Fat necrosis in the breast: a multimodality imaging review of its natural course with different aetiologies.

Fat necrosis of the breast is a commonly encountered condition in daily practice. It is a benign pathology, but it can have variable manifestations and patterns that may sometimes mimic malignancy, depending on its stage of evolution and its underlying cause. This review demonstrates the wide spectrum of appearances of fat necrosis on mammography, digital breast tomosynthesis (DBT), ultrasound, magnetic resonance imaging (MRI), computed tomography (CT), and positron-emission tomography (PET). Sequential follow-up images are included in some cases to illustrate the temporal change of the findings. The typical location and distribution of fat necrosis from a comprehensive list of aetiologies are discussed. Improved knowledge of the multimodality imaging features of fat necrosis could enhance diagnostic accuracy and clinical management, thus avoiding unnecessary invasive investigations.

Evaluation of performance of measurement of faecal α(1)-antitrypsin clearance and technetium-99m human serum albumin scintigraphy in protein-losing enteropathy.

BACKGROUND AND AIM: Our study aimed to compare the performance of faecal α(1)-antitrypsin clearance (AATC) and radiolabelled human serum albumin (HSA) scintigraphy in protein-losing enteropathy (PLE). METHODS: Patients studied by both AATC and technetium-99m ((99m)Tc)-labelled HSA scintigraphy were recruited and categorized into PLE and non-PLE groups based on clinical and laboratory findings. The performance of AATC and (99m)Tc-labelled HSA scintigraphy was evaluated using clinical diagnosis of PLE as a gold standard. RESULTS: 29 patients were recruited and 13 patients were considered to have definite PLE (PLE group). In the PLE group, all patients had a positive HSA scinigraphy and 10 (77%) had demonstrable positive tracing in the early phase. Conversely, only 6 of them (46%) had elevated AATC level (>13 m/day). Results of (99m)Tc-labelled HSA scan (but not AATC) showed significant agreement with the clinical diagnosis (κ 0.35, p = 0.013). (99m)Tc-labelled HSA scintigraphy carried higher sensitivity (100 vs. 46%) and negative predictive value (100 vs. 63%) compared to AATC in diagnosing PLE. The correlation between the results of these two investigations was only modest (κ 0.27, p = 0.04). The area under the receiver operating characteristic curve of AATC level showed no optimal diagnostic cut-off for PLE. CONCLUSION: (99m)Tc-labelled HSA scintigraphy was superior to AATC in diagnosing PLE.

Gene expression of the renin-angiotensin system in human kidney.

BACKGROUND: Immunocytochemical studies have revealed that all components of the renin-angiotensin system are widely distributed in human tissues yet the information on the gene expression of the renin-angiotensin system in various types of cell remains scarce. OBJECTIVE: We explored the presence of a local renin-angiotensin system in human kidney. METHODS: We sought to determine the presence of messenger RNA (mRNA) encoding for renin, angiotensinogen, and angiotensin converting enzyme (ACE) in cultured human glomerular cells and human umbilical vein endothelial cells using a two-step polymerase chain reaction. The gene expression of the renin-angiotensin system in normal human kidney and in diseased kidney was studied by in-situ hybridization using synthetic oligonucleotides. RESULTS: By using a two-step polymerase chain reaction, renin, angiotensinogen, and ACE mRNA were found in cultured mesangial and epithelial cells but only ACE mRNA was present in human umbilical vein endothelial cells. Renin mRNA was detected in juxtaglomerular granular cells and also in glomerular and tubular epithelia in normal kidney by in-situ hybridization. A similar tubular, but not mesangial, distribution was found with angiotensinogen and ACE mRNA. In contrast, stronger signals for renin, angiotensinogen and ACE mRNA were detected in mesangial and epithelial cells of kidney tissues from hypertensive patients and from patients with renal pathology characterized by mesangial proliferation (immunoglobulin A nephropathy, diabetes mellitus, or lupus nephritis). CONCLUSIONS: That gene expression of the renin-angiotensin system occurs in resident glomerular cells supports the hypothesis that there is a local renin-angiotensin system in human kidney. Our findings support the previous speculation that the renin-angiotensin system could be a local factor involved in the progression of chronic renal failure and consequent development of hypertension.

