Molecular diagnosis of Alport syndrome: the experience in Siena.
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Biomedical subjects
Publications and source records attributed to M Bruno.
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Preparative free-flow electrophoresis and aqueous two-phase polymer partition were used to obtain a plasma membrane-enriched fraction of adipocytes isolated from epididymal fat pads of the rat together with a fraction enriched in small vesicles with plasma membrane characteristics (thick membranes, clear dark-light-dark pattern). The electrophoretic mobility of the small vesicles was much less than that of the plasma membrane consistent with an inside-out orientation whereby charged molecules normally directed to the cell surface were on the inside. When plasma membranes and the small vesicle fraction were isolated from fat cells treated or not treated with 100 microU/ml insulin and the resident proteins of the two fractions analyzed by SDS-PAGE, the two fractions exhibited characteristic responses involving specific protein bands. Insulin treatment for 2 min resulted in the loss of a 90 kDa band from the plasma membrane. At the same time, a ca. 55-kDa peptide band that was enhanced in the plasma membrane was lost from the small vesicle fraction. The latter corresponded on Western blots to the GLUT-4 glucose transporter. Thus, we suggest that the small vesicle fraction with characteristics of inside-out plasma membrane vesicles may represent the internal vesicular pool of plasma membrane subject to modulation by treatment of adipocytes with insulin.
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We studied 47 adults (21 men, 26 women), with biopsy-proven celiac sprue and anti-endomysin antibody (EmA) positivity while untreated, to evaluate the usefulness of both serologic markers of celiac sprue [i.e., immunoglobulin A (IgA)-EmA and total Ig-anti-gliadin (AGA) antibodies] and of a detailed dietary inquiry in predicting the mucosal pattern after gluten withdrawal. A second biopsy was repeated 8-30 months after beginning a gluten-free diet, along with EmA and AGA determinations and the dietary inquiry. Both EmA and AGA were appraised by indirect immunofluorescence on monkey esophagus and rat kidney, respectively. Intestinal biopsy was graded according to Cooke's criteria. After gluten withdrawal, intestinal mucosa reverted to normal in only nine patients. Both EmA and AGA showed high positive but low negative predictive values on intestinal histologic outcome. The positive predictive value of admission of dietary lapses was 100%, whereas the negative predictive value was 39.1%. Neither serologic markers nor dietary inquiries are to be regarded as reliable predictors of intestinal outcome after a gluten-free diet. Biopsy remains the best means of ascertaining mucosal recovery.
Increased levels of pancreatic enzymes have been reported in patients with renal insufficiency and ascribed either to impaired urinary excretion or, in a few studies, to the presence of pancreatic damage. In the present study serum total amylase, pancreatic amylase, and lipase were evaluated in 63 patients with chronic renal insufficiency (CRF), in 98 patients on hemodialysis (HD), in 28 patients on continuous ambulatory peritoneal dialysis (CAPD), in 23 patients with renal transplantation (RT), and in 34 normal volunteers (C). Serum parathyroid hormone and triglyceride levels were also measured in the majority of patients. Ultrasound examination of the pancreas was performed in a select number of cases. Mean values of pancreatic enzymes were significantly higher in all the study groups in comparison with controls, but values exceeding three times the upper normal limit were detected only in HD patients, who also showed amylase and lipase levels significantly highly than those of CAPD and CRF subjects. Negligible amounts of pancreatic enzymes were detected in peritoneal fluid of CAPD patients. Significant correlations were found with serum creatinine in CRF, with parathyroid hormone in HD and CAPD, and with duration of treatment in HD. No pancreatic abnormalities were detected by ultrasound. In conclusion, very high levels of pancreatic enzymes are seen mainly in HD patients and might be related more to the metabolic derangement of long-term dialysis treatment than to the occurrence of acute pancreatic damage.
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Unexpectedly high early reinfection rates are reported in duodenal ulcer patients in whom Helicobacter pylori infection had been considered eradicated two months (T2) after appropriate therapy. Since some of these re-conversions to Helicobacter pylori positivity were probably recrudescences of a latent infection rather than reinfections, studies were performed to evaluate whether the type of antral gastritis could predict the infection outcome. In 142 eradicated patients at T2, endoscopies were repeated 6 (T6), 12 (T12) and 24 (T24) months after therapy to assess Hp status and to score antral gastritis. Re-conversion to Hp positivity occurred in 14.79% between T2/T6, in 5.40% between T6/T12 and 11.11% between T12/T24. The absence of active antral gastritis at T2 with its 87.31% negative predictive value was a fairly good marker of subsequently confirmed eradication (p = 0.017). It is suggested that, evaluation of antral gastritis soon after an eradicating course, could be a reliable parameter in assessing "true" Hp eradication.
Wegener's granulomatosis (WG) is a rare small vessels necrotizing and granulomatous systemic vasculitis which usually affects the respiratory-tract and the kidneys. Diagnosis is often difficult, but has become easier with antineutrophil cytoplasmic antibodies (ANCA) detection that may justify a more aggressive biopsy policy also in the elderly. Classic treatment with steroids and oral cyclophosphamide (CY) has proven to be of benefit, but side-effects are severe and frequent and the search for less toxic therapeutic schemes should be encouraged. We treated with intravenous pulses of CY (1 g/m2 monthly for 6 months, every two months for the following 6 and quarterly for another year) 5 of 7 patients with WG recently admitted to our institution. We obtained a quick, complete response in 4 of these patients, with no side effects, nor relapses, after a mean follow-up of 17 months. The only patient who did not respond was identified soon after the beginning of the treatment because of a poor reduction of ESR and could be shifted to oral administration of CY successfully. From our still limited experience CY intravenous pulses have proven to be safe and effective enough to advice its use as the first-choice treatment for WG.
