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Biomedical subjects

E Ranieri

Publications and source records attributed to E Ranieri.

At least 73 records · Page 4Linked to original sources
PubMed1991

Neonatal screening strategy for cystic fibrosis using immunoreactive trypsinogen and direct gene analysis.

OBJECTIVE: To assess the effectiveness of a two tier neonatal screening strategy for cystic fibrosis, which combines estimation of immunoreactive trypsinogen followed by direct gene analysis in dried blood spot samples collected at age 5 days. DESIGN: Prospective study of two tier screening strategy. The first tier of testing immunoreactive trypsinogen concentration was measured in dried blood spot samples from neonates aged 4-5 days. In the second tier direct gene analysis to detect cystic fibrosis mutations deltaF508 and deltaI506 was performed in those blood spot samples which produced the highest 1% of immunoreactive trypsinogen values. Direct gene analysis was also performed on blood spot samples from infants with suspected or confirmed meconium ileus, regardless of the immunoreactive trypsinogen value. SETTING: The South Australian Neonatal Screening Programme, operating from the department of chemical pathology at Adelaide Children's Hospital. Subjects--All 12,056 neonates born in South Australia between December 1989 and June 1990. No selection criteria were applied. INTERVENTIONS: All infants found to have two recognised cystic fibrosis mutations on direct gene analysis were referred directly for clinical management, and those with one recognised cystic fibrosis mutation were recalled for a sweat test; their families were given genetic counselling. MAIN OUTCOME MEASURES: Direct or exclusion of cystic fibrosis by sweat testing of neonates identified as being at high risk of cystic fibrosis on screening and of those at minimum risk but whose subsequent clinical history raised suspicion about the disease. RESULTS: Of the 12,056 infants screened, 11,907 (98.8%) were reported as "cystic fibrosis not indicated" on the basis of low immunoreactive trypsinogen values. Of the 148 (1.23%) infants with raised immunoreactive trypsinogen values and one (0.008%) with meconium ileus, 132 (1.09%) were reported as cystic fibrosis not indicated, four (0.033%) were identified as having cystic fibrosis, and 13 (0.108%) were recalled for sweat testing after direct gene analysis for the presence of the deltaF508 and deltaI506 cystic fibrosis mutations. No cases of affected infants are known to have been missed to date. CONCLUSION: The strategy of measurement of immunoreactive trypsinogen followed by direct gene analysis is a highly specific neonatal screen for cystic fibrosis, requiring only 2.8 families to be contacted for every case of cystic fibrosis diagnosed.

Cystic Fibrosis↗
PubMed1991

A maternal serum screen for trisomy 18: an extension of maternal serum screening for Down syndrome.

The feasibility of extending second-trimester maternal blood screening for Down syndrome so as to include screening for trisomy 18 was examined using stored maternal serum samples collected for neural tube-defect screening. There were 12 samples from trisomy 18 pregnancies and 390 controls. The median maternal serum concentration of alpha-fetoprotein, free alpha-subunit human chorionic gonadotrophin, free beta-subunit human chorionic gonadotrophin, intact human chorionic gonadotrophin, total estriol, unconjugated estriol, estradiol, human placental lactogen, and progesterone were lowered in those pregnancies affected by trisomy 18 when compared with unaffected pregnancies matched for racial origin, maternal age, gestational age, and sample-storage duration. At an estimated odds risk of 1:400, 83.3% of affected pregnancies were detected using an algorithm which combines the maternal age-related risk with the maternal serum concentrations of unconjugated estriol, free alpha-subunit human chorionic gonadotrophin, free beta-subunit human chorionic gonadotrophin, estradiol, and human placental lactogen. The associated false-positive rate was 2.6%. At high risk odds of 1:10, the detection rate was 58.3%, with an associated false-positive rate of 0.3%. beta-Subunit human chorionic gonadotrophin and unconjugated estriol were the most powerful discriminators. It is possible to incorporate into existing Down syndrome screening programs an algorithm for detecting trisomy 18 with high sensitivity and specificity.

Adult↗
PubMed1986

Preliminary evidence for a processing error in the biosynthesis of Gaucher activator in mucolipidosis disease types II and III.

