Method development for the characterization of biofuel intermediate products using gas chromatography with simultaneous mass spectrometric and flame ionization detections.

Accurate analytical methods are required to develop and evaluate the quality of new renewable transportation fuels and intermediate organic liquid products (OLPs). Unfortunately, existing methods developed for the detailed characterization of petroleum products, are not accurate for many of the OLPs generated from non-petroleum feedstocks. In this study, a method was developed and applied to the detailed characterization of complex OLPs formed during triacylglyceride (TG) pyrolysis which is the basis for generating one class of emerging biofuels. This method uses gas chromatography coupled simultaneously with flame ionization and mass spectrometry detectors (GC-FID/MS). The FID provided accurate quantification of carbonaceous species while MS enabled identification of unknown compounds. A programed temperature vaporizer using a 25 °C, 0.1 min, 720 °C min(-1), 350 °C, 5 min temperature program is employed which minimizes compound discrimination better than the more commonly utilized split/splitless injector, as verified with injections at 250 and 350 °C. Two standard mixtures featuring over 150 components are used for accurate identification and a designed calibration standard accounts for compound discrimination at the injector and differing FID responses of various classes of compounds. This new method was used to identify and quantify over 250 species in OLPs generated from canola oil, soybean oil, and canola methyl ester (CME). In addition to hydrocarbons, the method was used to quantify polar (upon derivatization) and unidentified species, plus the unresolved complex mixture that has not typically been determined in previous studies. Repeatability of the analytical method was below 5% RSD for all individual components. Using this method, the mass balance was closed for samples derived from canola and soybean oil but only ca. 77 wt% of the OLP generated from CME could be characterized. The ability to close the mass balance depended on sample origin, demonstrating the need for an accurate quantification method for biofuels at various stages of production.

Cardiac involvement in inflammatory bowel disease: role of acylcarnitine esters.

OBJECTIVES: Cardiovascular complications have been frequently described in Inflammatory Bowel Disease (IBD). Both Crohn disease and Ulcerative Colitis are characterized by malabsorption of some micronutrients, such as carnitine, which is a very important element for myocardial metabolism, being demonstrated that its deficiency correlates with heart involvement in coeliac disease. Aims of this study are to evaluate cardiac function in IBD patients asymptomatic for cardiovascular diseases and to correlate the cardiac data with the profile of carnitine esters plasma levels. MATERIALS AND METHODS: The study was carried out on 20 IBD patients by comparison with 18 sex- and age-matched clinically healthy controls. Personal and familial history, physical examination, standard electrocardiogram and echocardiogram were performed in all subjects. Complete panel of nutritional status parameters and serum levels of free carnitine and its esters were evaluated both in IBD patients and control subjects. RESULTS: Isovaleryl-carnitine, Tiglyl-carnitine, Octenoylcarnitine and Decanoyl-carnitine, were found to be significantly lower in IBD patients. Significant correlations were found between some carnitine esters and echocardiographic parameters although total and free carnitine were meanly more elevated in IBD. No statistically significant differences in echocardiographic parameters were found between IBD patients and control subjects. CONCLUSIONS: Deficiency of some isoforms of carnitine, especially those esterified with short chain fatty acids, may play an important role in cardiac involvement in course of IBD and could lead, over time, to dilated cardiomiopathy.

Henoch-Schönlein purpura in a patient with bowel bypass syndrome.

