Apmis

GREGERSEN, HENRIK; PEDERSEN, GITTE; SVENDSEN, NICOLAJ; THULSTRUP, ANE MARIE; SØRENSEN, HENRIK TOFT; CARL SCHØNHEYDER, HENRIK

doi: 10.1034/j.1600-0463.2001.d01-149.xpmid: N/A

The risk of multiple myeloma subsequent to an episode of serious pneumococcal infection has not been ascertained. We identified 328 episodes of community‐acquired pneumococcal bacteraemia and 77 episodes of pneumococcal meningitis in 227,000 persons over 40 years of age in the County of North Jutland, Denmark, in the period 1981 to 1996. The incidence rate of a subsequent diagnosis of multiple myeloma was determined by linkage to the Danish Cancer Registry. During 1,218 patientyears of follow‐up in the bacteraemia cohort, 7 cases of multiple myeloma were diagnosed compared with 0.13 cases expected (standardized incidence ratio (SIR) 53.5, 95% confidence interval 21.4‐111.4). During 444 patient‐years of follow‐up in the meningitis cohort, 4 cases of multiple myeloma were diagnosed compared with 0.05 cases expected (SIR 83.2, 95% confidence interval 22.6‐214.8). Patients who survive an episode of community‐acquired pneumococcal bacteraemia or meningitis are at increased risk of being diagnosed with multiple myeloma, but the absolute risk is low.

RIVASI, FRANCESCO; CASALI, BRUNO; NANETTI, ANNA; COLLINA, GUIDO; MAZZONI, ALDO

doi: 10.1034/j.1600-0463.2001.d01-138.xpmid: 11900050

Histoplasmosis, which is highly endemic in the United States, is rare in Europe, usually imported but sometimes autochthonous. In Africa, histoplasmosis capsulati coexists with “African histoplasmosis”, a characteristic skin infection caused by H. capsulatum var. duboisii. Histoplamosis due to H. capsulatum is one of the 12 secondary infections listed in the surveillance definitions of AIDS. We report the case of a 36‐year‐old black man with acquired immunodeficiency syndrome (AIDS) who was living in Italy but originally came from Ghana. Histoplasmosis was disseminated with fever and cutaneous manifestations. The diagnosis was demonstrated morphologically based on the presence of yeast, observed by light microscopy, in skin lesions and by identification of H. capsulatum var. capsulatum DNA by nested PCR from a paraffin sample. No clinical reports of histoplamosis capsulati in Ghana have been published until now. The present case stresses the role of immigration of subjects from outside Europe who have been infected in their native country.

GASTER, M.; BECK‐NIELSEN, H.; SCHRØDER, H. D.

doi: 10.1034/j.1600-0463.2001.d01-139.xpmid: 11900051

Primary satellite cell cultures have become an important tool as a model system for skeletal muscles. A common problem in human satellite cell culturing is fibroblast overgrowth. We combined N‐CAM (Leu19) immunocytochemical staining of satellite cells (Sc) with stereological methods to estimate the fraction of Sc in culture. Evaluation of different culture conditions allowed us to find proliferation conditions preferentially for Sc: a) Sc should be cultured on surfaces coated with ECM‐gel. b) Primary cell culture should be inoculated in DMEM supplemented with 10% fetal calf serum to increase cell adherence. c) Change of media to DMEM supplemented with 2% Ultroser‐G and 2% FCS after 24 h.d) Before subcultivation, cells should be preplated for 30 min. The fractional content of Sc in passage four when applying this method of cultivation was 0.82 +/‐ 0.07 (mean +/‐ SE, N=10). Our method enabled us to establish culture conditions which resulted in high Sc content despite several subcultivations. Estimation of the fractional cell content could be a useful tool for optimizing not only Sc‐culturing but all cultures initially containing more cell types.

GASTER, M.; KRISTENSEN, S. R.; BECK‐NIELSEN, H.; SCHRØDER, H. D.

doi: 10.1034/j.1600-0463.2001.d01-140.xpmid: 11900052

The aim of this study was to select an effective and stable protocol for the differentiation of human satellite cells (Sc) and to identify the optimal time period for the experimental use of differentiated human Sc‐cultures. In order to identify the differentiation conditions which give a good survival of myotubes and a high grade of differentiation, Sc‐cultures were induced to differentiate in media supplemented with either 2% fetal calf serum (FCS) 2% horse serum (HS) or 10% HS. Based on higher CK‐activities in cultures differentiating in FCS‐supplemented media compared to horse sera, fetal calf serum was chosen to induce differentiation. The ATP, DNA and protein content increased during the first 4 days after induction of differentiation and was followed by a period with minor changes. The maximal differences of ATP, DNA and protein between days 4–10 were evaluated and the differences in the three components were found to be less than 20% of the average value with a certainity of more than 0.9. Day 8‐myotubes were investigated morphologically and were found immunoreactive for fast myosin, and expressed areas with clear cross striation. We recommend the use of differentiated Sc‐cultures in the period from day 4 to 8 after induction of differentiation as only minor differentation‐related changes will take place in the cells during this period of time.