Activator protein (AP), which stimulated fibroblast sphingomyelinase activity, was isolated from the spleen of a patient with Gaucher's disease type I by the combined techniques of heat and alcohol denaturation, DEAE-cellulose column chromatography, gel filtration, preparative polyacrylamide-gel electrophoresis and decyl-agarose chromatography. Urea/sodium dodecyl sulphate (SDS)/polyacrylamide-gel electrophoresis showed two bands, one with an Mr of approx. 3,000 and the other with an Mr of 5,000-6,500. Similarly, SDS/polyacrylamide-gel electrophoresis performed in the absence of urea revealed the presence of two components, one of which adsorbed to a concanavalin A (Con A) column. Both components stimulated sphingomyelinase activity. On a non-denaturing polyacrylamide gel containing Triton X-100, four major components, two of which bound to Con A, were detected with the dye Stains-All. Cross-reacting material (CRM) to polyclonal Gaucher spleen AP antibodies was detected in normal fibroblasts and in fibroblasts from patients with sphingomyelinase and beta-glucocerebrosidase deficiency states (Niemann-Pick and Gaucher's diseases respectively). CRM in normal fibroblasts adsorbed to Con A columns and had the same mobility on SDS/polyacrylamide-gel electrophoresis as Con A-adsorbing Gaucher spleen AP. Normal AP was not observed in mucolipidosis type II (I-cell disease) fibroblasts; instead, extracts from these cells revealed the presence of two closely migrating bands with higher Mr values than normal fibroblast CRM. Furthermore, extracts of media from I-cell fibroblast cultures, but not from control or Gaucher fibroblast cultures, contained AP activity towards sphingomyelinase and beta-glucocerebrosidase. Fibroblasts from a patient with mucolipidosis type III (pseudo-Hurler polydystrophy) showed an intermediate pattern consisting of normal as well as the higher-Mr CRM. Our data provide evidence for the existence of AP in cultured skin fibroblasts and suggest that these proteins may be targetted to the lysosome by post-translational modification in a similar manner to that reported for lysosomal enzymes.

Cells, Cultured↗
PubMed1984

Studies on the activation of the enzymatic hydrolysis of sphingomyelin liposomes.

Two pH optima were observed for the hydrolysis of sphingomyelin liposomes by brain and fibroblast extracts; one at pH 4.2-4.5, the other at pH 7-8. The proportion of the acidic activity in fibroblasts was affected greatly by the culturing conditions. Both the acidic and neutral enzyme activities were deficient in Niemann-Pick Type A fibroblasts, suggesting that both were genetically related. Partially purified activators from normal as well as Gaucher disease spleen stimulated the hydrolysis of sphingomyelin, at both pH values, by fibroblast and brain extracts. After further purification by DE-52 and Sephacryl 200 column chromatography the Gaucher activator retained its ability to stimulate sphingomyelinase and was active as well towards beta-glucocerebrosidase and beta-galactocerebrosidase.

Animals↗
PubMed1984

Studies on the activation of sphingomyelinase activity in Niemann-Pick type A, B, and C fibroblasts: enzymological differentiation of types A and B.

Cultured skin fibroblast homogenates from patients with Niemann-Pick disease Type C, were able to degrade sphingomyelin liposomes at a normal rate. Fibroblasts from patients with Niemann-Pick disease Types A and B were less active (0.08-0.55 versus 0.96-2.93 nmol/h/mg). When fibroblasts were maintained in synthetic media (MCDB-104) devoid of fetal calf serum for a period of 21 days, sphingomyelinase activity measured at pH 3.8 increased in control and Niemann-Pick Type C (up to 15-fold) and in Niemann-Pick Type B (up to 3-fold) while Niemann-Pick Type A showed no significant increase in sphingomyelinase activity. Addition of a protein activator isolated from the spleen of a Type I Gaucher's disease patient stimulated a 2-7.5-fold increase in sphingomyelinase activity in normal, Niemann-Pick Type B and C fibroblasts, while under the same conditions the Niemann-Pick Type A fibroblast enzyme responded poorly. Our data show that the residual sphingomyelinase activity in Niemann-Pick Type A can be differentiated from that present in other phenotypic forms by its lack of response to the Gaucher activator. Furthermore, we can find no evidence to support the view that Niemann-Pick Type C sphingomyelinase differs from the normal enzyme in its response to Gaucher activator.

