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A Longo

Publications and source records attributed to A Longo.

At least 19 recordsLinked to original sources

DNA typing of HLA-DQ alleles by gene amplification of DQA and DQB variable exons: analysis of DQA/DQB haplotypes.

In the present report, we describe a DNA typing method that allows detection of all the polymorphic variants of DQA1 and DQB1 second exons. By the oligotyping procedure provided in this paper, we are able to identify 8 DQA1 and 13 DQB1 alleles and to type random individuals in any heterozygous combination. We provide the hybridization and washing temperatures for using either 32P labelled or non-radioactive probes. The discrimination power of this procedure, compared to serological and cellular techniques, is remarkable. Therefore, this typing method finds perfect application in transplantation immunology and it will be very helpful to optimize the matching of unrelated donors before BMT. It is apparent from our results that despite the linkage disequilibrium present between DQ and DR loci, a DR specificity may frequently be associated to different DQ haplotypes. This is the case for DR4, DR7, DR8, DR9, and DR13 specificities.

Alleles

[Influence of the use of Swan-Ganz catheter on the mortality from acute myocardial infarction].

OBJECTIVE: to evaluate the statistical significance of the haemodynamic examination making use of the Swan-Ganz catheter (S-G) on the hospitalary mortality of patients (pts) with acute myocardial infarction (AMI). CONCEPTION: Not randomized, not controlled and retrospective study. SETTING: Intensive coronary care unit (CCU). PATIENTS: 2562 pts with AMI sequentially admitted to a CCU were studied. METHODS: The 2562 pts were divided in two groups: Group A--involving 2117 pts not submitted to haemodynamic examination; Group B-constituted by 445 pts (17% of all pts) submitted to haemodynamic examination. All pts admitted to the study were distributed according to the four classes of Killip. The pts included in each Killip class were distributed by the groups A (not catheterized) and B (catheterized). Hospitalary mortalities of the pts included in the global A and B groups, and the mortalities of the patients included in the particular A and B groups in each Killip class were determined. To evaluate the statistical significance of the differences in distributions of the number of pts who survived and who died in each set of A and B groups square chi test was used. Values of p less than 0.05 were considered as statistical significant. RESULTS: The distributions of the pts who died and who survived during the stay in the hospital by the A and B groups, be it in the total group of patients be it in each Killip class, were not significant different (p greater than 0.05). CONCLUSIONS: The mortalities of pts with AMI submitted or not to haemodynamic examination with S-G catheter did not significantly differ even in each Killip class, which means that it was not demonstrated a benefit with the S-G catheter use in the pts with AMI. In the presence of this conclusion and considering that retrospective studies must be interpreted with circumspection, it seems necessary to accomplish a prospective randomized and controlled study for evaluation the risk/benefit relation of the haemodynamic examination with the S-G catheter in pts with AMI.

Catheterization, Swan-Ganz

Human allogeneic melanoma-reactive T-helper lymphocyte clones: functional analysis of lymphocyte-melanoma interactions.

Lymphocyte clones were isolated from CD4+ peripheral-blood lymphocytes (PBL) of melanoma (Me) patient 9923 (HLA-DR7, DQw2, w6), co-cultured for 30 days with autologous accessory cells, allogeneic Me (Me 1811) (HLA-DR7, DQw1, w2), IL-1 beta (2 U/ml) and IL-2 (15 IU/ml). The 55 clones tested displayed a CD3+, CD4+, CD8-, T-cell receptor (TCR) alpha/beta+, gamma/delta- phenotype. Twenty clones were assayed for proliferation in the presence of Me 1811 and B-lymphoblastoid cell line (LCL) 1811, both expressing HLA-class-I and -II (DR7 and DQw2 shared with patient 9923), intercellular adhesion molecule-1 (ICAM-1) and lymphocyte-function-associated antigen-3 (LFA-3) molecules. Eight clones were found to be reactive to Me 1811 but not to LCL 1811. Specificity analysis of these 8 clones revealed that each of them proliferated only to Me 1811, not to other 14 Me and 12 different LCL, suggesting recognition of melanoma-associated antigen (MAA) expressed on the stimulating Me. One clone (103) was analyzed in more detail. A wider specificity analysis showed that it reacted to Me 1811 but not to 10 other Me expressing or not HLA-DR7, 5 normal melanocyte cultures (2 of them typing HLA-DR7-positive when exposed to interferon-gamma--IFN-gamma), 4 tumors other than Me and 20 different LCL. Clones did not show proliferation in the presence of autologous Me cells. Clone proliferation in response to Me 1811 was significantly inhibited by monoclonal antibodies (MAbs) directed to CD3, TCR alpha/beta, TCR beta chain V12, CD4 and HLA-DR. Moreover, following stimulation with Me 1811, clone 103 showed increased surface expression of CD25 (IL-2 receptor) and CD71 (transferrin receptor) and produced significant amounts of IL-2 and IFN-gamma. The supernatant taken from co-culture of clone 103 with Me 1811 augmented the cytotoxicity of PBL 9923 and other allogeneic PBL against K562 and Me 1811. Thus, the lymphocyte clone 103 is a CD4+ Th clone which uses its CD3/TCR alpha/beta complex to recognize an MAA in conjunction with HLA-DR7. Availability of this type of reagent may prove useful to identify and characterize MAA recognized by T lymphocytes.

