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Biomedical subjects

G Pasquini

Publications and source records attributed to G Pasquini.

At least 19 recordsLinked to original sources
PubMed2001

Preoperative lymphoscintigraphy and sentinel lymph node biopsy in papillary thyroid cancer. A pilot study.

BACKGROUND AND OBJECTIVES: Lymphadenectomy for papillary thyroid cancer is a matter of debate. After showing its usefulness as a prognostic factor in both melanoma and breast cancer, the concept of sentinel lymph node biopsy was also recently applied to differentiated thyroid cancer. To date, all attempts to locate and remove the sentinel node were based on the intraoperative injection of a vital dye. The feasibility and the technical details of using preoperative lymphoscintigraphy coupled with intraoperative vital dye and gamma probe scanning were investigated and discussed. METHODS: Six patients diagnosed with papillary thyroid cancer were submitted to preoperative lymphoscintigraphy with (99m)Tc-labelled colloidal albumin at different dosages. The operation consisted in a total thyroidectomy with sentinel lymph node biopsy guided by intraoperative injection of a vital dye (Blu Patent V, 2.5%) and scanning with a hand-held gamma probe. Lymph node dissection was completed in the area in which the sentinel node was located. RESULTS: The sentinel node was identified using all the three methods in all cases (100%). Considering one of the methods alone, identification rates were 66, 50, and 83% for preoperative lymphoscintigraphy, vital dye, and probe scanning, respectively. One sentinel node was identified in four cases and two in the other two cases. The optimal dosage of the tracer appeared to be at 22 MBq. CONCLUSIONS: These results underline the necessity to use the combination of nuclear medicine imaging and lymphatic vital dye in order to enhance the identification rate of sentinel node also in thyroid cancer. It is now necessary to check the diagnostic accuracy of this procedure through a controlled trial involving a more extended lymph node dissection in the neck.

Adult↗
PubMed2000

TP53 mutations and mdm2 protein overexpression in cholangiocarcinomas.

Tumor suppressor protein p53 is a positive regulator of MDM2 gene expression and the mdm2 protein can bind to p53, preventing the transactivation of p53 responsive genes, thus mimicking TP53 mutation. The authors looked for alterations that could affect, directly and indirectly, p53 function in 13 patients with extrahepatic cholangiocarcinoma. Molecular analysis by single strand conformation polymorphism and DNA sequencing revealed that TP53 gene mutations occurred in only 2 of 13 cholangiocarcinomas. High levels of mdm2 protein were found, by immunohistochemical staining, in 61% of the cholangiocarcinomas and in almost all specimens (70%) displaying stabilized p53 protein in the absence and in the presence of TP53 mutations. The finding of co-overexpressed mdm2 and p53 proteins in cholangiocarcinomas indicates that they can upregulate the expression of mdm2 protein to a level sufficient for binding and accumulating p53 in a presumably inactive complexed form. The presence of TP53 mutations or upregulation of MDM2 gene expression in 9 of the 13 cholangiocarcinomas strongly supports that the impairment of the p53 pathway is an important and specific step in cholangiocarcinoma pathogenesis. At variance with other authors, no alteration of p16ink4/CDKN2 gene was observed in all 13 cholangiocarcinomas.

Aged↗
PubMed1998

Comparison of different methods of RNA isolation for plum pox virus detection by reverse transcription-polymerase chain reaction.

The diagnosis of plum pox virus (PPV) is still considered one of the most important aspects of the "sharka" problem. In fact, different studies demonstrated an uneven distribution of the virus in infected trees due to a high variability in virus concentration. These aspects complicate the PPV diagnosis. To date, biological, serological and molecular assays have been successively developed in order to obtain sensitive and efficient PPV detection techniques. In particular, the polymerase chain reaction (PCR) technique seems to be promising and can be considered the most sensitive and reliable one. Preparation of viral RNA is still a fundamental step in reverse transcription-PCR (RT-PCR) technique, especially when applied to large scale testing, i.e., for certification purposes. In order to find the most rapid and efficient procedure, we have compared three different procedures of extraction of viral RNA to be processed RT-PCR. Their common characteristics is their capacity to extract the RNA from a small amount of plant tissue without organic solvents in the extraction fluid. The procedures were as follows: an immuno-capture (IC) method using a specific antiserum, a silica-capture (SC) method using a non-specific matrix, and a simple and rapid RNA extraction (RE) method. They all were followed by one-tube RT-PCR. The obtained results show that all the three techniques allowed a successful amplification and detection of PPV in tested samples except the SC-PCR method which proved less effective. In fact, the IC-PCR and RE-PCR methods amplified and detected PPV in all isolates tested, while the SC-PCR method was able to reveal the presence of the virus in apricot and infected control samples only.