Changes of cytokine profiles during peritonitis in patients on continuous ambulatory peritoneal dialysis.

Continuous ambulatory peritoneal dialysis (CAPD) has emerged as an important dialysis treatment modality worldwide. One of the major complications is bacterial peritonitis, which may result in subsequent technique failure because of loss of peritoneal clearance or peritoneal fibrosis. Bacterial peritonitis leads to the release of proinflammatory cytokines from resident and infiltrating cells in the peritoneal cavity. We studied 35 patients undergoing CAPD with acute bacterial peritonitis. All patients treated with antibiotics for 2 weeks after the clinical diagnosis of peritonitis had a good recovery. Peritoneal dialysate effluent (PDE) was collected on days 1, 3, 5, 10, 21, and 42 after the start of treatment. Cell populations were monitored by flow cytometry. PDE levels of interleukin-1beta (IL-1), IL-6, transforming growth factor-beta (TGF-beta), and basic fibroblast growth factor (FGF) were measured by enzyme-linked immunosorbent assay. Gene transcription of TGF-beta in macrophages from PDE was measured by quantitative polymerase chain reaction. Bacterial peritonitis was associated with a sharp increase in total cell and neutrophil counts (400-fold) in PDE up to 3 weeks after peritonitis despite clinical remission (P < 0.0001). There was an increased absolute number of macrophages during the first 3 weeks despite the reduced percentage of macrophages among total cells in PDE compared with noninfective PDE. There was a progressive increase in the percentage of mesothelial cells or dead cells in the total cell population in PDE over the entire 6-week period. PDE levels of IL-1, IL-6, TGF-beta, and FGF increased markedly on day 1 before their levels decreased gradually. PDE levels of these cytokines or growth factors were significantly greater than those in noninfective PDE (n = 76) throughout the study period (P < 0.01). Similarly, TGF-beta complementary DNA (cDNA) molecules per macrophage were significantly greater than those of macrophages in noninfective PDE throughout this period (P < 0.01). There was no significant correlation between PDE levels of TGF-beta and TGF-beta cDNA molecules per macrophage, suggesting that peritoneal macrophages are not the only source of TGF-beta in PDE. We conclude there is an active release of proinflammatory cytokines and sclerogenic growth factors through at least 6 weeks despite apparent clinical remission of peritonitis. The peritoneal cytokine networks after peritonitis may potentially affect the physiological properties of the peritoneal membrane.

Dialysate hyaluronan concentration predicts survival but not peritoneal sclerosis in continuous ambulatory peritoneal dialysis.

Hyaluronan is an important component of extracellular matrix and plays a critical role in early phases of wound healing. Peritoneal mesothelium is a major site of hyaluronan production. Serum hyaluronan concentration has been shown to predict survival in maintenance hemodialysis patients. We hypothesize that mesothelial production of hyaluronan during the stable phase of continuous ambulatory peritoneal dialysis (CAPD) predicts the risk of peritoneal adhesion and mortality. We studied peritoneal dialysate effluent (PDE) hyaluronan levels from 116 stable CAPD patients. They were then followed-up for 3 years. During the follow-up period, there were 196 episodes of peritonitis in 78 patients. Tenckhoff catheter was removed in 31 episodes (15.8%). Tenckhoff catheter was reinserted successfully in 12 cases, and CAPD was resumed. Peritoneal adhesion developed in 16 cases. Three patients died before Tenckhoff catheter reinsertion was attempted. There was no difference in stable-phase PDE hyaluronan levels between patients who developed peritoneal adhesion and those who did not (159 +/- 63 versus 227 +/- 194 microgram/L, P = 0.27). Thirty-three patients died during the study period. Patients who died had significantly higher PDE hyaluronan concentration than survivors (272 +/- 194 versus 170 +/- 105 microgram/L, P < 0.01). Univariate analysis showed that increased PDE hyaluronan level was associated with a shorter patient survival (P < 0.001). There was no association between PDE hyaluronan level and serum albumin, protein nitrogen appearance, and percentage of lean body mass. Multivariate analysis confirmed that PDE hyaluronan level, serum albumin, and diabetic state were independent predictors of survival. We conclude that PDE hyaluronan level during stable phase of CAPD does not predict the risk of postperitonitis adhesion. However, it is a strong independent predictor of survival in CAPD patients.

