Tolerance induction ameliorates allograft vasculopathy in rat aortic transplants. Influence of Fas-mediated apoptosis

Based on successful induction of donor-specific unresponsiveness by alloantigenic stimulation in several animal models of acute rejection, we hypothesized that similar immune manipulations would also inhibit the evolution of chronic rejection and transplant vasculopathy. To induce immune tolerance, DA rats received a PVG heart allograft and were immunosuppressed with cyclosporine for 30 d. At day 100 the animals were challenged with a PVG aortic allograft after either 1 or 18 h of cold ischemia. 8 wk after the aortic transplantation, the grafts were investigated for morphological changes, infiltrating cells, apoptosis, and Fas-Fas ligand expression. Control allografts showed advanced transplant arteriosclerosis, whereas tolerance-induced aortic allografts displayed reduced neointimal formation, less medial atrophy, fewer apoptotic cells, and fewer Fas- and FasL-expressing cells. Prolonged ischemic storage time did not profoundly alter the morphological changes of the allografts. Fas expression was found in T cells, macrophages, vascular smooth muscle cells, and endothelial cells, whereas FasL was expressed mainly by T cells and macrophages. FasL mRNA expression was evident throughout the entire allograft wall. In conclusion, induction of allospecific tolerance can effectively prevent transplant arteriosclerosis. Cold ischemia damage does not abrogate the beneficial effect of tolerance, but creates a separate identity of mainly endothelial lesions. Furthermore, Fas-mediated apoptosis appears to be involved in the pathological lesions seen in chronic rejection.     

Rare causes of occult small intestinal bleeding, including aortoenteric fistulas, small bowel tumors, and small bowel ulcers

The most common cause for gastrointestinal bleeding of small bowel origin is angiodysplasia, followed by tumors of the small intestine, and various other causes, including small bowel ulcers and aortienteric fistulas. With the advent of improved diagnostic tests, including push and sonde enteroscopy, timely endoscopic diagnosis of these rare small bowel lesions has become possible, enabling the clinician to make better therapeutic decisions. This article focuses on the rare small bowel sources of intermittent and chronic gastrointestinal blood loss.     

Small bowel rejection in isolated small bowel transplantation and in multivisceral transplantation: a comparative study in a large animal model

31P NMR lineshapes of multilamellar liposomes composed mostly of a bilayer-spanning tetraether lipid are consistent with rapid axially symmetric motion about the bilayer normal. The residual chemical shift anisotropy of 36 ppm is comparable to that seen for diacylphosphatidylglycerol systems and suggests comparable headgroup motion. The lateral diffusion rates for Thermoplasma acidophilum total polar lipids in mutilamellar liposomes was measured by two dimensional exchange NMR as a function of temperature. At 55 degrees C, near the growth temperature, the rate of lateral diffusion, DL, is comparable to that of diester phospholipids in the Lalpha liquid crystalline phase, having a value of 2 x 10(-8) cm2/s. DL decreases with temperature reaching a value of 8-6 x 10(-9) cm2/s at 30 degrees C. The activation energy Ea for lateral diffusion is estimated to be 10 kcal/mol (approximately 42 kJ/mol). The lateral diffusion rates indicate that the tetraether liposomes have a membrane viscosity at 30 degrees C which is considerably higher than that of diester phospholipids in the liquid crystalline phase.     

Propulsion and mixing of small bowel contents after laparotomy in rat

Radioactive test substances were infused slowly into the duodenum of conscious rats via a permanent catheter starting 2, 12 and 24 h after a standardized laparotomy. Two differently labelled but otherwise identical test substances were used. The first test substance (125I-PVP) was infused for 4 h, the second (131I-PVP) for the remaining 1 h of the 5-hour infusion period. Immediately after the infusion the animals were killed, and the radioactivity emanating from each isotope was recorded from the excised bowel specimen. The bowel length passed by the border zone and the degree of overlap between the labels in this zone were taken as measures of propulsion and mixing, respectively. Propulsion and mixing were uninfluenced by laparotomy as measured 2--17, 12--17 and 24--29 h after laparotomy. The present findings indicate that laparotomy is not followed by a disturbance in the capability of the small bowel to transport and mix chyme, at least when no high demands with respect to chyme volume are required.     

