Accelerated healing of excisional skin wounds by PL 14736 in alloxan-hyperglycemic rats.

PL 14736 is a synthetic peptide, originally isolated from human gastric juice, that has anti-inflammatory and tissue-protective actions in experimental models of gastrointestinal inflammation. To investigate its possible benefit in poorly healing skin wounds, the effects of the topical application of PL 14736 in a gel formulation have been studied on full-thickness excisional wounds in rats, either healthy or made hyperglycemic by alloxan (175 mg/kg s.c.) 5 days previously. The effects of becaplermin gel (platelet-derived growth factor, PDGF-BB, Regranex, a standard therapy for diabetic foot ulcers, were investigated for comparison. Healing was evaluated for up to 7 days after wounding, using digital planimetry analysis, macroscopic scoring and histology. While healing was too rapid in healthy rats to observe enhancement by either treatment, in the hyperglycemic rats which exhibited delayed healing, PL 14736 (10-1,000 microg/wound) produced a dose-dependent acceleration of wound healing (determined by macroscopic scoring) equivalent at the highest doses to that observed with becaplermin. The beneficial effect on healing was associated with increased deposition of organized granulation tissue by day 7 for both PL 14736 and becaplermin, as determined histologically. PL 14736 tended to have a greater effect than becaplermin on the formation of granulation tissue containing mature collagen. Wound contraction, as measured by planimetry, was not significantly affected. In conclusion, topical PL 14736 produces a dose-dependent acceleration of deficient skin wound healing in hyperglycemic rats by facilitating granulation tissue formation, similar to the response seen with topical becaplermin, the standard therapy for diabetic skin wounds. PL 14736 may represent an alternative therapy for delayed wound healing, such as that seen with diabetic foot ulcers, without the proliferative concerns or immunogenicity associated with growth factors.

Immunohistochemical analysis of human serum sickness glomerulonephritis.

AIM: To analyze pathological and immunohistochemical characteristics of glomerulonephritis in human serum sickness. METHODS: Renal biopsy specimens from two female patients with serum sickness that ensued after application of anti-lymphocyte horse globulin for aplastic anemia were analyzed by light microscopy, immunofluorescence, and electron microscopy. To prove the depositions of foreign protein, frozen sections were incubated with fluorescein-conjugated anti-horse protein serum. Immunohistochemical analysis was performed on B5-fixed paraplast-embedded tissue or frozen acetone-fixed sections with the primary antibodies for molecules/cell markers CD35, CD43, CD45RO, CD68, CD2, lysozime, L26, and S100. RESULTS: Diffuse proliferating and necrotizing glomerulonephritis with crescents was found. There were coarse granular mesangial, subepithelial, subendothelial, and intramembranaceous deposits of mainly horse globulin, C3, and IgG. Most mesangium infiltrating cells were macrophages and T-lymphocytes. Electron microscopy revealed hypertrophy of podocytes, but immunohistochemistry did not show their normal CD35 (C3b-receptor) staining. Apart from epithelial cells, main crescent forming cells were macrophages and T-lymphocytes. Rare dendritic cells and abundant infiltration of macrophages, T-lymphocytes, and neutrophiles were found in the interstitium. CONCLUSION: In severe serum sickness, glomeruli and tubuli are destroyed beyond the range usually seen in other types of glomerulonephritis caused by immune complexes, except in cases with widespread crescents. Hypertrophy of podocytes and loss of their normal C3b-receptor staining has not yet been described in the literature. C3b-receptors on podocytes could play a role in pathogenesis of glomerular injury caused by immune complexes.

Heart metastasis of extraskeletal myxoid chondrosarcoma.

Isolated, remote heart metastasis of myxoid chondrosarcoma is extremely rare. We present an unusual case of isolated remote metastasis of extraosseus myxoid chondrosarcoma from the right ankle region to the right ventricle, its clinical course, and treatment in a 46-year-old woman. Although heart biopsy done before the surgery revealed myxoma, histopathologic diagnosis of the heart tumor was confirmed after its surgical resection from the right ventricle. Nine months after the surgery the patient was doing well but, seven months later, she died in the local hospital because of global heart failure. On the autopsy, the only metastatic lesion found was in the heart. The pericardium and heart muscle were infiltrated with the tumor, whereas all other organs were free from malignant dissemination.

