Evaluation of different approaches for fetal DNA analysis from maternal plasma and nucleated blood cells.
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DNA↗
Biomedical subjects
A Ferrari
Publications and source records attributed to A Ferrari.
Cardiac abnormalities in the acquired immunodeficiency syndrome. A prospective study with a clinical-pathological correlation in twenty-one adult patients.
OBJECTIVE: To evaluate the cardiac abnormalities and their evolution during the course of the acquired immunodeficiency syndrome, as well as to correlate clinical and pathological data. METHODS: Twenty-one patients, admitted to the hospital with the diagnosis of acquired immunodeficiency syndrome, were prospectively studied and followed until their death. Age ranged from 19 to 42 years (17 males). ECG and echocardiogram were also obtained every six months. After death, macro- and microscopic examinations were also performed. RESULTS: The most frequent causes of referral to the hospital were: diarrhea or repeated pneumonias, tuberculosis, toxoplasmosis or Kaposi sarcoma. The most frequent findings were acute or chronic pericarditis (42%) and dilated cardiomyopathy (19%). Four patients died of cardiac problems: infective endocarditis, pericarditis with pericardial effusion, bacterial myocarditis and infection by Toxoplasma gondii. CONCLUSION: Severe cardiac abnormalities were the cause of death in some patients. In the majority of the patients, a good correlation existed between clinical and anatomical-pathological data. Cardiac evaluation was important to detect early manifestations and treat them accordingly, even in asymptomatic patients.
Acquired Immunodeficiency Syndrome↗
Cardiac abnormalities in the acquired immunodeficiency syndrome. A prospective study with a clinical-pathological correlation in twenty-one adult patients.
OBJECTIVE - To evaluate the cardiac abnormalities and their evolution during the course of the acquired immunodeficiency syndrome, as well as to correlate clinical and pathological data. METHODS - Twenty-one patients, admitted to the hospital with the diagnosis of acquired immunodeficiency syndrome, were prospectively studied and followed until their death. Age ranged from 19 to 42 years (17 males). ECG and echocardiogram were also obtained every six months. After death, macro- and microscopic examinations were also performed. RESULTS - The most frequent causes of referral to the hospital were: diarrhea or repeated pneumonias, tuberculosis, toxoplasmosis or Kaposi sarcoma. The most frequent findings were acute or chronic pericarditis (42%) and dilated cardiomyopathy (19%). Four patients died of cardiac problems: infective endocarditis, pericarditis with pericardial effusion, bacterial myocarditis and infection by Toxoplasma gondii. CONCLUSION - Severe cardiac abnormalities were the cause of death in some patients. In the majority of the patients, a good correlation existed between clinical and anatomical-pathological data. Cardiac evaluation was important to detect early manifestations and treat them accordingly, even in asymptomatic patients.
Journal Article↗
Serum time course of naltrexone and 6 beta-naltrexol levels during long-term treatment in drug addicts.
The pharmacokinetics of naltrexone have been scarcely explored in patients during chronic treatment despite the observation that the pharmacological effect of the drug is related to its plasma concentrations. In this study we investigated the time course of serum levels of naltrexone and its active metabolite, 6 beta-naltrexol, in 13 heroin addicts (3 F, 10 M; age 22-32 years) in the 24 h after 100 mg of naltrexone orally. Six patients were studied once, at different times during chronic treatment, whereas in seven patients the study was done at the beginning and after 1 month of naltrexone treatment. Four of these patients also repeated the study after 3 months of naltrexone treatment. Serum naltrexone and 6 beta-naltrexol were assayed by GLC with a nitrogen-phosphorus detector. Our results showed large differences among patients in serum naltrexone and 6 beta-naltrexol levels. On the other hand, there were no differences in serum time course of both substances in the same patient over 3 months. Peak levels and AUCs of naltrexone were lower than those of 6 beta-naltrexol in ten addicts and higher than those of the metabolite in three patients. No significant differences in the apparent half-lives of the two drugs were detected among groups. These data are consistent with the occurrence of a decreased first-pass metabolism of naltrexone in three patients leading to a larger availability of an oral dose. The increased bioavailability of the drug is not very important for opioid receptor antagonist activity but may play a role in naltrexone treatment safety.
