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1.
Nutr Metab Cardiovasc Dis ; 29(4): 367-377, 2019 04.
Article in English | MEDLINE | ID: mdl-30795994

ABSTRACT

BACKGROUND AND AIMS: Adults with congenital heart disease (ACHD) are at risk of overweight and obesity, two major health problems, though underweight can be a negative prognostic factor too. Awareness of the body mass index (BMI) in ACHD is very limited. The present study describes the use and prevalence of BMI in Italian symptomatic hospitalized ACHD patients in relation to complexity by Bethesda system classification, diagnosis, sex and age. METHODS AND RESULTS: We classified 1388 ACHD patients, aged 18-69 years, on the basis of their BMI, and compared them to the Italian reference population. In our total ACHD population we found a significantly higher prevalence of underweight compared to the Italian reference population (6.34% vs 3.20%). ACHD women were more underweight than men. Underweight decreased with age. Overweight was significantly less frequent in the total ACHD population (26.73% compared to 31.70%) in the Italian reference population. Men were more likely to be overweight than women. In statistical terms obesity was similar in the Italian reference population (10.50%) and our ACHD population (9.58%). Both overweight and obesity increased with age. Results were comparable using a diagnostic anatomical-functional classification and the Bethesda system classification. CONCLUSIONS: In our cohort of ACHD the prevalence of underweight was double that of the Italian reference population. The prevalence of overweight was lower, while obesity was similar. Since BMI does not account for differences in body fat distribution, a future aim will be to quantify the visceral component of the adipose tissue in ACHD patients and examine their body composition in order to reflect their risk of acquired cardiovascular disease better, and either to maintain or achieve an adequate visceral component.


Subject(s)
Body Mass Index , Heart Defects, Congenital/epidemiology , Inpatients , Obesity/epidemiology , Thinness/epidemiology , Adiposity , Adolescent , Adult , Age Distribution , Aged , Female , Heart Defects, Congenital/diagnosis , Heart Defects, Congenital/physiopathology , Humans , Italy/epidemiology , Male , Middle Aged , Obesity/diagnosis , Obesity/physiopathology , Prevalence , Prognosis , Retrospective Studies , Risk Assessment , Risk Factors , Sex Distribution , Thinness/diagnosis , Thinness/physiopathology , Young Adult
2.
Minerva Anestesiol ; 77(6): 654-7, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21525834

ABSTRACT

This article presents the case of a pregnant woman affected by A/H1N1 flu progressed to ARDS requiring rescue therapy by VV ECMO. Due to the early gestational age, the patient was placed on ECMO before delivery. Four weeks after VV-ECMO placement, a Cesarean section was successfully performed while on ECMO support. One week after delivery, the patient was weaned from ECMO and at 8 weeks from admission she was discharged home without O2 support. The newborn is alive and was discharged at 40 days of age on neurological follow-up.


Subject(s)
Cesarean Section , Extracorporeal Membrane Oxygenation , Pregnancy Complications/therapy , Respiratory Distress Syndrome/therapy , Adult , Female , Humans , Pregnancy
3.
Eur J Surg Oncol ; 31(7): 715-20, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16005176

ABSTRACT

AIMS: To determine the axillary recurrence rate in breast cancer patients with negative sentinel lymph nodes (SLN) who did not undergo further axillary lymph node dissection (ALND), and to establish whether this conservative axillary approach leads to an increased recourse to diagnostic axillary biopsy during the follow-up period because of the clinical suspicion of nodal recurrence. METHODS: In 479 patients, operated on for early breast cancer between 1998 and 2002 in five institutions, SLN biopsy was negative and no further axillary surgery was performed. SLN was localized using subdermal injection with 30-50 MBq of 99m-Tc-colloidal albumin. Follow-up controls were performed at 6-monthly intervals. RESULTS: The mean number of SLNs removed was 1.4 per patient. Most patients (90.6%) were given adjuvant systemic therapy, based on the primary tumour characteristics. At a median follow-up of 35.8 months, no clinical axillary recurrence was found. No patient underwent surgical axillary biopsy for suspicious clinical or ultrasonographic findings. CONCLUSIONS: Our results confirm that SLN biopsy without ALND in SLN-negative patients with early breast cancer is not followed by clinically evident axillary recurrence in the short-term.


