Gene expression of the renal endothelin system in renal transplant recipients on cyclosporine A based immunosuppression.

BACKGROUND: Immunosuppressive therapy based on cyclosporine A (CsA) is potentially nephrotoxic, and each dose of CsA is followed by a transient increase in plasma endothelin (ET)-1. The aim of this study was to investigate the effect of CsA based immunosuppressive therapy on renal gene expression of the ET(A) and ET(B) receptor subtypes and preproET-1 in human transplant needle biopsies. METHODS: Twelve living donor renal transplant recipients, median age 51.5 years (range 24-63 years) were included in the study. Immunosuppressive therapy consisted of CsA, azathioprine, and prednisolone. Baseline renal cortical needle biopsies from the living donor kidneys were obtained just before nephrectomy. Follow-up biopsies were obtained from the same transplanted kidneys after 2-6 weeks of immunosuppressive therapy. We used a quantitative, competitive reverse transcriptase polymerase chain reaction assay to measure renal ET(A) and ET(B) receptor subtype mRNAs as well as preproET-1 mRNA levels in each of the biopsies. RESULTS: The renal ET system was not significantly altered by CsA-based immunosuppressive therapy. Median ET(A) mRNA level was 185 (range 35-244) at baseline, and 120 (11-189) amol/microg total RNA after CsA based immunosuppressive therapy (P=0.11). ET(B) mRNA level was 506 (209-1411) at baseline, and 463 (267-1609) amol/microg total RNA at follow-up (P=0.44) and preproET-1 mRNA level was 160 (112-392) before and 221 (187-361) amol/microg total RNA after immunosuppressive therapy based on CsA (P=0.58). CONCLUSION: This study indicates that 2-6 weeks of CsA-based immunosuppression neither significantly influences renal gene expression of the ET(A) or ET(B) receptor subtypes nor preproET-1 in living donor renal transplant kidneys.

Wernicke's encephalopathy in an autopsy material obtained over a one-year period.

In a material comprising 279 consecutive autopsies obtained over a one-year period there were four cases of Wernicke's encephalopathy (WE), of which three were inactive (chronic) and one was active (acute). The latter was not related to chronic alcoholism. Our findings show that WE may be regularly present in a routine autopsy series. In only one case was WE suspected clinically and none of the cases revealed macroscopic brain changes indicative of WE. We therefore suggest that sections of the mammillary bodies should be taken routinely to detect all possible cases of WE.

Large intramuscular spindle-cell lipoma. With review of the literature. A case report.

A case of a large spindle-cell lipoma in an intramuscular (subfascial) localization is reported. The tumour occurred in a 58-year-old male patient and was localized subfascially in his left deltoid muscle. The tumour measured 20 X 8 X 8 cm; its weight was 780 grams. After initial biopsy, the tumour was extirpated. Microscopically, the tumour tissue consisted of mature fat cells and spindle-cell areas in varying amounts, constituting a typical appearance of a spindle-cell lipoma. There were no histological signs of malignancy. Especially, no areas of liposarcoma differentiation were detected. The patient is well, with no signs of recurrence, one and a half years after the operation. We have reviewed the literature on spindle-cell lipoma, and we believe this is the first example of this neoplasm to be reported in an intramuscular (subfascial) localization.

Morphological studies of baseline needle biopsies from living donor kidneys: light microscopic, immunohistochemical and ultrastructural findings.

