 57% and proliferation rate decreased with 40% after HILP-TM, whereas the amount of apoptosis after HILP-TM more than doubled as before HILP-TM. The addition of IFN-gamma to HILP-TM did not influence the changes in tumor parameters and did not affect treatment response. A better clinical response to HILP-TM was correlated with high mitotic activity and low amount of apoptosis in tumor samples before HILP-TM. Patients with highly proliferative STS before and after HILP-TM had a relatively poor prognosis. Furthermore, patients who developed distant metastases after HILP-TM had a relatively high number of dividing cells in the tumor remnants after treatment.

Heterogeneity of rat liver and spleen macrophages in gadolinium chloride-induced elimination and repopulation.

Blockade of phagocytosis and selective elimination of macrophages (m phi s) are generally accepted procedures for gaining knowledge about the function of m phi s in vivo. This study demonstrates that intravenous injection of gadolinium chloride (GdCl3) not only blocks phagocytosis by rat liver m phi s (Kupffer cells) but also selectively eliminates the large m phi s situated in the periportal zone of the liver acinus. Repopulation of m phi s starts at 4 days after injection. During repopulation, m phi s are less vulnerable to GdCl3. When repopulation is complete, the new m phi s show the same vulnerability as the original ones. Splenic m phi s are less vulnerable to GdCl3 because only some of the red pulp m phi s transiently disappear. The white pulp m phi s are not affected. Repopulation occurs sooner than in liver. These results indicate that administration of GdCl3 is a suitable approach to studying the in vivo function of large Kupffer cells.

Enzyme histochemistry as a link between biochemistry and morphology.

The presented paper describes the role of enzyme histochemistry in cell biological investigations. In the first chapter a general discussion has been given about enzyme histochemistry as a connecting link between biochemistry and morphology. The methods available for determination of enzymes in a particular cell or cell compartment have been reviewed. In this respect the characteristics of enzyme histochemistry have been discussed. Furthermore, attention has been paid to the possibilities and limitations of enzyme histochemistry. In chapter two a comparison has been made between histochemically judged and biochemically determined enzyme activities. Some fundamental differences between the biochemical and the histochemical approach in cell biological investigations are dealt with. To correlate histochemically and biochemically determined enzyme activities, a description has been given of the application of histochemical methods on isolated fractions and sucrose-ficoll gradients of these fractions. Several experimental results are described concerning the question whether a relation exists between histochemically and biochemically determined activities of respectively alkaline phosphatase, glucose-6-phosphatase, 5'-nucleotidase and 3ss-hydroxysteroid dehydrogenase. From these results the conclusion could be drawn that in general a good correlation exists between histochemically judged activity per volume (area X thickness) and biochemically determined activity per gram tissue. In chapter three the role of enzymes as markers of cellular particles and as parameters of metabolic pathways is described. Histochemical methods are available for most marker enzymes. Only activities of key enzymes can be regarded as parameters of metabolic pathways. The distribution in sucrose-ficoll gradients of enzymes, regarded as markers of mitochondria, lysosomes, endoplasmic reticulum and plasma membranes has been given. The changes occur ing under different experimental conditions for a number of marker enzymes in rat liver are described. Attention has been given to the contibution of enzyme histochemistry in the study of the heterogeneity of mitochondria, the dual localization of some (lysosomal) enzymes, the complexity of the microsomal fraction, the function of the Golgi apparatus and the heterogeneity and function of plasma membranes. Based on these results and on literature findings the possible role of some marker enzymes in cell metabolism has been discussed. In chapter four problems coherent with species and sex differences in enzyme activities are described. The interpretation of histochemical and biochemical results in view of these differences is discussed. Enzymes characteristic for a given cell type -3ss-hydroxysteroid dehydrogenase in steroid producing cells, ATP-ase in liver plasma membrane surrounding the bile canaliculi - do show less variations between species and sexes than enzymes not directly involved in specialized functions...

Proliferative activity in human brain tumors: comparison of histopathology and L-[1-(11)C]tyrosine PET.