Subject(s)
Breast Neoplasms/pathology , Neoplasm Recurrence, Local , Sentinel Lymph Node Biopsy , Adult , Aged , Aged, 80 and over , Axilla , Case-Control Studies , Female , Follow-Up Studies , Humans , Lymph Node Excision , Middle Aged , Prognosis
4.
Biochemistry ; 39(50): 15454-61, 2000 Dec 19.
Article in English | MEDLINE | ID: mdl-11112531

ABSTRACT

Measurements of the H(+)/heme a, Cu(A) ratios for proton-electron coupling at these centers (redox Bohr effect) in CO-inhibited cytochrome c oxidase purified from bovine heart mitochondria, both in the soluble state and reconstituted in liposomes, are presented. In the soluble oxidase, the H(+)/heme a, Cu(A) ratios were experimentally determined upon oxidation by ferricyanide of these centers as well as upon their reduction by hexammineruthenium(II). These measurements showed that in order to obtain H(+)/heme a, Cu(A) ratios approaching 1, one-step full oxidation of both metal centers by ferricyanide had to be induced by a stoicheiometric amount of the oxidant. Partial stepwise oxidation or reduction of heme a and Cu(A) did produce H(+)/heme a, Cu(A) ratios significantly lower or higher than 1, respectively. The experimental H(+)/heme a, Cu(A) ratios measured upon stepwise reduction/oxidation of the metals were reproduced by mathematical simulation based on the coupling of oxido-reduction of both heme a and Cu(A) to pK shifts of common acid-base groups. The vectorial nature of the proton-electron coupling at heme a/Cu(A) was analyzed by measuring pH changes in the external bulk phase associated with oxido-reduction of these redox centers in the CO-inhibited oxidase reconstituted in liposomes. The results show that the proton release associated with the oxidation of heme a and Cu(A) takes place in the external aqueous phase. Protons taken up by the oxidase upon rereduction of the centers derive, on the other hand, from the inner space. These results provide evidence supporting the view that cooperative proton-electron coupling at heme a/Cu(A) is involved in the proton pump of the oxidase.


Subject(s)
Copper/chemistry , Electron Transport Complex IV/chemistry , Heme/chemistry , Animals , Cattle , Electron Transport Complex IV/metabolism , Electrons , Myocardium/enzymology , Oxidation-Reduction , Protons
5.
Biochemistry ; 39(21): 6373-9, 2000 May 30.
Article in English | MEDLINE | ID: mdl-10828951

ABSTRACT

A study is presented on the coupling of electron transfer with proton transfer at heme a and Cu(A) (redox Bohr effects) in carbon monoxide inhibited cytochrome c oxidase isolated from bovine heart mitochondria. Detailed analysis of the coupling number for H(+) release per heme a, Cu(A) oxidized (H(+)/heme a, Cu(A) ratio) was based on direct measurement of the balance between the oxidizing equivalents added as ferricyanide to the CO-inhibited fully reduced COX, the equivalents of heme a, Cu(A), and added cytochrome c oxidized and the H(+) released upon oxidation and all taken up back by the oxidase upon rereduction of the metal centers. One of two reductants was used, either succinate plus a trace of mitochondrial membranes (providing a source of succinate-c reductase) or hexaammineruthenium(II) as the chloride salt. The experimental H(+)/heme a, Cu(A) ratios varied between 0.65 and 0.90 in the pH range 6.0-8.5. The pH dependence of the H(+)/heme a, Cu(A) ratios could be best-fitted by a function involving two redox-linked acid-base groups with pK(o)-pK(r) of 5.4-6.9 and 7.3-9.0, respectively. Redox titrations in the same samples of the CO-inhibited oxidase showed that Cu(A) and heme a exhibited superimposed E'(m) values, which decreased, for both metals, by around 20 mV/pH unit increase in the range 6.0-8.5. A model in which oxido-reduction of heme a and Cu(A) are both linked to the pK shifts of the two acid-base groups, characterized by the analysis of the pH dependence of the H(+)/heme a, Cu(A) ratios, provided a satisfactory fit for the pH dependence of the E'(m) of heme a and Cu(A). The results presented are consistent with a primary involvement of the redox Bohr effects shared by heme a and Cu(A) in the proton-pumping activity of cytochrome c oxidase.