WEILE, BIRGITTE; GRODZINSKY, EWA; SKOGH, THOMAS; JORDAL, ROBERT; CAVELL, BERTIL; KRASILNIKOFF, PETER A.

doi: 10.1034/j.1600-0463.2001.d01-141.xpmid: 11900053

Aim: To disclose the prevalence of adult “silent” coeliac disease in Denmark and Sweden. Experimental design: 1573 Danish and 1866 Swedish healthy blood donors were screened for the presence of serum anti‐gliadin antibodies (AGA) by enzyme‐linked immunosorbent assay. AGA‐positive serum samples were further analysed for IgA anti‐endomysium antibodies (EmA) by indirect immunofluorescence microscopy. Main results: The Danish donor population had a higher mean age than the Swedish (41.4 years versus 37.6 years) and a higher proportion of females (41% versus 32%), and had a lower mean level of AGA (17.3 units versus 20.6 units). Sixty‐one (3.9%) Danish donors had AGA above the cut‐off limit, and four of these also had positive EmA tests. Sixty (3.2%) Swedish donors had AGA above the cut‐off limit, and five of these also had positive EmA. Coeliac pathology was proven by biopsy in all five coeliac disease‐suspected Swedish donors. No small intestinal biopsy was performed in the coeliac disease‐suspected Danish donors. Conclusions: Based upon the finding of EmA in AGA‐positive serum samples, silent coeliac disease may be suspected in 1 per 394 Danish blood donors (2.5 per 1,000). A similar rate was proven in 1 per 373 Swedish blood donors (2.7 per 1,000), indicating no major differences in the prevalence of adult silent coeliac disease between the two neighbouring countries.

ØSTERBY, R.; TAPIA, J.; NYBERG, G.; TENCER, J.; WILLNER, J.; RIPPE, B.; TORFFVIT, O.

doi: 10.1034/j.1600-0463.2001.d01-142.xpmid: 11900054

Renal biopsies were obtained from type 2 diabetic patients with elevated albumin excretion. The aim was to obtain quantitative structural data to correlate with clinical findings. Biopsies from 27 diabetic patients and 12 non‐diabetic cases were analysed. Stereological methods were applied by light‐ and electron microscopy. Diabetic patients showed quantitatively markedly expressed diabetic glomerulopathy, but also an increase in glomerular volume, in prevalence of new‐vessel formation at the vascular pole, prevalence of glomerular occlusion and in interstitial volume fraction. A significant correlation was not observed between the degree of interstitial and glomerular involvement. The glomerular hypertrophy is interpreted as a compensatory phenomenon, leading to preservation of filtration surface in the open glomeruli. Close correlation was seen between glomerulopathy and glomerular function, and also with the stage of retinopathy. New vessel formation at the vascular pole was most frequent in patients with proliferative retinopathy. Signs of non‐diabetic glomerulopathy were not observed, but various atypical ultrastructural changes accompanying the advanced stages are illustrated. Our present findings correspond to data from type 1 diabetic patients. It is emphasised that all compartments of the kidney are affected by the diabetic state. It is suggested that the interstitial and glomerular lesions are influenced by different factors.

NOORAFSHAN, A.; EBRAHIMPOOR, M. R.; SADEGHI, Y.

doi: 10.1034/j.1600-0463.2001.d01-143.xpmid: 11900055

Most research on diabetes mellitus has focused on physiological and biochemical aspects of the peripheral nervous system, whilst little work has been done on morphological changes of the neurons. In the present study the effects of diabetes mellitus on cervical and lumbar dorsal root ganglia (C7 and L5) were investigated using modern stereological methods. Twelve adult male Sprague‐Dawley rats were randomly divided into two groups. Each group contained six male rats. Diabetes was induced in the experimental group by intraperitoneal injection of 50 mg/kg streptozotocin. At the end of 6 weeks, the rats were fixed by whole body perfusion transcardially with a buffered formalin solution. The seventh cervical and fifth lumbar dorsal root ganglia were removed and immersed in buffered formalin. After tissue processing, the ganglia were embedded in cylindrical paraffin blocks. Isotropic uniform random sections were obtained using the orientator method. Sections (5 μm thick) were selected and stained with Heidenhain's azan. Volume of perikarya of A‐ and B‐cells and their nuclei was estimated using the nucleator method. Before estimating the mean volume, the cells were sampled using the physical disector and point sampling method. Measurements showed that mean perikaryal and nuclear volume of A‐ and B‐cells of dorsal root ganglia (C7 and L5) was reduced in diabetic rats (p<0.05). B‐cell mean perikaryal volume in diabetic rats and A‐ and B‐cell mean nuclear volume were reduced by 66% on average. The mean volume of A‐cell perikarya was affected less than the others (average 33%). In addition, the difference between the perikaryal and nuclear volume of the seventh cervical and fifth lumbar dorsal root ganglia was not statistically significant. The present study, using stereological techniques, demonstrates reduced perikaryal and nuclear volume of the seventh cervical and fifth lumbar dorsal root ganglia in diabetic rats.