UNLABELLED: To validate the protein synthesis rate (PSR) measured in human brain tumors using L-[1-(11)C]tyrosine (TYR) PET, the PSR was compared to histopathological parameters that reflect proliferation and protein synthesis. METHODS: We studied 20 patients who had a brain biopsy and who also underwent a PET study with TYR. Paraffin sections were stained with the monoclonal antibody MIB 1, targeted against the core antigen Ki-67, and nucleolar organizer regions (NORs) were measured as argyrophilic NORs (AgNORs). The TYR uptake was measured by PET, and with a kinetic model, the PSR was determined. RESULTS: PSR (nmol/ml/min) ranged from 0.44 to 1.99 (mean, 0.97), Ki-67 labeling indices (%) ranged from 0.9 to 33.5 (mean, 9.5) and AgNOR area (mm2/cm2) ranged from 0.13 to 0.85. No relationship was found between PSR and Ki-67 labeling index or AgNOR area. CONCLUSION: It seems that the PSR and proliferation, as measured by Ki-67, are independent processes. The role of the PSR is uncertain, but it is likely that it can be seen as a marker for the homeostasis of the cell.

Limited numbers of apoptotic cells in fresh paraffin embedded bone marrow samples of patients with myelodysplastic syndrome.

In myelodysplasia (MDS) the precise mechanism of ineffective erythropoiesis is not fully elucidated, but it is suggested that apoptosis may contribute to this process. We performed TdT-mediated dUTP-nick end labelling (TUNEL) staining of paraffin embedded bone marrow specimens to assess the amount of apoptotic cells in 21 MDS patients (7 RA, 3 RARS, 5 RAEB, 3 RAEB-T, 3 CMML) and five normal controls. In 10 MDS patients the TUNEL assay was performed in combination with immunostaining for Glycophorin-A (GpA) to determine apoptosis in the maturing erythroid compartment. To assess the proliferation of the bone marrow cells the expression of Ki-67 antigen was used as a marker. The mean apoptotic index (AI) in MDS patients was not increased (2.3 +/- 3.0% in MDS versus 4.8 +/- 1.2% in normal controls (P < 0.05)). Moreover, no significant difference in mean AI was observed in the GpA+ compartment between MDS and normal controls (0.8 +/- 0.2% versus 0.6 +/- 0.1%). In addition the different FAB-classifications and the different International Prognostic Scoring System (IPSS)-risk groups showed no significant differences between the subgroups. The expression of Ki-67, as marker for proliferative activity, in the GpA+ compartment from MDS did not differ significantly from normal controls (84.0 +/- 12.2% versus 79.9 +/- 20.2%). Our findings suggest that the observed increased apoptosis in in vitro culture assays is related to the detachment of the cells from the microenvironment leading to an increased susceptibility to apoptosis.

Nesidioblastosis and endocrine hyperplasia of the pancreas: a secondary phenomenon.

Diffuse endocrine cell proliferation (nesidioblastosis) and islet cell hyperplasia are considered causes of organic hyperinsulinism but have not been distinguished (by histometric or immunohistologic methods) from the normally variable pancreatic islet cell population during development and in adults. Therefore, in this study morphologic, immunohistologic (to detect insulin, glucagon, somatostatin, and pancreatic polypeptide), and morphometric features were evaluated in 1) normal pancreases (from fetal to adult; n = 49); 2) pancreases from patients with nesidioblastosis (n = 5); and 3) tumor-associated pancreases (TAP) from patients with insulin-producing islet cell tumors (n = 8). The study of normal postnatal development revealed that all features of fetal development remain present after birth and that the diagnosis of any diffuse endocrine disorder should therefore be based essentially on quantitative histometric parameters (total endocrine area, islet size distribution, distribution of each endocrine cell type). With these parameters endocrine cell hyperplasia was demonstrated in TAP from adults due to increased numbers of A and D cells. However, in the cases previously diagnosed as pathologic nesidioblastosis, all parameters were within the normal range. Thus, nesidioblastosis does not appear to be a pathologic entity. Careful re-examination of the pancreases, prompted by these data, revealed small islet cell tumors in three of these five cases. It is concluded that the endocrine pancreas can react rapidly, both morphologically and functionally, to changes in hormonal feedback, e.g., islet cell tumors. Therefore, the observation of a diffuse islet cell disorder in a patient with hyperinsulinism should not be considered an indication that an islet cell tumor is not present.

Quantitation of proliferation-associated markers Ag-NOR and Ki-67 does not contribute to the prediction of lymph node metastases in squamous cell carcinoma of the vulva.