Fifty-seven consecutive living donors (31 women and 26 men aged 20.7-72.3 years, mean 50.6 years) were subjected to needle biopsy during nephrectomy, immediately before removal of the kidney. By light microscopy, semiquantitative scoring (0-3) was performed for arteriosclerosis, arteriolar hyalinosis (hyalin arteriolosclerosis), glomerulosclerosis, interstitial mononuclear cell infiltration, and interstitial fibrosis/tubular atrophy. Whereas vascular changes were striking in many biopsies (arteriosclerosis grades 2-3: 28/54 cases, arteriolar hyalinosis grades 2-3: 15/55 cases), glomerular and tubulointerstitial changes were mostly low grade. The morphological changes tended to be more pronounced in middle-aged and older individuals, but, in particular, vascular changes were seen also in the younger age group. Immunofluorescence microscopy revealed glomerular granular staining for IgM in 52.7% of the cases, IgA in 9.1%), IgG in 1.8%, and C3 in 12.7%. The main ultrastructural finding was glomerulosclerosis; one case with diffuse glomerular IgA showed distinct dense deposits, and one case showed similar dense deposits without IgA deposition. Arteriolar wall deposition of C3 was found in 58.2% of the cases, and IgM in 10.9%. Especially C3 occurred both with arteriolar hyalinosis and in arterioles without light microscopic alterations. The observation of significant vascular changes in baseline biopsies is relevant especially in the differential diagnosis of chronic rejection and cyclosporine nephropathy. The immunohistochemical findings strongly indicate the occurrence of immunoglobulins and complement factor C3 in both glomeruli and arterioles without clinical or morphological signs of renal disease. The possible pathophysiological significance of such deposits remains, however, uncertain.

Postpartum renal failure: a complex case with probable coexistence of hemolysis, elevated liver enzymes, low platelet count, and hemolytic uremic syndrome.

BACKGROUND: Postpartum renal failure in previously healthy subjects is associated most often with preeclampsia and/or hypertension; hemolysis, elevated liver enzymes, low platelets (HELLP) syndrome, hemolytic uremic syndrome; or thrombotic thrombocytopenic purpura. Transient oliguria associated with preeclampsia is common, but renal failure is rare. Coexistence of HELLP and hemolytic uremic syndromes has been suggested, but histopathologic documentation of this combination has been scarce. CASE: A 30-year-old primigravida with severe preeclampsia at 35 weeks and 3 days' gestation presented with the development of HELLP syndrome and renal failure postpartum. Histopathologic lesions characteristic of hemolytic uremic syndrome were present in the kidney. CONCLUSION: Probable overlapping of HELLP and hemolytic uremic syndromes in pregnancy or postpartum should be taken into consideration when treating patients with these syndromes and associated complications, such as renal failure.

Neuronal cell death in the visual cortex is a prominent feature of the X-linked recessive mitochondrial deafness-dystonia syndrome caused by mutations in the TIMM8a gene.

The Mohr-Tranebjaerg syndrome (MIM 304700) and the Jensen syndrome (MIM 311150) were previously reported as separate X-linked recessive deafness syndromes associated with progressive visual deterioration, dystonia, dementia, and psychiatric abnormalities. In the most extensively studied Norwegian family, the Mohr-Tranebjaerg syndrome was reported to be caused by a one-basepair deletion (151delT) in the deafness/dystonia peptide (DDP) gene at Xq22. This gene has been renamed TIMM8a. We identified a stop mutation (E24X) in the TIMM8a gene segregating with the disease in the original Danish family with the Jensen syndrome, which confirms that the two disorders are allelic conditions. We also report abnormal VEP examinations and neuropathological abnormalities in affected males from the two unrelated families with different mutations. The findings included neuronal cell loss in the optic nerve, retina, striate cortex, basal ganglia, and dorsal roots of the spinal cord. The demonstration of mitochondrial abnormalities in skeletal muscle biopsies in some patients is compatible with the suggestion from recent research that the TIMM8a protein is the human counterpart of an intermembrane mitochondrial transport protein, Tim8p, recently characterized in yeast. The clinical and neuropathological abnormalities associated with mutations in the TIMM8a gene support that this X-linked deafness-dystonia-optic neuropathy syndrome is an example of progressive neurodegeneration due to mutations in a nuclear gene necessary for some, yet unknown mitochondrial transport function. We recommend sequencing the TIMM8a gene, thorough ophthalmological examination, and measuring visual evoked potentials in clinically suspected male patients with either progressive hearing impairment, dystonia, or visual disability in order to establish an early diagnosis and provide appropriate genetic counselling.