Increased production of hyaluronan by peritoneal cells and its significance in patients on CAPD.

Hyaluronan (HA) is a polysaccharide that forms a critical component of extracellular matrices. HA is present in high concentrations in tissues undergoing remodeling and morphogenesis, and it appears to have an important role in the early stages of wound healing. Here, we studied the level of HA in the peritoneal dialysate effluent (PDE) from 116 stable continuous ambulatory peritoneal dialysis (CAPD) patients. Longitudinal studies over a period of 6 weeks were performed in seven of these patients who developed peritonitis. The median HA level in PDE from these patients was 154.6 microg/L (range, 29.7 to 820.2 microg/L). Dialysate level of HA increased with age of the patients, but no such correlation was shown between HA level in PDE and duration of CAPD treatment or previous episodes of peritonitis. Patients with high or average peritoneal membrane transport of small solutes had a higher HA level in the PDE than those with a low peritoneal membrane transport (P = 0.046). A significant correlation was observed between PDE level of HA and interleukin-1beta (IL-1beta) or IL-6. The plasma level of HA in these patients was significantly greater than that of healthy controls (P < 0.0001), yet the plasma concentration of HA was only 85% that of the PDE concentration. In CAPD patients with peritonitis, there was a sharp increase in the PDE levels of HA, IL-1beta, and IL-6. These values decreased progressively with resolution of peritonitis. The changes in the PDE levels of HA closely followed those of IL-1beta or IL-6. In vitro [3H]-glucosamine incorporation studies suggest that the main bulk of HA is derived from synthesis by the peritoneal mesothelial cells, whereas the amount synthesized by macrophages is trivial. We conclude that elevated levels of HA found in the PDE of stable CAPD patients originate from increased synthesis by peritoneal mesothelial cells. This event may serve as a marker of regeneration and remodeling of the peritoneal lining.

Longitudinal study of peritoneal membrane function in continuous ambulatory peritoneal dialysis: relationship with peritonitis and fibrosing factors.

BACKGROUND: The peritoneal equilibration test (PET) is a useful assessment of peritoneal function in continuous ambulatory peritoneal dialysis (CAPD) patients. However, the natural course of longitudinal change in peritoneal transport is not well defined. PATIENTS: We studied 105 unselected CAPD patients. Average age at enrollment was 50.7 +/- 11.3 years. METHODS: A PET was performed at enrollment. Peritoneal transport was expressed as dialysate-to-plasma creatinine ratio at 4 hours (DIP). Fibrosing factors and mesothelial cell markers, including TGFbeta, epidermal growth factor (EGF), platelet-derived growth factor (PDGF), hyaluronan, and cancer antigen 125 (CA125), were measured in overnight peritoneal dialysate effluent (PDE). Patients were followed for two years. Peritonitis episodes were recorded. Severe peritonitis was defined as an episode that required catheter removal or antibiotic therapy for more than 3 weeks. After two years, 75 patients were still alive and on CAPD. RESULTS: The PET was repeated in 64 patients, of whom 35 were male and 9 had diabetes. The change in D/P over two years was represented as AD/P. No significant change in peritoneal transport was seen after two years (D/P: 0.56 +/- 0.12 vs 0.55 +/- 0.13). A centripetal pattern of change in D/P was observed. The deltaD/P had normal distribution and was inversely correlated with D/P at baseline (r = -0.427, p < 0.005). Both results suggest a regression-to-mean phenomenon. The deltaD/P had no significant correlation with the total number of peritonitis episodes (Spearman r = 0.052, p = 0.74), but after severe peritonitis, affected patients had higher deltaD/P than patients who experienced no severe infection (0.040 +/- 0.136 vs -0.032 +/- 0.120, p < 0.05). For patients with no episodes of severe peritonitis (n = 47), deltaD/P was weakly correlated with baseline TGFbeta level (r = -0.506, p < 0.01). No correlation was seen between the levels of other fibrosing factors and change in peritoneal transport. CONCLUSIONS: Our findings suggest that the centripetal change of peritoneal transport probably reflects a regression-to-mean phenomenon. Peritoneal transport increases after severe peritonitis. The role of TGFbeta levels in PDE with regard to longitudinal change in peritoneal transport requires further study.

