Dexamethasone promotes toxicity in U937 cells exposed to otherwise nontoxic concentrations of peroxynitrite: pivotal role for lipocortin 1-mediated inhibition of cytosolic phospholipase A2

Pretreatment with dexamethasone (Dex) was not toxic for U937 cells but caused a rapid lethal response upon subsequent exposure to otherwise nontoxic concentrations of peroxynitrite. This effect was not associated with enhanced formation of hydrogen peroxide taking place after peroxynitrite and was shown previously to play a pivotal role in the ensuing lethal response. Further analyses revealed that although Dex did not affect cytosolic phospholipase A(2) (cPLA(2)) expression, it markedly reduced the extent of arachidonic acid (AA) release mediated by peroxynitrite-dependent stimulation of cPLA(2). This event, as well as the enhanced toxicity, was abolished by mifepristone, a glucocorticoid receptor antagonist. The outcome of various approaches, using phospholipase A(2) inhibitors, cPLA(2) antisense oligonucleotide-transfected cells, and supplementation with exogenous AA, led to the demonstration that inhibition of cPLA(2) activity is causally linked to the increased susceptibility to peroxynitrite caused by Dex. Finally, the effects of Dex were shown to be mediated by enhanced expression of lipocortin 1 (LC1), a cPLA(2) inhibitory protein. These results indicate that Dex promotes toxicity in U937 cells exposed to otherwise nontoxic concentrations of peroxynitrite and that this event is causally linked to enhanced expression of LC1 leading to inhibition of cPLA(2). Thus, the increased lethal response arises because of LC1-dependent impairment of the AA-induced cytoprotective mechanism triggered by peroxynitrite.     

Differential associations of Head and Body Symptoms with depression and physical comorbidity in patients with cognitive impairment

OBJECTIVE: To test the hypothesis that physical symptoms referred to the head might be specifically associated with depression in patients with cognitive impairment. METHODS: Subjects were taken from those enrolled in 'The Mild Project' a prospective study on the natural history of mild dementia (Mini Mental State Examination > or = 18) and with a diagnosis of Alzheimer's disease, vascular dementia, and mild cognitive impairment. A total of 129 subjects were included in the study. Physical symptoms were assessed with a checklist investigating nine different body organs or apparati. Physical symptoms were grouped into those referred to the head (Head Symptoms: ear and hearing; eyes and sight; and head and face) and all the others (Body Symptoms). Depressive symptoms were assessed with the Geriatric Depression Scale (GDS) and physical comorbidity with Greenfield's Index of Disease Severity (IDS). RESULTS: The number of patients reporting one or more Head Symptoms linearly increased with increasing depression severity (Mantel-Haenszel test = 6.497, df = 1, p = 0.011), while the number of patients reporting one or more Body Symptoms linearly increased with increasing physical comorbidity (Mantel-Haenszel test = 4.726, df = 1, p = 0.030). These associations were confirmed in multivariate logistic regression models with adjustment for potential confounders (age, gender, education, cognitive performance, daily function, and diagnosis). CONCLUSIONS: Head Symptoms are specifically associated with depression while Body Symptoms with physical comorbidity, in patients with cognitive impairment. Recognizing these associations in individual patients may help clinicians decide whether to initiate or continue antidepressant therapy or whether to carry out physical instrumental investigations.     

Non-invasive diagnostic and functional evaluation of cardiac and pulmonary involvement in systemic sclerosis

BACKGROUND: The aim of the present study was to evaluate the role of non-invasive methods in the early detection of pulmonary and cardiac involvement in Systemic sclerosis (SSc) and to identify clinical and/or instrumental patterns of prognostic value. PATIENTS AND METHODS: Twenty female patients affected by SSc (8 with diffuse cutaneous SSc and 12 with limited cutaneous SSc) were enrolled in our study. Cardiac and pulmonary involvement (respiratory function tests and carbon monoxide lung diffusion [DLCO], chest radiography, high resolution computed tomography [HRCT] and lung perfusion magnetic resonance) were evaluated. RESULTS: All 18 patients studied with respiratory function tests showed a significant reduction of DLCO. HRCT was considerably more sensitive than traditional chest radiography (59% versus 28%; p<0.05). Lung perfusion MRI revealed normal findings in 15 patients. Abnormal lung perfusion MRI results were found only in 3 patients. Angina pectoris with electrocardiographic and scintigraphic ischemic changes, severe regional wall motion abnormalities and complex arrhythmias seemed to be associated with poor prognosis. CONCLUSION: Taken together these results indicate that a pulmonary involvement occurs both in limited and in diffuse cutaneous SSc patients and develops, in 83% of the cases, without any regional lung perfusion abnormality. Furthermore, cardiac involvement is detected in 65% of the cases as a consequence of a range of noxious events including myocardial ischemia, fibrosis and pressure overload which may result in ventricular dysfunction and arrhythmias. Lung perfusion MRI should be considered as a complementary diagnostic method for the functional evaluation of these symptoms in systemic sclerosis.     