Administration, Oral↗
The level of urokinase-type plasminogen activator receptor is increased in serum of ovarian cancer patients.
Ascites and serum of patients with ovarian carcinoma contain a soluble form of urokinase-type plasminogen activator receptor (uPAR). We now report that pro-uPA-Sepharose-purified uPAR from ascites of patients with ovarian carcinoma is the full-length molecule missing the glycosyl-phosphatidylinositol anchor, as determined by its amino acid composition. We next examined the significance of determining serum soluble uPAR (suPAR) levels in ovarian cancer patients using a specific ELISA and compared the results with serum concentrations of CA-125, an established diagnostic marker. Serum from pre- and postoperative ovarian cancer patients was assayed for suPAR and CA-125. The majority of the patients with ovarian cancer had enhanced preoperative serum levels of suPAR compared with healthy controls, but suPAR concentrations decreased after operation. Although uPAR was associated with most ovarian carcinomas, it appeared to be a less specific indicator for ovarian cancer than CA-125. On the other hand, suPAR was more specific for other types of solid tumors. Moreover, we have observed some cases of ovarian cancer that showed increase of suPAR but not of CA-125. The prognostic significance of serum suPAR assay for survival of ovarian carcinoma patients was evaluated using Cox's proportional hazards analysis. Our preliminary data show that high preoperative levels of suPAR were associated with worse survival of the patients, whereas CA-125 had no prognostic implications. This is the first report evaluating the assay of serum suPAR levels in ovarian cancer and analyzing its value as a tumor or prognostic marker.
Amino Acids↗
Chromogranin A and B and secretogranin II in prostatic adenocarcinomas: neuroendocrine expression in patients untreated and treated with androgen deprivation therapy.
BACKGROUND: Neuroendocrine (NE) expression in prostatic adenocarcinomas (PACs) has been related to an adverse clinical course, but the reported data are not unequivocal. METHODS: We immunostained a series of 64 PACs with three monoclonal antibodies raised against chromogranin A (CgA), chromogranin B (CgB), and secretogranin II (SgII). The patients were followed up for 18-88 months (mean 43 months, standard deviation +/- 20 months); 58 of them received preoperative androgen deprivation therapy for 3-6 months. RESULTS: Of the 64 PACs under study, 39 (approximately 61%) were immunoreactive to CgA, 51 (approximately 80%) to CgB, and 38 (approximately 59%) to SgII. We found a strict correlation between pronounced neuroendocrine differentiation and the most poorly differentiated tumors (P = 0.01 for CgA, P = 0.03 for CgB, and P = 0.05 for SgII), and relationship (approaching statistical significance only for CgB, P = 0.07) between Cgs/Sg expression and advanced (C and D) clinical stage. However, we failed to detect any correlation between chromogranin expression and clinical outcome. CONCLUSIONS: These results suggest that NE differentiation is a frequent event in PACs, especially in the most poorly differentiated. Nevertheless, as Cgs/Sg expression is not clearly related to advanced clinical stage and poor prognosis, our findings suggest that clinical staging and grading, rather than NE differentiation, remain the most powerful prognostic indicators in PACs.
Adenocarcinoma↗
Perfluorocarbon broncho-alveolar lavage and liquid ventilation versus saline broncho-alveolar lavage in adult guinea pig experimental model of meconium inhalation.