Subject(s)
Carbon Monoxide/chemistry , Carbon Monoxide/pharmacology , Electron Transport Complex IV/chemistry , Electron Transport Complex IV/metabolism , Heme/analogs & derivatives , Animals , Cattle , Electron Transport , Electron Transport Complex IV/antagonists & inhibitors , Heme/metabolism , Hydrogen-Ion Concentration , Kinetics , Mitochondria, Heart/enzymology , Models, Chemical , Oxidation-Reduction , Protons
6.
Pathologica ; 92(2): 72-7, 2000 Apr.
Article in Italian | MEDLINE | ID: mdl-10838871

ABSTRACT

Venice had a long tradition and great reputation in the study of anatomical science dating back to the 1300's. The "Serenissima" Republic favoured the study and practice of anatomy as part of medical professional formation. Before the construction of the anatomical theater of San Giacomo dell'Orio, which took place in 1671, anatomical dissections were performed in churches, convents, hospitals and private homes. Even though Venice was not a University seat, it boosted numerous Venetian anatomists, among whom Benedetti, Massa, Santorini, and the medical activity of illustrious professors at the nearby University of Padua such as Vesalio, Falloppio, Spigelio, Vislingio and Morgagni.


Subject(s)
Anatomy/history , Attitude to Health , Catholicism/history , History, 15th Century , History, 16th Century , History, 17th Century , History, 18th Century , History, Medieval , Humans , Italy , Religion and Medicine , Social Change/history
7.
Breast ; 9(3): 139-43, 2000 Jun.
Article in English | MEDLINE | ID: mdl-14731837

ABSTRACT

Axillary lymph node dissection (ALND) is an important tool in staging patients with breast cancer. However, this procedure has several sequelae and complications and improvement in early diagnosis has led to an increasing number of cases of ALND in which axillary nodes are found to be negative. Sentinel node (SN) biopsy appears to be a less invasive alternative to ALND. The aim of the present study was to assess whether SN is a reliable indicator for axillary staging. We studied 126 consecutive patients with T1-T2 breast cancer and clinically negative axilla. In each case, 30-70 MBq of 99mTC-labelled colloidal albumin was injected subdermally close to the tumour and SN was visualised by lymphoscintigraphy. Surgery was performed 24 h after injection and the SN was removed under the guidance of a gamma ray-detecting probe. ALND was then undertaken in all cases. A histopathologic examination of the SNs was then made and the findings compared with the status of the other axillary nodes. SNs were identified and biopsied in 115/126 patients (91.3%) and correctly predicted the axillary status in 110/115 cases (95.6%). In five cases (4.4%), SNs were found to be negative, but other axillary nodes were positive. Our data confirm that SN biopsy is a good method for staging the axilla in patients with breast cancer. However, before SN biopsy can replace ALND in daily clinical practice, some technical aspects must be standardized, and clinical trials are required in order to clarify the prognostic impact of false-negative cases.

9.
Hum Reprod ; 13(12): 3406-13, 1998 Dec.
Article in English | MEDLINE | ID: mdl-9886524

ABSTRACT

This study was undertaken with the aim of investigating the cytogenetic constitution of normal as well as abnormal spermatozoa and immature germ cells found in semen of normal men and infertile patients. A specific protocol of double in-situ hybridization for chromosomes 1 and 17 based on colorimetric detection of the hybridization signals (ISH) and brightfield microscopy analysis of cellular morphology was applied. Also the influence of paternal age on sperm aneuploidy was investigated. We found that, at least in the age range analysed (28-54 years) and for semen of good quality (total normal motile counts above 10 x 10(6)) (n = 17), paternal age has no influence on baseline rates of sperm aneuploidy. However, with decreasing semen quality (total normal motile sperm counts below 5 x 10(6)) (n = 6) significantly higher rates of sperm aneuploidy for autosomes 1 and 17 were scored (0.8 versus 1.43%) (P < 0.001). Regardless of the type of semen analysed, a number of morphologically abnormal spermatozoa were found to be hyperhaploid or diploid in a high percentage of cases (20 and 10% respectively). The same was found for immature germ cells (aneuploidy rate of 18%). We conclude that in infertile men with poor quality semen a direct relationship may exist between the impairment of the spermatogenesis process (as reflected by an increased production of morphologically and cytogenetically abnormal germ cells) and rates of baseline aneuploidy occurring in normal spermatozoa. Infertile couples undergoing assisted reproduction treatment need to be counselled about the risk of using spermatozoa which may carry higher rates of non-disjunction for different chromosomes. While sperm hyper- or hypohaploidy for some chromosomes (X,Y) implies counselling couples about the risk of abnormal phenotype in their offspring, most autosomal sperm aneuploidies probably translate only into lower rates of embryo fertilization and survival.