Adolescent↗
PubMed1983

The enkephalins are amongst the peptides hydrolyzed by purified acetylcholinesterase.

We have shown that purified acetylcholinesterase has the ability to hydrolyze a number of peptides including the physiologically occurring enkephalins. The enkephalins lost both the amino- and carboxyl-terminal amino acids, but several other peptides were not degraded. The enzyme was purified using an affinity chromatographic matrix that recognised one component of the active centre that is specific to cholinesterases, the anionic-binding site. The acetylcholinesterase was extracted from four tissues of diverse origin to minimise the risk of co-purifying a peptidase. The enzyme was essentially homogeneous on polyacrylamide gels, and there was only one protein that bound diisopropylfluorophosphate in the samples. The peptidase activity was not affected by the aminopeptidase inhibitor puromycin, but it was inhibited by acetylcholine at concentrations that also reduced the esterase activity. It was concluded that acetylcholinesterase also has the capacity for a novel type of hydrolysis of peptide bonds. The ability of acetylcholinesterase to hydrolyse naturally occurring compounds of different chemical nature, like esters and peptides, may help explain the long-standing puzzle of why the enzyme is more widely distributed than acetylcholine, once thought to be its sole natural substrate. The localization of the enzyme probably more accurately reflects the distribution of all its substrates, although their identity remains to be determined.

Acetylcholinesterase↗
PubMed1983

Sphingomyelinase in cultured skin fibroblasts from normal and Niemann-Pick type C patients.

The sphingomyelinase activities of extracts of normal cultured skin fibroblasts were compared with those obtained from Niemann-Pick disease Type A and B patients, and from a number of patients with a provisional clinical diagnosis of Niemann-Pick disease Type C. Even though fibroblasts from Type A and B patients were shown to be clearly deficient (less than 5% residual activity) the activity in Type C fibroblasts varied from approximately 50% of the lowest control value to normal activity. Isoelectric focussing of extracts of normal cultured skin fibroblasts on polyacrylamide gels revealed the presence of sphingomyelinase heterogeneity. However, no characteristic change in the behaviour of the enzyme in corresponding extracts from Niemann-Pick Type C cells was observed, suggesting that, if the defect in this condition is expressed fibroblasts, it does not manifest biochemically in the appearance or disappearance of a specific sphingomyelinase isoenzyme. Our data suggest that the heterogeneity observed in fibroblast extracts may simply reflect an interaction of the enzyme either with itself or with other hydrophobic components present in the cellular extracts.

Cell Line↗
PubMedDate not supplied

Ramipril inhibits in vitro human mesangial cell proliferation and platelet-derived growth factor expression.

Angiotensin-converting enzyme (ACE) inhibitors are antihypertensive drugs that have been shown to reduce proteinuria and to slow down the progression of renal function deterioration in different models of chronic glomerular disease. Major pathogenetic features of progressive glomerular injury leading to glomerulosclerosis are mesangial cell proliferation and platelet-derived growth factor (PDGF) expression. The aim of the present study was to evaluate the effect of ramipril, an ACE inhibitor, on these two potential therapeutic targets. Thus, the effect of ramipril on DNA synthesis, cell proliferation and PDGF A and B chain gene expression in fetal calf serum (FCS)-activated cultured human glomerular mesangial cells was investigated. DNA synthesis was evaluated by tritiated thymidine incorporation, cell proliferation by direct cell counting and cell viability by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT). PDGF A and B chain gene expressions were studied by Northern blot and RT-PCR, respectively. In a dose-dependent manner ramipril inhibited the FCS-induced DNA synthesis and cell proliferation. This effect was not dependent upon a toxic effect as demonstrated by MTT. The antiproliferative effect of ramipril was most likely independent of its ability to inhibit ACE present in the FCS and/or expressed by the cells, since a synthetic peptide that specifically inhibits ACE, at the same molar concentrations, did not inhibit FCS-stimulated DNA synthesis. Moreover, ramipril significantly reduced FCS-induced PDGF A and B chain gene expression. Finally, ramipril completely abolished the PDGF A and B chain gene expression induced by phorbol 12-myristate 13-acetate, a specific protein kinase C activator, suggesting a site of action downstream of this enzyme in the mitogenic signal transduction pathway. Our study would suggest that the modulatory action of ramipril on activated mesangial cell proliferation and PDGF expression is independent of its ability to inhibit ACE and could represent an additional mechanism in the renal protective effects of this drug.

