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Biomedical subjects

M Bertolotto

Publications and source records attributed to M Bertolotto.

At least 19 recordsLinked to original sources

Effect of different cytokines on mammaglobin and maspin gene expression in normal leukocytes: possible relevance to the assays for the detection of micrometastatic breast cancer.

In cancer patients, the ability to detect disseminated tumour cells in peripheral blood or bone marrow could improve prognosis and consent both early detection of metastatic disease and monitoring of the efficacy of systemic therapy. These objectives remain elusive mainly due to the lack of specific genetic markers for solid tumours. The use of surrogate tissue-specific markers can reduce the specificity of the assays and give rise to a clinically unacceptable false-positive rate. Mammaglobin (MAM) and maspin are two putative breast tissue-specific markers frequently used for detection of occult tumour cells in the peripheral blood, bone marrow and lymph nodes of breast cancer patients. In this study, it was evaluated whether MAM and maspin gene expression may be induced in the normal blood and bone marrow cells exposed to a panel of cytokines, including chemotactic factors (C5a, interleukin (IL)-8), LPS, proinflammatory cytokines (TNF-alpha, IL-1beta) and growth factors (IL-3, granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor). The experimental data show that all cytokines included in the panel, except for IL-8, were able to induce maspin expression; on the contrary, MAM gene was never induced. These results suggest that MAM is more specific than maspin and that the possible interference of cytokines should be taken into account in interpreting molecular assays for detection of isolated tumour cells.

Biomarkers, Tumor↗

The feasibility of urethral color ultrasound imaging in the diagnosis of female intrinsic sphincter deficiency: preliminary results.

STUDY DESIGN: Prospective study. OBJECTIVES: To evaluate the feasibility of color ultrasound imaging of the urethra in association with UPP to diagnose intrinsic sphincter deficiency (ISD). SETTING: Italy. METHODS: We studied a group of 13 normal female volunteers (mean age 29 years) during the estrogenic phase and a group of 15 patients (mean age 63.9 years) with ISD. All patients and normal volunteers underwent color ultrasound imaging using a 4-7 MHz convex broad band transducer. Translabial sagittal scans of the urethra were obtained. The color ultrasound parameters were optimized for detection of parenchymal slow flows. A subjective score of the degree of vascularization along the entire urethra was established as follows: (a) minimal or absent (1), (b) poor (2), (c) moderate (3), (d) good (4). Statistical analysis, using the non-parametric Mann-Whitney rank sum test, was carried out to determine differences of ultrasound scores between volunteers and patients. RESULTS: The statistical evaluation showed that the differences between the ultrasound scores in the two groups was statistically significant (P<0.001). CONCLUSION: We affirm that color ultrasound imaging of the urethra seems to be feasible and useful in association with UPP in the diagnosis of ISD even if this echographic investigation needs further observations.

Adult↗

Synovial fluid from patients with rheumatoid arthritis inhibits neutrophil apoptosis: role of adenosine and proinflammatory cytokines.

OBJECTIVE: In synovial fluid (SF) from patients with rheumatoid arthritis (RA), neutrophils are exposed to proinflammatory mediators endowed with either anti-apoptotic or pro-apoptotic properties. We investigated neutrophil apoptosis in the presence of SF from 11 RA patients. METHODS: SF was obtained from affected knees of 11 patients with RA. Human neutrophil apoptosis was evaluated by light microscopic examination and flow-cytometric analysis of annexin V binding. Immune complex-induced neutrophil activation was evaluated as superoxide anion production. Adenosine levels in SF were detected by chromatographic analysis and cytokine levels were studied by enzyme-linked immunosorbent assay. RESULTS: Spontaneous and immune complex-triggered neutrophil apoptosis was reduced by SF from eight out of 11 patients. Immune complex-induced neutrophil activation was unaffected by SF. The cytokines tested had no role in promoting the anti-apoptotic activity of SF. On the contrary, the anti-apoptotic activity of SF was found to depend on the presence of adenosine. Adenosine levels detected in the various samples of SF correlated significantly with the anti-apoptotic activity of the fluids and with the number of apoptotic neutrophils detected in the articular exudate. CONCLUSION: The microenvironment of rheumatoid SF is a proinflammatory milieu responsible for the in loco persistence of activated and long-surviving neutrophils. Adenosine plays a crucial role in this phenomenon, which is related to anti-apoptotic activity.

Adenosine↗

Colour Doppler ultrasonography of female urethral vascularization in normal young volunteers: a preliminary report.