Henoch-Schönlein purpura (HSP) is a common vasculitis being characterized by the classic tetrad of nonthrombocytopenic palpable purpura, arthritis or arthralgias, gastrointestinal and renal involvement. Antigen-antibody (IgA) complexes activate the alternative complement pathway, resulting in inflammation and small vessels vasculitis. We present the case of a 53 years old Italian woman with HSP who was previously hospitalized for purpura skin lesions of the lower legs and diarrhea; a skin biopsy showed a leukocytoclastic vasculitis with perivascular accumulation of neutrophils and mononuclear cells. She was treated with immunosuppressive therapy. After 8 months she was hospitalized again for a recurrent episode of purpura skin lesions of the lower legs. At age 49 she was affected by obesity (BMI = 41.6 Kg/m2), treated via a bilio-pancreatic diversion that led, within a year, to a BMI reduction (25 Kg/m2). We suppose that bariatric surgery played a role on the development of autoimmune phenomena and that the formation of immunecomplexes is secondary to the excess of intestinal bacterial antigens. A cyclic therapy with Paromomicine 500 mg twice daily and Metronidazole 250 mg twice daily was performed with a clear up of the clinical picture. In medical literature are described numerous complications which include arthritis, erythema nodosum-like lesions, eruptions and other skin manifestations in patients who have undergone jejunocolic bypass. This case report describes for the first time the presence of HSP in a patient with bowel bypass syndrome and it is also able to demonstrate the relationship between the intestinal bacterial overgrowth and the systemic autoimmune system.

Takotsubo syndrome and brachydactyly: a new heart-hand syndrome?

The Takotsubo syndrome (TTS) is a transient myocardic hypoadyskinesis so called for its typically aspect like the cages used by Japanese fisherman to trap octopus. Presently, several etiologies were considered for the development of TTS, the most accredited ones suggest that stress, adrenoreceptor hyperactivity and estrogens are the most important causes of transient morphofunctional alterations of left ventricle. We describe the case of 60 year-old woman with a chest pain occurred after a quarrel. Because of this, she went to the Emergency Department (ED) where T wave flat in lead D-I, increase of CK-MB mass and Troponin I were found. Subsequently, a coronary-ventriculography was performed and a diagnosis of Takotsubo syndrome was made. Also a bilateral IV toe brachydactyly was noted. In medical literature, there are many syndromes described about the association between skeletal alteration of the hands and heart abnormalities (heart-hand syndrome). The Authors wonder if this case can belong to this kind of complex malformations as well as if a molecular-genetic trait can link the two clinical aspects.

A singular case of granulomatous lesions of unknown significance.

The acronym used for granulomatous lesion of unknown significance, (GLUS), was first employed in 1990, to describe epithelioid-cell granulomas occasionally observed in biopsies from liver, lymph-nodes and other tissues without any apparent cause than thorough diagnostic examinations. Authors describe a case of a 51-year-old woman who was admitted to hospital because of fever, nodules in the liver and spleen, skin ulcerative lesions on the legs. The patient had a very long medical history; recurrent fever, liver and spleen enlargement have been lasting since 1975. Laboratory data showed erythrocyte sedimentation rate increase (125 mm 1 hour), anemia, (Hb=8.1 g/dL), an increase of alkaline phosphatase activity (328.4 mU/mL NV=80-275) and also an increase of policlonal gamma-globulins; patient's total proteins was 6.8 g/dL while their gamma-globulins were 29.5%; IgG=2 260 mg/dL (NV=751-1 560 mg/dL), IgM=277 mg/dL (NV=46-230 mg/dL), IgA=405 mg/dL (NV=70-400 mg/dL). One culture of the blood and a needle aspiration specimen of liver nodule was positive for Staphylococcus coagulase negative. This case cannot be regarded as GLUS-syndrome because of the long time duration of the disease and other clinical features. The dramatic, even if transient, liver nodules improvement obtained by Linezolid therapy, and the well-being obtained by prolonged monociclin use, make the authors hypothesize that Staphylococcus coagulase negative could be the etiological agent of this granulomatous clinical picture.

Plasma glucose level decreases as collateral effect of fermented papaya preparation use.

