Cardioprotective effects of fullerenol C(60)(Oh)(24) on a single dose doxorubicin-induced cardiotoxicity in rats with malignant neoplasm.

The therapeutic utility of the anthracycline antibiotic doxorubicin is limited due to its cardiotoxicity. Our aim was to investigate the efficacy of fullerenol C(60)(OH)(24) in preventing single, high-dose doxorubicin-induced cardiotoxicity in rats with malignant neoplasm. Experiment was performed on adult female Sprague Dawley rats with chemically induced mammary carcinomas. The animals were sacrificed two days after the application of doxorubicin and/or fullerenol, and the serum activities of CK, LDH and alpha-HBDH, as well as the levels of MDA, GSH, GSSG, GSH-Px, SOD, CAT, GR, and TAS in the heart, were determined. The results obtained from the enzymatic activity in the serum show that the administration of a single dose of 8 mg/kg in all treated groups induces statistically significant damage. There are significant changes in the enzymes of LDH and CK (p < 0.05), after an i.p. administration of doxorubicin/fullerenol and fullerenol. Comparing all groups with untreated control group, point to the conclusion that in the case of a lower alpha-HBDH/LDH ratio, results in more serious the liver parenchymal damage. The results revealed that doxorubicin induced oxidative damage and that the fullerenol antioxidative influence caused significant changes in MDA, GSH, GSSG, GSH-Px, SOD, CAT, GR, and TAS level in the heart (p < 0.05). Therefore, it is suggested that fullerenol might be a potential cardioprotector in doxorubicin-treated individuals.

Sixteen novel mutations identified in COL4A3, COL4A4, and COL4A5 genes in Slovenian families with Alport syndrome and benign familial hematuria.

Alport syndrome (ATS) and benign familial hematuria (BFH) are type IV collagen inherited disorders. Mutations in COL4A5 are generally believed to cause X-linked ATS, whereas mutations in COL4A3 and COL4A4 genes can be associated with the autosomal-recessive and -dominant type of ATS or BFH. In view of the wide spectrum of phenotypes, an exact diagnosis is sometimes difficult to achieve. This study involved screening each exon with boundary intronic sequences of COL4A3, COL4A4, and COL4A5 genes by optimized polymerase chain reaction-single-stranded conformational polymorphism analysis in 17 families with ATS and in 40 families diagnosed as having BFH. Twelve different mutations were found in the COL4A5 gene in ATS patients, comprising nine missense mutations, a splice site mutation, a mutation causing frameshift, and a nonsense mutation. One of the missense mutations (p.G624D) was present not only in one family with ATS but also in five families with suspected BFH. Three heterozygous mutations in the COL4A3 gene (two missense and one frameshift) and four heterozygous mutations in COL4A4 (two splice site, one in-frame deletion, and one missense) were identified in patients with BFH. Sixteen mutations are to the best of our knowledge new and private.

De novo diabetic nephropathy on renal allografts.

BACKGROUND: Posttransplant diabetes mellitus (PTDM) is a well-recognized clinical problem following renal transplantation. Long-term risks of PTDM are similar to those of diabetes mellitus in general population. The aim of our study was to identify de novo diabetic nephropathy (DN) in our group of patients with PTDM. METHODS: Thirty-four patients with PTDM were reviewed retrospectively. Light microscopy, immunofluorescence, and electron microscopy techniques were performed in 10 of 21 patients with graft biopsy. RESULTS: Five patients (four women, one man), aged 47.4 years (range, 29 to 58), four of whom received cadaveric grafts, were found to have de novo DN. Their serum creatinine was 211.4 micromol/L (range, 140 to 294). Three patients were slightly proteinuric (0.3 to 0.5 g/L). PTDM was diagnosed 2.4 months after transplantation (range, 1 to 6). Histologic diagnosis of de novo DN was made, on average, 52.6 months after transplantation (range, 8 to 115), and 50.2 months (range, 2 to 114) after PTDM. De novo DN presented as diffuse diabetic glomerulosclerosis in four patients and nodular diabetic sclerosis in one patient, and combined with transplant glomerulopathy in all five patients. The mean graft survival time for this group of patients was equivalent with a control group. Although the difference in slopes of serum creatinine between the studied groups was clinically relevant, it was not statistically significant. CONCLUSION: In view of our findings, when histologic de novo DN was found in 5 out of 10 patients, one could conclude that de novo DN could be a frequent complication of PTDM.