Recurrent angiolymphoid hyperplasia with eosinophilia mimicking temporal arteritis associated with nephrotic syndrome.

We report on a middle-aged Caucasian male who presented with nephrotic syndrome that on 2 consecutive recurrences was accompanied by a pulsating tumor suggesting temporal arteritis. Renal biopsies showed features of a low-grade mesangial-proliferative glomerulonephritis. The resected tumor in the temporal region revealed a lesion consistent with angiolymphoid hyperplasia with eosinophilia (ALHE), with moderate inflammatory involvement of the temporal artery. The patient was successfully treated with oral prednisolone in addition to removal of the tumor, but has remained steroid-dependent. To our knowledge, only 2 cases of ALHE and nephrotic syndrome have been reported so far in non-Japanese individuals [Altman et al. 1995, Sonkodi et al. 1987], and we are not aware of any previous case combining these features while simultaneuosly mimicking temporal arteritis.

Glomerular monocyte/macrophage influx correlates strongly with complement activation in 1-week protocol kidney allograft biopsies.

BACKGROUND: The specific role of monocytes/macrophages (MO) in kidney graft rejection is not yet fully elucidated. In a recent protocol biopsy study of living-donor recipients, we demonstrated massive capillary influx of MO, associated with severe complement activation and acute rejection (AR) 1 week after transplantation [Sund et al.]. To gain further insight into the possible relationship between MO and complement activation, we analyzed glomerular and interstitial MO in these biopsies. METHODS: Twenty-seven protocol biopsies were stained with antibodies to calprotectin (L1) and CD68 as markers for MO. Cells were counted as an average number per glomerulus and as an average number per defined visual field in the interstitium. Polymorphonuclear leukocytes (PMN) were counted in glomeruli and interstitium by light microscopy. Baseline specimens from 10 of the patients served as controls. The results were compared with data on deposition of complement from the foregoing study, and with histopathologic and clinical data on AR. RESULTS: Cases with diffuse C4d deposition in peritubular capillaries consistent with acute antibody-mediated rejection (AbAR) (n = 4) had significantly higher numbers of intraglomerular MO than the other protocol biopsies (L1: median 20.7 vs 3.6, p = 0.0002; CD68: median 10.1 vs. 2.0, p = 0.0008). With a cut-off of 10 L1-positive and 6 CD68-positive MO, the specificity for the diagnosis of AbAR was 96% and 91%, respectively. The number of interstitial MO was significantly higher in patients with AR than in those without, but in contrast to glomerular MO, interstitial MO could not discriminate between complement positive and negative AR. The number of glomerular and interstitial PMNs was significantly higher in the AbAR group than in the other protocol biopsies. CONCLUSIONS: The strong correlation between complement activation and early glomerular influx of MO in the kidney allograft suggests a causal relationship between these 2 events. At 1 week after transplantation, a number of 10 L1-positive and 6 CD68-positive MO per glomerulus indicates AbAR.

Morphological and functional renal effects of long-term low-dose cyclosporin A treatment in patients with rheumatoid arthritis.

