De novo collapsing glomerulopathy in renal allografts

BACKGROUND: Collapsing glomerulopathy is a recently described form of glomerular injury characterized by capillary collapse and visceral epithelial hypercellularity associated with nephrotic range proteinuria and a rapid, progressive decline in renal function. The lesion has rarely been described in allografts. METHODS: We reviewed 892 allograft biopsies from a population of 1079 recipients who received renal transplants between 1978 and 1996. RESULTS: Five cases of de novo collapsing glomerulopathy were identified (0.6% of biopsies; 3.2% since 1993). None occurred before 1993. The patients were 31 to 66 years of age and they presented 6 to 25 months after transplantation. The 24-hr urinary protein ranged from 1.8 to 11.8 g. All patients and donors were negative for the human immunodeficiency virus and had no risk factors for human immunodeficiency virus infection. Diffuse or focal, global or segmental collapse of glomerular capillaries, swelling and hypercellularity of the visceral epithelium, hyaline arteriolosclerosis, and interstitial fibrosis were characteristic histologic features. Two cases had concomitant glomerular immune complex deposits. Progressive decline in allograft function occurred within 2-24 months after diagnosis, culminating in return to dialysis in all patients. CONCLUSION: Collapsing glomerulopathy can arise in renal allografts as a de novo disease. Although its pathogenesis remains to be clarified, it is important to distinguish this lesion in allografts as it can be associated with rapidly progressive graft failure.     

Capture and expansion of bone marrow-derived mesenchymal progenitor cells with a transforming growth factor-beta1-von Willebrand's factor fusion protein for retrovirus-mediated delivery of coagulation factor IX

Mesenchymal stem cells give rise to the progenitors of many differentiated phenotypes, including osteocytes, chrondocytes, myocytes, adipocytes, fibroblasts, and marrow stromal cells, which are capable of self-renewal and undergo expansion in the presence of transforming growth factor-beta1 (TGF-beta1). The present study was designed to test the concept that mesenchymal progenitor cells could be selected and expanded by virtue of their intrinsic physiologic responses to TGF-beta1. Human bone marrow aspirates were initially cultured, under low serum conditions, in collagen pads or gels impregnated with a genetically engineered TGF-beta1 fusion protein bearing an auxiliary von Willebrand's factor-derived collagen-binding domain (TGF-beta1-vWF). Histologic examination of TGF-beta1-vWF-supplemented collagen pads from 8-day cultures revealed the selective survival of a population of mononuclear blastoid cells. The TGF-beta-responsive cells were expanded to form stromal/fibroblastic colonies by serum reconstitution, and further to form osteogenic colonies upon supplementation with osteoinductive factors. In comparative studies, both marrow-derived progenitor cells and mature stromal cells were transduced with a retroviral vector bearing a human factor IX construct. Both the transduced progenitor cells and mature stromal cells expressed the factor IX transgene at levels comparable to those reported for human fibroblasts. Transplantation of murine progenitor cells bearing the human factor IX vector into syngeneic B6CBA mice resulted in detectable circulating levels of the human factor IX antigen. Taken together, these data demonstrate a novel physiologic approach for the selection of mesenchymal precursor cells followed by mitotic expansion, transduction, and transplantation of these progenitor cells with retroviral vectors bearing therapeutic genes.     

Alterations in gene expression and signal transductions in human melanocytes and melanoma cells

The development of techniques to cultivate human primary melanocytes in vitro has provided the technical foundation for understanding the biology of this cell. Human melanocytes require various growth factors and agents for proliferation in vitro. These compounds activate two major signal transduction pathways: a calcium- and phospholipid-dependent (protein kinase C or PKC) pathway and a cyclic AMP (cAMP)-dependent (protein kinase A or PKA) pathway. Alterations in these signal transduction pathways coupled with changes in specific genes (protooncogenes, growth factors, and tumor suppressor genes) have been observed in human melanoma cells compared with normal melanocytes. Our own work indicates that loss in the expression of the PKC beta II isotype is a common, if not universal, alteration that occurs early in human melanocyte transformation. In this review, we concentrate on alterations in the signal transduction pathways in human melanocytes and melanoma cells and delineate how an understanding of these changes may allow us to understand the molecular mechanisms involved in human melanocyte transformation.     

Regionalization of infection prevention and control services in a publicly funded health care system

In 1994 health services in the Edmonton region were consolidated into an integrated network called Capital Health. Infection control professionals in the region met to develop a vision for the future of infection control; the tasks were to prepare a proposal for a regional program, develop indicators for outcome measurements, and standardize guidelines and products. Although regionalization of infection control is a complex process, we have had success with a proactive approach led by infection control professionals.     

