Immuno-metabolic impact of the multiple sclerosis patients' sera on endothelial cells of the blood-brain barrier.

BACKGROUND: Multiple sclerosis (MS) is an autoimmune disease which results from the invasion of the brain by activated immune cells across the endothelial cells (ECs) of the blood-brain barrier (BBB), due to loss of immune self-tolerance. Many reports define the metabolic profile of immune cells in MS, however little is known about the metabolism of the BBB ECs during the disease. We aim to determine whether circulating factors in MS induce metabolic alterations of the BBB ECs compared to a healthy state, which can be linked with disruption of BBB integrity and subsequent immune cell extravasation. METHODS AND RESULTS: In this report, we used an in vitro model to study the effect of sera from naïve-to-treatment, relapsing-remitting MS (RRMS) patients on the human brain microvascular endothelium, comparing effects to age/sex-matched healthy donor (HD) sera. Our data show that RRMS serum components affect brain endothelial cells by impairing intercellular tightness through the down-modulation of occludin and VE-cadherin, and facilitating immune cell extravasation through upregulation of intercellular adhesion molecules (ICAM-1) and P-glycoprotein (P-gp). At a metabolic level, the treatment of the endothelial cells with RRMS sera reduced their glycolytic activity (measured through the extracellular acidification rate-ECAR) and oxygen consumption rate (oxidative phosphorylation rate-OCR). Such changes were associated with the down-modulation of endothelial glucose transporter 1 (GLUT-1) expression and by altered mitochondrial membrane potential. Higher level of reactive oxygen species released from the endothelial cells treated with RRMS sera indicate a pro-inflammatory status of the cells together with the higher expression of ICAM-1, endothelial cell cytoskeleton perturbation (stress fibres) as well as disruption of the cytoskeleton signal transduction MSK1/2 and ß-catenin phosphorylation. CONCLUSIONS: Our data suggest that circulating factors present in RRMS patient serum induce physiological and biochemical alterations to the BBB, namely reducing expression of essential tightness regulators, as well as reduced engagement of glycolysis and alteration of mitochondrial potential. As these last changes have been linked with alterations in nutrient usage and metabolic function in immune cells; we propose that the BBB endothelium of MS patients may similarly undergo metabolic dysregulation, leading to enhanced permeability and increased disease susceptibility.

Immunometabolic biomarkers of inflammation in Behçet's disease: relationship with epidemiological profile, disease activity and therapeutic regimens.

Behcet's disease (BD) is a systemic inflammatory disease with a still unclear pathogenesis. Although several inflammatory molecules have been studied, current biomarkers are largely insensitive in BD and unable to predict disease progression and response to treatment. Our primary aim was to explore serum levels of soluble CD40 L (sCD40L), soluble intracellular adhesion molecule (sICAM-1), monocyte chemoattractant protein-1 (MCP-1), myeloperoxidase (MPO), leptin, resistin, osteoprotegerin (OPG), soluble type 1 tumour necrosis factor receptor (sTNFR), interleukin (IL)-6 and serum amyloid A (SAA) serum concentration in a cohort of 27 BD patients. The secondary aim was to evaluate potential correlations between the putative circulating biomarkers, demographic profile of patients, the status of disease activity, the specific organ involvement at the time of sample collection and different therapeutic regimens. Serum concentrations of sTNFR (P = 0·008), leptin (P = 0·0011), sCD40L (P < 0·0001) and IL-6 (P = 0·0154) were significantly higher in BD patients than in HC, while no difference was found in MCP-1, MPO and resistin serum levels. Moreover, we observed significantly higher sTNFR serum concentrations in BD patients presenting inactive disease than HC (P = 0·0108). A correlation between sTNFR and age was also found, with higher levels in patients over 40 years than HC (P = 0·0329). Although further research is warranted to elucidate the role of circulating biomarkers, some of that may contribute to the understanding of the physiopathology processes underlying BD activity and damage as well as to provide useful tools for prognostic purposes and a personalized treatment approach.

Patients with type 3 severe von Willebrand disease are not protected against atherosclerosis: results from a multicenter study in 47 patients.