A key prognostic parameter for vulvar carcinoma is the presence of lymph node metastases. Determination of proliferation markers has been suggested as a method to predict lymph node metastases in several tumor types. If this were true in vulvar carcinomas, reduced surgical therapy for patients with low-risk vulvar carcinoma could be considered. The authors analyzed whether the proliferation-associated markers silver nucleolar organizer region (Ag-NOR) and Ki-67 are predictors for inguinofemoral lymph node metastases in women with vulvar carcinoma. The authors also analyzed whether these proliferation markers are interrelated. Data were obtained from samples of 145 patients with T1/T2 squamous cell carcinoma of the vulva who were treated with vulvectomy and bilateral lymphadenectomy. None of these patients received preoperative therapy, and the invasion depth of the tumors was more than 1 mm. The median age was 71 years. The group consisted of 67 patients with differentiation grade 1, 64 with grade 2, and 18 with grade 3; 22% (15 of 67) of the patients with grade 1, 45% (29 of 64) with grade 2, and 43% (six of 14) with grade 3 had lymph node metastases. Formalin-fixed, paraffin-embedded sections were stained for proliferation markers Ag-NOR and MIB-1 (an equivalent of Ki-67 for fixed material). Both parameters were scored at the tumor stroma interface. Ag-NOR number and areas were quantified by interactive image analysis and Ki-67 index was scored microscopically with a grid. No relation was found between Ki-67 or Ag-NOR and lymph node metastases. A relation was found between Ki-67 and mitotic index (MI), but not between Ag-NOR and MI or Ki-67 index. Therefore, it is questionable whether Ag-NOR is, indeed, a marker for proliferation. The authors conclude that quantitation of Ki-67 and Ag-NOR does not contribute to the prediction of inguinofemoral lymph node metastases in squamous cell carcinoma of the vulva.

Efforts to improve interobserver agreement in histopathological grading.

The present study aimed to assess the interobserver variation in histopathological grading of 88 slides by four experienced pathologists who, before the start of the study, reached consensus about the method of grading through a joint session behind a discussion microscope. The results were compared with two previous studies, the first assessing the observer variation under normal conditions, the second doing the same after theoretical agreement about which morphological characteristics were relevant for grading. In the present study the pathologists first made a provisional diagnosis on low power view, followed by a final diagnosis on high power view, scoring various morphological characteristics as in the second study. The unweighted and weighted group kappa values for the final diagnosis were 0.41 and 0.71 respectively in the present study (after the consensus meeting), compared to 0.33 and 0.70 respectively in the second study (after only theoretical consensus), and 0.28 and 0.55 in the first study (without consensus). We conclude that a consensus meeting appears to be useful to improve the interobserver agreement.

Interobserver variation in histopathological grading of cervical dysplasia.

In order to assess the variability among histopathologists in grading cervical dysplasia, four experienced histopathologists examined the same set of 106 biopsy specimens and assigned them to one of five diagnostic categories. These were: no dysplasia, mild dysplasia, moderate dysplasia, severe dysplasia and carcinoma in situ. The histopathologists did not discuss the grading criteria beforehand. There was considerable disagreement among the pathologists: unweighted group kappa 0.28, weighted group kappa 0.56. It appeared that all grades of dysplasia were equally difficult to distinguish from adjacent categories. Various explanations for this interobserver variation are put forward.

Sources of interobserver variation in histopathological grading of cervical dysplasia.

The present study aimed to assess where the interobserver variation in grading cervical dysplasia stems from. Four experienced pathologists examined 93 histological slides, after they agreed on which morphological characteristics should be considered relevant for grading. They scored 6 morphological characteristics for each slide and assigned it to a degree of dysplasia. Compared to a previous study, the interobserver variation showed a statistically significant improvement: the weighted group kappa value increased from 0.55 to 0.69. For the scores of the individual characteristics considerable interobserver variation was observed: weighted group kappa values ranged from 0.28 to 0.49. The pathologists slightly differed in which characteristics they considered most important for their grading. The agreement on the degree of dysplasia turned out to be better than the agreement on the morphological characteristics on which this diagnosis is based. In the discussion, a few explanations for this paradoxical finding are put forward.

Relationship between ovarian production of estrone, estradiol, testosterone, and androstenedione and the ovarian degree of stromal hyperplasia in postmenopausal women.