Urinary mRNA expression of ACE and ACE2 in human type 2 diabetic nephropathy.

AIMS/HYPOTHESIS: The interplay of ACE and type 2 ACE (ACE2) has been recognised as playing an important role in the tissue renin-angiotensin system within the kidney. In the present study, we measured urinary mRNA expression of ACE and ACE2 in patients with type 2 diabetic nephropathy. METHODS: We studied 50 patients with diabetic nephropathy: 26 were being treated by ACE inhibitor (ACEI) alone (ACEI group), the other 24 by ACEI and angiotensin-receptor blocker (ARB) (ACEI+ARB group). mRNA expression of ACE and ACE2 was measured by real-time quantitative RT-PCR at 0 and 12 weeks. All patients were then followed for 56 weeks. RESULTS: Proteinuria correlated significantly with urinary ACE (r=0.454, p=0.001) and ACE2 expression (r=0.651, p<0.001). Urinary ACE2 expression correlated with estimated GFR (r= -0.289, p=0.042). In the ACEI group, there was a significant inverse correlation between the rate of GFR decline and urinary ACE2 expression at baseline (r= -0.423, p=0.031) as well as at 12 weeks (r= -0.395, p=0.046). In contrast, there was no significant correlation between the rate of GFR decline and urinary ACE2 expression at baseline or at 12 weeks in the ACEI+ARB group. The rate of GFR decline did not correlate with the baseline urinary ACE expression of either group. CONCLUSION/INTERPRETATION: There was a relationship between urinary mRNA expression of ACE2 and the degree of proteinuria. The physiological implication and possibility of clinical application of quantifying urinary ACE2 expression require further study.

Effect of anti-DNA autoantibodies on the gene expression of interleukin 8, transforming growth factor-beta, and nitric oxide synthase in cultured endothelial cells.

Our study aims to determine whether anti-dsDNA exerts any effect on the gene expression of IL-8 or TGF-beta in cultured HUVEC. Both cytokines have angiogenic effect on endothelial cells. IgG was purified from 19 patients with SLE and from 19 healthy controls. Anti-dsDNA-depleted polyclonal IgG was also prepared from serum IgG of lupus patients by affinity chromatography with DNA cellulose column. Compared with either control IgG or anti-dsDNA-dep-IgG, HUVEC incubated with anti-dsDNA-containing-IgG expressed higher levels of IL-8 mRNA (p = 0.0001) and TGF-beta 1 mRNA (p = 0.0014). We demonstrated a significant increase in the percentage of cells with fragmented DNA in HUVEC incubated with anti-dsDNA-containing-IgG compared with those incubated with anti-dsDNA-dep-IgG, supporting the notion that anti-dsDNA may exert a direct apoptotic effect on cultured endothelial cells. Our study provides in vitro evidence that anti-dsDNA could play an important pathogenetic role in inducing inflammatory injury of vascular endothelium in SLE.

[Anesthetic considerations in complete obstruction of major veins of upper trunk--report of two cases].

This article presents two cases about complete obstruction of major veins of upper trunk. One was a 45 year-old woman who suffered from complete obstruction of superior vena cava due to bronchogenic carcinoma. Venous bypass grafting from left innominate vein to right atrium was performed. The other was a 41 year-old man. Both of his right and left, internal and external jugular veins were completely obstructed after radical laryngectomy and radiotherapy due to hypopharyngeal carcinoma. Venous bypass grafting from cerebral transverse sinus to right atrium was performed. In both cases, general anesthesia with neurosurgical anesthetic concepts were performed with continuous blood pressure monitoring, CVP monitoring, pulse oximetry, end-tidal CO2 monitoring, blood gas analysis, urine output and EKG monitoring. The outcome was satisfactory in both cases.

Increased ascitic level of hyaluronan in liver cirrhosis.