Herpes simplex virus gD and virions accumulate in endosomes by mannose 6-phosphate-dependent and -independent mechanisms

Herpes simplex virus (HSV) glycoprotein D (gD) is modified with mannose 6-phosphate (M6P) and binds to M6P receptors (MPRs). MPRs are involved in the well-characterized pathway by which lysosomal enzymes are directed to lysosomes via a network of endosomal membranes. Based on the impaired ability of HSV to form plaques under conditions in which glycoproteins could not interact with MPRs, we proposed that MPRs may function during HSV egress or cell-to-cell spread (C. R. Brunetti, R. L. Burke, B. Hoflack, T. Ludwig, K. S. Dingwell, and D. C. Johnson, J. Virol. 69:3517-3528, 1995). To further analyze M6P modification and intracellular trafficking of gD in the absence of other HSV proteins, adenovirus (Ad) vectors were used to express soluble and membrane-anchored forms of gD. Both membrane-bound and soluble gD were modified with M6P residues and were localized to endosomes that contained the 275-kDa MPR or the transferrin receptor. Similar results were observed in HSV-infected cells. Cell fractionation experiments showed that gD was not present in lysosomes. However, a mutant form of gD and another HSV glycoprotein, gI, that were not modified with M6P were also found in endosomes in HSV-infected cells. Moreover, a substantial fraction of the HSV nucleocapsid protein VP6 was found in endosomes, consistent with accumulation of virions in an endosomal compartment. Therefore, it appears that HSV glycoproteins and virions are directed to endosomes, by M6P-dependent as well as by M6P-independent mechanisms, either as part of the virus egress pathway or by endocytosis from the cell surface.     

Lactose maldigestion is not an impediment to the intake of 1500 mg calcium daily as dairy products

BACKGROUND: A National Institutes of Health consensus conference concluded that a daily calcium intake of 1500 mg reduces the severity of osteoporosis. Because dairy products are the main natural source of dietary calcium, a diet providing 1500 mg Ca must contain large quantities of dairy products. However, it is widely believed that the lactose content of these products will not be tolerated by persons with lactose maldigestion (approximately 30% of the adult US population). OBJECTIVE: We evaluated the symptoms of lactose maldigestion and digestion when the diet was supplemented with dairy products providing 1300 mg Ca/d. DESIGN: Sixty-two women (31 with lactose maldigestion and 31 without) were studied in a double-blind, randomized protocol. Symptoms were compared during 1-wk periods when the diet was supplemented with 480 mL (2 cups) milk, 56 g cheese, and 240 mL yogurt provided as conventional products (34 g lactose/d) or as lactose-hydrolyzed products (2 g lactose/d). RESULTS: Women who digested lactose reported no significant difference in symptoms between the 2 treatment periods. Women with lactose maldigestion reported significantly increased flatus frequency and subjective impression of rectal gas during the period of high lactose intake; however, bloating, abdominal pain, diarrhea, and the global perception of overall symptom severity were not significantly different between the 2 treatment periods. CONCLUSION: The symptoms resulting from lactose maldigestion are not a major impediment to the ingestion of a dairy-rich diet supplying approximately 1500 mg Ca/d.     

Characteristics of tick, Rhipicephalus appendiculatus, glands distinguished by surface lectin binding

Incubation of fluorescein-conjugated and biotinylated lectins with Rhipicephalus appendiculatus salivary glands revealed differences between the basal laminae of the haemocoelic surfaces of the three acinal types. There were some similarities in the lectin binding characteristics of the surfaces of type II and type III acini when Con A, PNA and TVA were applied, indicating the presence of galactose, mannose and glucose moieties on the acinal surfaces. The binding of BPA, HPA and HAA lectins, specific for galactosyl and glycosyl ligands, which occurred only on the surface of type III acini, was moderate to intense. The remaining galactose-reactive lectins (GMA, DBA and VVA) also bound only to type III acini and the level of binding was weak to moderate. Although the relationship between the haemocoelic surface of the R. appendiculatus salivary gland acini and Theileria kinetes is not known, the consistent differences in the surface carbohydrate composition of the acini confirm the existence of differences in the basic physiology of the acini which may correlate with the specific susceptibility of the type III acinus to Theileria parva infection.     

Structural Dependence on Sintering Temperature of Lead Zirconate-Titanate Solid Solutions

Pb(Zr_0.525Ti_0.475)O₃ piezoceramics, both unmodified and doped with 2 wt% Bi₂O₃ or Nb₂O₅, were prepared by the usual techniques, using sintering temperatures from 900° to 1250°C. The microstructural data showed that the sintering temperature which produces minimum porosity is altered by the oxide additions. X-ray diffraction demonstrated the coexistence of both ferroelectric phases. The lattice parameter measurements showed that the tetragonal and rhombohedra1 unit cells of the two ferroelectric phases depend on the sintering temperature.     

Internal pancreatic fistula: surgical treatment

The authors present 13 cases of internal pancreatic fistula, of which 11 were secondary to a chronic pancreatitis and two were caused by an abdominal trauma. Beside the clinical picture, the diagnosis was anticipated by the high amylase levels present in the fluid obtained by paracentesis or thoracocentesis. The diagnosis was confirmed by the radiological analysis of the pancreatic duct system, when an endoscopic retrograde pancreatography was performed in seven patients, one pancreatography was carried out during surgery in five cases, and one patient underwent an injection of hydrosoluble contrast in the pleural cavity. The treatment was a latero-lateral pancreaticojejunoanastomosis in five cases, associated with a corpora-caudal pancreatectomy in four patients; a cephalic duodenopancreatectomy was performed in one case. Two patients underwent a cystoenteroanastomosis, while the option chosen in the last four cases was an external drainage. One patient refused to undergo surgical treatment. Operation mortality was null. The conclusion was that an adequate surgical treatment results in the occlusion of the internal pancreatic fistula and, furthermore, allows for the definitive resolution of underlying pancreatic affection.