In-vivo and in-vitro anti-inflammatory effect of Echinacea purpurea and Hypericum perforatum

Echinacea purpurea (L.) Moench and Hypericum perforatum (L.) were evaluated for their anti-inflammatory activity against carrageenan-induced paw oedema in mice. Each drug was administered orally to mice at 30 and 100 mg kg(-1), twice daily. Only the higher dose significantly inhibited, time dependently, the formation of oedema, evaluated as area under the curve (echinacea P < 0.01; hypericum P < 0.05). Western blot analysis showed that in-vivo treatment with these extracts could modulate lipopolysaccharide (LPS) and interferon-gamma induced cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expression in peritoneal macrophages. In particular, treatment with 100 mg kg(-1) hypericum inhibited both iNOS and COX-2 expression, whereas treatment with 100 mg kg(-1) echinacea down-regulated only COX-2 expression. The present study suggests that the anti-inflammatory effect of these extracts could be in part related to their modulation of COX-2 expression.     

Restoration of human beta-globin gene expression in murine and human IVS2-654 thalassemic erythroid cells by free uptake of antisense oligonucleotides

Correct human beta-globin mRNA has been restored in erythroid cells from transgenic mice carrying the human gene with beta-globin IVS2-654 splice mutation and from thalassemia patients with the IVS2-654/beta(E) genotype. This was accomplished in a dose- and time-dependent manner by free uptake of morpholino oligonucleotide antisense to the aberrant splice site at position 652 of intron 2 in beta-globin pre-mRNA. Under optimal conditions of oligonucleotide uptake, the maximal levels of correct human beta-globin mRNA and hemoglobin A in patients' erythroid cells were 77 and 54%, respectively. These levels of correction were equal to, if not higher than, those obtained by syringe loading of the oligonucleotide into the cells. Comparison of splicing correction results with the cellular uptake of fluorescein-labeled oligonucleotide indicated that the levels of mRNA and hemoglobin A correlate well with the nuclear localization of the oligonucleotide and the degree of erythroid differentiation of cultured cells. Similar but not as pronounced results were obtained after the oligonucleotide treatment of bone marrow cells from IVS2-654 mouse. The effectiveness of the free antisense morpholino oligonucleotide in restoration of correct splicing of IVS2-654 pre-mRNA in cultured erythropoietic cells from transgenic mice and thalassemic patients suggests the applicability of this or similar compounds in in vivo experiments and possibly in treatment of thalassemia.     

Neurosteroid secretion in panic disorder

Evidence that neurosteroids have anxiolytic effects in animal models of anxiety has suggested that alterations of neurosteroid secretion might be implicated in the pathogenetic mechanisms of anxiety disorders in humans. In 25 female patients with panic disorder (PD) and 11 healthy female controls, we measured plasma concentrations of progesterone (PROG), pregnenolone (PREG), allopregnanolone (3alpha,5alpha-tetrahydroprogesterone=3alpha,5alpha-THPROG), dehydroepiandrosterone (DHEA) and tetrahydrodeoxycorticosterone (3alpha,5alpha-THDOC) during a drug-free month and during the following month of paroxetine therapy. The neurosteroids were measured during the early follicular phase, the mid-luteal phase and the premenstrual phase of both months (days 7, 22 and 27 from the beginning of the cycle). Significantly higher levels in patients than controls were found in PROG during the mid-luteal phase of both months, PREG in the premenstrual phase in the drug-free month, 3alpha,5alpha-THPROG during the follicular phase of the drug-free month and during the premenstrual phase of the therapy month, and 3alpha,5alpha-THDOC during the premenstrual phases of both months. DHEA levels did not differ in patients and controls. These results suggest that neurosteroids in PD are hypersecreted, possibly as an attempt to counteract the anxiogenic underlying hyperactivity of the hypothalamo-pituitary-adrenal axis and to improve a reduced GABA(A) receptor sensitivity.     