Activity against Plasmodium falciparum of cycloperoxide compounds obtained from the sponge Plakortis simplex

There is an urgent need to discover new antimalarials, due to the spread of chloroquine resistance and the limited number of available drugs. In the last few years, artemisinin, the endoperoxide sesquiterpene lactone derived from Artemisia annua, and its derivatives proved to be very active against Plasmodium falciparum. These compounds are characterized by an endoperoxide pharmacophore that is critical for their antimalarial activity. There are several reports, from our group and others, that marine organisms can be another natural source of stable cyclic peroxides, with selective antifungal or antibacterial activity. With the aim of identifying new bioactive molecules, we evaluated in vitro the antimalarial activity of the major cycloperoxides extracted from the sponge Plakortis simplex. The six-membered endoperoxide compounds plakortin and dihydroplakortin, but not the five-membered cycloperoxide plakortide E, inhibited the growth of cultured P. falciparum parasites, both chloroquine-sensitive D10 strain and chloroquine-resistant W2 strain. The IC50 values were similar for both compounds and in the range of 1263-1117 nM against D10, and 735-760 nM against W2, using the colorimetric parasite lactate dehydrogenase assay. The activity of plakortin and dihydroplakortin was significantly higher against chloroquine-resistant than chloroquine-susceptible parasites, following a pattern similar to that of artemisinin, although they were 50-fold less active. Moreover, plakortin and dihydroplakortin showed an additive effect when used in combination with chloroquine. These results support further studies on cycloperoxides of marine origin to characterize their mechanism of action and identify/synthesize new compounds with stronger antimalarial activity.     

MRI lung perfusion 2D dynamic breath-hold technique in patients with severe emphysema

BACKGROUND: Our aim was to prospectively evaluate a semi-quantitative pulmonary perfusion MR technique using a breath-hold 2D dynamic sequence in patients with severe pulmonary emphysema. MATERIALS AND METHODS: Thirty patients with severe emphysema were studied with pulmonary perfusion MRI. Results were compared with those obtained through lung scintigraphy, considered as the gold standard technique. We used Fast Field Echo (FFE) pulse sequences in the coronal and sagittal planes, with paramagnetic contrast agent administration. Ten healthy volunteers were studied as the control group. Three thoracic radiologists independently reviewed all the subjects, evaluating the site and entity of perfusional defects. Peak intensity and apparent mean transit time were calculated. RESULTS: MRI showed high sensitivity (86.7%) and good specifity (80.0%) in detecting perfusional defects. We observed lower peak signal intensities in emphysematous regions. CONCLUSION: Lung perfusion MR is a potential alternative to Nuclear Medicine in the evaluation of severe pulmonary emphysema.     

The treatment of large supratentorial arachnoid cysts in infants with cyst-peritoneal shunting and Hakim programmable valve

OBJECTIVE: This retrospective case series examines 7 infants with large supratentorial arachnoid cysts who underwent cyst-peritoneal shunting and insertion of a Hakim programmable valve. Comparing pre- and postoperative clinical data, neuroradiological and regional cerebral blood flow (rCBF) findings we evaluated the efficacy of the surgical procedure. METHODS: Infants, ranging in age from 1 to 55 days (mean age 29.5 days), were assessed pre- and postoperatively by neurological examination, developmental profile and neuroimaging. RESULTS: Post procedure, all patients showed a significant reduction in the cyst/brain ratio on neuroimaging (p<0.001), 6 had a normal developmental profile (p<0.001) and 5 cases showed a significant amelioration of clinical symptoms and neurological signs. Two patients underwent preoperative SPECT scans, which showed hypoperfusion in the area surrounding the cyst; this decreased rCBF also improved post shunting. CONCLUSIONS: Large supratentorial arachnoid cysts in infants can be successfully treated with cyst-peritoneal shunting and insertion of a Hakim programmable valve. This is the first study specifically aimed at evaluating the long-term results of these conditions.     