Histology, immunohistochemistry, and electron microscopy of small round cell tumors of bone

Small round cell tumors (SRCTs) of the bone make up a family of primary bone sarcomas with morphologically, biologically, and clinically specific features. Among them, Ewing's sarcoma (ES) is the most common entity, but several varieties such as atypical ES, large cell ES, and ES with neuroectodermal differentiation (peripheral primitive neuroectodermal tumor of the bone or neuroepithelioma of the bone) have been identified recently. Histology and electron microscopy together with the variable expression of several epitopes (as shown by immunohistochemistry, mainly HBA/71 [Mic2 antigen]) provide the basis for characterizing the group within the context of neuroectodermal-derived neoplasms. A number of other ES-like tumors with small round cells, mimicking those previously described, have been characterized; Askin's tumor of the thoracopulmonary region will be considered as an ES similar to those already described, but within a particularly anatomic location. On the other hand, the presence of an endothelial appearance within a poorly differentiated neoplasm may be present in some ES-like SRCTs (atypical ES with endothelial features). The differential diagnosis with other sarcomas defined by small round to spindle cell contours might prove difficult. Particular attention must be paid to small cell osteosarcoma and mesenchymal chondrosarcoma. Likewise, "primitive sarcoma of bone" is considered in this study because it is a very rare neoplasm differing from the formerly discussed types; its pluripotentiality provides this tumor a blastemic character and a multiphenotypic expression. Malignant non-Hodgkin's lymphoma is an unusual presentation when primary to the bone, previous to any other anatomic location. Several subtypes have been considered within a histology that encompasses that seen in lymph nodes.     

Retardation of human drug-resistant HL-60 cell in G1 phase and induction of sensitive cell to apoptosis by cyclosporine A

To further study the relationship between resistance to apoptosis and drug resistance in harringtonine-resistant HL-60 cells (HR20), cyclosporine A (CsA) 20, 10 micrograms.ml-1 was shown to induce the sensitive HL-60 cells to apoptosis, showing a typical DNA "ladder" band. But the same concentrations of CsA retarded the HR20 cells in G1 phase and could not induce the cells to apoptosis. The cellular daunorubicin accumulation increased when HR20 cells were treated with low concentration of CsA and the reversal of drug resistance by CsA was unrelated to the retardation of cell cycle progression. High phosphorylation of about 50 kDa protein occured when HR20 cells were treated with CsA 10 micrograms.ml-1. The results domonstrate that cyclosporine A retarded the harringtonine-resistant HL-60 cells in G1 phase but induced HL-60 cells to apoptosis, and the retardation was unrelated to drug resistance.     

Clinicopathologic features of neuroendocrine carcinomas of the stomach: appraisal of small cell and large cell variants

OBJECTIVE: To clarify the clinicopathologic features of neuroendocrine carcinomas (NECs) of the stomach, we reviewed 56 cases of surgically treated gastric carcinomas with a solid growth or with areas containing patterns characteristic for neuroendocrine (NE) tumors. METHODS: Of the 56 cases reviewed, we selected 33 cases of NEC based both on histologic patterns common to NE tumors and on histochemical/immunohistochemical results. RESULTS: The average age of patients with NEC was 69.8 years (range, 44-92 years). The majority of patients were men (male-female ratio, 23:10). The most frequently affected site was the upper third (46%) of the stomach. Grossly, 9 cases of NEC were fungating and 24 were ulcerated. As compared with 23 patients with non-NEC tumors, the patients with NECs had a worse prognosis. Histologically, NECs had a variety of histologic patterns, including solid, organoid, trabecular, pseudoglandular, spindle cell, and rosettelike. Based on both cell size and morphologic features, we subdivided NECs into 2 variants, namely, small cell NEC and large cell NEC. Our series included 12 cases of small cell NEC and 21 cases of large cell NEC. CONCLUSIONS: Compared with small cell NECs, large cell NECs had a higher mitotic count, larger polygonal cells, a lower nuclear-cytoplasmic ratio, coarser nuclear chromatin, and more frequent conspicuous nucleoli. Large cell NEC was an aggressive tumor with a very poor prognosis (median survival time, 15.2 months; 1-year survival rate, 58%), which approached that for small cell NEC.     

99Tcm-MAG3: a sensitive indicator for evaluating perfusion and rejection of renal transplants

Radionuclide renography has a role in evaluating perfusion of transplanted kidneys. In the course of rejection, cortical perfusion decreases before urinary excretion changes. Based on the facts that 99Tcm-MAG3 has different pharmacokinetics and shows a higher kidney-to-background count ratio than 99Tcm-DTPA, we postulated that 99Tcm-MAG3 was a sensitive and reproducible agent to measure cortical perfusion of transplanted kidneys. To clarify the feasibility of using 99Tcm-MAG3 to measure the cortical perfusion index (CPI), sequential renography was performed using 99Tcm-DTPA and 99Tcm-MAG3 in 14 patients with stable renal transplants, who had changes in serum creatinine concentration of less than 50% between the two studies. The CPI was calculated with 99Tcm-DTPA and 99Tcm-MAG3 and these were then compared and correlated with concurrent serum creatinine concentration. The CPI with 99Tcm-MAG3 was 1.43 times that with 99Tcm-DTPA in patients with changes in serum creatinine concentration equal to or less than 20%, and regression analysis revealed that the difference in CPI was larger in patients with more severely decreased renal perfusion than in patients with normal or mildly decreased renal perfusion. This preliminary study has indicated that the CPI with 99Tcm-MAG3 is a sensitive index for detecting changes in renal function, and thus is a feasible indicator of cortical perfusion when evaluating the rejection of transplanted kidneys.     