OBJECTIVE: This study compares perfluorocarbon broncho-alveolar lavage (PFC-BAL) with isotonic saline broncho-alveolar lavage (saline-BAL) in an experimental model of meconium aspiration in adult guinea pigs. DESIGN: Prospective controlled experimental study. PATIENTS AND METHODS: Ten male guinea pigs were given 1 ml of human meconium, diluted to 10% in isotonic saline, via tracheostomy and then artificially ventilated. After stabilization, five animals (control group) underwent BAL with 10 ml/ kg isotonic saline solution. After bronchial suctioning, a further BAL with 2 ml/kg saline was performed. The other five animals (study group) underwent BAL with 100 ml/kg of PFC (RIMAR 101). Bronchosuction was effected at 5 min and then a BAL with PFC 2 ml/kg was performed. Both groups received conventional mechanical ventilation during the lavage procedures. Within 20 min all the saline-BAL treated animals died, whereas the PFC-BAL treated animals survived and were then treated with total liquid ventilation (TLV) by gravity. MEASUREMENTS AND RESULTS: After meconium inhalation in both groups, a large alveolar-arterial oxygen difference, hypercarbia, severe acidosis and tachycardia were noticed. In PFC treated animals, an improvement in blood gases was noted and acid-base balance remained stable compared to saline-BAL treated animals. No haemodynamic change was observed during or after PFC-BAL, while during saline-BAL there was evidence of bradycardia, hypotension and respiratory failure, which led to the death of the animals. The histological lung sections in the PFC-BAL group showed evidence of normal alveolar expansion with a minimal presence of meconium debris in the small bronchioles. In saline-BAL treated animals, the lung structure appeared severely compromised with the presence of meconium in bronchioles and alveoli, intra-alveolar oedema and haemorrhagic areas. CONCLUSIONS: This study confirms the detrimental effects of meconium aspiration and the impairment of lung function following saline-BAL. By contrast, PFC-BAL and TLV by gravity seemed to prevent absorption of meconium, facilitate its removal, improve gas exchange and reduce lung barotrauma.
Acute Disease↗
Multiple testing in fetal gender determination from maternal blood by polymerase chain reaction.
Fetal male DNA can be identified in maternal blood by polymerase chain reaction (PCR) amplification of Y-specific sequences. This technology has not reached a satisfactory accuracy and reproducibility in fetal gender determination because of the very low concentration of fetal cells. Our purpose was to evaluate the possibility of improving the reliability of this test by setting up a repeated amplification system. We amplified, by nested PCR of the Y-specific sequence DYS14, 137 DNA samples extracted from maternal peripheral blood (93 from male-bearing and 44 from female-bearing pregnancies ranging from the 6th to the 36th gestational week). Each maternal DNA sample was tested doubly, in two different PCR sessions, with a total of four amplifications. We obtained discordant results in the four amplifications in 82/137 (60%) samples. The best interpretation of these discordant results was obtained by applying a positivity cutoff of at least two positive amplifications for considering a DNA sample as belonging to a male-bearing pregnancy. We obtained a sensitivity of 83%, a specificity of 93%, a positive predictive value of 96% and a negative predictive value of 72% in fetal male gender diagnosis. By applying this quadruple testing system, we significantly improved PCR accuracy and predictive values compared with single and double testing of the same samples. We conclude that, for future investigations of fetal DNA retrieved from maternal blood, the application of a quadruple testing system is better than the single PCR test.
DNA↗
Results of the routine use of a modified endoprosthesis to drain the common bile duct after laparoscopic choledochotomy.
BACKGROUND: One hundred eighty-one patients were submitted to laparoscopic common bile duct exploration. METHODS: A transcystic approach was used in 147 patients, choledochotomy in 14, and both in 20. The indications to perform a choledochotomy included stones larger than 20 mm, stones proximal to the cystic duct entrance, and cases in which the transcystic duct approach proved impossible or unsuccessful. RESULTS: The common bile duct was drained by a T-tube in four patients, by laparoscopic sphincterotomy in one, by laparoscopic choledochoduodenostomy in one, and by a 10 Fr endoprosthesis in 28. The stent placement was technically feasible in all patients but one. The biliary drainage was adequate. Mean hospital stay was 2.1 days. Complication was limited to one umbilical infection and one self-limited biliary leak. CONCLUSIONS: The procedure proved to be technically simple, safe, and efficient, and resulted in a low morbidity rate and short hospital stay.