GULMANN, CHRISTIAN; ØSTERBY, RUTH; BANGSTAD, HANS‐JACOB

doi: 10.1034/j.1600-0463.2001.d01-144.xpmid: 11900056

Aim: To determine the long‐term changes of the juxtaglomerular apparatus in incipient diabetic nephropathy. Methods: Three renal needle biopsies were performed on 15 young type 1 diabetic patients with microalbuminuria; at baseline and after an average of 2.4 and 8.2 years. Using light microscopy, 1 μm serial sections of the plastic‐embedded biopsies were investigated and volumes of the juxtaglomerular apparatus and glomerulus and areas of the macula densa and lumina of the afferent and efferent arterioles were measured. Results: From baseline to second follow‐up there was a significant decrease in JGA relative to glomerular volume. There was an increase in luminal area of the efferent arteriole which was paralleled by (non‐significant) changes in the afferent arteriole. Conclusion: Over a period of 8.2 years JGA size remained stable, but decreased relative to glomerular size. Also, an increase in luminal area was noted in efferent arterioles. This may be due to increased single nephron blood flow secondary to nephron loss.

AVGERINOS, DIMITRIOS V.; BJÖRNSSON, JOHANNES

doi: 10.1034/j.1600-0463.2001.d01-145.xpmid: 11900057

Background. During the past few decades, hospital autopsy rates have steadily declined throughout the Western world. This decline is mainly attributed to the introduction of advanced diagnostic techniques. Despite technological developments, discrepancy rates between clinical diagnoses and autopsy findings remain high. Few studies have addressed discrepancy rates exclusively with regard to malignant neoplasms. In the present study, we reviewed the records of 3,118 autopsies performed at Mayo Clinic during a 6‐year period (1994–1999) and identified clinically undiagnosed malignancies found at autopsy and clinically diagnosed cancers not confirmed at postmortem examination. Materials and Methods. Autopsy protocols, provisional and final anatomic diagnoses, and data from the Mayo Autopsy Pathology Quality Assurance program were reviewed in an attempt to identify discrepancies between clinical diagnoses and autopsy findings regarding malignant neoplasms. Results. In 3,118 autopsies performed at Mayo Clinic between 1994 and 1999, a malignant tumor was identified in 768 cases (25%). In 128 of 3,118 cases (4.1%), the malignancy was not diagnosed clinically. In 14 of 3,118 cases (0.45%), autopsy failed to confirm a clinically diagnosed cancer. A review of the literature is presented. Conclusions. Autopsy remains an effective tool for the confirmation and refutation of clinical diagnostic findings regarding malignant neoplasms.

JÓNASDÓTTIR, O.; PETERSEN, J.; BENDTZEN, K.

doi: 10.1034/j.1600-0463.2001.d01-146.xpmid: 11900058

Wegener's granulomatosis (WG) is a systemic inflammatory disease with vasculitis as the key feature. Abnormal expression of tumour necrosis factor α (TNFα) is considered of prime pathogenic importance in several inflammatory diseases. The effects of TNFα are mediated by TNF receptors (TNF‐R), and these receptors are often found in soluble forms (sTNF‐R), which can modulate TNFα actions. To evaluate the clinical importance of the TNF family of cytokines, the serum levels of TNFα, TNFβ, now termed lymphotoxin (LTα), and sTNF‐R1 and sTNF‐R2 were measured by ELISA in 8 patients with WG during active disease and during immunosuppressive treatment, and in 11 healthy controls in parallel. Serum concentrations of TNFα were undetectable in all except two controls (18%) and three patients with WG (37%). After 7 days of therapy, six of the WG patients had measurable TNFα levels. Examination of the relative amounts of TNFα and sTNF‐R indicated that TNFα was mostly bound to its soluble receptors. In WG, the serum levels of sTNF‐R1 and sTNF‐R2 were dramatically increased (p<0.01), with little or no variation during treatment. While the IL‐1β levels did not deviate significantly from controls, the IL‐1ra levels were significantly elevated in the WG patients throughout the study period (p<0.01).