OBJECTIVE: To assess urethral vascularization in healthy young women, using colour Doppler ultrasonography. SUBJECTS AND METHODS: Eleven healthy young women volunteers (mean age 33.6 years, range 24-46) with no pelvic floor disorders and no history of incontinence were assessed. The subjects underwent colour Doppler ultrasonography using a 4-7 MHz convex broadband transducer. Translabial sagittal scans of the urethra were taken when the subjects had a full bladder, both during the oestrogenic and progestogenic phases. The colour Doppler ultrasonography parameters were optimized to detect slow flows in the anterior and posterior distal, middle and proximal urethra. A rank-sum nonparametric test was used to assess differences between the resistive indices. RESULTS: The statistical evaluation showed a significant difference in the resistive index only in the anterior urethra, between the distal and middle plus proximal urethra, in both the progestogenic (P = 0.002) and oestrogenic (P = 0.0127) phases. CONCLUSIONS: This study confirmed that the vascularization of the urethra plays an important role in the 'seal' effect, which is considered one of the most important factors in urethral closure. There was a significantly greater resistive index in the anterior proximal urethra than in the middle and distal urethra. These findings suggest that the seal effect is related to the existence of a rich venous urethral vascularization, involved in the mechanism of urethral closure. Colour Doppler ultrasonography of the urethra seems to be feasible and useful for understanding the mechanism of the vascular component in female continence.

Adult↗

[Spiral-CT with multiplanar reconstructions (MPRS) in the evaluation of ureteral neoplasms: preliminary results].

PURPOSE: Urothelial tumors of the upper urinary tract (renal pelvis and ureters) are rare: the most common of these are Transitional Cell Carcinomas (TCC), usually localized in the bladder (75% of cases), whereas only 7-10% arise in the renal pelvis and 2,5-5% in the ureter. Multicentricity is not infrequent and synchronous or metachronous tumors have been reported to develop in 11-13% of patients with upper urinary tract cancer and 2-4% of those with bladder cancer. The presence of additional lesions can change patient management. Clinically these tumors may present with hematuria or flank pain, but sometimes there are no specific clinical findings. Radiological studies play an important role in tumor detection and staging and in follow up. The diagnosis is usually performed by IVU and US, while CT is generally employed in the staging. Our purpose was to verify the role of Spiral CT with Multiplanar Reconstructions (MPRs) in the evaluation of ureteral tumors. MATERIAL AND METHODS: Nine patients with ureteral neoplasms were examined with a single detector spiral-CT, using the following parameters: 120 kV, 200 mA; 5 mm collimation, pitch 1.4, slice reconstruction every 5 mm; image acquisition 2-5 minutes after injection of 120 cc of contrast medium (300 mgI/ml). All the exams were transferred to a separate workstation to obtain curvilinear MPRs. RESULTS: Spiral CT on the axial plane visualized 4 urothelial tumors as concentric ureteral wall thickening; 3 as intraluminal soft tissue masses, with involvement of almost the entire urinary tract, as far as the bladder in 1 case. In other 2 patients there were synchronous lesions which presented both patterns (wall thickening and intraluminal mass). In all cases the lesions exhibited moderate enhancement after c.m. injection. Medium- to high-grade hydronephrosis above the tumors was always present. On MPRs both ureteral involvement and dilated, urine-filled upper urinary tract were clearly represented on coronal and sagittal oblique planes; these images allowed to visualize the craniocaudal extension of short or long lesions, and to detect multicentricity. DISCUSSION: The radiological diagnosis of ureteral tumors is usually performed by IVU, US and CT. IVU allows to visualize the tumors as filling defects or irregular narrowing of the ureteral lumen. IVU however is unable to show the real extent of the tumor in obstructive disease or in nonfunctioning kidney. US visualizes the lesion as an echoic mass in the lumen, although it is sometimes difficult to differentiate neoplastic lesions from other filling defects and to visualize the entire ureteral tract. CT is generally employed in the staging of ureteral tumors. CT findings are usually very useful for distinguishing ureteral neoplasms from other filling defects based on their attenuation and enhancement characteristics. Spiral CT with MPRs allows to obtain panoramic views of the urinary tract; these images are more effective than those of IVU, because of their capability to show the longitudinal extension of the lesion and the presence of multicentric tumors. Furthermore with MPRs it is possible to visualize the ureteral tract distal to the lesion, overcoming the limitations of IVU in nonfunctioning kidney or obstructive disease. CONCLUSIONS: Spiral CT with MPRs is useful in the evaluation of ureteral tumors as it is capable of visualizing the whole extension of the lesion and the presence or absence of multicentric lesions, which can change patient management.

