Effect of olive leaves feeding on phenolic composition and lipolytic volatile profile in goat milk.

The aim of this study was to evaluate phenolic composition, antioxidant potential, and lipolytic events in raw milk obtained from goat fed a dietary supplementation with olive leaves (OL), a by-product of the olive oil production chain. For this purpose, 30 Saanen goats were randomly allocated into 2 groups of 15 goats each: the control group received a standard diet that was prepared by taking into account the nutritional needs of lactating goats, whereas the experimental group (EG) was fed with an OL-supplemented diet (10% on a dry matter basis). At the end of the 30 d of the trial, the individual milk samples were collected and immediately analyzed for total phenolic content and antioxidant activity (AOA). Subsequently, the individual phenolic compounds have been identified and quantified through an ultra-high-performance liquid chromatography system and a characterization of free fatty acids released in milk has been performed. The results showed a positive effect of dietary OL supplementation in improving total phenolic content and AOA; furthermore, 19 phenolic compounds, including phenolic acids, flavonoids, simple phenols, and secoiridoids, have been identified in EG milk. In addition to this, a reduced accumulation of free fatty acids has been found in EG milk, and this finding leads us to hypothesize an inhibitory action of the identified phenolic compounds toward the enzymes responsible for lipolytic events. The use of the molecular docking approach verified the interactions, defining a fairly interesting framework for cinnamic acid, which should be able to noncovalently bind these enzymes, interfering with the recruitment of the substrate and therefore, slowing down their hydrolytic activity. In any case, this information will be subjected to in vitro evaluations for an accurate characterization of the biochemical mechanisms that can be established in milk naturally enriched with bioactive compounds.

Chronic urticaria and blastocystis hominis infection: a case report.

We report a case of a 45 year old woman which fulfilled the criteria of chronic urticaria (remitting and relapsing bouts of erythematous and pruriginuos lesions without angioedema, lasted four months). Cutaneous manifestations were not related to a specific inducing factor, had no benefit from antihystamine and steroid drugs and were associated sometimes with mild gastroentric disorders. Patient was submitted to extensive clinical, laboratory and intrumental investigations which permit to exclude many conditions: allergy to inhalants, food, insects and drug adverse reactions, autoimmune urticaria, autoimmune diseases, neoplastic and infectious diseases. Finally coprocolture disclosed the presence of Blastocystis hominis in stool samples thus permitting to associate urticaria to parasitic infection. Both cutaneous manifestations and mild abdomen disturbs disappeared after appropriate treatment. Despite the high diffusion the aetiopathogenesis of chronic urticaria remains often undefined. A large number of parasites have been correlated with urticaria but few data exist as regards Blastocystis hominis infection; then our findings may add evidence to the role of this parasite in inducing chronic urticaria. Considering that Blastocystis hominis is a modest pathogen for humans, the mechanism is probably the typical one of cutaneous allergic hypersensitivity; antigen parasites induce the activation of specific clones of Th2 lymphocytes, the release of related cytokines and the consequent IgE production.

Microvascular assessment in Behçet disease: videocapillaroscopic study.

The aim of this study was to evaluate microvascular assessment in patients with Behcet disease (BD) by means of an intravital videocapillaroscopic study. Sixteen BD patients were compared with an equivalent group of healthy subjects matched for age and sex. Videocapillaroscopy (VCP) was performed in peripheral areas and in conjunctiva, and morphological and quantitative parameters were assessed. In both areas VCP showed several morphological alterations (microaneurysms, megacapillaries, desertification areas) detectable in a high percentage of patients; quantitatively we found significant changes of incisuring and sludging score, of capillary loop intermediate branch length (in peripheral areas) and of arteriole/venule diameter (in conjunctiva). Therefore, vessel involvement included both the number and the whole vessel structure and was seen both in peripheral and conjunctival areas when the two different vascular beds of micro- and paramicrocirculation were examined. We conclude that an important rearrangement of microcirculation is detectable in BD and that VCP may have diagnostic and prognostic value, providing qualitative and quantitative information able to define the systemic extension of vascular damage and the degree of vessel wall alteration.

POEMS syndrome with vascular lesions: a role for interleukin-1beta and interleukin-6 increase--a case report.

The authors describe the case of a 60-year-old man with POEMS syndrome associated with vascular lesions. The patient had osteosclerotic myeloma IgA (lambda), polyneuropathy, endocrinopathy, and skin changes. Subsequently, he developed gangrene of the lower limbs with no response to heparin therapy. The humoral study showed thrombocythemia, high levels of interleukin-1beta (IL-1beta) and IL-6 and of some coagulative/fibrinolytic and endothelial factors (von Willebrand factor, plasmin-antiplasmin complexes, plasminogen activator, and endothelial adhesion molecule ICAM-1). The authors suggest that these factors, induced by the increased levels of cytokines, could be responsible for microvascular damage, gangrene, and heparin resistance.