Nerve fibre composition of the palmar cutaneous branch of the median nerve and clinical implications.

Fifteen fresh human cadaver hands were dissected, using x2.8 loupe magnification, to study the subcutaneous innervation at the site of the incision (in the line with the radial border of the ring finger) for standard open carpal tunnel decompression. Subcutaneous nerve branches were detected and traced proximally to determine their origin. Morphometric analysis of nerve cross sections from the site of the incision and from the main nerve trunk proximal to cutaneous arborisation was performed using light and transmission electron microscopy and a computer-based image analysis system. At the site of the incision, the ulnar sub-branch (US) of the palmar cutaneous branch of the median nerve (PCBMN), which innervates the skin over the hypothenar eminence, was found in 10 of 15 cases. Branches from the ulnar side were not detected. The main trunk of PCBMN consisted on average of 1000 (SD 229) myelinated axons arranged in 1-4 fascicles. In the US of the PCBMN there were on average 620 (SD 220) myelinated axons, 80% of them smaller than 40 microm(2) i.e. thin myelinated axons, and on average 2037 (SD 1106) unmyelinated axons, arranged in 1-3 fascicles. The ratio of the number of myelinated axons in the US and the main trunk of the PCBMN was on average 63% (SD 19%). Frequency distribution of cross-sectional areas of myelinated axons shows no significant difference between the US and the main nerve trunk of the PCBMN. The importance of incision trauma to subcutaneous innervation of palmar triangle is emphasised and possible mechanisms of scar discomfort are discussed.

Interstitial capillary in normal and in transplanted kidneys: an ultrastructural study.

Knowledge about the normal structure and pathology of interstitial capillary is limited. Splitting and multilayering of the basal membrane (BM), as a marker of chronic rejection, has been published in association with transplant glomerulopathy. The authors investigated the ultrastructural features of the interstitial capillary basal membrane in normal (15 biopsies) and in transplanted kidneys (27 biopsies from 21 patients), expressing transplant glomerulopathy (8 biopsies from 6 patients), acute tubulo-interstitial rejection (9 biopsies from 6 patients), and recurrent or de novo glomerulonephritis (10 biopsies from 8 patients). All biopsies were fixed in 1% OsO4, embedded in Epon, and examined by electron microscope. Measurements of the interstitial capillary BM were made. The BM of interstitial capillary of intact kidney was a homogenous continuous structure, 88 nm in width on average. Thickening with diffuse multilayering of BM was most intensive in patients with transplant glomerulopathy, and much less intensive in patients with acute tubulointerstitial rejection and in patients with recurrent or de novo glomerulonephritis. These findings may provide the first information about the morphology of the normal basal lamina of interstitial capillary and support the diagnostic value of interstitial capillary changes in chronic rejection.

Correlation among WHO classes, histomorphologic patterns of glomerulonephritis and glomerular immune deposits in SLE.

In addition to the conventional World Health Organization (WHO) classification of lupus glomerulonephritis (GN), various concomitant approaches have been introduced in the evaluation of renal biopsies of patients with systemic lupus erythematosus (SLE) in order to increase the impact of biopsies on the decision concerning the most appropriate therapy as well as for establishing the prognosis. Three hundred and seventy kidney tissue samples from 267 SLE patients were analysed using standardised light, electron and immunofluorescence microscopic techniques. In 155 patients, a comparative clinical follow-up study and statistical analysis were performed. The study highlighted the heterogeneity of WHO classes IV and III, which include 5 and 6 different conventional histomorphologic types of GN, respectively. Mixed membranous and proliferative GN associated with "full-house" mesangial-transmembranous immune deposits, demonstrated in more than one third of our SLE cases, appears to be diagnostically most characteristic. Immune deposits distributed in the glomeruli in five different patterns, obviously play a major role in the pathogenesis of various WHO classes and histomorphologic types of lupus GN. Additional mechanisms related to the occurrence of antiphospholipid antibodies and antineutrophil cytoplasmic antibodies are suggested to contribute to the histomorphologic heterogeneity of WHO class III and IV lupus GN, particularly to the development of thrombotic, necrotising and crescentic glomerular lesions. In the present study, a statistically significant association was demonstrated between increasing mean values of the activity index and glomerular deposit distribution patterns labeled by subendothelial deposits. Furthermore, a significant correlation was established between an increasing risk of developing renal failure and increasing mean values of the chronicity index. Differences in the increasing risk of developing renal failure between groups with different histomorphologic types of GN and different immune deposit distribution patterns were not statistically significant. The surprisingly high renal survival rate of more than 80% noted in lupus patients with predominantly necrotising crescentic GN during the mean follow-up period of 40 months appears to be related to the more aggressive treatment of those patients. Our study confirmed a significant role of the WHO classification of lupus GN in the decision concerning the most appropriate treatment and prognostication. An increasing risk of irreversible renal failure in patients with WHO class IV lesions in contrast to those of WHO class III and in contrast to those of the category incorporating all other WHO classes was shown to be statistically significant.