BACKGROUND: The results of a number of studies in pigs and mice suggest that absence of von Willebrand factor (vWF) protects against the development of atherosclerosis. We studied whether patients with a complete deficiency of vWF (type 3 von Willebrand disease [vWD]) develop fewer atherosclerotic vessel wall changes than healthy controls. METHODS AND RESULTS: This study included 47 individuals with type 3 vWD and 84 healthy controls. Early atherosclerotic changes were assessed by measuring the thickness of the intima-media in the carotid and femoral arteries by B-mode ultrasonography. Advanced atherosclerotic changes were quantified by summing the maximal thickness of atherosclerotic plaques in the carotid and femoral arteries and were expressed as a plaque score. Established risk factors were determined to adjust for possible differences between the groups. We found no substantial difference in intima-media thickness between vWD patients and controls (adjusted difference for carotid artery 0.007 mm, 95% CI -0.022 to 0.036 mm; femoral artery 0.069 mm, 95% CI -0.056 to 0.19 mm). Similar proportions of patients and controls had atherosclerotic plaques (19% and 17%, respectively). No difference was found in the plaque score between groups (adjusted difference -0.22 mm, 95% CI -0.69 to 0.26). Among vWD patients, we found no effect of treatment with vWF concentrates on intima-media thickness or plaque score. CONCLUSIONS: The results of this study indicate that vWF does not play a substantial role in human atherogenesis.

Gemcitabine/cyclophosphamide/5-fluorouracil/folinic acid triplet combination in anthracycline- and taxane-refractory breast cancer patients: a Southern Italy Cooperative Oncology Group phase I/II study.

We sought to define the recommended dose of cyclophosphamide (CTX) for subsequent phase II assessment when combined with fixed doses of gemcitabine (Gemzar; Eli Lilly and Company, Indianapolis, IN) and 5-fluorouracil/folinic acid in metastatic breast cancer patients previously treated with anthracyclines and taxanes. Patients age 70 or less, with an Eastern Cooperative Oncology Group performance status 0 to 2, were enrolled. Patients received gemcitabine 1,000 mg/m(2), 5-fluorouracil 425 mg/m(2), folinic acid 100 mg/m(2), and escalating doses of CTX (in 100-mg/m(2) increments), starting at 500 mg/m(2), on days 1 and 8 every 3 weeks. Since March 1999, 46 patients, with a median age of 51 years (range, 38 to 74 years), entered the trial in seven cohorts. Cyclophosphamide dose escalation was stopped at 600 mg/m(2) when three of six patients experienced dose-limiting toxicity (one each with grade 3 thrombocytopenia, grade 3 neutropenia, and persistent grade 2 neutropenia), and then continued with granulocyte colony-stimulating factor support. The CTX dose of 800 mg/m(2) was proven safe and was chosen for phase II study. Two complete and 15 partial responses provided an overall response rate of 37% (95% confidence interval, 23% to 51%). Gemcitabine/CTX/5-fluorouracil/folinic acid is well tolerated by metastatic breast cancer patients pretreated with anthracyclines/taxanes, up to a CTX dose of 800 mg/m(2). The phase II study is ongoing. Semin Oncol 28 (suppl 10):50-56.

Pharmacokinetics of monoclonally-purified and recombinant factor VIII in patients with severe von Willebrand disease.

A monoclonally-purified factor VIII (FVIII) concentrate, containing little von Willebrand factor (vWF), was infused to 11 patients with severe von Willebrand disease and unmeasurable levels of plasma vWF. In comparison with the historical data obtained infusing hemophiliacs in the same conditions, monoclonally-purified FVIII had a significantly shorter half-life and faster clearance from plasma but similar in vivo recovery and volume of distribution. Two additional patients with severe von Willebrand disease were also infused with recombinant FVIII totally devoid of vWF. Half-life was very short and in vivo recovery low, with a larger volume of distribution than for monoclonally-purified FVIII. We conclude that in patients with severe von Willebrand disease the small amounts of vWF contained in the monoclonally-purified FVIII concentrate are not sufficient to stabilize infused FVIII, nor to support the normal circulation of endogenous FVIII that these patients produce at a normal rate.

Phase I study of mitoxantrone, raltitrexed, levofolinic acid and 5-fluorouracil in advanced solid tumours.