POEMS syndrome with vascular lesions and renal carcinoma - possible role of cytokines.

We describe here the case of a 60 years man with POEMS syndrome associated with renal tumor and vascular lesions. The patient had osteosclerotic myeloma IgA-lambda, polyneuropathy, endocrinopathy and skin changes. In addition, he developed renal clear cell carcinoma and gangrena of lower limbs. The humoral study showed thrombocytosis, high levels of IL-1beta and IL-6 and of some coagulative/fibrinolytic and endothelial factors (von Willebrand factor, plasmin-antiplasmine complexes, plasminogen activator). We suggest the hypothesis that these factors are capable of determining some manifestations of POEMS syndrome.

Evaluation of some immune functions in a patient affected by common variable immunodeficiency using luminescent techniques.

Common variable immunodeficiency is a primary immunodeficiency characterized by a failure of antibody synthesis, whose fundamental immunologic abnormality is still unknown. In our study, we evaluated some immune functions using chemiluminescence in a 32-year-old woman affected by common variable immunodeficiency. In particular, we showed an impairment of her lymphomonocyte proliferative response which was evaluated using a method based on the bioluminescent measurement of ATP. Besides, we found a reduction of her lymphomonocyte IL2 and IL4 production: the IL4 production was evaluated through an ELISA method, whereas the IL2 activity was determined by its ability to support the IL2-dependent murine T-cell line (CTLL) proliferation which was established through a method based on the bioluminescent measurement of ATP. Finally, we evaluated both yeast-induced and fMLP-induced polymorphonuclear and monocyte oxidative metabolism through a luminol-amplified chemiluminescence; these functions were within normal values. Therefore, in our patient affected by common variable immunodeficiency, we demonstrated an impairment of cellular immunity, which might contribute to the pathogenesis of the disease.

The in vitro effect of Pidotimod on some immune functions in cancer patients.

There are several reports concerning an impairment of cellular immune response in patients affected by malignant disease. The aim of this study was to evaluate the in vitro effect of Pidotimod, a synthetic biological response modifier, on some immune functions in 14 cancer patients. In particular, we showed that these subjects had a significantly reduced peripheral blood mononuclear cell (PBMC) proliferation both in response to PHA and to Con A in comparison with a group of healthy subjects. Besides, they showed a significantly reduced PBMC IL2 production, which was evaluated both through an ELISA method and a biological assay. The in vitro addition of increasing concentrations of Pidotimod (10, 25 and 50 ug/ml) was able to enhance PBMC proliferation and IL2 production significantly. However, in spite of the addition of Pidotimod, both immune functions in our neoplastic patients did not reach normal values.

Changes of some immune functions after percutaneous transluminal coronary angioplasty (PTCA).

This study aimed to evaluate some aspects of the immune response in 10 cardiopathic patients during the execution of percutaneous transluminal coronary angioplasty (PTCA) by obtaining blood samples from coronary sinus. In particular we considered some PMN functions as well as lysosomal release and oxidative metabolism evaluated as chemiluminescence and superoxide anion (O2) production. We also studied serum levels of complement C3 and C4, lymphocyte populations (CD3, CD4, CD8, CD19, CD16) and plasmatic determinations of interleukin 2 (IL2). After PTCA, we found a decrease of total count of blood lymphocytes, whereas the number of neutrophils remained unchanged. The decrease involved to a similar extent the lymphocyte subsets CD3, CD4 and CD8, whereas CD19 and CD16 were unchanged. The plasmatic levels of IL2 did not show any significant modification. Concerning PMN, their chemiluminescence was significantly increased after PTCA as compared to basal values: this response was promptly detectable in isolated PMN, both without and with stimulation with fMLP. Similarly superoxide anion production, both spontaneous and stimulated, was increased in PMN suspensions after PTCA, even if this increase did not reach statistical significance. As regards circulating levels of lysosomal enzymes, we found a significant increase of plasmatic levels of elastase, whereas the serum determinations of lysozyme and betaglucuronidase did not change. Concerning the complement system, we found a significant decrease of complement fractions C3 and C4. In conclusion, our results showed certain changes in some humoral and cellular systems; in particular the neutrophil activation through the release of proteolytic enzymes and the generation of oxygen radicals could increase the damage to vessel walls and activate other systems having a negative effect in the ischaemia-associated consequences.

The binding of human serum transferrin to its specific receptor reconstituted into liposomes.