Adolescent↗
Soluble intercellular adhesion molecule-1 in ovarian follicles: production by granulosa luteal cells and levels in follicular fluid.
OBJECTIVE: To determine the concentration of the soluble form of intercellular adhesion molecule (ICAM)-1 in granulosa luteal cell-conditioned media and in follicular fluid (FF). DESIGN: Granulosa cells and FF samples were obtained at the time of oocyte retrieval for IVF. In 10 women, a total of 33 fluids were obtained from individual follicles, whereas in 70 women, the follicular aspirates were pooled. SETTING: Clinica "L. Mangiagalli" and Reproductive Center, San Raffaele Hospital, Milan, Italy. PATIENT(S): Eighty women referred for IVF for tubal factor or male factor infertility. INTERVENTION(S): Women underwent ovarian hyperstimulation. MAIN OUTCOME MEASURE(S): Soluble ICAM-1 was measured by an ELISA, and its levels were correlated with follicular size, the number of retrieved oocytes, and the number of follicles with a diameter of >15 mm. RESULT(S): The concentration of soluble ICAM-1 in granulosa luteal cell-conditioned media was 17.8 +/- 1.8 ng/5 x 10(5) cells. Interleukin-1beta can stimulate soluble ICAM-1 release in a dose-dependent manner. A significant positive correlation was demonstrated between levels of soluble ICAM-1 in pooled FF and the number of retrieved oocytes or the number of follicles with a diameter of >15 mm. CONCLUSION(S): Soluble ICAM-1 can be released by granulosa luteal cells and can be detected in FF after ovarian hyperstimulation. Levels of soluble ICAM-1 in FF correlate directly with some indices of ovarian function.
Cells, Cultured↗
The management of paratesticular rhabdomyosarcoma: a single institutional experience with 44 consecutive children.
PURPOSE: We report a single institutional experience with 44 consecutive children treated from 1976 to 1996 for paratesticular rhabdomyosarcoma. MATERIALS AND METHODS: After primary surgical excision, 27 cases were classified as Intergroup Rhabdomyosarcoma Study (IRS) group I, 1 as group II, 8 as group III and 8 as group IV. Except for the first 5 patients, retroperitoneal node dissection was not a required staging procedure and was performed only for radiologically suspicious node metastases. Adjuvant chemotherapy was administered to all patients. Additional radiotherapy was given when nodal involvement or residual disease was documented. RESULTS: The 5-year event-free survival rate was 77% for the entire series of patients, 93% for those in groups I and II, 62.5% in group III and 37.5% in group IV with a median followup of 11 years (range 2 to 20). CONCLUSIONS: Our report confirms the excellent prognosis of localized paratesticular rhabdomyosarcoma. Due to the accuracy of computerized tomography potential morbidity of lymphadenectomy, low rate of retroperitoneal recurrence and presumed efficacy of chemotherapy for controlling microscopic disease, retroperitoneal lymph node dissection seems unnecessary for localized disease.
Adolescent↗
Sclerosing stromal tumor of the ovary. Report of eight cases and review of the literature.
In this article, we describe eight cases of sclerosing stromal tumors (SST) of the ovary and review the literature. We could not demonstrate unequivocal hormonal activity in any of the cases, although suggestive evidence for it has been reported in the literature in the form of clinical, histologic, electron microscopic and immunohistochemical findings.
Adolescent↗
No evidence of WT1 involvement in a Burkitt's lymphoma in a patient with Denys-Drash syndrome.