Complete remission of cryoglobulinemic glomerulonephritis (HCV-positive) after high dose interferon therapy.

We report the case of a 64-year old woman with hepatitis C virus infection, mixed cryoglobulinemia type II (IgG + IgM kappa) and cryoglobulinemic glomerulonephritis. The patient was treated with the standard dose of recombinant interferon alpha-2b (3 million units 3 times a week) for one year, resulting in complete clinical remission and undetectable levels of serum hepatitis C virus RNA. AST and ALT normalized and proteinuria decreased from 2.78 to 0.98 g/day. However, a relapse occurred when therapy was stopped. Additional therapy with interferon-alpha (5 million units 3 times a week for 9 months) resulted again in quick and prolonged remission. The clinical course of our patient showed sustained clinical and virologic response after high-dose interferon-alpha treatment confirming the usefulness of interferon alpha in treatment of patients with cryoglobulinemic glomerulonephritis. Our observation is in agreement with others, suggesting that recommended standard dosage and duration of initial treatment with interferon alpha should be re-evaluated. Although our patient had sustained virologic and clinical response after interferon alpha monotherapy, recent studies clearly support combination therapy of interferon alpha and ribavirin for treatment of chronic HCV infections.

Histopathological features of gastritis before and after treatment for Helicobacter pylori.

AIM: To assess five main histological features of gastritis in gastric mucosa colonized with Helicobacter pylori before and after the treatment. METHODS: Histologic assessment of H. pylori-associated gastritis was performed according to the Sydney classification before and after the treatment in 97 patients. Two additional parameters - the presence of lymphocytic aggregates and coccoid forms of bacteria - were also analyzed. Helical and coccoid forms of H. pylori were detected by immunohistochemistry in biopsies after the treatment. RESULTS: Whereas acute epithelial damage was quickly repaired, some of the local responses to bacteria, e.g., lymphoid aggregates and intestinal metaplasia, persisted after treatment. Higher H. pylori and cocci density was found before and after treatment in patients with intestinal metaplasia (p=0.020). Correlation between H. pylori and mucosal atrophy was found only after treatment (p=0.009). Immunohistochemical staining was more sensitive in detecting of H. pylori than Giemsa staining (p=0.007) in cases where, using only Giemsa staining, it was not possible to distinguish coccoid forms of H. pylori from other cocci. CONCLUSION: After treatment, H. pylori-associated gastritis showed reduction of acute and chronic inflammation, but lymphoid aggregates and intestinal metaplasia persisted. Immunohistochemistry of different forms of H. pylori may be a valuable technique in monitoring the success of the treatment.

Analysis of renal allograft frozen-sections.

Frozen-section analysis of renal allograft biopsy specimens is performed to permit modification of therapy during acute events in transplanted patients. Over a two-year period, out of 110 renal allograft biopsies 66 were analyzed on a frozen-section. The authors compared diagnoses obtained on a fresh-material frozen-section with those following profound specimen analysis. The accurate diagnosis was achieved in 61 (92.4%) cases, acute rejection being predominant (53 cases). Missdiagnoses referred to oxalosis (1 case), acute rejection (2 cases), acute pyelonephritis (1 case) and cortical infarction (1 case). The authors conclude that a frozen-section analysis is a useful tool in cases requiring fast decision.

Pathology of the human mesangium in situ.

Mesangial cells play an important role in the development and progression of human glomerular disease. This article summarizes some important aspects of mesangial properties and behaviour in situ. Intrinsic mesangial cells express alpha-smooth muscle actin and are best characterized as myofibroblasts or glomerular pericytes. The main integrin receptor in the mesangium is the alpha 1 beta 1 integrin. The beta 2 and beta 3 integrins have not been detected. Mesangial cells in situ fail to react with many monoclonal antibodies which stain human mesangial cells in culture, including leukocyte activation antigens. Prominent reactions in glomerular disease are mesangial expansion and progressive glomerular sclerosis, which are preceded by or associated with mesangial cell hypertrophy and/or proliferation. Mesangial enlargement is accompanied by an altered integrin expression and an abnormal composition of extracellular mesangial matrix. From the numerous autocrine and paracrine mediators identified in vitro which stimulate or inhibit mesangial cell growth and extracellular matrix synthesis, up to now only a few factors have been shown to be present in selected human glomerulopathies. These include platelet derived growth factors and platelet derived growth factor receptor beta, transforming growth factors beta, interleukin 1 beta, tumor necrosis factor alpha, and interleukin 6. Further identification of such mediators in situ will improve our understanding of pathological glomerular processes, particularly with respect to the multifunctional properties of the mesangial cell.