Subject(s)
Aneuploidy , Chromosomes, Human, Pair 17 , Chromosomes, Human, Pair 1 , Infertility, Male/genetics , Infertility, Male/pathology , Spermatozoa/pathology , Adult , Humans , In Situ Hybridization , Male , Spermatogenesis/genetics
10.
Biochimie ; 80(10): 821-36, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9893941

ABSTRACT

In the last few years, evidence has accumulated supporting the applicability of the cooperative model of proton pumps in cytochrome systems, vectorial Bohr mechanisms, to heme-copper oxidases. The vectorial Bohr mechanism is based on short- and long-range protonmotive cooperative effects linked to redox transitions of the metal centers. The crystal structure of oxidized and reduced bovine-heart cytochrome c oxidase reveals, upon reduction, the occurrence of long-range conformational changes in subunit I of the oxidase. Analysis of the crystal structure of cytochrome c oxidase shows the existence of hydrogen-bonded networks of amino acid residues which could undergo redox-linked pK shifts resulting in transmembrane proton translocation. Our group has identified four proteolytic groups undergoing reversible redox-linked pK shifts. Two groups result in being linked to redox transitions of heme a3. One group is apparently linked to CuB. The fourth group is linked to oxido-reduction of heme a. We have shown that the proton transfer resulting from the redox Bohr effects linked to heme a and CuB in the bovine oxidase displays membrane vectorial asymmetry, i.e., protons are taken up from the inner aqueous space (N), upon reduction, and released in the external space (P), upon oxidation of the metals. This direction of proton uptake and release is just what is expected from the vectorial Bohr mechanism. The group linked to heme a, which can transfer up to 0.9 H+/e- at pHs around neutrality, can provide the major contribution to the proton pump. It is proposed that translocation of pumped protons, linked to electron flow through heme a, utilizes a channel (channel D) which extends from a conserved aspartate at the N entrance to a conserved glutamate located between heme a and the binuclear center. The carboxylic group of this glutamic acid, after having delivered, upon electron flow through heme a, pumped protons towards the P phase, once reprotonated from the N phase, moves to deliver, subsequently, to the binuclear center chemical protons consumed in the conversion of the peroxy to ferryl and of the latter to the oxy intermediate in the redox cycle. Site-directed mutagenesis of protolytic residues in subunit I of the aa3-600 quinol oxidase of Bacillus subtilis to non-polar residues revealed that the conserved Lys 304 is critical for the proton pumping activity of the oxidase. Crystal structures of cytochrome c oxidase show that this lysine is at the N entrance of a channel which translocates the protons consumed for the production of the peroxy intermediate. Inhibition of this pathway, by replacement of the lysine, short-circuits protons from channel D to the binuclear center, where they are utilized in the chemistry of oxygen reduction.


Subject(s)
Electron Transport Complex IV/metabolism , Heme/analogs & derivatives , Proton Pumps/chemistry , Animals , Cattle , Copper/metabolism , DNA Mutational Analysis , Heme/physiology , Hydrogen-Ion Concentration , Liver/enzymology , Mitochondria/enzymology , Models, Chemical , Models, Molecular , Myocardium/enzymology , Oxidation-Reduction , Rats
11.
Gynecol Endocrinol ; 11(5): 321-6, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9385531

ABSTRACT

This study aimed to standardize the clomiphene citrate test (CC-t) in our laboratory while comparing two different protocols of controlled ovarian stimulation in poor responders. One hundred and forty-four patients scheduled for assisted reproductive techniques were submitted to the CC-t within 3 months before starting stimulation; 133 underwent controlled ovarian stimulation with a blocking protocol. Poor responders in the first cycle (n = 30) were subsequently treated with a flare-up protocol. Although it was not statistically significant, more patients reached oocyte retrieval with the flare-up protocol. In the completed cycles, more gonadotropin ampules (55 +/- 15 vs. 34 +/- 13; p < 0.001) and more stimulation days (12.6 +/- 1 vs. 11.6 +/- 1.2; p < 0.005) were needed in the blocking than in the flare-up protocol. No difference was observed in peak 17 beta-estradiol levels, preovulatory follicles, oocytes retrieved or pregnancy rate between the two protocols. According to the threshold values, established on CC-t of patients who obtained a clinical pregnancy (n = 44), the incidence of abnormal results was 10%. All but one patient with abnormal CC-t were poor responders during the first stimulation cycle. The flare-up protocol did not improve the ovarian response in these patients.