Human placental transferrin receptor (HPTR), purified following a procedure based on affinity chromatography step, was reconstituted by the detergent dialysis method into various kinds of phosphatidylcholine vesicles and the receptor ability to bind 125I-labelled human serum transferrin (HST) was then evaluated. In our experimental conditions, the binding of the labelled protein to its specific receptor showed several features, in particular: (1) in cholesterol/1-alpha-dipalmitoylphosphatidyl choline (CHO/DPPC) liposomes, a positive cooperatively of the transferrin binding resulted at the lowest cholesterol/phospholipids (C/P) ratio; 1-alpha-dioleylphosphatidyl choline (DOPC) and phosphatidic acid (PA) containing liposomes showed an opposite binding curve trend; (2) the apparent dissociation constant (K'd) did not change significantly as a function of the lipid composition, being always around 1.00 x 10(-6) M; (3) the encapsulation capacity of liposomes decreased from 27% to about 13% with increasing amounts of cholesterol and was around 20% in the presence of DOPC or PA; about 8-13% of this receptor was found to be functional; (4) receptor-loaded liposomes treated with polyclonal anti-HPTR rabbit antibodies showed a remarkable binding decrease for transferin. All these results seem to point out the crucial role played by the environment in the binding behaviour of the transferrin receptor.

Structural analysis of seminal and serum human transferrin by second derivative spectrometry and fluorescence measurements.

Denaturation of human seminal transferrin (HSmT) compared with human serum transferrin (HSrT) was followed to check structural differences between these two proteins. Second derivative UV spectroscopy indicated that treatment with 6 M guanidine hydrochloride (Gnd.HCl) induced greater structural changes in HSrT than in HSmT and, in particular; (i) the exposure value of tyrosinyl residues was almost 2.5-fold higher in native HSmT than in native HSrT; and (ii) a much more pronounced movement of tryptophanyl residues toward a higher polar environment could be noticed in HSrT after incubation with denaturing agent. Fluorescence measurements showed that: (i) a shift of the maximum emission wavelength of HSmT occurred (maximum emission was centered at 333 nm instead of 323 nm as for HSrT; excitation = 280 nm); (ii) the intrinsic tryptophan fluorescence intensity of HSmT increased after 36 hr in the range of 1.5-4.0 M of denaturant, whereas an opposite behavior was found for HSrT in the range 0.0-2.0 M; and (iii) the wavelength maximum of fluorescence emission changed in a biphasic manner for HSrT and, conversely, under the same experimental conditions, HSmT gave a linear and parallel increase of fluorescence emission after 1 and 36 hr. We can conclude that this different behavior of HSmT with respect to HSrT might be due mainly to the fact that both the number and the exposure of tyrosinyl and tryptophanyl residues are different. Lately, these effects are discussed in relationship with the fact that HSmT contains less than half disulphide bridges than HSrT.

Hemodynamic and hemorheologic effects of i.v. Dilevalol in hypertensive patients.

The authors evaluated the effect of Dilevalol infusion on blood pressure, heart rate, central hemodynamics, and rheologic parameters in hospitalized inpatients affected with mild or moderate hypertension. After a dose-finding phase and a washout period of one week, 10 patients aged fifty to seventy-two-years (median 61.5) were given either a single dose of Dilevalol 60 mg or placebo, and seven days later they underwent the other treatment, according to a single-blind, crossover design. Central hemodynamic measurements were performed by means of M-mode echocardiography, and hemorheologic parameters were evaluated by means of strain-gauge plethysmography. The maximal increase in lower extremity flow at rest had been obtained with the infusion of 60 mg Dilevalol during dose-finding, and so this dose was chosen for the second part of the study. The infusion of Dilevalol significantly increased rest flow and decreased blood viscosity, but the changes in central, parameters were not considered clinically relevant, although statistically significant. Blood pressure decreased without significant changes in heart rate. Thus, the acute administration of Dilevalol reduced blood pressure, without affecting heart rate and central hemodynamics, confirming the vasodilating effect of the drug. A significant improvement was also shown on blood viscosity in these hypertensive patients.

Pharmacokinetics and pharmacodynamics of slow-release theophylline during treatment with nimesulide.

The pharmacodynamic and pharmacokinetic interactions were studied between nimesulide, a recently introduced non-steroidal anti-inflammatory drug, and theophylline, another highly protein-bound drug, in patients who were receiving slow-release theophylline for a chronic airflow-obstruction and who also needed anti-inflammatory treatment. A good tolerability was demonstrated of the two drugs association and there was an absence of pharmacodynamic interaction, as shown by lung function parameters, assayed before and after the coinciding nimesulide association. The pharmacokinetics of nimesulide and 4-hydroxy-nimesulide (its active metabolite) were not modified, in agreement with data shown by other authors. On the contrary, there was a slight alteration of theophylline pharmacokinetics, yet neither clinically nor biologically significant, probably due to an enzymatic induction.

Clinical study on pharmacological interaction between nimesulide and warfarin.