Laminin localization in enterocytic basement membrane of rat small bowel grafts. A light and electron microscopic study

In vitro laminins stimulate numerous biological effects, such as cell migration, proliferation, attachment and differentiation. In vitro laminins influence immunocompetent cells and in vivo possibly play an important role in graft rejection. To establish how laminins could be involved in the regulation of acute rejection of small bowel allografts (with and without immunosuppression), we investigated laminin distribution in rat small bowel allografts four days after transplantation, i.e., before the onset of histological signs of rejection, using antibodies against alpha1, beta1, gamma1 chain of laminin-1. In immunosuppressed allografts, the ultrastructure of the enterocytic basement membrane appeared normal, but no laminin staining was seen in this membrane, although basement membranes of intramural blood vessels and muscle cells were normally stained. In non-operated immunosuppressed rats, laminin staining was clearly reduced in the enterocytic basement membrane, demonstrating that cyclosporin A is able to affect this membrane. Since only rats in which laminin is altered survive, this laminin alteration in the enterocytic basement membrane presumably plays an important role in overcoming the acute rejection.     

Eccentric polyposis of the small bowel. A possible radiologic sign of plexiform neurofibromatosis of the small bowel and its mesentery

Eccentric polyposis was demonstrated on the small bowel series in a patient with Von Recklinghausen's disease. This proved to represent plexiform neurofibromatosis of the small bowel and its mesentery. This unique radiographic appearance may be a radiologic sign of plexiform neurofibromatosis.     

Induction of Fas-mediated apoptosis in a human renal epithelial cell line by interferon-gamma: involvement of Fas-mediated apoptosis in acute renal rejection

To examine the role of the Fas ligand/Fas (FasL/Fas) system and apoptosis in renal allograft rejection, we analyzed FasL/Fas expression and apoptosis in 63 renal allograft biopsy specimens obtained from 56 renal transplant recipients in Tokyo, Japan. Sixteen biopsy specimens were diagnosed (Banff criteria) as acute rejection (AR), 17 as AR plus chronic rejection, 10 as borderline stages, and 20 as no rejection (NR). Immunohistochemically, Fas antigen was highly expressed in the renal tubular epithelium in tissues from patients with rejection. The mean number of Fas-positive tubular epithelium was significantly higher in biopsy specimens with AR than in those with NR, but FasL expression was highly expressed in infiltrating lymphocytes in the interstitium of allografts with cellular rejection. The mean number of FasL-positive infiltrating lymphocytes was significantly higher in specimens with AR than in those with NR or borderline stages. For detection of apoptotic cells, the specimens were subjected to terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling, which showed that the mean number of tubular epithelial cells positive for this labeling was highest for the specimens with AR, the number being significantly higher than in those with AR plus chronic rejection or with NR. Thus, Fas expression on epithelial cells might actively trigger their apoptosis by the interaction between Fas and FasL. Studies of human normal renal-derived cell lines (RPTEC 2601 [epithelial] and NHMC 5155 [mesangial]) showed that both constitutively expressed Fas (but not FasL) mRNA. After pretreatment with interferon-gamma, Fas-induced apoptosis was observed in the RPTEC 2601 line, but without interferon-gamma pretreatment, anti-Fas-mediated apoptosis was not seen. Under identical conditions, the NHMC 5155 line was resistant to anti-Fas-mediated apoptosis regardless of interferon-gamma pretreatment. This suggested that AR might be associated with increased apoptosis in the renal tubular epithelial cells mediated by the Fas/FasL system.     

NK/T细胞淋巴瘤细胞凋亡和细胞增殖特征及意义

Objective:The aim was to study the features and clinical significance of cell apoptosis and proliferation of NK/T cell lymphoma. Methods:TdT-mediated dUTP nick end labeling and immunohistochemical Streptavidin-peroxidase method were used to study cell apoptosis and the expression of proliferation cell nuclear antigen in 25 NK/T cell lymphoma and 10reactive lymphoid tissues. Results:Apoptotic index (AI) and proliferative index (PI) averaged (1.92% ± 0.86%) and (41.48%± 5.10%) respectively in the 25 NK/T cell lymphomas and (6.70% ± 1.89%) and (20.10% ± 2.77%) in the 10 reactive lymphoid tissues. Compared with reactive lymphoid tissues, AI was significantly reduced in NK/T cell lymphoma (t = 10.80, P < 0.01)while PI significantly increased (t = 12.39, P < 0.01). In addition, in NK/T cell lymphoma, AI and PI were positively related (r = 0.69, P < 0.01). Conclusion:In NK/T cell lymphoma, cell apoptosis is reduced while cell proliferation increased. The imbalance between cell apoptosis and cell proliferation is closely related to the development and progression of NK/T cell lymphoma.     