PURPOSE: We have recently evaluated the combination of raltitrexed, levofolinic acid (LFA) and 5-fluorouracil (5-FU) in advanced head and neck and colorectal cancer, and we have shown that this combination is well tolerated and has clinical activity. Clinical combination studies have shown that raltitrexed and anthracyclines can be combined at full doses without unexpected toxicities. Based on these observations, we started a phase I study of mitoxantrone plus raltitrexed administered on day 1, followed by LFA and 5-FU on day 2 in patients with advanced solid tumors. PATIENTS AND METHODS: Mitoxantrone was given at a starting dose of 6 mg/m2, raltitrexed at a fixed dose of 3 mg/m2, LFA at a fixed dose of 250 mg/m2, and 5-FU at a starting dose of 750 mg/m2. Mitoxantrone and 5-FU doses were subsequently escalated alternately up to dose-limiting toxicity. Treatment was repeated every 14 days. RESULTS: Four dose levels were tested in 18 patients. All three patients treated at the fourth dose level had grade 4 neutropenia after the first cycle. Therefore, this level was defined as the maximum tolerated dose and the dose level immediately below (mitoxantrone 7 mg/m2 and 5-FU 900 mg/m2) was selected for further evaluation. Neutropenia was the main toxic effect. Nonhaematologic side effects were mild. One complete response and five partial responses (all but one in patients with head and neck cancer) were observed, for an overall response rate of 33% (95% confidence interval, 13% to 59%). CONCLUSIONS: Mitoxantrone, raltitrexed and 5-FU can be combined at doses which are close to those used in monotherapy. The observed activity is encouraging, especially in the subset of patients with head and neck cancer.

Increased placental production of leukotriene B4 in gestational hypertension.

Production of some lipoxygenase and cyclooxygenase derivatives of arachidonic acid was measured in placental tissue obtained from women with gestational hypertension and with normal pregnancies. The levels of leukotriene B4 were about five times higher in placentas from hypertensive women and also raised thromboxane A2 and reduced prostaglandin E2 levels were observed. Prostacyclin production was lowered only in women with more severe hypertension, in association with the highest measured levels of leukotriene B4 and thromboxane A2. It is suggested that increased placental levels of leukotriene B4 and thromboxane A2 appear already in mild gestational hypertension, while depression of prostacyclin may occur only at more severe stages of gestational hypertensive disease.

Neoadjuvant chemotherapy for intermediate/high-grade soft tissue sarcomas: five-year results with epirubicin and ifosfamide.

BACKGROUND: Neoadjuvant chemotherapy for intermediate/high-grade soft tissue sarcomas (STS) may provide some advantages for facilitating the surgical resection of the tumor and for disease control. However its role as induction therapy before surgery should still be proved. PATIENTS AND METHODS: Twenty-one patients with intermediate/high-grade STS and tumor size > or = 5 cm were consecutively treated from 1997 to 2001 with neoadjuvant chemotherapy based on epirubicin 60 mg/m2/day on days 1 and 2 and ifosfamide 1.8 gr/m2/day on days 1 through 5 every three weeks. Evaluation of objective tumor response and toxicity were carried out according to WHO criteria. RESULTS: Nine partial responses were documented; stable disease in 11 patients, progressive disease in one patient. Apart from nine cases of grade 4 neutropenia, the treatment was generally well-tolerated. Twelve patients underwent conservative and limb salvage surgery. CONCLUSION: This therapeutic approach seems to be effective in facilitating surgery. Neutropenia was the most significant toxicity but it was preventable or medically treatable with G-CSF support.

Influence on response to therapy and overall survival of mediastinal involvement by Hodgkin's disease.

To assess the prognostic significance of mediastinal involvement of Hodgkin's disease, 91 patients with stage I to III disease treated at our Institute were reclassified according to size of mediastinal disease and other clinical and therapeutic characteristics. Complete remission (CR) was achieved in 46 of 67 (81%) patients without mediastinal involvement, and in 16 of 17 (94%) patients with small mediastinal masses, but only in 10 of 17 (59%) patients with large masses (P less than 0.05). Twenty-seven of 32 (84%) patients treated with irradiation alone and 26 of 28 (93%) patients treated with combined modality therapy reached a CR, whereas such a result was obtained only in 19 of 31 (61%) patients who received MOPP chemotherapy alone (P less than 0.01). In particular, none of the patients with large masses had a CR when treated with chemotherapy alone, whereas no differences in response to therapy were found between patients with large vs. small or no masses when irradiation or combined treatments were utilized. Since combined treatment seems to reach a high proportion of CR and to prevent extranodal relapse, further randomized clinical trials are needed to decide its routine utilization in patients with poor prognostic factors such as large mediastinal adenopathies.

[Early evaluation using PET-FDG of the efficiency of neoadjuvant radiochemotherapy treatment in locally advanced neoplasia of the lower rectum]. / Valutazione precoce con PET-FDG della efficacia del trattamento radiochemioterapico neoadiuvante nelle neoplasie del retto inferiore localmente avanzate.