Antigens, CD

Determination of N-acetylcysteine in human plasma by gas chromatography-mass spectrometry.

An automatic mass spectrometric method for the quantitation of N-acetylcysteine (NAC) in human plasma has been developed. NAC was extracted from plasma with ethyl acetate and derivatized in two steps with 2-propanol and pentafluoropropionic anhydride. The volatile derivative obtained was ideal for gas chromatographic-mass spectrometric analysis. Data obtained by analysing the plasma of healthy volunteers to whom 600 mg of NAC had been orally given are reported.

Acetylcysteine

Growth factors in the human prostate.

Recent studies have focused on the potential role of local polypeptide growth-regulating factors in the etiology of benign prostatic hyperplasia (BPH) and prostatic carcinoma. In our studies we confirmed the presence of specific receptors for epidermal growth factor (EGF) in prostatic tissues from patients affected by BPH. In addition, we demonstrated that specific receptors for insulin-like growth factor type I (IGF-I) are present in BPH tissues. In order to identify a possible interaction between androgens and these growth-regulating factors, we investigated the effect of testicular suppression-induced androgen withdrawal on both EGF and IGF-I receptor concentrations in prostatic tissue from patients affected by BPH treated with a long-acting luteinizing hormone-releasing hormone analog. Both EGF and IGF-I binding capacities were significantly increased after treatment. This finding suggests that in vivo IGF-I and EGF receptor levels may be under negative androgenic regulation, indicating a potential role for these growth-regulating factors in the mechanism of response to the castration-induced regression of androgen-dependent prostatic tissue. Moreover, preliminary studies indicate that in human BPH prostatic tissue multiple IGF-binding proteins (IGF-BP) are present. This finding suggests a possible role of IGF-BP in modulating IGFs biological activities at the prostate level.

Aged

Insulin-like growth factor-I receptors in human hyperplastic prostate tissue: characterization, tissue localization, and their modulation by chronic treatment with a gonadotropin-releasing hormone analog.

Insulin-like growth factor I (IGF-I) receptors were characterized in membranes obtained from prostate tissue of patients affected by benign prostatic hyperplasia (BPH) before and after treatment with a GnRH agonist analog. Binding of [125I]IGF-I to membranes obtained from untreated patients was specific and time and temperature dependent. Analysis of the binding data yielded two classes of binding sites, one of high affinity (Kd, 10(-11) mol/L) and one of lower affinity (Kd, 10(-9) mol/L). BPH membrane preparations were affinity-cross linked to labeled IGF-I, and then subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Analysis by autoradiography revealed one labeled protein with an apparent Mr = 300K under nonreducing conditions and two labeled protein with Mr = 270K and Mr = 130K under reducing conditions. Excess unlabeled IGF-II reduce both of them, whereas the same excess of IGF-I completely abolished them. In membrane preparations of prostatic tissues from patients affected by BPH and treated for 2 months with a GnRH agonist analog, the binding capacities of both binding sites were significantly higher than those of BPH tissue from untreated patients, whereas binding affinities were unchanged. The IGF-I receptor in BPH prostate tissue of untreated patients was mainly localized in the basal layer of the epithelium, as demonstrated by immunohistochemical staining, whereas in the tissue from treated patients positive staining was found also in the glandular epithelium. These results demonstrate that: 1) specific binding sites for IGF-I are present in prostatic tissue from patients with BPH, 2) androgen deprivation increases their binding capacities and seems to modify their epithelial localization.

Aged

Specific recognition of human CD3-CD16+ natural killer cells requires the expression of an autosomic recessive gene on target cells.