Subject(s)
Clomiphene , Fertilization in Vitro , Gamete Intrafallopian Transfer , Ovulation Induction/methods , Adult , Buserelin/administration & dosage , Chorionic Gonadotropin/therapeutic use , Clomiphene/therapeutic use , Desogestrel/administration & dosage , Embryo Transfer , Estradiol/blood , Ethinyl Estradiol/administration & dosage , Female , Follicle Stimulating Hormone/blood , Follicle Stimulating Hormone/therapeutic use , Humans , Infertility/therapy , Menotropins/therapeutic use , Pregnancy , Triptorelin Pamoate/therapeutic use
12.
FEBS Lett ; 414(2): 414-8, 1997 Sep 08.
Article in English | MEDLINE | ID: mdl-9315731

ABSTRACT

The vectorial nature of redox Bohr effects (redox-linked pK shifts) in cytochrome c oxidase from bovine heart incorporated in liposomes has been analyzed. The Bohr effects linked to oxido-reduction of heme a and CuB display membrane vectorial asymmetry. This provides evidence for involvement of redox Bohr effects in the proton pump of the oxidase.


Subject(s)
Electron Transport Complex IV/chemistry , Electron Transport Complex IV/metabolism , Mitochondria, Heart/enzymology , Animals , Carbon Monoxide , Cattle , Copper , Heme/analogs & derivatives , Heme/metabolism , Hydrogen-Ion Concentration , Kinetics , Liposomes , Oxidation-Reduction
13.
Pathologica ; 89(4): 397-404, 1997 Aug.
Article in Italian | MEDLINE | ID: mdl-9471608

ABSTRACT

Three methods are currently available for defining the estrogen receptor (ER) status in breast carcinomas. These include biochemical ligand binding assay (LBA), immunohistochemical (IC) and mRNA in situ hybridization (ISH) semiquantitative analysis. There is still a debate as to which method should be considered the "gold standard" to define ER status. To address this topic we evaluated the above three methods in a series of 43 breast neoplasms. The results of IC (on fixed sections with ER1D5 immunostaining) and LBA assay showed moderate correlation (p = 0.004), as there were 8 (18.6%) discrepant results. ISH was extremely sensitive, with 92% of positive results. ISH results did not correlate with either IC or LBA. In view of the relative lack of reciprocal correlation among the three methods, it is difficult to define which is the most accurate system to be used to localize ER in breast carcinomas. The easiest and least expensive method probably should be used. The immunohistochemical approach has the advantage of being easily applicable to routinely fixed material, to cytological specimen and is very useful in small lesions identified with mammography only.


Subject(s)
Breast Neoplasms/chemistry , Carcinoma/chemistry , Estrogens , Immunoenzyme Techniques , In Situ Hybridization , Neoplasm Proteins/analysis , Neoplasms, Hormone-Dependent/chemistry , Radioligand Assay , Receptors, Estrogen/analysis , Antibodies, Monoclonal/immunology , Evaluation Studies as Topic , Female , Humans , Neoplasm Proteins/genetics , Neoplasm Proteins/immunology , RNA, Messenger/analysis , RNA, Neoplasm/analysis , Receptors, Estrogen/genetics , Receptors, Estrogen/immunology , Reproducibility of Results
14.
Mol Hum Reprod ; 3(5): 431-8, 1997 May.
Article in English | MEDLINE | ID: mdl-9239728