Fruit↗
PubMed1998

Detection of plum pox virus in apricot seeds.

Twelve different apricot selection trees from a germplasm collection naturally infected with plum pox virus (PPV) were chosen to investigate the role of seeds in the epidemiology of this dangerous pathogen. All the considered plants showed typical symptoms on leaves and fruits and were positive in enzyme-linked immunosorbent assay (ELISA). The virus was characterized by immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) followed by restriction fragment length polymorphism (RFLP) analysis with RsaI enzyme as a PPV-D isolate. The presence of PPV was checked on fully ripe seeds and seedlings. One half of the seed stock was analysed immediately by ELISA and IC-RT-PCR tests: the cotyledons containing also the embryo were separated from the teguments. The other half of the seed stock was germinated and maintained in an insect-proof screenhouse over a 2-year period. PPV was detected by ELISA only in the seed coat while by IC-RT-PCR also in cotyledons. Seedlings from infected seeds did not show any typical symptoms and were PPV-negative in serological and molecular assays. So far, the presence of PPV in seeds seems to play no role in its epidemiology.

Blotting, Western↗
PubMed1997

Distinct mdm2/p53 expression patterns in liposarcoma subgroups: implications for different pathogenetic mechanisms.

Recent findings have indicated that TP53 inactivation in sarcomas may result from mutation and/or deletion of the TP53 gene or, alternatively, from binding to the MDM2 gene products. To investigate further a possible role of the two genes in sarcomas, 24 large and deep-seated lipomas and 74 liposarcomas of various subtypes were analysed for mdm2 and p53 overexpression by immunocytochemistry. Nineteen cases of the same series were also molecularly analysed for both MDM2 gene amplification and TP53 mutations, and a further ten cases for non-random chromosomal abnormalities. In the retroperitoneal well-differentiated-dedifferentiated (WD-DD) group, 15/16 WD and 8/8 DD liposarcomas displayed the mdm2+/p53+ phenotype, consistent with MDM2 gene amplification in the absence of TP53 mutations. In the non-retroperitoneal WD-DD group, 5/11 WD liposarcomas also retained the mdm2+/p53+ phenotype whereas all DD liposarcomas showed an immunophenotype and, when assessed, a genotype consistent with mutant TP53. Null mdm2 immunophenotype, coupled with evidence of a specific chromosome translocation t(12;16), was constantly observed in both the usual and the cellular subtypes of myxoid liposarcoma, three cases of which also showed TP53 alterations at the genetic or protein level. Neither mdm2 nor p53 overexpression was observed in the lipomas. The results show the existence of three main pathogenetically distinct groups of liposarcoma. The first retroperitoneal WD-DD group, which represents a novel class of tumours within a single histological category of sarcoma, where MDM2-mediated inactivation of p53 could be related to the pathogenetic mechanism. The second is the non-retroperitoneal WD-DD group, where the TP53 mutations appear to correlate with the dedifferentiation process. The third is the myxoid group, which is characterized by its own unique cytogenetic profile and never shows any involvement of TP53 or MDM2 genes. As for diagnostic significance, the absence of mdm2 and p53 reactivity in lipomas seems to represent a useful marker for differential diagnosis from lipoma-like WD liposarcomas.

Adolescent↗
PubMed1994

Epstein-Barr virus genomes in undifferentiated and squamous cell nasopharyngeal carcinomas in Italian patients.