AIM: Fermented Papaya Preparation (FPP) is a natural healthy drug that has been commercially sold in Japan and Philippines. This nutricetive, bio-normalizer product has antioxidant action, inhibitory effect on oxidative DNA damage and tissue injury, being a potent OH scavanger. The wide use of FPP, expecially by elderly people, made us note an unknown collateral effect, i.e., blood sugar level dropping signs especially in the afternoon. The aim of the present work was to scientifically verify the possibility that individuals, who are taking the nutriceutical FPP, might have a decrease of plasma sugar levels. MATERIALS AND METHODS: For this purpose, 50 subjects, divided in two groups, were enrolled. The first group was made of 25 patients: 13 females and 12 males affected by type-2 diabetes mellitus under treatment with the oral antidiabetic drug, glybenclamide. The control group included 25 clinically-healthy subjects: 16 females and 9 males, matching in age. All subjects were given 3 grams of FPP daily, during lunch, for two months. RESULTS: The results of this study confirmed the empirical experience that FPP use can induce a significant decrease in plasma sugar levels in both healthy subjects and type 2 diabetic patients. This hypoglycaemic effect, associated with clinical signs, induced the diabetic patients to reduce the dosage of their antidiabetic oral therapy (in one patient the therapy was really suspended). CONCLUSIONS: In accordance with these results, the FPP administration is suggested as an adjuvant drug to join the oral antidiabetic therapy in type 2 diabetes meltus.

Do hypertension and diabetes mellitus influence the site of atherosclerotic plaques?

BACKGROUND: The concomitant occurrence of atherosclerotic plaques in carotid, coronary and peripheral vessels has been described in a number of studies. A few studies were, on the contrary, done for determining the role of hypertension and/or type 2 diabetes mellitus for the occurrence of the atherosclerotic plaques in different anatomical sites. Moreover these studies deal with atherosclerotic lesions that are generally considered, without differentiating their morphology as a function of the underlying disease, territory, and risk factors. Primary aim of this study is, thus, to verify whether the two most common causes for atherosclerotic disease, i.e., hypertension and type 2 diabetes mellitus, may influence the site of appearance of the atherosclerotic plaque. A second aim is to verify if the anatomical site of the plaque influences plaque morphology and vulnerability. PATIENTS AND METHODS: A retrospective study of 244 patients affected with type 2 diabetes mellitus or hypertension was performed; 114 subjects were affected by moderate-severe and drugs-treated hypertension (Group A); 55 were affected by type 2 diabetes mellitus in treatment with oral antidiabetic drugs (Group B); 75 were diagnosed as affected by the association hypertension and diabetes (Group C). The inclusion criteria were: exhaustive images of the cardiovascular system (coronary angiography, colour Doppler ultrasound of lower limb arteries and carotid arteries, transthoracic Doppler echocardiography ) and a serum lipid profile (total serum cholesterol, HDL-cholesterol, LDL-cholesterol, triglycerides). Three different anatomical sites: carotid axis, ilio-femoral arteries and coronary district, were considered. In each site a plaque lesion-classification was performed to describe the morphology of the plaque. RESULTS: In patients with hypertension, carotid district seems to be the preferential site of onset of atherosclerotic plaques even if a statistical significant association between the two conditions was not found. Statistical evaluation didn't show significant association between different risk factors and coronary district too. On the opposite, a significant association (p < 0.001) between diabetes and the presence of atherosclerotic plaques into lower limb district was found. A very significant association (p < 0.001) between type 2-diabetes and the presence of non-ulcerative plaques was found too. CONCLUSIONS: Our study underlines the relationship between vessel plaques localization and concomitant risk factors for atherosclerosis and suggests a possible difference in plaque morphology and biological behaviour related to different anatomical site.

Carnitine deficiency in patients with coeliac disease and idiopathic dilated cardiomyopathy.