This article describes the pharmacological interaction between nimesulide, a recently introduced non-steroidal anti-inflammatory drug, and warfarin, an indirect anticoagulant. The aim of the study was to demonstrate if nimesulide could potentiate the activity of this anticoagulant drug, as previously shown by some authors. Ten patients, who were taking 5 mg/day of warfarin, were treated with nimesulide 100 mg twice a day, for seven days: the association of the two drugs did not alter, in a statistical way, neither prothrombin time, nor partial thromboplastin time, nor fibrinogenemia, nor bleeding time. The findings showed that, in a short-term treatment, there was no bleeding risk in combining warfarin with nimesulide.

Effect of cefodizime (HR 221) on immunological defects induced by surgical stress.

Cefodizime, a new aminothiazolylcephalosporin, has been shown to possess immunomodulating activity in many experimental models in vivo and in vitro. The in-vivo effect of the drug was evaluated in a model represented by the surgical patient, being surgical practices usually associated with an immunological impairment involving many aspects of the immune response. Two groups of ten subjects were treated respectively with cefodizime (2 g i.v. daily) and another cephalosporin (ceftriaxone) at the same dosage. Aspecific immunity (complement activity, neutrophil phagocytosis, chemiluminescence and superoxide anion production) and cell-mediated reactivity (lymphocyte subpopulations and E-rosette-forming cells) were evaluated before, and at predetermined intervals after, surgery and antibiotic treatment. In the control group an important immunological derangement is observed, involving both lymphocytes and neutrophil functions and complement system. The treatment with cefodizime displays a positive effect with a significant improvement of impaired functions. The effect of the drug particularly influences neutrophil phagocytosis, explored with both the NBT test and determinations of chemiluminescence, and the complement system, through both the classic and the alternative pathways. A slight effect is observed on lymphocyte functions.

[The effects of vinburnine on the blood rheology parameters and on the oxyhemoglobin dissociation curve and 2,3-diphosphoglycerate. In vitro research]. / Effetti della vinburnina sui parametri emoreologici e sulla curva di dissociazione ossiemoglobinica e 2-3 difosfoglicerato. Ricerche in vitro.

The authors investigate the protective action of vinburnine on impairment of blood haemorheology and on metabolic parameters due to in vitro ageing of blood samples. In each sample the following parameters were measured: blood viscosity and filterability, p50 and 2,3-DPG. Vinburnine is able to induce a slight improvement of haemorheological parameters and of p50 compared to controls. After incubation for 4 hr, a reduced impairment of the mentioned parameters was observed in comparison with controls. Some hypotheses are discussed about the mechanisms of action of the drug.

[The effects of the acute administration of vinburnine on blood rheology, oxygen transport and regional circulation under normal conditions and in hypoxemia. In vivo research]. / Effetti della somministrazione acuta di vinburnina sulla reologia ematica, trasporto dell'ossigeno e sulla circolazione distrettuale in condizioni normali e di ipossiemia. Richerche in vivo.

Vinburnine has been shown to be effective in avoiding chronic ischaemic tissue injury. Many studies have demonstrated the antihypoxic and oxygenation properties of this drug. In our study we have evaluated in two groups of patients the effects on blood rheology, oxygen transport, regional circulation and ergospirometric data of 40 mg vinburnine infused in a systemic vein. We have selected in the first group 10 patients with low hemoglobin oxygen affinity (p50 28.6 +/- 1.37 mmHg), in the second one 7 patients with normal values (p50 26.6 +/- 0.84 mmHg). In the two groups of the study we evaluated nutritional blood flow by a plethysmographic method, blood gas analysis [correction of hemogasanalitic] parameters in arterial and venous blood, haemoglobin oxygen affinity expressed as p50 and erythrocytic 2,3-DPG, hemorheologic and ergospirometric parameters. Data were evaluated in basal conditions and at the end of the infusion. The improvement of rheological properties, hemodynamic and metabolic data in the two groups of the study seems to confirm the oxyphoretic properties of this drug.

PPIX induced photohemolysis of erythrocytes partially-depleted of cholesterol.

The protoporphyrin IX (PPIX)-sensitized hemolysis of erythrocytes depleted of cholesterol was investigated. From 20% to 30% of the total membrane cholesterol was removed from cells by incubation with old autologous plasma or by means of interaction with L-alpha-phosphatidylcholine dipalmitoyl (DPPC) liposomes. As expected, after this treatment, the cells show an overall increase in membrane fluidity revealed by means of specific fluorescent probes. The same cells are more susceptible to the photohemolysis induced by PPIX excited by visible light, but gave no lysis in the absence of the sensitizer. As a consequence, the primary oxidative damage which is produced during irradiation can be possibly assigned to the phospholipidic and/or proteic moiety instead of the steroidal moiety.