BACKGROUND AND PURPOSE: Preoperative chemoradiation allows downstaging of locally advanced rectal cancer and in selected patients also a sufficient downsizing to ensure sphincter preservation. Selection of patients warranting a preoperative approach is improved by magnetic resonance imaging (MRI) which is able to define the involvement of mesorectal circumferential margin. Similarly it would be crucial to define the response to chemoradiation during the treatment but traditional morphologic imaging techniques may fail in differentiating neoplastic tissue from scarring. PET-FDG has been successfully used in the detection of metastatic colorectal cancer allowing imaging of deposits as small as 0.5 cm and may have a role in evaluating early response to chemoradiation. METHODS: In the present study, in patients with T3-T4 rectal cancer undergoing preoperative chemoradiation PET-FDG and flow cytometry analysis on endoscopic biopsy specimen have been performed before, during and after preoperative chemoradiation. RESULTS: Chemoradiation treatment has been successful in terms of downsizing and downstaging of the tumor. PET-FDG was able to demonstrate local response at only ten-fifteen days after the beginning of neoadjuvant therapy, also identifying non responding patients. CONCLUSIONS: FDG-PET may have a role in defining the response to chemoradiation and modulate the treatments strategy in patients with advanced rectal cancer.

Computed tomography and second-look surgery in ovarian cancer patients. Correlation, actual role and limitations of CT scan.

Computed tomography (CT scan) was performed on 58 clinically disease-free ovarian cancer patients. The scans were correlated with the results obtained at a subsequent second-look laparotomy. The sensitivity was 0.47, the specificity 0.87, diagnostic accuracy 0.63, positive predictive value 0.84 and negative 0.53. Undetected microscopic disease was classified as a false-negative result. Sensitivity was poor for omental, mesenteric and peritoneal implants and for bowel infiltration, good for lymphnodal involvement and abdominal mass and decisively good for intrahepatic and plenic metastases of ovarian cancer. Due to a still high false-negative rate a normal CT scan does not provide sufficiently accurate diagnostic information to replace a second-look laparotomy. But on the other hand, due to a high specificity, the usefulness of CT can be limited to approximately 27% of patients, with true-positive findings, who might have been saved surgical reexploration. Adjunct studies such as immunoscintigraphy with radiolabelled monoclonal antibodies and measurement of tumor markers further increase its diagnostic accuracy.

Tc-99m sestamibi imaging in the diagnostic assessment of patients with lymphomas: comparison with clinical and radiological evaluation.

Tc-99m MIBI imaging has been used in nuclear oncology, but its role in detecting lymphomas has not been widely investigated. In this study, 31 patients with lymphomas (20 non-Hodgkin's and 11 Hodgkin's) underwent Tc-99m MIBI whole-body imaging. A total of 74 tumor lesions were detected in 25 patients, while the remaining 6 patients were disease-free. The diagnostic accuracy of MIBI imaging for lesion detection was 85%. A total of 11 unknown tumor lesions in 3 patients were discovered on MIBI scans. Tumor size was significantly (p = 0.01) higher in lesions with increased MIBI uptake (3.5 +/- 2.0 cm) compared with those with no uptake (1.8 +/- 1.0 cm). No false positive MIBI findings were observed. The accuracy of MIBI scintigraphy in patients with Hodgkin's disease was lower (72%) compared to that of patients with non-Hodgkin's lymphomas (94%). However, this difference was not related to tumor type, but to lesion size. In fact, tumor size was significantly (p = 0.02) lower in lesions of patients with Hodgkin's disease (2.5 +/- 1.3 cm) compared to those of patients with non-Hodgkin's lymphomas (3.7 +/- 2.2 cm). MIBI imaging may be useful in patients with lymphomas for detecting tumor lesions and, hence, may be considered an alternative to gallium scanning, providing better imaging quality. However, the intense Sestamibi activity in the lower chest and abdomen as well as tumor size may limit the diagnostic sensitivity of this radionuclide technique in patients with lymphomas.

[Relationship between symptomatology of acute ischemic heart disease and glucose metabolism balance]. / Rapporti tra sintomatologia della cardiopatia ischemica acuta e controllo glicometabolico.

Diabetes mellitus is one of the most important cardiovascular risk factors. An increased prevalence of silent or paucisymptomatic myocardial ischaemia has been described in diabetic patients. The authors examined the relationship between glucose metabolism balance and acute ischemic heart disease symptoms in 174 patients: 46 diabetics and 128 non diabetics. Diagnosis of diabetes mellitus was made during admission to hospital in four patients. No differences of strength, length, type, site and radiation of pain between 46 diabetic patients and 128 non diabetics were found. As regards the other symptoms, the authors found that only dyspnoea and palpitations were prevalent in diabetic patients with ten-year or more-disease length. On the other hand, the prevalence of these two symptoms was not different between non diabetics and diabetics with less than ten-year-disease length.