BACKGROUND AND AIMS: Idiopathic dilated cardiomyopathy (IDCM) and coeliac disease (CD) are two pathological conditions which may lead, by different mechanisms, to malabsorption of various micronutrients, including carnitine, active in cardiac metabolism. The aim of the present investigation was primarily to evaluate differences in serum concentrations of total carnitine between IDCM patients and patients with IDCM associated with CD and then also to evaluate, in the latter, the effect of a gluten-free diet on serum concentrations of total carnitine. METHOD AND RESULTS: Serum carnitine was determined by enzymatic spectrophotometric assay in three groups of individuals: group A, 10 patients (5 males, 5 females), mean age 46.5+/-10.8 years, presenting isolated IDCM; group B, 3 patients (2 males, 1 female), mean age 34+/-8 years, with IDCM+CD; and group C, 10 healthy subjects (5 males, 5 females), mean age 38.6+/-11.1 years. All patients with IDCM belonged to class NYHA I-II. Mean concentrations of total serum carnitine in the group of patients with isolated IDCM (group A) were found to be lower than in the controls (group C). The concentrations in patients with IDCM associated with CD (group B) were lower than in the control group and also lower than in the isolated IDCM (group A). After 2 years on a gluten-free diet, patients presenting IDCM associated with CD showed a progressive increase in mean serum carnitine levels compared to values observed prior to the diet. CONCLUSIONS: Patients presenting IDCM associated with CD show a greater decrease in serum total carnitine levels than patients presenting the isolated form of IDCM. A gluten-free diet, in these patients, leads to a progressive increase in serum levels of this substance.

[Sodium channels and multiple sclerosis]. / Canali del sodio e sclerosi multipla.

Multiple Sclerosis is a chronic demyelinating disease of the central nervous system of undetermined etiology. Damage of myelinated fibers leads to block of conduction of impulses. In myelinated axons sodium channels are expressed at high density and they play a very important role in the conduction of nervous impulse. In myelinated fibers affected by Multiple Sclerosis substantial variations of sodium channels pattern occurs. These variations can help to explain pathophysiological and clinical aspects of Multiple Sclerosis and open a new way to approach and, probably, treat this disease.

[A five year follow-up of an HHV-8 related lymphoma in an HIV-negative elderly patient]. / Follow-up a 5 anni di un caso di Linfoma Primitivo delle cavità correlato all'HHV-8, in un paziente anziano HIV negativo.

Primary Effusion Lymphoma (PEL) associated with the Human Herpes Virus 8 (HHV-8), is a rare type of non-Hodgkin lymphoma. It mainly strikes HIV-positive men. Five-year follow-up of a case of PEL HHV-8 related in HIV negative elderly man, is described. The patient was admitted to our Department for dyspnea, thoracic pain and persistent slight temperature. Chest radiography showed a left pleural effusion. Cytomorphological, immunohistochemical and molecular assays performed on pleural fluid, demonstrated the presence of a PEL HHV-8 related, in absence of Epstein-Barr virus (EBV) infection in the tumour cells. Serologic test for HIV (ELISA) resulted negative. Chest TC, taken after thoracentesis, showed marked thickening of diaphragmatic pleura. Because patient's age and general conditions, no chemotherapy was performed. Five years after diagnosis, clinical examination and chest tomographyshowed resolution of the described syndrome; particularly chest TC showed complete disappearance of diaphragmatic pleura thickening. This biological behaviour is unusually for PEL: medical literature shows that this lymphoma has a very poor prognosis; this case-report suggests, as already proposed from some authors, that PEL, in HIV-negative EBV-negative patients, is a distinct clinical entity, with a different clinical behaviour.

[Evaluation of C3c, C4 and C1-esterase inhibitor (C1-INH) during unstable angina]. / Valutazione del C3c, del C4 e dell'inibitore della C1q-esterasi (C1-INH) in corso di angina instabile.