Tubulo-interstitial involvement in lupus nephritis with emphasis on pathogenesis.

Glomerular lesions in lupus nephritis have been extensively studied in recent decades, but much less attention has been paid to the tubulo-interstitial compartment. The aim of this study was to contribute to the understanding of the pathogenesis of tubulo-interstitial lesions in lupus nephritis by analysing their incidence, character, and their associations. One hundred and ninety kidney biopsies of 190 patients fulfilling American Rheumatology Association (ARA) criteria of systemic lupus erythematosus (SLE) were examined by traditional light, immunofluorescence and electron microscopy. Interstitial inflammatory infiltration and tubulo-interstitial immune deposits concurred in 72 cases (37.9%). Their frequency was the highest in WHO class IV lupus glomerulonephritis. By multivariate analysis, the intensity of interstitial inflammatory infiltration correlated best with the percentage of renal corpuscules with extracapillary crescents and the extent of interstitial fibrosis. On immunohistochemical assessment, the inflammatory infiltrate was found to be composed of CD45RO positive T lymphocytes (191.3/mm2), CD68 positive macrophages (101.7/mm2) and CD45RA positive B lymphocytes (17.2/mm2). For all cell types the median value was higher in cases with extracapillary crescents, and did not correlate with presence and intensity of tubulo-interstitial immune deposits. Infiltration showed the tendency of periglomerular distribution, especially around glomeruli showing extracapillary proliferation and destruction of the capsular basal membrane. Rare S100 positive cells were only found in the interstitium. Tubulo-interstitial lesions estimated semiquantitatively correlated with the degree of proteinuria. Our findings suggest that tubulo-interstitial deposits do not play a major role in the pathogenesis of tubulo-interstitial lesions. The formation of interstitial cell infiltrates appears to be greatly influenced by the development of extracapillary crescents, perhaps by direct transmission of the severe inflammatory process to the adjacent interstitium. The composition of the infiltrate, including antigen presenting cells may signalize an additional involvement of cell-mediated immune mechanisms acting against so far hypothetical tubular epithelial neoantigens.

Fingerprint and other organised deposits in lupus nephritis.

A prominent feature of lupus glomerulonephritis is extracellular, predominantly homogeneous electron dense deposits. Fingerprint-like deposits have been reported in 6 to 10% of cases. On electron microscopy, we studied the frequency and characteristics of organised deposits in 227 kidney tissue samples obtained by biopsy in 185 patients with systemic lupus erythematosus (SLE). Fingerprint forms of deposits were demonstrated in 34 biopsies of 32 patients (17.3%). In the control group of 626 kidney biopsies of patients with primary renal and systemic diseases other than SLE, no fingerprint deposits were found. In 227 kidney biopsy samples, fingerprint deposits were found to be associated with mesangial (8.8%), mesangial-subendothelial (3.8%), subepithelial (28.6%), mesangial-subepithelial (11.1%) and mesangial-transmembranous (19.4%) glomerular deposit distribution patterns. They were demonstrated more often at different locations along the peripheral capillary glomerular basal membrane (77.4%) than within the mesangial matrix (43.7%). In extraglomerular locations, fingerprint deposits were present in the interstitium in 8.8%, along the tubular in 23.5% and peritubular capillary basal membrane in 20.5%, in the wall of the arterioles in 64.7% and in the juxtaglomerular apparatus in 18.2% of biopsies. Organised fingerprint deposits consisted of semicircular dark and light lines, each with a diameter of about 10 to 15 nm. Unilaterally, spiky processes at a periodicity of 10 to 15 nm were seen. Among 14 of 185 SLE patients with cryoglobulinemia, fingerprint deposits were demonstrated in only 2 patients. We conclude that fingerprint deposits are characteristic, diagnostically relevant for SLE and represent morphologically a homogeneous group of organised deposits unrelated to cryoglobulins. In 3 SLE patients, 20 to 100 nm tubules and in 2 SLE patients, 10 and 18 nm Congo red negative fibrils were found. By their morphology and their structural characteristics, the tubules and fibrils resemble the tubules in primary immunotactoid glomerulopathy and fibrils in primary fibrillary glomerulonephritis.