Hyaluronan (HA) is a polysaccharide that forms a critical component of extracellular matrixes. It is present in high concentrations in tissues undergoing remodeling and morphogenesis. Serum HA is elevated in patients with chronic liver disease, and this has been considered to be caused by impaired degradation by the liver endothelial cells. We studied the level of HA in the ascitic fluid and plasma from 27 patients with cirrhotic ascites. These values were compared with peritoneal dialysate effluent (PDE) and plasma from 33 patients with uremia who were undergoing continuous ambulatory peritoneal dialysis (CAPD). The median HA levels in ascitic fluid and plasma from our 26 patients with cirrhosis were significantly higher than corresponding PDE and plasma values from the 33 CAPD patients (p < 0.0001). The median peritoneal/plasma ratios of creatinine, albumin, and immunoglobulin G in either cirrhotic or CAPD patients were less than unity. In contrast, the median peritoneal/plasma ratios of HA in both groups of patients exceeded one with a higher peritoneal/plasma ratio of HA in patients with cirrhosis (p = 0.0035). A significant correlation was observed between the ascitic level of HA and interleukin-1beta, interleukin-6, or transforming growth factor-beta. Our in vitro cell culture studies revealed that HA is synthesized by both mesothelial cells and macrophages. We observed an additive effect in the synthesis of HA by mesothelial cells when the macrophage-conditioned medium was added to the RPMI culture medium. We conclude that a high level of HA is found in ascites from patients with cirrhosis. Our results strongly suggest that simultaneous increased synthesis of HA by the peritoneal cells and a reduction of degradation by liver endothelial cells occur in these patients with cirrhosis with ascites. This event of increased HA synthesis may be contributory to remodeling and regeneration of the peritoneal lining.

[Surgical and anesthetic mortality in Mackay Memorial Hospital 1988-1989].

The mortality associated with 52128 anesthetics administered over two years (1988-1989) at Mackay Memorial Hospital, Taipei, was reported. The frequency of death to which anesthesia contributed was 0.4/10000 (2 cases in 52128 anesthetics). The total mortality rate from surgery within one week was 0.2% (105 cases in 52128 anesthetics). Anesthetic deaths were responsible for 1.9% of the total mortality. There were 25 cases of cardiac arrest with 18 fatal cases (7 cases were recovered after cardiopulmonary resuscitation). The causes of cardiac arrest during anesthesia included anesthetic factors (24%), surgical factors (24%) and patients' pathological factors (52%).

Is routine preanesthetic hemoglobin test necessary in minor pediatric surgery?

The value of preanesthetic assessment of anemia and analysis of the hemoglobin level prior to a minor pediatric surgery has been recently questioned in some reports. This study was to retrospectively analyse 8859 pediatric patients who underwent minor surgery in the period from January 1987 to December 1990 in our hospital. They were all ASA class I-II in physical status with age ranging from one month to 19 years. Those patients with their hemoglobin values determined at other laboratories or hospitals in spite of our recognition and those suspected of having an immune or oncologic disease were excluded from this study. The mean hemoglobin value of the patients under study was 12.99 +/- 0.82 g/dl. 0.62% of the patients (55) were found to have hemoglobin values less than 10 g/dl which were similar to the results obtained by Wood et al (0.7%) in 1981 and Roy et al (0.5%) in 1990. Among the 55 anemic patients, 41 (74.5%) were at the age between 2 to 4 months (within the physiologic anemic period of infancy). Sampling of blood for routine preanesthetic hemoglobin determination which caused discomfort and pain was often rejected by pediatric patients and struggle for escape also upset the children very much. Based on the results from our analysis, we suggest that in healthy pediatric patient scheduled for minor surgery routine hemoglobin test could be excluded. Hemoglobin test is selectively performed in a patient is anemic or under suspicious circumstances. The value and shortcomings of selective hemoglobin test before surgery require further evaluation.

[Pulmonary lavage for alveolar proteinosis. A case report].

Pulmonary alveolar proteinosis is characterized by an amorphous lipoproteinaceous material filling the acini of the lung, resulting in hypoxemia. At present, the most effective treatment for alveolar proteinosis is lung lavage. Here, we report the first case in Taiwan treated with this method. A 38-year-old male with a 6-month history of cough and increasing breathlessness was transferred from another hospital. On admission, his arterial oxygen tension was 51 mmHg when breathing room air. Chest X-ray films showed diffuse infiltration of both lung fields. Transbronchoscopic lung biopsy was consistent with alveolar proteinosis, therefore the patient was submitted to pulmonary lavage. Under general anesthesia, the patient was intubated with a left-sided double lumen endotracheal tube, and lavage of the right lung was repeated with 0.9% saline until the washing became clear. At the end of the procedure, the patient was extubated after making strong respiratory efforts. Several days later, the left lung was lavaged using the same technique. The mechanical washing of amorphous material from alveoli by pulmonary lavage is of great benefit in relieving hypoxemia. Judicious use of lung lavage is effective and safe for patient with alveolar proteinosis.