BACKGROUND: We previously reported the case of a patient affected with Denys-Drash syndrome (DDS), who developed disseminated EBV-related Burkitt's lymphoma (BL) after kidney transplantation. Here, we describe the molecular characterisation of the WT1 gene in the constitutional and tumour DNA of this patient. PATIENTS AND METHODS: WT1 exons 2 to 10 were sequenced in constitutional and tumour DNAs. By Southern blotting the latter was also investigated for the presence of gene rearrangements. Gene expression analysis in tumour cells was performed by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: A germline missense mutation affecting one of the zinc finger domains of the gene, and previously reported in other DDS cases, was observed. No alterations of the constitutionally wild-type WT1 allele and no expression of the gene were observed in BL cells. A small group of BLs from other paediatric patients showed a variable expression of WT1. CONCLUSIONS: Our findings indicate that WT1 is unlikely to be involved in the onset of BL in our case. However, a possible role of the gene in at least a subset of these lymphoproliferative diseases may be suggested.
Abnormalities, Multiple↗
Secondary haematological neoplasm after treatment of adult acute lymphoblastic leukemia: analysis of 1170 adult ALL patients enrolled in the GIMEMA trials. Gruppo Italiano Malattie Ematologiche Maligne dell'Adulto.
Between 1983 and 1994 the incidence of secondary haematological neoplasms (SHM) was evaluated in 1170 new cases of ALL enrolled in the GIMEMA trials. Of the 942 patients who achieved complete remission (CR); seven developed a SHM: four AMLs and three NHLs. The median latency from onset of ALL and of secondary haematological neoplasm was 69 months for AML and 61 months for NHL. Three out of four patients with secondary AML were unresponsive to the new chemotherapy and died, whereas the fourth patient achieved a new CR. Among the three NHL cases, two patients are presently alive in CR, whereas the third patient was refractory to chemotherapy and died. The relative risk of haematological malignancy among the GIMEMA trials population, as compared to that of the Italian Cancer Registries, was 15.25-fold higher, and the actuarial estimated cumulative proportion of ALL patients with a secondary haematological neoplasm at 5 and 10 years were 0.59% and 3.63% respectively. The incidence of adult ALL who developed a SHM, although apparently lower than in the paediatric ALL series, was higher when compared to the normal population. The difference between paediatric and adult ALL is probably due to the lack of craniospinal radiotherapy and to the lower doses of epipodoxiphyllotoxins used in adult trials. The higher percentage of childhood ALL with a prolonged event-free survival could result in an increase of secondary neoplasms in these cases, which suggests that secondary haematological neoplasms in adult ALL patients are real, although rare, events.
Adolescent↗
Endogenous dopaminergic activity in Child-Pugh A cirrhosis: potential role in renal sodium handling and in the maintenance of clinical compensation.
BACKGROUND: We studied the main determinants of aldosterone secretion in a group of 20 patients with biopsy-proven Child-Pugh A cirrhosis without previous ascites or diuretic consumption. METHODS: We evaluated the plasma levels of adrenocorticotrophic hormone (ACTH), active renin and aldosterone (both supine at 07.00 h and after 30 min of upright posture),and active renin and aldosterone responses 30 min and 60 min after the administration of metoclopramide, a dopamine DA2 antagonist (10 mg e.v.). Nine normal subjects were also submitted to the metoclopramide stimulation test. RESULTS: Compared with control subjects, the patients showed significantly greater incremental aldosterone responses both 30 min and 60 min after metoclopramide (+30 min: 157.5+/-73.3 vs. 83.5+/-32.2 pg mL(-1), P< 0003; +60 min: 142.1+/-87.2 vs. 36.8+/-39.0 pg mL(-1), P < 0-001). We found significant positive correlations between amplitude of aldosterone response 30 min after metoclopramide and 24-h urinary fractional excretion of sodium (r=0.61, P < 0.01) and basal morning aldosterone levels (r=0.69, P < 0.001). CONCLUSIONS: The higher incremental aldosterone responses observed after metoclopramide in cirrhotic patients are expressions of increased dopaminergic activity in these patients compared with control subjects. Moreover, the correlation we found between the degree of dopaminergic activity and 24-h urinary fractional excretion of sodium suggests a role for endogenous dopamine as a relevant mediator of natriuresis in cirrhosis, at least in patients with compensated disease.