We analyzed the recently defined ability of CD3-CD16+ cells to specifically recognize and lyse normal allogeneic target cells (PHA-induced blasts). The susceptibility to lysis by a given alloreactive natural killer (NK) clone ("1 anti-A") was expressed by PHA blasts derived from 9 of 38 random donors analyzed. In all instances, the specific lysis of "susceptible" target cells was greater than 35% while that of "nonsusceptible" targets was less than 6% at an E/T cell ratio of 5:1. In addition to 1 anti-A, A anti-1 specific CD3-CD16+ clones could also be isolated from the reverse MLC combination. The relationship existing between lysis of normal allogeneic cells or tumor cells by the same CD3-CD16+ effector cell has been investigated: 1 anti-A specific CD3-CD16+ clones lysed PHA blasts of three of six cancer patients, while they lysed fresh tumor cells (ovarian carcinoma) from all six patients. The type of inheritance of the character "susceptibility to lysis" was analyzed in representative families. This analysis revealed that the character is inherited in an autosomic recessive fashion, and it is therefore different from MHC. We further investigated the type of segregation of the opposite character "resistance to lysis" (which is inherited in a dominant mode). The finding that this character segregated in all donors expressing given MHC haplotypes indicated that the gene regulating the expression of the NK-defined alloantigen is present on chromosome 6.

Antibodies, Monoclonal

Is multi-infarct dementia representative of vascular dementias? A retrospective study.

Multi-infarct dementia (MID) indicates a dementia disorder primarily caused by multiple cerebral infarcts. Since other pathogenetic mechanisms cause vascular dementia we evaluated clinical, CT scan and CSF neurochemical parameters of 134 MID and 67 PVD (probable vascular dementia) patients. We found no differences with regard to the presence of major risk factors. Only TIA/stroke episodes and focal neurological signs were significantly more frequent in MID than in PVD cases, an anticipable result on the basis of MID definition. CT scan findings showed a prevalence of subcortical with respect to cortical lesions in both groups, with a higher frequency in MID patients. Subjects with deep infarcts more frequently showed TIA/stroke episodes and diabetes mellitus. No differences were detectable in CSF monoamine metabolite levels. We conclude that in the majority of vascular dementias subcortical damage seems to have a major pathogenetic role.

Aged

[Rhythmic profiles of the late phase of acute myocardial infarction as a function of age].

OBJECTIVES: To evaluate the incidence of ventricular arrhythmias in the late phase of acute myocardial infarction (AMI) and to compare it with the following clinical parameters: age, sex, AMI localization, ventricular function (Killip classes), maximal creatinokinase (CK max) and the presence of sinus tachycardia. DESIGN: Prospective study, during a period of 31 months, of a non-selected group of patients with AMI. SETTING: Coronary Care Unit (UTIC-Arsénio Cordiero). PATIENTS: Non-selected group of 153 patients with acute myocardial infarction who survived the second week of disease. MATERIAL AND METHODS: 24-hour Holter ECG performed between the 4th and the 25th day of AMI. The patients were divided into two groups according to the hourly frequency of premature ventricular beats (PVB): less than 3 per hour (PVB less than 3/h) and 3 or more per hour (PVB greater than or equal to 3/h). RESULTS: PVB greater than or equal to 3/h occurred in 36 patients (24%). There was no differences in the occurrence of ventricular arrhythmias between sex, AMI localization, AMI size evaluated by CK max, and the presence of sinus tachycardia. Patients in Killip class III had more ventricular arrhythmias (67%) than patients in Killip class I (23%) (p less than 0.005), in Killip class II (18%) (p = 0.007), and in Killip IV (0%) (p = 0.017). In patients with serious left ventricular failure (classes III + IV) the ventricular arrhythmias were not significantly higher (40%) than in patients without serious left ventricular failure (classes I + II) (22%) (chi 2 = 2.5; p less than 0.25 NS). Patients with less than 41 years old had less PVB greater than or equal to 3/h (4%) than patients between ages 41-69 (24%) (p less than 0.05), and than patients over 70 years old (47%) (p = 0.00075). CONCLUSIONS: The majority of patients (76%) showed a low risk rithmic profile (PVB less than 3/h) in the late phase of AMI. Among all parameters the age of the patients was the one best related to the occurrence of ventricular arrhythmias. Sex, AMI localization, AMI size, and the presence of sinus tachycardia were not related to the presence of PVB. A slight tendency was found in patients with heart failure to have more PVB. On the other hand the elder group carried a statistically significant risk factor for a higher occurrence of ventricular premature beats.

Adult