[Deep venous thrombosis. Evaluation of a non-invasive diagnostic procedure based on compression ultrasonography and measurement of D-dimer plasma levels]. / Trombosi venosa profonda. Validazione di una procedura diagnostica non invasiva basata su ultrasonografia con compressione associata a dosaggio dei D-dimeri plasmatici.

BACKGROUND: Clinical diagnosis of deep venous thrombosis (DVT) of the leg is unreliable. An accurate diagnosis is important for therapeutic decision since anticoagulant treatment, though potentially dangerous, is useless in case of a false positive diagnosis, whereas a false negative diagnosis may lead to withdrawal of an extremely necessary anticoagulation. Contrast venography is still recognized as the gold standard method for the diagnosis of DVT, but in recent years a variety of accurate non-invasive methods has been developed. The ultrasound compression sonography (CUS) is considered a simple non invasive test highly sensitive and specific for proximal DVT in symptomatic outpatients, though non adequately sensitive and specific for isolated calf DVT. Plasma D-dimer levels (DD, fibrin degradation products) have a high negative predictive value for DVT. The aim of this study, performed in outpatients with suspected leg DVT, was to validate, versus venography, a non-invasive, easy to perform and fast diagnostic procedure based on a combination of CUS and D-dimer test. End points of the procedure were: confirmation or exclusion of proximal DVT; suspicion of isolated calf DVT in which case the test would be repeated in a few days to detect any possible proximalization of thrombosis. MATERIALS AND METHODS: Sixty-eight consecutive outpatients, 37 male, with clinically suspected first episode of leg DVT were eligible and examined with CUS, DD test and venography. RESULTS: The results showed that the diagnostic procedure under examination has a high sensibility and specificity for DVT. CONCLUSIONS: It can thus be recommended as routine diagnostic procedure in symptomatic outpatients with suspected DVT reserving venography special cases only.

Orthopaedic treatment and passive motion machine: consequences for the surgical treatment of clubfoot.

The efficacy of orthopaedic treatment and its influence on clubfoot surgery has never been truly demonstrated. In the unsorted mass of clubfeet treated, it is difficult to determine exactly how effective orthopaedic treatment is for severely affected feet. If properly performed, perfectly synchronized, and supported by a Kinetec machine, such treatment can noticeably reduce the rate of operation and, when operation is still required, reduce its extent. In grade II soft > stiff feet with scores of 5-10, Kinetec-supported orthopaedic treatment is extremely effective. Operation is required in 32% of cases only, and posterior surgery is often sufficient. Lateral release, in this category, is never required. In grade III stiff > soft feet, with scores of 10-15, the efficacy of orthopaedic treatment associated with the Kinetec machine is far from negligible and operation most often includes posterior and medial release (PMR), variably associated with plantar release. Lateral release is exceptional (15%), and operation is necessary in 75% of cases. In grade IV stiff = stiff feet, with scores of 15-20, orthopaedic treatment with the Kinetec machine has a true, though limited, effect. In this category, operation is necessary in 90% of cases. Lateral release is performed in 50%. In the postoperative period, orthopaedic treatment combined with use of the Kinetic machine must be continued. Orthopaedic treatment coordinated with use of the machine has considerably shortened the duration of plaster cast immobilization; 2 months when operation included posterolateral-medial (PLMR) release or PMR, and only 1 month when operation was posterior release (PR). The machine has noticeably changed the results and has indisputably influenced operation on the whole.

[Subconjunctival Loiasis: case reports and review of cases described in Italy]. / Filariasi sottocongiuntivale da Loa loa: revisione della letteratura di casi clinici descritti in Italia.

Filariasis or Loiasis is an eye infection caused by a parasite belonging to the species Loa loa, also known as "African eye worm" since it is present in the rainforests belt of western and central Africa. The purpose of this study is the description of a clinical case presented to our department. A 40-years-old Congolese woman, in the third month of pregnancy, complained a foreign body sensation in the right eye for which she was submitted to complete ophthalmologic examination. Slit-lamp examination revealed, in the infero-temporal subconjunctival space, the presence of a live whitish worm that moved slowly in the thickness of the tissues. Place a clinical suspicion of subconjunctival filariasis, were performed parasitological and serological analysis; laboratory test showed peripheral blood eosinophilia and the blood film examination was negative for the presence of circulating microfilariae. The worm was surgically removed under topical anesthesia and the histological examinations of the parasite confirmed that it was an adult male worm belonging to the species Loa loa. Due to the large increase of migratory flows to Europe, number of reports of loiasis is becoming more frequent; the knowledge of this emerging pathogens is essential for diagnosis and treatment planning.