To evaluate a twelve-month effect of Helicobacter pylori eradication, 258 consecutive out-patients with H. pylori related active duodenal ulcer were given a ten-day eradicating treatment. After healing no maintenance antiulcer medication was given. On entering the study and then 2, 6 and 12 months after the completion of therapy patients were scored for symptoms and underwent endoscopy to assess the presence of duodenal ulcer and to score antrum and corpus gastritis. Statistical analysis was performed by means of the chi 2 test. Histological eradication, defined as the inability to detect H. pylori six months after the completion of the eradication course, was proved in 85 subjects while the 123 non-eradicated ones were considered as the control group. Ulcer relapse rate and ulcer-like symptoms were significantly less frequent among eradicated than non eradicated throughout the follow-up. As compared to non eradicated, gastritis significantly improved among eradicated in both antrum and corpus. H. pylori eradication may be recommended since, by reducing ulcer relapse rate and related symptoms, there is no need for further antiulcer maintenance therapy with a significant drop in socioeconomic costs.
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Aerial parts of Centaurea napifolia furnished the sesquiterpene lactones cnicin, 4'-acetoxycnicin, melitensin, dehydromelitensin, two known esters of dehydromelitensin, lappaol and a lappaol isomer, and a 1,2-diacylated glucose.
Surfactant protein A (SP-A) was modified by covalent linkage with polylysine of average M(r) 21 kD ([Lys]21kD-SP-A) and utilized to transfect human airway epithelial cells (H441) in vitro. Transfection of H441 cells was more efficient with [Lys]21kD-SP-A than with polylysine-DNA or unmodified SP-A-DNA complexes. Transfection with [Lys]21kD-SP-A was effective at a protein-to-DNA molar ratio of 400:1 and in the presence of an exogenous surfactant preparation, Survanta. Transfection with [Lys]21kD-SP-A was reduced in the presence of unmodified SP-A consistent with the concept of a receptor mediated uptake of protein-DNA complexes. Increased transfection efficiency correlated with decreasing diameter of the [Lys]21kD-SP-A-DNA complexes, and these complexes bound to the cell surface and pseudopodia of H441 cells. Transfection was enhanced by co-incubation with replication-deficient adenovirus. Cotransfection by [Lys]21kD-SP-A-DNA and [Lys]10kD-SP-B resulted in an additive level of reporter gene (CAT) expression. [Lys]21kD-SP-A-DNA is likely to be useful as a component of a surfactant-based DNA delivery system for transfection of airway cells.
We compared the diagnostic accuracy of a new immunological marker of celiac sprue (CS), the antijejunum antibody (JAB), with that of antigliadin (AGA) and antiendomysium (EmA) antibodies. One hundred untreated adults with biopsy-proven CS, 52 healthy controls, and 57 patients with inflammatory bowel disease, lymphoma of the small bowel, Whipple's disease, and irritable bowel syndrome were investigated. Only JAB and EmA were detected at a similar titer in all patients with untreated CS but in no controls (100% sensitivity and specificity). Sensitivity of AGA was, respectively, 55% for IgA and 78% for Ig class, with a 100 and 82% specificity. The differences in frequencies between both EmA and JAB with IgA and IgG AGA were highly significant. We conclude that JAB and EmA provide a reliable noninvasive screening test for clinically significant gluten-sensitive enteropathy. The lower cost of IgA-JAB is a major advantage, owing to the different availability of the lower third of the esophagus and jejunum from primates. The sensitivity and specificity of the two tests are almost identical, but we find interpreting EmA easier than JAB especially when the titer is low.
To date, only a few studies of gastric emptying in Crohn's disease have been published in the literature. This might be clinically important because slow-release drug formulations are increasingly available for treatment. Studies in children with gastric involvement suggest that gastric emptying may be delayed in this condition. We studied the scintigraphic gastric emptying of 21 adult patients with nonobstructive Crohn's disease and without gastric or duodenal involvement by the disease, compared with that of 20 healthy volunteers. Overall, no significant differences were found, but subgroup analysis showed that symptomatic patients [half-time (t1/2) 133 +/- 75.9] and those with colonic involvement (t1/2 127.2 +/- 64) had a significantly (p < 0.01) delayed gastric emptying over controls (t1/2 85.5 +/- 15.4). Such a difference was also observed between symptomatic and asymptomatic patients (p < 0.05). We conclude that gastric emptying is slowed in symptomatic patients with nonobstructive Crohn's disease and in those with colonic involvement. This may have therapeutic implications.
Algal extracts of Anabaena planctonica from Lake Mulargia in Italy were tested for toxins by mouse bioassay, the Microtox system, and GC-MS and HPLC chromatography. Anatoxin-a was identified by GC-MS after derivatization with pentafluorobenzyl bromide. Hepatotoxins were also present and are perhaps related to the microcystins present in other species of Anabaena.