ABSTRACT

The purpose of the study was to analyse the frequency of sex-chromosome numerical abnormalities in human spermatozoa of infertile men by using a standardized experimental protocol of double target in-situ hybridization (ISH). The experiments were performed on decondensed sperm heads from 15 infertile patients (six cases of unexplained infertility and nine cases of severe oligoasthenoteratozoospermia). Three men of proven fertility were used as controls. The probes employed recognized the centromeric regions of human X chromosome and the long arm of the Y chromosome. In a smaller number of cases, additional experiments of double ISH were performed using centromeric probes for chromosomes 1 and 17. Signal detection was based on protocols of enzymatic cytochemical reactions. A total of 24,508, 24,679 and 42,285 cells were scored in the control, unexplained infertility and severe male factor groups of patients respectively. In all the patients in the ISH efficiency result was approximately 98%. In controls, unexplained infertility and severe male factor patients, the frequency of morphologically normal sperm cells carrying an abnormal chromosome constitution (XX or YY or XY or > 2 sex chromosomes signals) was 0.86, 0.75 and 1.35% respectively. The value of this last group of patients (severe male factor) was significantly higher than in the other two groups of patients (P < 0.008). The same findings were made using the autosomic probes. Our preliminary data support the possibility of an increased risk from paternal origin sex chromosome aneuploidies in children born after intracytoplasmic sperm injection (ICSI). Further investigations of the cytogenetic constitution of spermatozoa from severe male factor patients is warranted.


Subject(s)
Aneuploidy , Infertility, Male/genetics , Spermatozoa/abnormalities , Spermatozoa/ultrastructure , X Chromosome , Y Chromosome , Case-Control Studies , Cytoplasm , Fertilization in Vitro/adverse effects , Fertilization in Vitro/methods , Humans , In Situ Hybridization, Fluorescence/methods , Infertility, Male/therapy , Male , Microinjections , Risk Factors
15.
Biochim Biophys Acta ; 1318(1-2): 255-65, 1997 Jan 16.
Article in English | MEDLINE | ID: mdl-9030268

ABSTRACT

A study is presented of co-operative redox-linked protolytic reactions (redox Bohr effects) in soluble cytochrome-c oxidase purified from bovine-heart mitochondria. Bohr effects were analyzed by direct measurement, with accurate spectrophotometric and potentiometric methods, of H+ uptake and release by the oxidase associated with reduction and oxidation of hemes a and a3. CuA and CuB in the unliganded and in the CN- or CO-liganded enzyme. The results show that there are in the bovine oxidase four protolytic groups undergoing reversible pK shifts upon oxido-reduction of the electron transfer metals. Two groups with pKox and pKred values around 7 and > 12 respectively appear to be linked to redox transitions of heme a3. One group with pKox and pKred around 6 and 7 is apparently linked to CuB, a fourth one with pKox and pKred of 6 and 9 appears to be linked to heme a. The possible nature of the amino acids involved in the redox Bohr effects and their role in H+ translocation is discussed.


Subject(s)
Electron Transport Complex IV/metabolism , Mitochondria, Heart/enzymology , Animals , Cattle , Endopeptidases/metabolism , Hydrogen-Ion Concentration , In Vitro Techniques , Kinetics , Ligands , Oxidation-Reduction , Proton Pumps/metabolism , Solubility
16.
Clin Chem ; 42(10): 1634-8, 1996 Oct.
Article in English | MEDLINE | ID: mdl-8855147

ABSTRACT

The determination of tumor markers in urine samples has been proposed as an effective diagnostic tool in bladder cancer. The aim of the present investigation was to validate in urine samples the assay of the CYFRA21.1 cytokeratin-related marker, the serum concentrations of which showed promising diagnostic utility in patients with bladder cancer. First-voided urine samples were collected from patients with different malignancies. CYFRA21.1 was assayed with a commercially available enzyme immunoassay (Boehringer Mannheim). Different centrifugation patterns, the use of different buffers and nonionic detergents, and pH variations were evaluated. We demonstrated that: (a) cells and cell debris contain a large amount of CYFRA21.1 and must be eliminated by centrifugation; (b) storage at -20 degrees C causes amorphous precipitate, which may aspecifically bind CYFRA21.1; (c) the latter behavior may be prevented by diluting fresh urine samples with phosphate buffer with nonionic detergent added; (d) pH variations within the range 4.9-8.2 do not significantly affect CYFRA21.1 assay results. Provided that samples are diluted with buffer containing nonionic detergent, the CYFRA21.1 assay showed good precision and accuracy characteristic in urine samples. We therefore propose a standard protocol for the collection of urine samples for CYFRA21.1 assay. In a preliminary clinical evaluation, CYFRA21.1 concentrations in 16 patients with primary bladder cancer were higher than in healthy subjects. In the urine collected in the follow-up of patients treated for bladder cancer, CYFRA21.1 tended to be higher in relapsed patients than in those without evidence of disease. These preliminary data induced us to extend the clinical trial to establish the actual role of this assay in routine use.