Aged↗

US and MR imaging of peripheral nerves in leprosy.

OBJECTIVE: To analyze peripheral nerves with ultrasonography (US) and magnetic resonance imaging (MR) in leprosy and assess the role of imaging in leprosy patients. DESIGN AND PATIENTS: Fifty-eight nerves with abnormal clinical features or electromyograms were examined in 23 leprosy patients by means of gray-scale US, Doppler US and MR imaging. Image analysis included: measurement of nerve cross-sectional area; assessment of nerve structure and MR signal intensities; identification of nerve compression within osteofibrous tunnels; detection of endoneural color flow signals and Gd-DTPA enhancement. Correlations were made with clinical findings and a control group of 20 subjects. Fourteen nerves in active reversal reaction were followed up after therapy. RESULTS: Leprosy nerves were classified into three groups based on imaging appearance: group I consisted of 17 normal-appearing nerves; group II, of 30 enlarged nerves with fascicular abnormalities; group III, of 11 nerves with absent fascicular structure. Group II nerves were from patients subjected to reversal reactions; 75% of patients with group III nerves had a history of erythema nodosum leprosum. Nerve compression in osteofibrous tunnels was identified in 33% of group II and 18% of group III nerves. Doppler US and MR imaging were 74% and 92% sensitive in identifying active reactions, based on detection of endoneural color flow signals, long T2 and Gd enhancement. In 64% of cases, follow-up studies showed decreased color flow and Gd uptake after steroids and decompressive surgery. CONCLUSIONS: US and MR imaging are able to detect nerves abnormalities in leprosy. Active reversal reactions are indicated by endoneural color flow signals as well as by an increased T2 signal and Gd enhancement. These signs would suggest rapid progression of nerve damage and a poor prognosis unless antireactional treatment is started.

Adolescent↗

Characterization of unifocal liver lesions with pulse inversion harmonic imaging after Levovist injection: preliminary results.

The aim of this study was to evaluate capabilities of pulse inversion harmonic imaging (PIHI) in characterization of unifocal liver lesions. We evaluated with PIHI (HDI5000, ATL, Bothell, Wash.) and spiral CT 46 consecutive patients with a single liver lesion identified by fundamental US [7 hepatocellular carcinomas (HCC), 2 cholangiocarcinomas, 7 focal nodular hyperplasias (FNH), 17 hemangiomas and 13 metastases]. The PIHI was performed before and 30 s, 2 and 4 min after bolus administration of Levovist (2.5 g, 300 mg/ml). Scans were digitally stored and reviewed using a dedicated software. Hepatocellular carcinoma was hyperechoic on 30-s scan, and hypoechoic (n = 5) or isoechoic (n = 2) on 2-min scan. Cholangiocarcinoma had inhomogeneous persistent enhancement. Focal nodular hyperplasia was hyperechoic (n = 5) or isoechoic (n = 2) on 30-s scan, hyperechoic (n = 4), isoechoic (n = 2) or slightly hypoechoic (n = 1) on 2-min scan. Large hemangioma revealed peripheral enhancement on 30-s scan which extended centripetally on 2-min scan. Small hemangioma appeared isoechoic on 2-min scan in all but two cases in which they were hypoechoic on 2-min scans and hyperechoic on 4-min scan. Metastasis was hypoechoic on all scans, 70% with rim enhancement. Similar changes in enhancement pattern have been observed at spiral CT. The 30-s and the 2-min scans revealed a conclusive importance in characterization of HCC, cholangiocarcinoma, and large hemangioma. The 2-min scan often furnished enough information for characterization of small hemangioma and metastasis. The 4-min scan allowed characterization of two hemangiomas which appeared hypoechoic on 2-min scans. In the other cases it did not provide further information. Diagnosis of FNH is usually reached with Colour Doppler US; PIHI should be used when colour Doppler is biased by artefacts or when colour Doppler findings are not characteristic. Our results seem to show that PIHI could be a valuable alternative diagnostic approach to spiral CT for unifocal liver lesion characterization. This hypothesis needs to be confirmed with an increased number of lesions.

Adult↗

[Radial fascio-cutaneous flap of the forearm and myocutaneous gracilis muscle flap in urologic surgery: surgical anatomy and techniques].