The effect of [Arg8]vasopressin on electrogenic chloride secretion in cultured rat epididymal epithelia.

Primary cultured rat efferent ductal epithelia and cauda epididymal epithelial were mounted in Ussing chambers to study the effect of arginine vasopressin (AVP) on chloride secretion in the male excurrent duct. The regional differences in the signal transduction pathways involved were also investigated. In both the efferent duct and the cauda epididymidis, basolateral addition of AVP resulted in a dose-dependent increase in the short-circuit current (Isc), which was mediated via V1 receptor. Replacement of ambient Cl- with gluconate or pretreatment of a Cl- channel blocker, diphenylamine-2-carboxylate (apical, 1 mM), completely abolished the response, whereas addition of amiloride had no effect on the Isc. Pretreating the epithelia of the efferent duct with indomethacin (apical, 5 microM) or forskolin (basolateral, 1 microM), but not thapsigargin (apical, 1 microM) or trifluoperazine (apical, 20 microM), significantly inhibited the AVP response (P < 0.001). By comparison, pretreating the epithelia of the cauda epididymidis with any of the four agents significantly reduced the AVP-evoked response. These results suggested that the stimulation of chloride secretion by AVP in the efferent duct and the cauda epididymidis is mediated by prostaglandin synthesis and involves adenosine 3',5'-cyclic monophosphate (cAMP) as a second messenger. In the cauda epididymidis, calcium, in addition to cAMP, may play a role in mediating the AVP-induced response.

Growth factors in continuous ambulatory peritoneal dialysis effluent. Their relation with peritoneal transport of small solutes.

Recent studies reveal conflicting results on the change of solute transfer with time on continuous ambulatory peritoneal dialysis (CAPD) and recurrent peritonitis. Herein, we performed a cross-sectional study of 76 patients on CAPD to examine their peritoneal permeability by measuring the dialysate to serum ratio of creatinine (D/P) and the mass transfer area coefficients of creatinine (MTACCr) or glucose (MTACGlu). Transforming growth factor-beta1 (TGF-beta1), platelet-derived growth factor (PDGF), and epidermal growth factor (EGF) were measured in the dialysate by ELISA. TGF-beta1 mRNA in peritoneal macrophages were determined by a quantitative polymerase chain reaction. We failed to observe any correlation between the duration on dialysis and the peritoneal permeability in those patients with no previous peritonitis. Frequency of peritonitis episode did not affect the MTACCr, MTACGlu, or D/P. The MTACCr correlated well with MTACGlu (r = 0.78, p = 0. 001) and with D/P (r = 0.98, p < 0.0001). No inverse correlation was demonstrated between dialysate PDGF or EGF and the peritoneal permeability. A positive correlation was demonstrated between the dialysate TGF-beta1 and MTACCr, MTACGlu or D/P (r = 0.64, 0.54, and 0.64 respectively, p < 0.001). The dialysate TGF-beta1 levels in patients with low D/P (

Upregulation of adhesion molecule expression on endothelial cells by anti-DNA autoantibodies in systemic lupus erythematosus.