[Significance of histopathologic analysis of skin lesions in scleromyxedema. Light microscopy, electron microscopy, immunohistochemistry and immunofluorescence microscopy]. / Bedeutung der histopathologischen Analyse der Hautläsionen bei Skleromyxödem. Lichtmikroskopie, Elektronenmikroskopie, Immunhistochemie und Immunfluoreszenzmikroskopie.

Scleromyxedema (SM) may be considered as a possible disease entity in the differential diagnosis of scleroderma. Clinical data and the results of light, immunohistochemical, immunofluorescence and electron microscopic study of skin biopsies taken from a 53-year old patient with SM are reported. In the patient with SM in which abnormal serum paraprotein was not identified, the skin biopsy showed mucinous material in the dermis and proliferation of fibroblasts accompanied by mild dermal sclerosis. Immunofluorescence showed scanty granular IgG along the epidermal basement membrane and IgG and C1q focally along the connective tissue fibres in the dermis of clinically involved skin. In addition to clinical findings, detailed skin biopsy studies including contemporary techniques can contribute to the diagnosis of the disease.

The significance of histologic analysis of skin lesions in porphyria cutanea tarda. Light microscopy, electron microscopy, immunohistochemical and immunofluorescence analysis.

Porphyria cutanea tarda (PCT) is one of several entities in the differential diagnosis of scleroderma. We report a 62-year-old man with PCT diagnosed since two decades. Clinical data and the results of light microscopy, immunohistochemistry, immunofluorescence and electron microscopy of skin biopsies are presented. The biopsy revealed sclerosis of the dermis mainly due to increased collagens I and III, and accumulation of collagen IV, which had caused the vessel wall to thicken. Immunofluorescence for detection of immune reactants was negative. It was concluded that the histomorphology of PCT of long duration may be similar to that of scleroderma. Nevertheless, in addition to clinical findings, detailed skin biopsy studies including contemporary techniques can contribute to the differentiation of these diseases.

Contribution to the pathogenesis of radiation-induced injury to large arteries.

We report a case of a 35-year-old man who died of a brain infarct 20 months after radiotherapy for carcinoma of the tonsil with metastases to the cervical lymph nodes. Histology revealed mild atherosclerosis, necrotizing vasculitis, and occlusive thrombosis of the internal carotid artery. Significant changes were observed in the vasa vasorum: swelling and detachment of the endothelium, subendothelial oedema, hyaline change, fibrinoid necrosis of the vessel walls with mononuclear cellular infiltration, accompanied by focal haemorrhages and chronic inflammation in the periadventitial soft tissue. We believe that these changes of the vasa vasorum and necrotizing vasculitis are causally related and that vasculitis represents focal ischaemic necroses with inflammatory reaction. Our findings support the hypothesis, based on experimental studies, that injury to the vasa vasorum is an important mechanism in the development of radiation-induced vasculopathy of large arteries. They also suggest an evolution of the injury to the vasa vasorum and periadventitial tissue from the early lesions described in our patient, to late stages resulting in dense periadventitial fibrosis as reported previously. We suggest that injury to the vasa vasorum and the consequent ischaemic lesions of the arterial wall are morphological features distinguishing radiation-induced arterial injury from spontaneous atherosclerosis.

Transmission electron microscopic analysis of trabecular meshwork in secondary glaucoma after intravitreal silicone oil injection.