To explore the possible nephrotoxic effects of low-dose cyclosporin A (CyA) treatment, we analyzed the data from 10 patients, aged 35-66 years (mean 51.8 years), who had a clinical diagnosis of rheumatoid arthritis and no known kidney disease. The study protocol included consecutive kidney biopsies and a pretreatment biopsy in all cases. A second biopsy was taken after 5-20 months (mean 17.8 months) of treatment and, in seven patients, a third biopsy was performed after 30-46 months (mean 38.6 months). Evaluation of the kidney biopsies included a semiquantitative estimation of different histological parameters as well as assessment of a chronicity index (CI). Transmission electronmicroscopic examination was performed on all biopsies. There was a significant reduction of glomerular function at the time of both the second (n = 10; p < 0.01) and third (n = 7; p < 0.05) biopsies. Five patients showed an increase in CI on the second biopsy and five on the third biopsy in comparison to pretreatment values. Only one patient showed a progressive increase in CI on three consecutive biopsies. The mean CI increased on both the second (n = 10) and third (n = 7) biopsies compared with baseline, but there was no increase on the third biopsy from the second. The morphological findings were, as a rule, slight or moderate, and focal interstitial fibrosis, tubular atrophy and arteriolar hyalinosis were the most consistent findings. Although even low-dose CyA treatment may be nephrotoxic and may induce morphological alterations in the kidney, such changes do not occur in all patients and may not necessarily be progressive.(ABSTRACT TRUNCATED AT 250 WORDS)

Small cell pleomorphic T-cell lymphoma presenting with cutaneous vasculitis.

The association between cutaneous vasculitis and lymphoproliferative disease has been increasingly recognized. We report a female patient who presented with cutaneous vasculitis which was due to a small cell pleomorphic T-cell lymphoma. In spite of aggressive therapy, she died two years after onset of disease. The clinical picture of vasculitis associated with lympho-proliferative disease is addressed with particular emphasis on symptoms and signs suggesting malignancy rather than vasculitis appearing in concert with well-defined connective tissue disorders.

Refractory cardiogenic shock and complete heart block after unsuspected verapamil-SR and atenolol overdose.

A 57-year-old female presented with complete heart block and then developed refractory hypotension despite temporary pacing. Moderate left ventricular dysfunction with focal wall motion abnormalities, as well as severe hypoxemia, were demonstrated. However, neither significant coronary disease nor evidence for pulmonary embolus or other lung disease could be determined. Hemodynamic stabilization was achieved with the use of an intra-aortic balloon pump and multiple high-dose pressor agents. A retrospective diagnosis of toxic verapamil-SR and atenolol ingestion was confirmed, and the patient gradually recovered. The relevant literature is reviewed and various treatment approaches are discussed.

Vascular relationships in right colectomy for cancer: clinical implications.

AIMS: The study aim was to provide data on pattern and length of crossing of the ileocolic artery (ICA) and right colic artery (RCA) with the superior mesenteric vein (SMV). METHODS: Specimens from 30 fresh human cadavers underwent corrosion casting. Methylacrylate was injected into the SMV and superior mesenteric artery (SMA). Length of crossing was measured with a scaleable ruler and copper wire. Values are mean (SD; range). RESULTS: ICA was present in all specimens and crossed posterior to the SMV in 19 (63.33%) of 30 specimens. Length of crossing was 17.01 (7.84; 7.09-42.89) mm. RCA was present in 19 (63.33%) of 30 specimens. RCA crossed anterior to SMV in 16 (84.21%) of 19 specimens. Length of crossing was 20.63 (8.09; 6.3-35.7) mm. CONCLUSIONS: ICA was always present, crossed posterior to SMV in 60% of specimens with a crossing length of 17 mm. RCA was present in 63% of specimens, crossed anterior to the SMV in 84% of specimens with a crossing length of 20 mm. Clinical implications include arterial length left behind with main nodes, arterial bleeding and safety of laparoscopic access.

Intramuscular hemangioma in the oral region: report of three cases.

The occurrence of intramuscular hemangioma in an intraoral or perioral localization is rare, and a thorough knowledge of these tumors is necessary for adequate diagnosis and treatment. Three cases are presented with discussion, and their histopathology and differential diagnosis are discussed. An adequate primary excision is necessary to avoid recurrence.

Actin dynamics at the living cell submembrane imaged by total internal reflection fluorescence photobleaching.