OBJECTIVE: To study the relationship between ovarian production of estrone (E ), estradiol (E2), testosterone (T), and androstenedione (A) and the ovarian degree of stromal hyperplasia in postmenopausal women. DESIGN: In 18 postmenopausal women, the ovarian vein hormone levels of E1, E2, T, and A were compared with the degree of ovarian stromal hyperplasia. The degree of stromal hyperplasia was assessed by histological analysis (group 1: atrophic ovaries, n = 8; group 2: slight stromal hyperplasia, n = 8; group 3: moderate or severe stromal hyperplasia, n = 2). RESULTS: The ovarian levels of E1 and E2 did not correlate with the ovarian degree of stromal hyperplasia. The ovarian levels of A in group 3 were significantly higher than those in groups 1 and 2 (p < 0.02 and p < 0.01, respectively). The ovarian levels of T in group 3 were significantly higher than those in group 1 (p < 0.01) but did not differ significantly from those in group 2. CONCLUSIONS: The amount of stromal hyperplasia in postmenopausal ovaries is correlated with the ovarian vein levels of A and T. Morphological characteristics of the postmenopausal ovary determine the local (pelvic) endocrine status and may play a role in the etiology of hormone-dependent diseases of the internal genitals.

Intranodal and extranodal tumour growth in early metastasised non-small cell lung cancer: problems in histological diagnosis.

AIM: To question the observer reliability or agreement of reports on the intranodal and extranodal tumour growth patterns in early metastasised non-small cell lung cancer (NSCLC). METHODS: In a pilot study original histological sections of mediastinal lymph node metastases from NSCLC obtained by lymph node dissection (n = 82) or by mediastinoscopy (n = 62) were examined and classified independently by three pathologists as extranodal, intranodal, or indefinite. After clear criteria for these growth patterns had been defined sections were re-examined and recategorised one year later. Interobserver agreement was examined for both investigations. RESULTS: In the dissected lymph nodes the kappa value improved significantly from 0.52 (moderate agreement) at the first investigation to 0.72 (good agreement) at the second. In the mediastinoscopic lymph node biopsy specimens an increase in kappa value from 0.50 at the first to 0.67 at the second examination was found, although this improvement was not significant. In mediastinoscopic biopsy specimens a very high proportion of tissue samples showed indefinite tumour extension. CONCLUSION: Good reproducibility of intranodal and extranodal growth patterns in the histological examination of mediastinal lymph node metastases can be achieved, provided that pathologists use strictly defined criteria. In mediastinoscopic biopsy specimens it is often impossible to differentiate between intranodal and extranodal tumour growth.

Detection of HPV-16 DNA by PCR in histologically cancer free lymph nodes from patients with cervical cancer.

The prognostic value of detection of human papillomavirus (HPV) type 16 DNA in histologically cancer free lymph nodes was assessed in left obturator lymph nodes from cervical cancer patients with HPV-16 positive primary tumours. HPV-16 DNA was detected by polymerase chain reaction in 12 of 35 patients with histologically cancer free lymph nodes. Of these 12 patients, only one developed a recurrence, suggesting HPV-16 DNA detection in cancer free lymph nodes has no prognostic value.

Proliferation rate of colonic mucosa in normal subjects and patients with colonic neoplasms: a refined immunohistochemical method.

An increased colonic epithelial proliferation rate and an increase of the cryptal proliferative zone are probable markers of increased susceptibility to colonic cancer. In this study an immunohistochemical method using 5-bromo-deoxyuridine (BrdUrd) to measure the proliferation rate of colonic mucosa in vitro was used. Fresh endoscopic colonic biopsy specimens were incubated with BrdUrd and then processed for immunohistochemistry using a monoclonal antibody. Essential procedures with respect to the equal distribution of nuclei stained with BrdUrd in the biopsy specimens proved to be the cutting of the specimens before incubation and the use of a microwave oven at the beginning of incubation. The use of the procedure of the running average showed that 12 length cut crypts are sufficient to determine reliably the proliferation rate, expressed as the labelling index (LI). This was determined in the biopsy specimens of 10 subjects without organic colonic disease, eight patients with adenomatous colonic polyps, and in six patients with (recent) colonic carcinoma. Mean LI in the controls was significantly lower than in patients with colonic polyps and in those with colon cancer. It is concluded that this method is promising for screening persons at risk for colon cancer and will be of great potential in performing dietary intervention studies in these subjects.

Analysis of cryolesions in the uterine cervix: application techniques, extension, and failures.