Adrenocorticotropic Hormone↗
Non-variceal upper gastrointestinal bleeding and Forrest's classification: diagnostic agreement between endoscopists from the same area.
BACKGROUND AND STUDY AIMS: The lack of uniformity in defining the stigmata of hemorrhage in patients with bleeding ulcers is suggested by the wide range among published studies in prevalence and rebleeding rates for the same stigmata. Moreover there is, in published trials of endoscopic hemostasis, little standardization of definitions of stigmata of hemorrhage. The aim of this study was to assess the interobserver agreement among endoscopists from the same area (Piedmont and Valley of Aosta). PATIENTS AND METHODS: A workshop for 47 expert endoscopists was organized in order to evaluate their agreement in the diagnosis of stigmata of recent hemorrhage, according to Forrest's classification. During the meeting 25 videotapes from endoscopic examinations of patients with recent non-variceal bleeding were shown to the 47 endoscopists, who were asked to classify every lesion. RESULTS: The overall and beyond chance interobserver agreement was calculated by means of the kappa statistic. The overall agreement among endoscopists was highly significant (p < 0.001, kappa=0.60), while the beyond chance agreement varied from excellent to good for lesions with active bleeding (kappa=0.76 and kappa=0.61 for FIA and FIB lesions respectively), whereas for lesions with stigmata of recent hemorrhage kappa varied from 0.44 to 0.49. CONCLUSIONS: These data suggest the need for better knowledge of endoscopic criteria, in order to evaluate the results of endoscopic therapy and to assess new treatments.
Education↗
Feto-neonatal ultrasonography to detect renal abnormalities: evaluation of 1-year screening program.
To determine the accuracy of antenatal renal pelvis measurement as a screening tool for renal anomalies we reviewed the antenatal ultrasounds performed at our institution over 1 year. Renal pelvis dilation was found in 3.8% of pregnancies followed by our obstetrics department. In nine (17.6%) infants renal anomalies were detected after birth, the remaining cases remained unchanged or resolved over a 1-year postnatal follow-up. In our series dilations of less than 10 mm in utero resolved in the first months after birth while dilations greater than 15 mm were all associated with malformations of the genitourinary tract. The choice of the most appropriate follow-up for dilations between 10 and 15 mm remains an issue because these dilations can herald renal anomalies.
Dilatation, Pathologic↗
Polycystic ovary syndrome: anomalies in progesterone production.
The underlying cause of anovulation and miscarriage in polycystic ovary syndrome (PCOS) is unknown. Progesterone may play an important role in oocyte fertilization and embryo implantation. Therefore, in this study we analyse the endocrine function of luteinizing granulosa cells to synthesize progesterone in vivo and in vitro in PCOS and normal patients participating in an in-vitro fertilization programme. Human luteinizing granulosa cells were obtained from 10 patients with normal ovaries (controls) and 10 patients with PCOS by follicular aspiration of individual follicles of each patient and pooled in an attempt to obtain three groups: cells from follicle sizes < or =10,>10< or =15 and > or =16. Serum concentrations of oestradiol and progesterone on the day of human chorionic gonadotrophin (HCG) injection were significantly higher (P < 0.01 and P < 0.05) in PCOS patients than in controls. After HCG stimulation, in-vitro progesterone production was enhanced in granulosa cells of the control group and concentrations increased with follicular size as expected. However, the concentration of progesterone of PCOS patients did not increase with follicular size and there was a significant difference between normal and PCOS groups in follicles >10< or =15 mm (P < 0.05) and > or =16 mm (P < 0.01). Oestradiol production was increased in follicles > or =16 mm in both groups, although this did not reach significance. In summary, it seems that PCOS granulosa cells demonstrate an abnormal capacity to synthesize progesterone in vivo and in vitro. The understanding of granulosa cell function in PCOS may explain the anovulation and miscarriage that occurs in these patients.
Abortion, Spontaneous↗