All the concepts and principles commonly espoused in plastic surgery are very useful in urologic reconstructive operations too. Hypospadia's repair, neo-bladder reconstruction, microsurgery of the seminal way require as certain rules as an absolute respect for anatomy, sparing of the finest tissue vascularization and tension free sutures. Pedicled skin flaps harvesting and utilisation are techniques typical of plastic surgery but are also largely used in urologic adult and paediatric surgery. They are adopted for urethral, penile and corpora cavernosa reconstruction. Pedicled flaps are utilised for the closure of large skin defect in case of complicated wound or when an urinary fistula is present, especially after radiotherapy. A perfect knowledge of the flap nourishment and of the method of harvesting is crucial if the best results must be obtained. In our work we describe the surgical technique for the utilisation of gracilis muscle and forearm flap. Special care is taken to the anatomical description.

Adult↗

Spontaneous apoptosis in neutrophils is associated with downregulation of HLA Class I and is prevented by ligation of Class I.

In many types of cells, ligation of human leukocyte antigens (HLA) Class I molecules with specific mAbs results in the transduction of signals that trigger different cell functions. We have investigated the effects of Class I ligation in human neutrophils. After several hours in culture, neutrophils split spontaneously into two subpopulations, one with normal and the other with reduced levels of Class I. The latter subpopulation displayed high binding capacity for Annexin V, showed a hypodiploid peak, electrophoretic DNA fragmentation, and morphological features of apoptotic cells. The addition of drugs known to delay apoptosis (GM-CSF or cAMP) resulted in a reduction of Class I modulation. Furthermore, ligation of surface Class I with F(ab')2 fragments of the anti-Class I mAb W6/32 resulted in a delay in the progression of apoptosis. These data indicate that this surface Class I molecule is a marker of age-related apoptosis, and the ligation of these molecules results in the transduction of a signal that inhibits apoptosis. Thus, the downregulation of HLA Class I molecules in aging neutrophils prevents their halting the apoptotic process.

Adult↗

[Kidney obstruction: potential use of ultrasonography and Doppler color ultrasonography].

Ultrasonography is the first imaging approach in evaluation of patients with urinary obstruction. Presence of hydronephrosis, urinomas and inflammatory complications can be assessed. Moreover, the level and the cause of obstruction are often identified. The major limits of ultrasonography are poor visibility of the lumbar portion of the urinary tract and lack of specificity in detection of pyelocaliectasis. These limits are partially overcome with Doppler studies. Several factors can bias diagnostic accuracy of colour Doppler ultrasonography in evaluating renal obstruction; in particular, partial or intermittent obstruction cause minimum hemodynamic changes in patients with renal colic. Sensitivity of colour Doppler ultrasonography to detect obstruction decreases following medical treatment with NSAID and increases following hydration and diuretics administration. Diuretic ultrasound, colour Doppler evaluation of ureteral jets and evaluation of the twinkling artefact of stones are useful imaging techniques as well. Tissue harmonic imaging is a new imaging technique available in most latest generation US equipments. Evaluation of the kidney and of the urinary tract improves using this technique due to artefact reduction and increased contrast resolution.

Anti-Inflammatory Agents, Non-Steroidal↗

Detection of liver metastases with pulse inversion harmonic imaging: preliminary results.

The aim of this study is to evaluate capability of contrast enhanced ultrasonography (US) using pulse inversion harmonic imaging (PIHI) to detect liver metastases in comparison to fundamental B-mode ultrasound and spiral CT. Thirty-six consecutive patients with known malignancies and sonographically proved or suspicious liver metastases have been examined with fundamental B-mode US, with PIHI 2', 4' and 6' after Levovist injection and with four phase spiral-CT. Presence, conspicuity and number of lesions have been evaluated comparing PIHI with fundamental B-mode US and spiral-CT. A strong grey-scale enhancement of the liver parenchyma has been observed 2' and 4' after Levovist injection. The optimum parenchymal enhancement and contrast difference between liver and metastases was observed during the 2' measurements. PIHI revealed more lesions than fundamental B-mode US in 56 % of patients, while in 39 % and in 5 % revealed respectively the same number and fewer lesions. PIHI and spiral-CT were in agreement in 67 % of patients, while in 22 % and 11 % PIHI revealed respectively more and fewer lesions. PIHI accurancy presents restrictions in anterior superficial and in deep liver areas, whereas it may be superior to spiral-CT in studying sub-diaphragmatic liver regions.

Aged↗

Penile sonography.

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Blood Flow Velocity↗

Cefoperazone prevents the inactivation of alpha(1)-antitrypsin by activated neutrophils.