Dendritic cells in glomerulonephritis.

Renal biopsies (n = 45) from patients with various forms of glomerulonephritis (GN), comprising mesangial IgA-GN (n = 25), focal glomerular sclerosis (n = 13) and acute GN (n = 7), were examined by double staining immunocytochemistry (APAAP, streptavidin-peroxidase) using unconjugated monoclonal antibodies (Ab) against--(i) the CD1b antigen expressed on dendritic cells (DCs), (ii) the invariant chain (Ii), and (iii) biotin-conjugated Ab against HLA-DR. In normal control kidneys (n = 7) without interstitial inflammation, CD1b-positive DCs were not detected. Glomerular endothelial cells and a few cells in mesangial areas showed double staining with the Ab against HLA-DR in Ii. In GN without active interstitial inflammation (n = 9), CD1b-positive DCs were not found. In biopsies with interstitial inflammation (n = 36) CD1b-positive DCs were found interspersed among other inflammatory cells. In seven of the biopsies showing IgA-GN DCs were seen in the vicinity of those glomeruli that exhibited either crescents or glomerular sclerosis with splitting of Bowman's capsule. In proximal tubular epithelial cells de novo expression of HLA-DR/Ii-chain was only seen when DCs were present. We conclude that in different forms of GN: (i) CD1b-positive DCs play an important role in the development of interstitial inflammation, and (ii) their presence may be related to the de novo coexpression of HLA-DR/Ii in tubular epithelial cells, possibly mediated through the production of interferon gamma and other cytokines.

[Histologic criteria for the diagnosis of superficially spreading melanoma in patients with proven metastases]. / Histoloski kriteriji za dijagnozu superficijalno sirecih melanoma u bolesnika kod kojih su dokazane metastaze.

The majority of authors agree that the prognosis of malignant melanoma depends on the depth of extension of malignant melanocytes into the dermis. A number of studies were performed to determine histologic characteristics of primary malignant melanoma that already metastasized. Their goal was to predict the biological behaviour of malignant melanoma by the histological picture. The study was designed to establish histological characteristics of superficial spreading melanoma. To prove the malignant biological behaviour, only superficial spreading melanomas with metastases in lymph nodes were analysed. The aim of the study was to see whether the melanoma's biological behaviour could be predicted on the basis of its histological picture. A total of 39 cases of superficially spreading malignant melanoma from the Department of Pathology, University Hospital Center Rebro, Zagreb, were analysed. The sections were taken from 5 different places, fixed in 10% purified formalin, paraffin embedded and stained with hematoxylin-eosin. In some cases the sections were additionally stained with Fontana to prove melanin. The sections were analysed by light microscopy. The following histological characteristics were analysed: 1. poor circumscription of the intraepidermal melanocytes; 2. individual melanocytes extending laterally; 3. number of atypical melanocytes above the basal membrane; 4. marked variation in the size and shape of melanocytic nests; 5. tendency of melanocytic nests towards confluence; 6. presence of melanocytes with nuclear atypia; 7. absence of maturation of melanocytes with descent into the dermis; 8. intradermal necrosis and degeneration of melanocytes; 9. lymphocytic infiltrate; 10. desmoplasia and ulceration. There were 22 male (56.4%) and 17 female (43.6%) patients.(ABSTRACT TRUNCATED AT 250 WORDS)

[Primary neuroendocrine carcinoma of the skin (Merkel cell tumor)]. / Primarni neuroendokrini karcinom koze (tumor Merkelovih stanica).

Three cases of neuroendocrine carcinoma of the skin studied by light and electron microscopy and by immunohistochemical methods, are presented. It is generally accepted that these tumors originate from Merkel's cells. Some consider that they belong to the group of APUD-omas. Positive findings of epithelial (EMA, CAM 5.2) and neuroedocrine marker (NSE) in these three cases support the hypothesis of neuroendocrine differentiation in a neoplasm of epithelial origin.