Chronic hypoxia: a model for cyanotic congenital heart defects

OBJECTIVE: The postoperative course of cyanotic children is generally more complicated than that of acyanotic children. A possible reason is reoxygenation injury at the beginning of cardiopulmonary bypass. In this study we tested the hypothesis that reoxygenation of chronically hypoxic hearts is worse than that of normoxic hearts. METHODS: Two groups of rats (n = 9 each) were exposed to either room air (fraction of inspired oxygen, 0.21%) or chronic hypoxia (fraction of inspired oxygen, 0.10%) for 2 weeks. Hearts were then isolated and perfused for 30 minutes with hypoxic buffer (oxygen saturation, 10%), followed by 30 minutes of reoxygenation (oxygen saturation, 100%). RESULTS: In hypoxic rats hematocrit values, hemoglobin concentrations, and red cells were higher (69% +/- 6% vs 40% +/- 6%, 219 +/- 14 vs 124 +/- 12 g/L, and 10.30 +/- 0.6 vs 6.32 +/- 0.5/microL/1000, respectively; P <.0001); the amount of ingested food was less (22.3 +/- 4.8 vs 30.7 +/- 3.9 g/d, P <.001), as was the amount of ingested water (21.0 +/- 3.1 vs 50.4 +/- 14.6 mL/d, P <.0001); and body weight was lower (182 +/- 14.2 vs 351 +/- 40.1 g, P <.0001), as was heart weight (1107 +/- 119 vs 1312 +/- 128 mg, P <.005). The heart weight/body weight ratio was higher (6.10 +/- 0.8 vs 3.74 +/- 0.1 mg/g, P <.0001). Systolic and diastolic functions, not different during the hypoxic baseline period, were more impaired in hypoxic than in normoxic hearts after the reoxygenation, whereas coronary resistance remained lower. During the hypoxic perfusion, the venous partial pressure of oxygen remained low in both groups, whereas during reoxygenation, partial pressure of oxygen was higher in hypoxic hearts, with a lower (P <.01) oxygen uptake. During hypoxic baseline adenosine triphosphate turnover, lactate production and lactate turnover were lower in hypoxic hearts (P <.005, P <.0001, and P <.0001, respectively). CONCLUSIONS: Body and blood values are severely affected by chronic hypoxia, and the cardiac effects of uncontrolled reoxygenation after chronic hypoxia are more severe than after acute hypoxia.     

Captopril enhances transforming growth factor (TGF)-beta1 expression in peripheral blood mononuclear cells: a mechanism independent from angiotensin converting enzyme inhibition? A study in cyclosporine-treated kidney-transplanted patients

BACKGROUND: Angiotensin (Ang) II blockade has been shown to prevent the development of renal injury in immunologically mediated diseases, but the mechanism whereby it exerts its protective effect has not been clearly defined. Transforming growth factor (TGF)-beta1 is a multifunctional cytokine with a potent immunomodulatory activity that has the potential to counteract many of the pro-inflammatory effects apparently evoked by the activation of renin-angiotensin system (RAS) in immune cells. METHODS: We set up an ex vivo and in vitro model to evaluate the effect of the angiotensin converting enzyme inhibitor (ACEi) captopril on the gene and protein expression of TGF-beta1 in human peripheral blood mononuclear cells (PBMC). RESULTS: In 20 kidney transplant recipients chronically treated with cyclosporine (CsA), 1-month treatment with captopril increased TGF-beta mRNA by 120% and TGF-beta1 protein release by 140% upon stimulation of PBMC with phytohemagglutinin (PHA) and phorbol myristate acetate (PMA) ( P<0.01). PBMC from healthy controls, when exposed in vitro to 5 microM captopril, showed a significant increase of TGF-beta1 release, whereas the ACEi enalapril failed to modify the expression of the cytokine. Ang II (100 pM) strongly inhibited TGF-beta1 synthesis by PBMC, and such effect was completely abolished by the addition of 200 ng/mL CsA, as well as by 1 micrpM losartan. Thus, captopril enhances TGF-beta1 gene and protein expression by PBMC by way of a mechanism independent, at least in part, from ACE inhibition, while CsA abrogates the inhibition of TGF-beta1 expression induced by Ang II. CONCLUSION: Collectively, these findings support the utility of combined treatment with captopril and CsA in the multitherapeutic management of organ transplant and, possibly, a strategy to decrease the dose of the calcineurin inhibitor in kidney-transplant recipients.     

Postsplenectomy immunodepression and its implications: an underestimated problem

The most important sequelae of splenectomy is immunity depression. This study, conducted in three phases, was aimed at confirming this clinical condition. Data from our phase 1 study clearly show that patients undergoing splenectomy for trauma are in a critical condition because of a latent immunodeficiency shown by skin tests (ST) and in vitro evaluation of the aspecific immune activity. Because the in vitro study of the unspecific immunity that we used seems to be more expensive and complicated than ST, the aim of the phase 2 study was to compare the efficacy and the limits of the two assays (ST versus in vitro study) in detecting the immunodeficiency status of the splenectomized patient. The aim of the phase 3 study was to ascertain whether postsplenectomy immunodeficiency could be a consequence of an altered equilibrium between the lymphocyte subpopulations T helper (Th)1/Th2, evaluated by serum dosage of interferon-gamma and interleukin-4.     

Phase I study of gemcitabine and liposomal doxorubicin in relapsed ovarian cancer

Twenty-three patients were enrolled in a phase I study conducted to determine the maximum tolerated doses (MTD) of combined liposomal doxorubicin (CAE) and gemcitabine (GEM) in relapsed ovarian cancer patients. A total of 82 courses are evaluable, with a median number of three cycles administered per patient (range 2-8). GEM was administered on days 1 and 8 by 30-min intravenous infusion immediately after CAE given by 60-min intravenous infusion on day 1; cycles were repeated every 21 days. The starting doses were CAE 20 mg/m(2) and GEM 600 mg/m(2). Following dose levels were 20/800; 20/1,000; 30/800; 30/1,000; 35/800, and 35/1,000 for CAE and GEM, respectively. The MTD was reached at dose level 5, with febrile neutropenia and thrombocytopenia as dose-limiting toxicities. After the MTD, granulocyte-colony stimulating factor was administered in 15% of cycles. Non-hematological toxicity was mild and manageable. All patients are so far evaluable for response. Among them, 5 partial responses (21.7%; 95% confidence interval, CI: 4.9-38.5), 5 disease stabilizations (21.7%, 95% CI: 4.9-38.5) and 13 progressions (56.6%, 95% CI: 36.4-76.8) have been registered. These results warrant further research in a phase II study.     