Subject(s)
Antigens, Neoplasm/urine , Biomarkers, Tumor/urine , Urinary Bladder Neoplasms/urine , Buffers , Centrifugation , Detergents , Freezing , Humans , Hydrogen-Ion Concentration , Immunoenzyme Techniques , Keratin-19 , Keratins , Neoplasm Recurrence, Local/urine , Reproducibility of Results , Sensitivity and Specificity , Urologic Diseases/urine
17.
Hum Reprod ; 11(8): 1638-43, 1996 Aug.
Article in English | MEDLINE | ID: mdl-8921108

ABSTRACT

A brightfield microscopical in-situ hybridization (ISH) technique was applied to semen samples of two 47,XYY males, one 46,XY/47,XXY/XXY male with fertility problems, and two normal 46,XY men, who served as controls. The use of a standardized nuclear DNA decondensation method, together with double-target ISH and morphological staining, allowed an accurate study of the sex chromosomal content and morphology of spermatozoa. In the males carrying an extra sex chromosome, we detected X- and Y-bearing spermatozoa in a ratio which did not differ significantly from the 1:1 ratio found in normal males. Aneuploidy for the sex chromosomes was found in approximately 15% of the spermatozoa of both XYY males and in 3% of the XXY male. The most striking finding was the relatively low percentage of spermatozoa in these patients, with an average of 65% in the XYY males and 84% in the XXY male. The other cells represented immature germ cells (IGC), including spermatogonia and spermatocytes arrested at various stages of spermatogenesis. Apparently, in XYY or XXY men, these IGC are shed into the semen to an increased extent as compared to normal, fertile men. The sex chromosome constitution of these IGC was heterogeneous. However, the finding that the majority of spermatozoa in semen of 47,XYY and 47,XXY males carried a single sex chromosome strengthens the hypothesis that a 46,XY germ cell line must be present, apparently with a proliferative advantage over the 47,XYY or 47,XXY cells.


Subject(s)
Chromosome Aberrations/diagnosis , Semen/physiology , Adult , Cellular Senescence , Chromosome Aberrations/physiopathology , Chromosome Disorders , Humans , In Situ Hybridization , Male , Sex Chromosomes/physiology , Spermatozoa/cytology , Spermatozoa/physiology
18.
Biochemistry ; 35(33): 10800-6, 1996 Aug 20.
Article in English | MEDLINE | ID: mdl-8718871

ABSTRACT

A study is presented of the factors affecting the H+/e- stoichiometry of the proton pump of mitochondrial cytochrome c oxidase, isolated and reconstituted in phospholipid vesicles (COV). Under level flow conditions, i.e., in the absence of a transmembrane delta muH+, the H+/e- ratio, obtained from spectrophotometric measurements of the initial rates of electron flow and H+ release specifically elicited by cytochrome c, varied from around 0 to 1, depending on the actual rate of electron flow through the oxidase. At steady state the H+/e- ratio for the oxidase was specifically depressed by the transmembrane delta pH. The study of the H+/e- ratio of the pump was complemented by an analysis of the redox pattern of cytochrome c, CuA, and heme a. From both sets of results and recent structural data from other groups, it is concluded that the dependence of the H+/e- ratio on the rate of electron flow through the oxidase and transmembrane delta pH is associated with the possible occurrence of two electron transfer pathways in cytochrome c oxidase, a coupled one (cyt c-->CuA-->heme a-->heme a3-CuB) and a decoupled one (cyt c-->CuA-->heme a3-CuB). The contributions of the two pathways, differently affected by kinetics and thermodynamic factors, will determine the actual H+/e- ratio of the pump. A possible role of heme a in the proton pump and the physiological implication of the variable H+/e- ratio in the oxidase are discussed.