Although undifferentiated carcinoma (UC) and squamous cell carcinoma (SCC) of the nasopharynx are regarded as two distinct histopathologic and clinical entities, it is unclear whether, like UC, SCC carries Epstein-Barr virus (EBV) genomes. We used the polymerase chain reaction (PCR) on paraffin-embedded biopsy specimens to test for the presence of EBV DNA in 20 cases of UC and 9 cases of SCC. Multiple copies of the viral genome were regularly detected in all UCs; however, of the nine cases of SCC, seven had no detectable EBV DNA and two contained viral genomes in a low copy number. In parallel, a marked difference in the serum levels of anti-EBV antibodies between patients with UC and SCC was found. Our findings provide evidence for the specific association of EBV with UC in Italian patients and prove by means of a highly sensitive molecular technique that SCC is occasionally related to EBV DNA. Because of the absence of EBV DNA in most cases of SCC and the minimal viral DNA copy number in the few EBV-associated cases of SCC, a different pathway of oncogenic transformation and growth of the nasopharyngeal epithelium is suggested for SCC and UC.

Antibodies, Viral↗
PubMed1992

HPV DNA in intraepithelial neoplasia and carcinoma of the vulva and penis.

Surgical specimens of 15 patients with early and 12 patients with advanced squamous cell carcinoma of the vulva and the penis were examined for the presence of human papillomavirus (HPV) type 6, 11, 16, and 18 DNA by Southern blotting (SB) and polymerase chain reaction (PCR) analysis. By SB, HPV type 16 DNA was detected in all early carcinomas and 2 of 12 cases of advanced squamous cell carcinoma (ISCC) of the vulva and penis. PCR revealed HPV DNA in four additional cases of vulvar and penile ISCC negative by SB. Three cases contained HPV16 and one HPV18. Two cases of vulvar and penile Buschke-Löwenstein (BL) tumor with malignancy and one case of vulvar verrucous carcinoma were also examined by both techniques. While BL tumors were associated with DNA of HPV6 or 11, no HPV association was found for verrucous carcinoma. Our results confirm that the detection rate of HPV DNA in early vulvar and penile carcinomas is much higher than in invasive carcinomas. In addition, we have shown that in the lower genital tract, 50% of cases of ISCC are HPV16 correlated. The absence of HPV DNA (types 6, 11, 16, and 18) in the remaining 50% of cases of ISCC thus suggests that vulvar and penile ISCC may have more than one pathogenetic pathway.

Base Sequence↗
PubMed1992

Human papillomavirus genomes in male urethral cells.

Sixty-four samples of urethral cells from male sexual partners of women with genital human papillomavirus (HPV) infection were analyzed for the presence of HPV types 6, 11, 16, and 18 by polymerase chain reaction (PCR) followed by slot blot hybridization. Additional samples from 37 of these subjects were analyzed for the presence of viral cytopathic effects by conventional cytology. By PCR, HPV DNA was detected in 21% (14/64) of samples. By cytology, 16% (6/37) of the samples showed cellular changes consistent with HPV infection. Polymerase chain reaction and cytology results were concordant for presence and absence of HPV in 5 and 28 cases, respectively. Three additional HPV-positive cases were obtained with PCR in the cytologically negative samples. The cytologic abnormalities were found to be associated with the presence of both low-risk HPV types and meatal acetoreactivity. On the contrary, HPV DNA positivity by PCR was unrelated to viral type and peniscopic findings. Urethral HPV infection was detected by PCR in 30% of males with visible penile lesions and in 18% of those without. These results indicate that PCR analysis of urethral samples is a helpful adjunct to cytology for the detection of HPV DNA in absence of cytologic evidence of infection.

Base Sequence↗
PubMed1989

Production of parathyroid hormone-related protein by a rat parathyroid cell line.

Parathyroid hormone-related protein (PTHrP), the peptide associated with humoral hypercalcemia of malignancy, has been identified in fetal and adult parathyroid glands. We here report a sub-clone of a rat parathyroid cell line which secretes a single peptide species corresponding in size to PTHrP(1-84). Biological activity of the secretion product was blocked by a specific antiserum against PTHrP, but not by parathyroid hormone (PTH) antiserum. Secretion of PTHrP by these cells was regulated by extracellular calcium in the physiological range. A single messenger RNA species for PTHrP was identified, though PTH mRNA could not be shown in these cells. Hybrid CAT genes containing 700-1000 bp of 5'-flanking DNA from the human PTH or PTHrP genes were transfected into these cells, and the PTHrP gene was expressed at 10-fold higher levels than the PTH gene. These cells thus provide a valuable model system for investigation expression of PTHrP in a non-transformed cell line.