Foetal rat pancreatic transplantation: posttransplantation development of foetal pancreatic iso- and allografts and suppression of rejection with mycophenolate mofetil (MMF) and cyclosporine based immunesuppression

CONTEXT: High-dose iodine 131 is the treatment of choice in the United States for most adults with hyperthyroid disease. Although there is little evidence to link therapeutic (131)I to the development of cancer, its extensive medical use indicates the need for additional evaluation. OBJECTIVE: To evaluate cancer mortality among hyperthyroid patients, particularly after (131)I treatment. DESIGN: A retrospective cohort study. SETTING: Twenty-five clinics in the United States and 1 clinic in England. PATIENTS: A total of 35 593 hyperthyroid patients treated between 1946 and 1964 in the original Cooperative Thyrotoxicosis Therapy Follow-up Study; 91 % had Graves disease, 79% were female, and 65% were treated with (131)I. MAIN OUTCOME MEASURE: Standardized cancer mortality ratios (SMRs) after 3 treatment modalities for hyperthyroidism. RESULTS: Of the study cohort, 50.5% had died by the end of follow-up in December 1990. The total number of cancer deaths was close to that expected based on mortality rates in the general population (2950 vs 2857.6), but there was a small excess of mortality from cancers of the lung, breast, kidney, and thyroid, and a deficit of deaths from cancers of the uterus and the prostate gland. Patients with toxic nodular goiter had an SMR of 1.16 (95% confidence interval [CI], 1.03-1.30). More than 1 year after treatment, an increased risk of cancer mortality was seen among patients treated exclusively with antithyroid drugs (SMR, 1.31; 95% CI, 1.06-1.60). Radioactive iodine was not linked to total cancer deaths (SMR, 1.02; 95% CI, 0.98-1.07) or to any specific cancer with the exception of thyroid cancer (SMR, 3.94; 95% CI, 2.52-5.86). CONCLUSIONS: Neither hyperthyroidism nor (131)I treatment resulted in a significantly increased risk of total cancer mortality. While there was an elevated risk of thyroid cancer mortality following (131)I treatment, in absolute terms the excess number of deaths was small, and the underlying thyroid disease appeared to play a role. Overall, (131)I appears to be a safe therapy for hyperthyroidism.     

Apoptosis and necrosis: mechanisms of cell death induced by cyclosporine A in a renal proximal tubular cell line

BACKGROUND: The mechanisms of cyclosporine (CsA)-induced nephrotoxicity are not fully understood. While hemodynamic changes may be involved in vivo, there is also some evidence for tubular involvement. We previously showed direct toxicity of CsA in the LLC-PK1 renal tubular cell line. In the current study we examined mechanisms (apoptosis or necrosis) of cell death induced by CsA in the LLC-PK1 renal proximal tubular cell line. The possible role of the Fas (APO-1/CD95) antigen-Fas ligand system in the mediation of CsA-induced cell death was also investigated. METHODS: Cells were treated with CsA (0.42 nM to 83 microM) for 24 hours and alterations in DNA and protein synthesis and membrane integrity were examined. Flow cytometry was used to investigate: (i) alterations in the DNA content and cell cycle; (ii) the forward (FSC) and side (SSC) light scattering properties (indicators of cell size and granularity, respectively); (iii) the externalization of phosphatidylserine (PS) as a marker of early apoptosis using FITC-annexin V binding; and (iv) expression of the apoptotic Fas protein. DNA fragmentation in apoptotic cells was also determined by the TUNEL assay. RESULTS: CsA (all doses) caused a block in the G0/G1 phase of the cell cycle as indicated by a decrease in DNA synthesis and supported by an increase in the % of cells in the G0/G1 phase with concurrent decreases of those in the S and G2/M phases. The effect on protein synthesis appeared to be much less. Lower doses of CsA (4.2 nM) caused the appearance of a "sub-G0/G1" peak, indicative of reduced DNA content, on the DNA histogram that was paralleled by a reduction in cell size and an increased cell granularity and an increase in FITC-annexin V binding. DNA fragmentation was evident in these cells as assessed using the TUNEL assay. Higher doses of CsA increased cell size and decreased cell granularity and reduced membrane integrity. Expression of Fas, the cell surface molecule that stimulates apoptosis, was increased following low dose CsA exposure. CONCLUSIONS: These results indicate that CsA is directly toxic to LLC-PK1 cells with reduced DNA synthesis and cell cycle blockade. The mode of cell death, namely apoptosis or necrosis, is dose dependent. Fas may be an important mediator of CsA induced apoptosis in renal proximal tubular cells.