Subject(s)
Electron Transport Complex IV/metabolism , Mitochondria, Heart/enzymology , Animals , Cattle , Electron Transport , Hydrogen-Ion Concentration , Intracellular Membranes/enzymology , Intracellular Membranes/metabolism , Kinetics , Mitochondria, Heart/metabolism , Oxidation-Reduction , Protons
19.
Breast Cancer Res Treat ; 37(3): 267-76, 1996.
Article in English | MEDLINE | ID: mdl-8825138

ABSTRACT

Four different methods to measure in parallel the erbB2 protein expression (p185neu) were evaluated in order to: a) compare two enzyme immunoassays with the immunohistochemical assays (IHC) and western blotting (WB) and b) extrapolate eventual relationships between erbB2 and biological parameters. Tissue samples from 248 patients with primary breast cancer were consecutively assayed. We used two different cut-off levels for WB, ELISA, and EIA, defined as follows: 1) the highest level of expression of non malignant tissue was chosen as the discriminant threshold between 'low' and 'elevated' samples: 2) the elevated group was further subdivided into two subgroups: 'intermediate' and 'high', according to their median value. According to the first cut-off, the results were considered 'elevated' in about 52% of cases with the three biochemical methods, while using the second cut-off the percentage lowered to about 26%. Considering this cut-off, the concordance rates between the paired biochemical methods ranged between: 78.4% (WB vs EIA), 93% (ELISA vs EIA), and 82.6% (ELISA vs WB). The comparison between biochemical and immunohistochemical methods gave these concordance rates: 82% (WB vs IHC), 90.5% (ELISA vs IHC), and 85.5% (EIA vs. IHC). According to the first cut off level, 27.5% of tumor samples showed IHC detectable p185 levels, in agreement with other immunohistochemical studies. The relationship between high erbB2 and estrogen and progesterone receptors showed an inverse association. No relationship was found between erbB2 and axillary lymph node positivity or tumor size. In short, the results of the four methods seem generally well correlated; nevertheless, it appears that different methodological approaches of measuring p185neu are not completely equivalent, and there is a need for an authoritative standardization and quality control for clinical applications.


Subject(s)
Breast Neoplasms/chemistry , Receptor, ErbB-2/analysis , Blotting, Western , Breast Neoplasms/pathology , Female , Humans , Immunoenzyme Techniques , Immunohistochemistry , Receptors, Estrogen/analysis , Receptors, Progesterone/analysis
20.
Obstet Gynecol ; 85(6): 1011-6, 1995 Jun.
Article in English | MEDLINE | ID: mdl-7770246

ABSTRACT

OBJECTIVE: To evaluate the clinical usefulness of a computer based system that stores and analyzes high-resolution digital colposcopic images. METHODS: Three hundred patients referred to our institutions for evaluation of abnormal cervical cytology underwent colposcopy with development of digital images. The first 70 patients were used to standardize the system. The images obtained from 188 evaluable patients were analyzed and scored by software according to internationally recognized colposcopic criteria. The results were compared with traditional colposcopic diagnoses and with corresponding histology results using kappa statistics for inter-observer agreement and McNemar test for significance. RESULTS: The exact concordance rate between computer-aided impressions and histology (85.1%, kappa = 0.77) was significantly higher (P < .001) than that observed between traditional colposcopic findings and histology (66%, kappa = 0.40). The computer-assisted colposcopy was much more accurate than traditional colposcopy in diagnosing high-grade lesions (91.2 versus 61.8% of exact concordance, P < .001) as well as normal histology (74.1 versus 34.5%, P < .001), but not significantly different when evaluating low-grade lesions (89.6 versus 86.5%). CONCLUSION: The integration of computer imaging and colposcopy can improve the colposcopic diagnostic accuracy. An inexperienced colposcopist may benefit from computerized support to obtain the most appropriate histologic specimen, and eventually access to distant consultation via modem or through on-line services. An additional advantage is the ability to develop a space-saving permanent record of digitized images readily available to review a patient's cervical history or perform effective programs of quality control in colposcopy.


Subject(s)
Colposcopy/methods , Diagnosis, Computer-Assisted , Uterine Cervical Dysplasia/pathology , Uterine Cervical Neoplasms/pathology , Adolescent , Adult , Confidence Intervals , Female , Humans , Middle Aged , Pilot Projects , Predictive Value of Tests , Sensitivity and Specificity
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