Adenylyl Cyclases↗
PubMed1983

[Hemorrheological changes in peripheral obliterative arterial diseases].

The relationships between blood rheology and peripheral arterial disease are reviewed. The connections between abnormal blood rheology and peripheral arterial disease are discussed considering the opinions of the enthusiastic and skeptic rheologists. A prospective for future investigations is also proposed.

Arterial Occlusive Diseases↗
PubMed1983

[Relation of blood viscosity, plasma viscosity and hematocrit].

The results from 398 consecutive measurements of blood viscosity, plasma viscosity and hematocrit were submitted to a statistical analysis. Hematocrit appeared to be the main determinant of blood viscosity, even though its influence was not so strong as illustrated in previous investigations. The correlation between blood viscosity and hematocrit appeared strictly connected with the red cell amount of the blood sample, being higher when the latter exceeded its physiological range; this correlation disappeared when red blood cell amount was strongly reduced, while the correlation between plasma viscosity and blood viscosity had an opposite behaviour. From these results we can conclude that correlations between hematocrit and plasma viscosity with blood viscosity have opposite trends and that a reciprocal interference is often present.

Blood Viscosity↗
PubMed1983

[A small subpopulation of rigid erythrocytes changes the erythrocyte filtration test].

By means of the filterability test of Reid et al., the influence of a subpopulation of rigid erythrocytes (5% and 2.5%, respectively) was studied using Nucleopore polycarbonate membranes with pore size of 5 mu and diameter of 13 mm. The addition of 5% and 2.5% of rigid red cells delays the erythrocyte filtration times and alters the red cell deformability index. The meaning of the obtained data is discussed.

Erythrocyte Indices↗
PubMed1983

[The lack of correlation between erythrocyte filtration parameters and other hemorrheological parameters].

The findings of 300 consecutive measurements of erythrocyte filterability, whole blood viscosity, plasma viscosity, hematocrit, mean corpuscular volume and white blood cell count have been statistically analyzed in order to find a possible correlation between these parameters. No correlation between erythrocyte filterability, whole blood viscosity, plasma viscosity, hematocrit, mean corpuscular volume and white blood cell count was found. Some hypotheses are enunciated to explain such absence of correlation. The meaning of erythrocyte filtration test is still unclear.

Blood Viscosity↗
PubMed1983

[Hemorrheological changes induced by submaximal exertion in subjects with insulin-dependent diabetes mellitus].

The influence of an acute physical exercise on hemorheological parameters in insulin-dependent diabetes mellitus in not actually known. We have examined the behavior of hemorheological parameters and other laboratory findings such as blood glucose, blood lactic acid, O2 and CO2 venous pressure and venous pH before and after a submaximal physical exercise standardized by ECG monitoring and by means of an 'oxygen consumption computer'. The work load was significantly lower in diabetic patients than in normal control subjects. On the other hand, changes in hemorheological parameters were more evident in diabetic patients. The significance of these findings is also discussed.

Adolescent↗
PubMed1977

[Blood viscosity and triglyceridemia. Findings using a co-axial cylindrical viscosimeter at low "shear rates"].

Examination of blood viscosity at low shear rates using a co-axial cylinder viscometer showed a significant difference between the means of values observed in hypertrigliceridemic patients compared with that of control subjects. This result differs from what has been reported by most workers although generally greater shear rates have been used. Calculation of the "r" coefficient and plotting of the regression line for each shear rate showed that there is no linear correlation between blood viscosity and triglyceridaemia, whose variations occur quite independently. It is suggested that the absence of a correlation between the two parameters examined may depend on various factors, of which the most important are those pertaining to the rheological properties of red blood cells and to the structure and chemical and physical characteristics of the triglyceride molecule and of the lipoproteins and chylomicrons which transport them.

Blood Viscosity↗