PURPOSE: We analyzed morphological changes in trabecular meshwork in glaucoma developing after intravitreal silicone oil injection with transmission electron microscopy (TEM). METHODS: Specimens obtained from 2 patients after surgical trabeculectomy were fixed in McDowell fixative in one patient and in buffered formaldehyde in the other, dehydrated and embedded in Epon. Ultrathin sections were made and stained with lead citrate and uranyl acetate for TEM. RESULTS: In specimens mainly connective tissue sheets with disorganized and narrowed intertrabecular spaces were observed. At higher magnification the collagen sheets were composed of interstitial collagen and some of them were covered by amorphous basal membrane structures resembling material. Cellular remnants were present along the intertrabecular spaces. No other cells or clearly evident silicone oil were noted. CONCLUSION: Changes such as decrease in cell content, fibrosis and build-up of basement membrane-like material have been described by other authors. They are not specific and can be seen in many secondary glaucomas. No macrophages and foreign-body granulomas in response to silicone oil were present. The pathological changes in angle structures are probably caused by other mechanisms and are not induced by silicone oil itself.

Immunotactoid glomerulopathy with unusually thick extracellular microtubules and nodular glomerulosclerosis in a diabetic patient.

It has recently been suggested that immunotactoid glomerulopathy be separated from much more common fibrillary glomerulonephritis by ultrastructural features of highly organized immune deposits containing tubules of more than 30 nm in diameter. We report and discuss the results of a light, immunofluorescence and electron microscopic study of a needle renal biopsy from a 75-year-old, non-insulin dependant diabetic female presented with nephrotic syndrome, hypertension and a progressive renal failure. A unique coexistence of nodular glomerulosclerosis, as traditionally ascribed to diabetes with a peculiar type of immunotactoid glomerulopathy was confirmed by the exclusion of amyloidosis, monoclonal gammopathies, systemic autoimmune diseases and cryoglobulinemia. Mesangial, scattered subepithelial and segmentally prominent subendothelial immune deposits were found highly organized in mostly parallel arrays of 40 to 91 nm thick tubules. The average thickness of 67 nm exceeds the average diameter of tubules in all other 11 published cases of immunotactoid glomerulopathy to date. By immunofluorescence, predominantly capillary wall, thick, ribbon-like glomerular deposits contained IgG, IgM, kappa and lambda light chains of equal intensity, C3, C4 and fibrin related antigens. Mild to moderate glomerular cell proliferation associated with nodular sclerosis has been assumed to be causally related to immunotactoid deposits.

Renal function, protein excretion, and pathology of Balkan endemic nephropathy. III. Light and electron microscopic studies.

This study was performed to evaluate histomorphological features of BEN in 50 kidney biopsies from patients who met the epidemiologic, clinical and laboratory criteria for BEN. This is the first such study reported in detail. The patients were divided into three groups based on the DTPA clearance values: group 1, greater than 99 ml/min, group 2, 51 to 99 ml/min, and group 3, 29 to 50 ml/min. All patients in all groups had an increase in proteinuria consisting of proteins less than 25,000 daltons. Multifocal interstitial sclerosis spreading from the superficial into the deep cortex was found in 49 (98%), tubular atrophy in 48 (96%), and global glomerular sclerosis with microvascular hyalinosis/sclerosis of sclerotic and atrophic changes were significantly increased when compared to age-related standards. An accelerated aging process may be assumed to occur in BEN. More peculiar additional findings with much lower incidence and extent included multifocal vascular and glomerular capillary changes resembling the chronic form of thrombotic microangiopathy group of diseases. These findings, together with the presence of arteriolar hyalinosis and tubulointerstitial sclerosis seen in patients with cyclosporine nephrotoxicity suggest that the mechanism of toxicity may be similar to BEN. We conclude that the histopathology is predominantly tubulointerstitial sclerosis without infiltrates. The combination of the histology, tubular proteinuria, geographic distribution, familial occurrence, and the remarkable association with papillary transitional cell carcinoma of the renal pelvis and ureters, qualifies BEN as a unique disease.

Synovial sarcoma of the soft palate in a child: a light and electron microscopic study.

An extremely rare case of synovial sarcoma localized in the soft palate of a 9-year-old boy is reported, describing all details noted during light and electron microscopic observation. All features characteristic of this mesenchymal tumour were evident in the light microscopy. Electron microscopy also confirmed the diagnosis. The text includes full particulars of the therapy administered.