Angiotensin-Converting Enzyme Inhibitors↗
PubMedDate not supplied

Fine needle aspiration cytology of squamous cell carcinoma arising in a thyroglossal duct cyst. A case report.

BACKGROUND: Carcinoma arising in a thyroglossal duct cyst is a rare entity. Malignancy generally is revealed only after surgical excision of a thyroglossal duct lesion because of the lack of specific physical findings. CASE: A 68-year-old male presented with a squamous cell carcinoma arising in a midline cyst in the neck. The malignancy had been discovered because of an enlarged metastasized lymph node in the left side of the neck. Fine needle aspiration cytology (FNA) showed cells with marked nuclear atypia and keratinizing cells. CONCLUSION: To our knowledge, this is the 10th squamous cell carcinoma reported and, among these, the first case diagnosed preoperatively by FNA cytology. A firm diagnosis at the preoperative stage is important with this neoplasm to plan extensive surgical intervention.

Aged↗
PubMedDate not supplied

The role of alpha-smooth muscle actin and platelet-derived growth factor-beta receptor in the progression of renal damage in human IgA nephropathy.

BACKGROUND: The degree of tubulointerstitial damage can be considered a better indicator of renal function outcome in IgA nephropathy (N) than the extent of glomerular sclerosis. MATERIALS AND METHODS: To investigate the pathogenetic mechanisms of interstitial injury in IgAN, we used immunohistochemistry and in situ hybridization to evaluate the glomerular and tubolointerstitial expression of PDGF-beta receptor (R) and alpha-smooth muscle actin (SMA), two markers of mesenchymal cell activation, and correlated these findings with the histopathologic and clinical features of the disease. We studied 155 IgAN patients, divided into three groups based on the histological findings (mild, moderate and severe histological lesions). RESULTS: In normal kidneys and in patients with mild histological lesions, the interstitial areas showed scattered peritubular cells positive for PDGF-betaR and alpha-SMA, with a distribution resembling the capillary network. In the glomeruli several cells (mainly in the mesangial area) stained for PDGF-betaR, but only very few cells were positive for alpha-SMA. Alpha-SMA and PDGF-betaR staining, as expected, was also observed in vascular smooth muscle cells. Compared to patients with mild histological lesions, alpha-SMA expression was strikingly increased in patients with moderate to severe lesions, particularly in areas of tubulointerstitial fibrosis. In these patients, PDGF-betaR gene and protein expression, at the tubulointerstitial level, paralleled that in alpha-SMA. Both signals were significantly correlated with the interstitial damage (interstitial infiltrate and fibrosis). Interestingly, these patients showed a different pattern of distribution of alpha-SMA and PDGF-betaR in the glomeruli: PDGF-betaR expression was upregulated, whereas no changes were seen in alpha-SMA staining. In addition, glomerular PDGF-betaR staining was significantly correlated with mesangial cell proliferation, while alpha-SMA was not. Image analysis showed that 40.2+/-10.3/1,000 microm2 of interstitial cells were positive to both PDGF-betaR and alpha-SMA, but only 2.8+/-1.8/1,000 microm of glomerular cells expressed both signals. CONCLUSIONS: Our study supports the hypothesis that interstitial PDGF-betaR and alpha-SMA positive cells may play a key role in the pathogenesis of tubulointerstitial damage.

Actins↗
PubMedDate not supplied

[Gene expression of the MAGE-1, -3, and -6 antigens in hepatic carcinoma].

MAGE is a family of genes specifically associated to human melanoma, but also found in hepatocellular carcinoma (HCC). In this study we evaluated the expression of such genes in 41 HCC patients, their correlation with pathological and clinical aspects of cancer, and the impact on prognosis. MAGE are expressed in most of HCC samples (28/41), no correlation was found with type and stage but they may be used as potential target for IT.

Aged↗
PubMedDate not supplied

[Role of immunity and tolerance in renal diseases].