Safety and efficacy of two protocols for sedation in pediatric oncology procedures

Invasive procedures, such as the lumbar puncture, can cause anxiety and pain in children undergoing treatment for acute lymphoblastic leukemia (ALL). We investigated the safety and efficacy of two different protocols for pain relief in 20 children with ALL undergoing lumbar puncture. Protocol A was composed of an association between propofol and alfentanil. Protocol B consisted in the combination of propofol and ketamine. Vital and behavioural parameters, sedation and pain scores were recorded at different times during and after the procedure. All patients showed a satisfactory sedation and analgesia. We found a statistically significant difference of vital parameters between protocol A and protocol B, while there were no significative differences between sedation scores and the other parameters evaluated. Patients in protocol A showed a higher incidence of major side effects, such as respiratory depression. Our results show that both protocols are effective to obtain a good sedation and analgesia in children with ALL undergoing lumbar puncture, but the association between propofol and ketamine appears more safe due to the lower incidence of side effects.     

Prognostic value of estrogen receptor status can be improved by combined evaluation of p53, Bcl2 and PgR expression: an immunohistochemical study on breast carcinoma with long-term follow-up

Steroid receptor analysis is the only widely accepted prognostic/predictive marker in breast cancer (BC) treatment. In the present study we evaluated the prognostic role of ER/PgR with p53 and Bcl2, in a series of 277 BC (153 pN1/2, 122 pNO, 2 pNx) with a long-term follow-up (67 months for DFS, 75 months for OS). Our results, besides confirming the usefulness of ER immunohistochemical expression as a prognostic marker, showed that PgR expression alone had a borderline/not significant prognostic value in the whole series (p=0.08 for DFS and p=0.09 for OS), while showed to be prognostic in N+ cases (p=0.02 for DFS and p=0.03 for OS). PgR prognostic value, however, was not independent at the multivariate analysis. By combining ER with PgR, p53 and Bcl2, we showed that ER/p53 and ER/Bcl2 phenotypes had a better discriminant role than ER/PgR phenotype. The ER+/p53+ phenotype was at higher risk of relapse/death as compared with ER+/p53- phenotype. Conversely ER-/p53+ phenotype showed the most unfavourable prognosis. Similar results could be observed concerning ER/Bcl2 phenotypes. Our study showed that the combined evaluation of ER/PgR weakly enhanced the prognostic/predictive power of ER status alone. On the contrary, the combined evaluation of ER/p53 and ER/Bcl2, improved this prognostic/predictive capability and allowed the separation of ER positive and ER negative cases into subgroups with different prognosis.     

Variants of CARD15 are associated with an aggressive clinical course of Crohn's disease--an IG-IBD study

BACKGROUND: Three major variants of the CARD15 gene confer susceptibility to Crohn's disease (CD). Whether or not these variants correlate with specific clinical features of the disease is under evaluation. AIM: We investigated the possible association of CARD15 variants with specific clinical characteristics, including the occurrence of anti-Saccharomyces cerevisiae antibodies (ASCA) and antineutrophil cytoplasmic antibodies (ANCA), in a large cohort of inflammatory bowel disease (IBD) patients and their unaffected relatives. METHODS: Three hundred and sixteen CD patients (156 with positive family history), 408 ulcerative colitis (UC) patients (206 with positive family history), 588 unaffected relatives, and 205 unrelated healthy controls (HC) were studied. Single nucleotide polymorphisms (SNPs) R702W, G908R, and L1007finsC of the CARD15 gene were investigated and correlated to age at diagnosis, gender, family history, localization, extraintestinal manifestations, previous resective surgery, stenosing/fistulizing pattern, ANCA, and ASCA. RESULTS: Compared to HC, the frequencies of all three variants in CD were significantly increased: 8.7% versus 4.1% for R702W (p < 0.006), 7.3% versus 2.7% for G908R (p < 0.002), 9.3% versus 0.7% for L1007finsC (p < 0.00001). At least one risk allele was found in 38.2% (p < 0.0001, compared to HC), 13.7% (NS), and 15.1% of CD, UC, and HC, respectively. The L1007finsC risk allele was also significantly increased in unaffected relatives of familial (9.5%; p < 0.00001), and sporadic CD (9%; p < 0.00001), compared to HC (0.7%). Sixteen healthy relatives, carriers of two risk alleles, were asymptomatic after 5-8 yr of follow-up. CD carriers of at least one variant were younger (p= 0.03), more likely to have ileal localization (p= 0.0001), stenosing pattern (p= 0.01), previous resective surgery (p= 0.0001), and presence of ASCA (p= 0.0001). No difference in SNPs frequency between familial and sporadic cases of CD was found. CONCLUSION: In our population, both familial and sporadic CD patients carrying at least one major variant of CARD15 had an aggressive clinical course.