Although reversible chemistry is crucial to dynamical processes in living cells, relatively little is known about relevant chemical kinetic rates in vivo. Total internal reflection/fluorescence recovery after photobleaching (TIR/FRAP), an established technique previously demonstrated to measure reversible biomolecular kinetic rates at surfaces in vitro, is extended here to measure reversible biomolecular kinetic rates of actin at the cytofacial (subplasma membrane) surface of living cells. For the first time, spatial imaging (with a charge-coupled device camera) is used in conjunction with TIR/FRAP. TIR/FRAP imaging produces both spatial maps of kinetic parameters (off-rates and mobile fractions) and estimates of kinetic correlation distances, cell-wide kinetic gradients, and dependences of kinetic parameters on initial fluorescence intensity. For microinjected rhodamine actin in living cultured smooth muscle (BC3H1) cells, the unbinding rate at or near the cytofacial surface of the plasma membrane (averaged over the entire cell) is measured at 0.032 +/- 0.007 s(-1). The corresponding rate for actin marked by microinjected rhodamine phalloidin is very similar, 0.033 +/- 0.013 s(-1), suggesting that TIR/FRAP is reporting the dynamics of entire filaments or protofilaments. For submembrane fluorescence-marked actin, the intensity, off-rate, and mobile fraction show a positive correlation over a characteristic distance of 1-3 microm and a negative correlation over larger distances greater than approximately 7-14 microm. Furthermore, the kinetic parameters display a statistically significant cell-wide gradient, with the cell having a "fast" and "slow" end with respect to actin kinetics.

Can a diagnosis of renal allograft rejection be based on histology alone?

Acute renal allograft rejection is suspected by the clinician when the serum creatinine value increases in a patient for no other particular cause. A renal allograft biopsy may confirm the diagnosis. This report describes 2 patients with stable serum creatinine; however, protocol biopsy showed acute rejection changes according to the Banff criteria. No anti-rejection treatment was started and their graft function remained stable for 6 months. These two cases focus on the fact that renal allograft rejection should first of all be regarded as a clinical diagnosis which could be substantiated by histological findings.

The histone-like protein H-NS acts as a transcriptional repressor for expression of the anaerobic and growth phase activator AppY of Escherichia coli.

The transcriptional activator AppY is required for anaerobic and stationary-phase induction of the cyx-appA and hya operons of Escherichia coli, and expression of the appY gene itself is induced by these environmental conditions. The sequence of the appY gene and its promoter region is unusually AT rich. The nucleoid-associated protein H-NS has a DNA-binding specificity for intrinsically curved AT-rich DNA. Using a single-copy transcriptional appY-lacZ fusion, we have shown that appY gene expression is derepressed in hns mutants during aerobic exponential growth. In the hns mutant, growth phase and growth rate regulation under aerobic conditions was maintained, while ArcA-dependent anaerobic induction was greatly diminished. Judged by two-dimensional gel electrophoresis, the appY promoter fragment exhibits the features characteristic of curved DNA. Gel retardation assays showed that purified H-NS protein bound with high affinity to two different segments of the appY promoter region. The role of H-NS in the AppY regulatory cascade is discussed and compared with its function in the regulatory cascades of the AppY homologs CfaD and VirF.

Cell membrane orientation visualized by polarized total internal reflection fluorescence.