Dendritic cells (DC) have been recognized as the most efficient antigen presenting cells that have the capacity to initiate naive T-cell response in vitro and in vivo. During their differentiation and maturation pathways, DC can efficiently capture, process and present antigens for T-cell activation. These characteristics make DC an attractive choice as the cellular adjuvant for can-cer vaccines. In humans, two DC subsets, myeloid DC (mDC) and plasmacytoid DC (pDC), have been characterized and have distinct origins and functions: mDC are involved in the induction of the Th1 response, while pDC regulate immunity and initiate adaptive antiviral immune responses. Advances in DC generation, loading and maturation methodologies have made it possible to generate clinical grade vaccines for various human trials. Several studies have shown that tumor antigen-loaded DC vaccination is safe and promising for the treatment of cancer. We are investigating the use of autologous tumor lysate-pulsed mature DC in renal cancer patients with metastasis. DC can play a central role in the development of T-cell tolerance, and its maintenance in the periphery is critical for the prevention of autoimmunity. DC are likely to have a role in the pathogenesis of autoimmunity and are, to date, the only APC capable of provoking autoimmune disease. Systemic lupus erythematosus (SLE), a systemic autoimmune disease with multi-organ involvement with autoreactive T and B cells, could be due to DC alterations, and pDC have a potential role in this disease. Given their pivotal role in controlling immunity, DC are logical targets for treating both cancer and autoimmune diseases.

Autoimmunity↗
PubMedDate not supplied

The role of cytology in differentiation of breast lesions.

The introduction of cytologic examination in to the diagnostic procedure has made it possible to define breast lesions better as early as the preoperative stage. However, there are interpretative problems depending on the nature of the lesions that make a histologic examination necessary. The cytology of nipple discharge or FNA of breast lesions would permit the best possible selection of the cases, indicating not only whether surgery is necessary, but also the basis on which it should be performed (outpatient under local anesthesia or inpatient under general anesthesia). The definitive histologic diagnosis of 447 cases submitted to surgery under general anesthesia and of 379 who were operated under local anesthesia has been correlated with the relative cytologic examination. The sensitivity and specificity of the cytologic examination are respectively 97.7% and 98.8%.

Adult↗
PubMedDate not supplied

The integration of diagnostic tests and the role of outpatient surgery in the management of breast disease.

The aim of the earliest possible diagnosis of breast carcinoma achieved through the integration of clinical, mammographic, sonographic and cytologic data has determined an increased detection of breast lesions and a greater accuracy in their description. Nevertheless, the nature of some of these lesions cannot be well-defined because of their biological characteristics and their physical, radiological or cytological aspects. The need to obtain a definitive diagnosis in any case has given rise to a paradoxical increase in histologic examinations: in specific conditions and for certain kinds of lesions, outpatient surgery under local anesthesia could very well represent a supplementary tool in the diagnostic and therapeutic strategy for the management of breast lesions. The data here reported concern 397 breast lesions for which it was thought appropriate to complete the diagnostic procedure with the surgical excision on an outpatient basis under local anesthesia. Excluding the 3 breast lymphomas and the 19 loco-regional recurrences, for which the purpose of outpatient surgery was therapeutic, at the histologic examination 35% benign lesions, of which 91 associated with an increased risk of breast cancer, were found, as well as 11 in situ and 10 invasive carcinomas (false negatives), with a therapeutic option error of 2.5%. Now that one of the principal aims of public management is to put a check on health expenditures, outpatient surgery would contribute to obtaining the correct balance between proper management of breast lesions and low cost/effective ratio.

Adult↗
PubMedDate not supplied

Primary lymphoma of the breast: a case report.

The clinical features of a patient with a primary lymphoma of the breast are herein reported. The diagnosis was reached by histological examination after outpatient surgery. Surgical resection was followed by cytostatic treatment and locoregional radiotherapy. This case report is an example where the integration of physical and mammographic examination together with ultrasonography (and cytology), in the context of diagnostic procedures, induced us to perform a surgical excision on an outpatient basis. This line of action allowed us, in one step, to arrive at both the definitive diagnosis and the appropriate choice of therapy. Thus we believe that this diagnostic procedure should be carried out whenever a breast lesion, thought to be "probably benign", is found by physical or mammographic exam in a peri-postmenopausal woman.

Breast Neoplasms↗