High-dose cytarabine as consolidation treatment for patients with acute myeloid leukemia with t(8;21)

Seventeen patients affected by acute myeloid leukemia (AML) with t(8;21) were prospectively programmed to receive three courses of high-dose cytarabine (HDARA-C) as post-remission therapy. The median age was 39 years and in all cases t(8;21) was the only karyotypic abnormality. Complete remission (CR) was achieved in 14 out of 17 cases (82%) and, after first consolidation with NOVIA regimen (intermediate dose ARA-C plus mitoxantrone), all patients received the three planned courses of HDARA-C (3g/m(2) q12h on days 1, 3, 5). There were two documented infections, while all patients experienced fever of unknown origin (FUO). Nonhematological toxicity was mild. Thirteen out of 14 patients are in continuous CR after a median follow-up of 44 months. One patient relapsed at 16 months and, following CR2 achievement, underwent allogeneic transplantation; he died 3 months later while in CR from acute graft versus host disease (GVHD). Survival at 5 years is projected at 79%. Our data confirm the efficacy of repeated courses of HDARAC for patients with t(8;21) AML.     

Motion sickness and postoperative vomiting in children

BACKGROUND: Motion sickness is considered an important risk factor for postoperative nausea and vomiting in children. The aim of this study was to verify the impact of motion sickness on the incidence of vomiting after routine surgery in children, and to compare the incidence of vomiting, after combined regional/general anaesthesia, using either halothane or sevoflurane. METHODS: We prospectively studied 420 children (369 males and 51 females) who received general anaesthesia and inguinal field block for common paediatric surgery. The children were randomly allocated into one of two groups (halothane or sevoflurane). In the 200 children in the first group (H), general anaesthesia was induced and maintained with halothane, whereas in the 220 children in the second group (S), anaesthesia was induced and maintained with sevoflurane. RESULTS: There were 79 children with a prior history of motion sickness (MS+) and 341 without such a history (MS-). In the MS+ population, the incidence of vomiting was similar in both H and S groups, being around 33%. However, repeated episodes of vomiting in MS+ children were more frequent when halothane was used. In the MS- group, the incidence of vomiting was significantly greater in the H group (19%) than in the S group (8%). CONCLUSIONS: In the postoperative period, we found that MS+ children vomit more than MS- children, regardless of the inhalation anaesthetic used. However, MS- children displayed a higher incidence of vomiting when halothane was used rather than sevoflurane.     

Low-dose oral methotrexate and cyclophosphamide in metastatic breast cancer: antitumor activity and correlation with vascular endothelial growth factor levels.

BACKGROUND: Anticancer chemotherapy is thought to be effective by means of direct cytotoxicity on tumor cells. Alternative mechanisms of efficacy have been ascribed to several common anticancer agents, including cyclophosphamide (CTX), methotrexate (MTX), anthracyclines and taxanes, postulating an antiangiogenic activity. PATIENTS AND METHODS: We evaluated the clinical efficacy and impact on serum vascular endothelial growth factor (VEGF) levels of low-dose oral MTX and CTX in patients with metastatic breast cancer. MTX was administered 2.5 mg bd on days 1 and 2 each week and CTX 50 mg/day administered continuously. RESULTS: Sixty-four patients were enrolled, 63 were evaluable: Eastern Cooperative Oncology Group (ECOG) performance status 0-1, > or =2 sites of metastatic disease (n = 50 patients), progressive disease at study entry (n = 51), 1 regimen for metastatic disease (n = 32) and > or =2 regimens (n = 20). Among the 63 evaluable patients, there were two complete remissions (CR), 10 partial remissions (PR) for an overall response rate of 19.0% (95% CI 10.2% to 30.9%) and an overall clinical benefit (CR+ PR+ stable disease >24 weeks) of 31.7% (95% CI 20.6% to 44.7%). Grade > or =2 leucopenia was registered in only 13 patients. The median serum VEGF level for the subgroup of patients on treatment for at least 2 months decreased with treatment from 315 pg/ml (95% CI 245 to 435) at baseline to 248 pg/ml (95% CI 205 to 311) at 2 months (P <0.001). Both responders and non-responders showed similar reductions in serum VEGF (P = 0.78). After 6 months patients still on treatment had a median VEGF level of 195 pg/ml (95% CI 96 to 355), which was significantly lower than the median baseline values (P = 0.001). CONCLUSIONS: Continuously low-dose CTX and MTX is minimally toxic and effective in heavily pretreated breast cancer patients. A drop in VEGF was associated with the treatment and so alternative hypotheses, other than that of direct toxicity on tumor cells, must be favored when trying to explain the anticancer effect.