In living cells, variations in membrane orientation occur both in easily imaged large-scale morphological features, and also in less visualizable submicroscopic regions of activity such as endocytosis, exocytosis, and cell surface ruffling. A fluorescence microscopic method is introduced here to visualize such regions. The method is based on fluorescence of an oriented membrane probe excited by a polarized evanescent field created by total internal reflection (TIR) illumination. The fluorescent carbocyanine dye diI-C(18)-(3) (diI) has previously been shown to embed in the lipid bilayer of cell membranes with its transition dipoles oriented nearly in the plane of the membrane. The membrane-embedded diI near the cell-substrate interface can be fluorescently excited by evanescent field light polarized either perpendicular or parallel to the plane of the substrate coverslip. The excitation efficiency from each polarization depends on the membrane orientation, and thus the ratio of the observed fluorescence excited by these two polarizations vividly shows regions of microscopic and submicroscopic curvature of the membrane, and also gives information regarding the fraction of unoriented diI in the membrane. Both a theoretical background and experimental verification of the technique is presented for samples of 1) oriented diI in model lipid bilayer membranes, erythrocytes, and macrophages; and 2) randomly oriented fluorophores in rhodamine-labeled serum albumin adsorbed to glass, in rhodamine dextran solution, and in rhodamine dextran-loaded macrophages. Sequential digital images of the polarized TIR fluorescence ratios show spatially-resolved time-course maps of membrane orientations on diI-labeled macrophages from which low visibility membrane structures can be identified and quantified. To sharpen and contrast-enhance the TIR images, we deconvoluted them with an experimentally measured point spread function. Image deconvolution is especially effective and fast in our application because fluorescence in TIR emanates from a single focal plane.

[Disseminated fungal infections in neonates--risk factors, treatment and course]. / Disseminerte soppinfeksjoner hos nyfødte--risikofaktorer, behandling og forløp.

During 10 1/2 months in 1990/91 eight premature babies and one mature baby with an intra-abdominal disease had disseminated Candida albicans infections. The incidence in premature newborns was 9% (8/92 patients). Risk factors such as respirator therapy, the use of broad spectrum antibiotics, supplemental parenteral nutrition and central intravascular catheters were frequently seen. Four patients survived the fungal infection. These included three of five babies treated with amphotericin B 0.5 mg/kg/day. Two patients who received fluconazole 3 mg/kg/day died after three days. In one patient the diagnosis was obtained post-mortem, and one patient with possible fungemia survived without therapy. The treatment of these patients depends on optimal fungal cultures and good co-operation between paediatricians and microbiologists.

[Clinical significance of early kidney biopsy]. / Betydningen av tidlig nyrebiopsi.

We use the unexpected results of five kidney biopsies to discuss how early biopsy in renal disease can change the therapy and correct the diagnosis of the disease. The first patient was a 73 year-old male diabetic who had osteomyelitis and developed rapidly progressive glomerulonephritis. The next patient was a 72 year-old man who was treated for cardiac failure and increasing serum creatinine. The kidney biopsy revealed rapidly progressive glomerulonephritis. The third patient developed acute renal failure after an episode with vomiting. Here the histological diagnosis was acute renal failure and parenchymatous renal disease could be ruled out. The next patient was a 13 year-old girl. She had proteinuria (5-6 g/d) and hypertension (200/140 mm Hg). After four months, serum creatinine was 200 mumol/l. She was then biopsied, and we found membranoproliferative glomerulonephritis type 1. After the diagnosis was established she was treated with immunosuppression and her condition improved. The last patient was a 55 year-old male diabetic. He developed nephrotic syndrome and the histological diagnosis of the kidney biopsy was membranous glomerulonephritis stage 1. Six months after the kidney biopsy we found carcinoma of the lung. This underlines the importance of the fact that 10% of membranous glomerulonephritides are tumour associated.

Induction of tumor-specific immunity in mice by immunization with reconstituted tumor membrane liposomes containing recombinant B7-2 (CD86).

There has been considerable interest in developing experimental vaccines using genetically modified tumor cells expressing cytokines or costimulatory molecules to enhance immunogenicity. The authors investigated an alternative approach of using protein transfer rather than gene transfer to introduce costimulatory molecules rapidly into tumor membranes. Immunization with a single dose of reconstituted tumor membrane liposomes containing purified recombinant B7-2 (CD86) induced tumor rejection in mice challenged with syngeneic tumors, including the poorly immunogenic AG104A fibrosarcoma. These findings support the possibility that cell-free vaccines composed of reconstituted tumor membrane liposomes containing additional immunostimulatory proteins may offer a practical and safe alternative to genetically modified tumor cells for treating human cancer.