The mechanism of vasculopathy in systemic lupus erythematosus (SLE) remains unclear and the evidence for a direct pathogenic role of anti-double-stranded DNA antibodies (anti-dsDNA) is not strong. Our study aims to determine whether anti-dsDNA exerts any effect on the expression of adhesion molecules on endothelial cells. IgG was purified from 17 patients with SLE (median anti-dsDNA titer, 404 IU/ml) and from 9 healthy controls (median titer 16 IU/ml). Anti-dsDNA-depleted polyclonal IgG (anti-dsDNA-dep-IgG) (median anti-dsDNA titer 17 IU/ml) was prepared from sera of these patients with SLE by affinity chromatography with DNA cellulose column. Expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin on human umbilical vein endothelial cells (HUVEC) cultured with either control IgG or anti-dsDNA were compared by flow cytometry. The levels of adhesion molecules in the supernatant of cultured HUVEC were assessed by sandwich ELISA. Compared with either control IgG or anti-dsDNA-dep-IgG, HUVEC incubated with anti-dsDNA expressed a significantly higher mean fluorescence intensity of ICAM-1 and in VCAM-1 and a higher supernatant concentration of ICAM-1 and VCAM-1 but not E-selectin. At the same time, ICAM-1 mRNA was also raised with increased neutrophil adherence in HUVEC incubated with anti-dsDNA. Pretreatment of HUVEC with native DNA or histone before incubation with anti-dsDNA did not increase the expression of adhesion molecules. Our study provides in vitro evidence that anti-dsDNA could play an important pathogenic role in inducing inflammatory injury of vascular endothelium in SLE.

Regional differences in bioelectrical properties and anion secretion in cultured epithelia from rat and human male excurrent ducts.

Bioelectrical properties and anion secretion in cultured epithelia from different regions of rat and human male excurrent ducts were studied by measuring the short-circuit currents (ISC). In all regions of the rat excurrent duct, Cl- secretion accounts for over 90% of the basal ISC, although the magnitude varied in different regions. Cl- secretion was found to be mediated by a Cl-/HCO3- exchanger, an Na+/H+ exchanger, and an Na+/K+/2Cl- symport located on the basolateral side of the epithelial cells. Forskolin, an activator of adenylate cyclase, and ionomycin, a Ca2+ ionophore, were used to investigate the relative importance of cAMP and Ca2+ as intracellular messengers regulating Cl- secretion in different regions. It was found that in both species, the forskolin-evoked ISC response was larger in the proximal end (efferent duct/caput epididymidis [rat/human, respectively]) than in the distal end (cauda/corpus epididymidis). The response to ionomycin in the rat cauda epididymidis (distal end) was larger than that in the efferent duct (proximal end); on the other hand, no significant difference in the ionomycin-induced ISC was observed in the caput and the corpus regions from the human epididymis. Our results indicate that while the cAMP- and Ca(2+)-dependent pathways are both involved in regulating Cl- secretion in all regions along the male excurrent ducts in both species, a regional difference exists with respect to the relative importance of the two regulatory pathways involved in Cl- secretion along the male reproductive tract.

Increased release of von Willebrand factor antigen from endothelial cells by anti-DNA autoantibodies.

OBJECTIVE: To determine whether antibodies to double stranded DNA (anti-dsDNA) have a pathogenic role in systemic lupus erythematosus (SLE). METHODS: IgG was purified from 17 patients with SLE (median anti-dsDNA titre 1212 IU/ml) and nine healthy controls (median titre 40 IU/ml). Anti-dsDNA depleted polyclonal IgG (median anti-dsDNA titre 17 IU/ml) was also prepared from sera of the 17 patients by affinity chromatography on a DNA cellulose column. Binding to antiendothelial cell antibodies (AECA) and expression of von Willebrand factor (VWF) antigen by cultured human umbilical vein endothelial cells (HUVECs) were studied by flow cytometry. RESULTS: The percentage of HUVECs binding to AECA or expressing VWF was greater for cells incubated with IgG from patients with SLE than for cells incubated with control IgG, though values did not reach statistical significance; nevertheless, HUVECs incubated with IgG from patients expressed a greater mean fluorescence intensity with AECA (p = 0.0001) and greater VWF expression (p = 0.019). Both the fluorescence intensity and percentage of HUVECs binding to AECA or expressing VWF were significantly greater in HUVEC incubated with IgG containing anti-dsDNA than in those incubated with anti-dsDNA depleted IgG. The concentration of VWF in the supernatant was significantly increased in HUVECs incubated with IgG containing anti-dsDNA compared with control IgG or anti-dsDNA depleted IgG. Pretreatment of HUVECs with native DNA before incubation with IgG from lupus patients did not increase binding to AECA, or expression or release of VWF. CONCLUSIONS: Our study provides in vitro evidence that antibodies to DNA have a pathogenic role in the induction of inflammatory injury of the vascular endothelium in SLE.