At sites of neutrophilic inflammation, tissue injury by neutrophil elastase is favored by phagocyte-induced hypochlorous acid-dependent inactivation of the natural elastase inhibitor alpha(1)-antitrypsin. In the present study, cefoperazone prevented alpha(1)-antitrypsin inactivation by neutrophils and reduced the recovery of hypochlorous acid from these cells. Moreover, the antibiotic reduced the free elastase activity in a neutrophil suspension supplemented with alpha(1)-antitrypsin without affecting the cells' ability to release elastase. These data suggest that the drug inactivates hypochlorous acid before its reaction with alpha(1)-antitrypsin, thereby permitting the antiprotease-mediated blockade of released elastase. In conclusion, cefoperazone appears to have the potential for limiting elastase-antielastase imbalances, attenuating the related tissue injury at sites of inflammation.

Cefoperazone↗

Changes of renal resistive index in response to hydration and diuretic administration in normal subjects and in patients with small ureteral stone.

The renal resistive index has been measured before and after hydration and administration of diuretics in persons with normal kidneys and in kidneys with small ureteral stone, either obstructing or nonobstructing, to assess induced flow changes and to identify features differentiating obstructing from nonobstructing stones. In normal kidneys the resistive index was normal (mean, 0.62+/-0.03); no changes in the resistive index occurred within 15 to 60 min after hydration alone, whereas the resistive index rose within 15 min after hydration plus administration of diuretics and then returned to initial values within 30 min. In both cases the resistive index decreased below basal values after 75 to 90 min. Similar changes were observed in kidneys with a nonobstructing ureteral stone. In kidneys with an obstructing ureteral stone the resistive index was higher than in normal subjects (mean, 0.73+/-0.02, P<0.01), increased further within 15 min after hydration and administration of diuretics (P<0.01), and remained higher than basal values during the following 90 min. In conclusion, different resistive index changes have been observed in response to hydration and diuretics in normal and obstructed kidneys. Duplex Doppler sonography and diuresis duplex Doppler sonography seem promising diagnostic tools to identify obstructing stones.

Diuretics↗

Activation of neutrophil respiratory burst by cytokines and chemoattractants. Regulatory role of extracellular matrix glycoproteins.

OBJECTIVE AND DESIGN: We investigated the in vitro responsiveness of neutrophils adherent to fibronectin (FN) and laminin (LM), toward natural pro-inflammatory and/or phagocyte-activating agents. MATERIALS AND METHODS: Neutrophils from normal volunteers were layered on polystyrene wells precoated or not with FN and/or LM and tested for their ability of responding to eleven pro-inflammatory mediators by evaluation of superoxide anion (O2-) production and adherence. Results, expressed as mean +/-1SEM, were evaluated by non-parametric analyses (Mann-Whitney U-test or Kruskal-Wallis non-parametric ANOVA analysis) RESULTS: Precoating polystyrene wells with LM or FN prevented the plastic-induced neutrophil (O2-) production. Among eleven agents, tumor necrosis factor-alpha (TNF, 3.0+/-0.3 nmoles (O2-)/5 x 10(4) neutrophils/180 min, p < 0.001), granulocyte-macrophage colony stimulating factor (GM-CSF, 2.1+/-0.3 nmoles (O2-)/5 x 10(4) neutrophils/180 min, p < 0.05) and formyl-peptides (fMLP, 2.5+/-0.5 nmoles (O2-)/5 x 10(4) neutrophils/180min, p < 0.01) caused massive (O2-) production by neutrophils adherent to FN. None of the mediators was capable of triggering (O2-) production by neutrophils adherent to LM. LM, mixed with FN to coat wells, caused a dose-dependent inhibition of the oxidative burst triggered by TNF (IC50 LM: 0.84+/-0.03 microg, mean+/-1 SEM), GM-CSF (IC50 LM: 0.36+/-0.16micro/g, mean+/-1SEM) and fMLP (IC50 LM: 0.54+/-0.008 microg, mean+/-1 SEM). To the contrary, fMLP (85.5+/-27.7%), TNF (163.1+/-67.5%), and GM-CSF (121.8+/-66.4%) caused a significant augmentation of neutrophil adherence to LM, suggesting that LM-mediated inhibition of neutrophil oxidative metabolism does not depend on the concomitant LM-induced inhibition of neutrophil adherence. Finally, neither solid-phase FN nor LM affected (O2-) production by neutrophils in response to immune complexes. CONCLUSIONS: Extracellular matrix glycoproteins dictate the response of neutrophils to soluble mediators but not to immune complexes. This appears to be a biologically meaningful mechanism to localise the risk of cellular reactions to mediators that are able to diffuse easily from tissue sites of generation and become widely distributed in body fluids during inflammatory diseases.

Cells, Cultured↗