Although cryosurgery is regularly used for treatment of cervical intraepithelial neoplasia (CIN), there are few data concerning freeze technique and attendant extension of the cryolesion. This study evaluated how to create cryolesions extensive enough to eradicate the CIN lesion completely. The way the extension of the cryolesion was influenced by type of probe, anatomical position in the cervix, shape of the external os, and freeze time was analyzed. Furthermore, we examined whether localization of the cryolesions corresponded with the CIN III location. Cryosurgery was applied to the cervix of 64 women the day before hysterectomy was performed for benign disease. Four types of probes were tested and freezing was done with a double freeze cycle. After extirpation of the uterus, slides were cut at the 3-, 6-, 9-, and 12-o'clock positions. With a computerized graphic tablet, the depth and linear extension of the cryolesion were measured morphometrically. After short freeze times, it appeared that an adequate lesion was present in only 67.4% of the slides. The large cone probe gave the best results; the small flat cervix probe the worst. At the 3- and 9-o'clock positions, a significantly higher percentage of inadequate lesions was found (60.8 and 65.3%, respectively). This proved to be due primarily to the extensive vascular supply at those positions. Longer freeze times gave an excellent result within all slides, even at the 3- and 9-o'clock positions. The topographic position of the cryolesion corresponded completely in all cases with that of the CIN III lesion.(ABSTRACT TRUNCATED AT 250 WORDS)

Minimum extension and appropriate topographic position of tissue destruction for treatment of cervical intraepithelial neoplasia.

Minimum extension and topographic position of tissue destruction for treatment of cervical intraepithelial neoplasia (CIN) is determined by the extension and the localization of the pathologic epithelium. In 65 cone specimens, we studied the depth of CIN II crypt involvement and the linear extent and topographic position of the CIN III lesions. The topographic position of the CIN III lesion was related to a reference point R, the most caudal point of the ectocervix. The mean maximum depth of CIN III crypt involvement appeared to be 1.6 +/- 1.0 mm, and the mean linear extent of the CIN III lesion was 7.4 +/- 3.7 mm. The distal border of the CIN III lesion was located at a mean distance of 8.2 +/- 4.4 mm from the reference point R, and the proximal border at a mean distance of 13.3 +/- 3.7 mm. Taking the mean + 2 SD values as directives (97.7% of the population) suggests that in almost all patients, the depth of crypt involvement did not exceed 3.6 mm; the linear extent of the CIN III did not exceed 14.8 mm. Furthermore, this implies that in almost all patients, the CIN III lesion was located between 0.6 mm distally (mean - 2 SD) and 20.7 mm proximally (mean + 2 SD) from the reference point R. Based on these results, we conclude that minimum local tissue destruction for treatment of CIN should have a depth of 4 mm over a distance of 15 mm, and should be localized at least between 1 mm distally and 21 mm proximally from the most caudal point of the ectocervix.

Meningeal carcinomatosis as presenting symptom of a gallbladder carcinoma.

Meningeal carcinomatosis as presenting symptom is rare. We report a patient who presented with an isolated abducens nerve palsy and an unsteady gait. At autopsy the primary tumor proved to be a gallbladder carcinoma. Only one similar case has been reported. Several aspects are discussed, including the route by which the tumor cells could have reached the subarachnoid space.

Malignant lymphoma of the testis.

Five patients with primary non-Hodgkin's lymphoma of the testis were studied. The lymphomas had an intermediate grade of malignancy according to the Working Formulation. One patient was in clinical stage IEA, 3 in clinical stage IIEA and one in clinical stage IVB. Central nervous system (CNS) infiltration occurred in 3 patients. The survival was very poor and ranged from 9 days to 5 1/2 years after diagnosis, median 7 1/2 months. In order to improve prognosis aggressive (3 or more cytostatics) chemotherapy is recommended to follow orchidectomy and staging. As for other lymphomas of intermediate- and high-grade malignancy, a more effective systemic therapy is needed for the younger patients, but in the elderly (greater than 70 years) the regimen and the dose intensity have to be adapted to their tolerance. Prophylactic radiation of the opposite uninvolved testis is not recommended, but frequent scrotal ultrasonography during the first 2 years after diagnosis appears essential. Prophylactic intrathecal chemotherapy means an overtreatment for patients in clinical stages I and II, but is indicated in relapsing patients and in clinical stages III and IV.

Clinical and pathological aspects of benign and malignant squamous epithelium of the corpus uteri; a report of two cases.

Two patients with primary squamous cell carcinoma of the uterus are described. In both patients, the disease was at an advanced stage when the diagnosis was made. Although endometrial malignancies with squamous elements account for 10-30% of endometrial carcinomas, primary squamous cell carcinoma of the uterus is extremely rare. Up to now fewer than 30 cases have been reported. The pathogenesis, morphogenesis and aetiology of squamous epithelium in the corpus uteri is discussed. Finally, some clinical aspects of the primary squamous cell carcinoma of the corpus uteri are reviewed.