BACKGROUND: The complement system plays an important role in the physiopathology of acute myocardial infarction (AMI) taking part in myocardial damage and reperfusion injury. The aim of this study is to investigate the plasmatic levels of some complement components (C3c, C4 and C1-INH) during unstable angina (C1-INH) and their different concentrations in relation to the different myocardial areas affected by ischemia. METHODS: The plasmatic levels of C1-INH, C3c and c4 in 30 patients affected by unstable angina, and those of 22 clinically healthy subjects (control group) were evaluated (Nefelometer Behering). The patients were divided into four groups according to the different myocardial area affected by ischemia (anterior, antero-lateral, lateral or inferior ischemia), RESULTS: No statistically significant differences were found in plasmatic levels of C3c, C4 and C1-INH between the group of patients and the control group. There is a statistically significant difference between the C1-INH levels of the patients with inferior ischemia and the plasmatic concentrations of the whole patients' group (p<0,01), the control group (p<0,01) and the group of patients with lateral ischemia (p<0,02). CONCLUSIONS: There seems to be a different activation of the complement system during unstable angina, in relation to the different myocardial area affected by ischemia.

[Circadian rhythm of serum levo-carnitine: parameters]. / Ritmo circadiano della levo-carnitina sierica: parametri ritmometrici per programmare una eventuale cronoterapia.

OBJECTIVE: The present study investigates the circadian rhythm (CR) of levo-carnitine (L-c) in systemic venous blood, in order to detect rhythmometric parameters that can be used for programming an eventual chronoterapy with such a molecule. MATERIALS AND METHODS: The L-c CR was investigated in 10 clinically healthy subjects (5 M; 5 F; mean age: 26.02 +/- 1.11 yrs), who were diurnally active and nocturnally resting. Blood samples were taken at 06:00; 08:00; 12:00; 18:00; 20:00; 24:00, not juxtaposed to breakfast, lunch and dinner. The serum concentrations of L-c were assayed via a spectrophotometric method. RESULTS: A nychtohemeral variability in circulating levels of L-c was observed, with a peak in the afternoon. Such a intradiem variability was validated to have the properties of a significant CR (MESOR = 33.37 mumol/l with a SEM of 1.19 mumol/l; amplitude = 6.31 mumol/l with 95%CL ranging from 3.58 mumol/l to 9.69 mumol/l; acrophase at 15:52 h:min with 95%CL ranging from 13:28 h:min to 17:00 h:min). CONCLUSIONS: It is important to remark that the validation of a CR for the systemic serum levels of L-c was obtained in diurnally-active/nocturnally resting subjects. Such validated rhythmometric properties are parameters that can be used for programming an eventual chronotherapy, considering that the molecule L-c is used for treating its various types of primary and secondary deficiency.

[Relationship among VIP plasma levels, esophageal dysfunction, and microcirculation in systemic sclerosis]. / Rapporti fra livelli plasmatici di VIP, disfunzione esofagea e del microcircolo in corso di sclerosi sistemica.

BACKGROUND: Recent studies suggest that esophageal dysmotility occurring in systemic sclerosis might be caused by neurotransmitter levels decrease. The aim of the present study is to value VIP plasma levels, and to relate them with the pressure of the inferior esophageal sphincter (IES) and the capillaroscopy score in a group of patients affected by Systemic Sclerosis (SSc). METHODS: Eleven subjects affected by SSc (eight male and three female, age from 30 to 72 years old) have been studied through esophageal manometry, capillaroscopy and VIP plasma levels evaluation. Fifteen healthy volunteers, as control group, have been enlisted. RESULTS: Our results show a decrease of VIP plasma levels in patients with SSc compared with control group. The difference between two groups has statistical significance (p < 0.01). Capillaroscopy has shown remarkable microcirculatory impairment and the esophageal manometry proved a decreased IES pressure. The scores of capillaroscopy, VIP plasma levels and pressures of IES have been compared and it has been observed that there is a relationship between VIP plasma level and pressure of IES. CONCLUSIONS: VIP plasma levels decrease enhances the role of the autonomic disorder in SSc and may contribute to produce the alteration of vascular tone as well as the gastroenteric musculature dysfunction.

Human herpes virus-8 associated primary effusion lymphoma of the pleural cavity in HIV-negative elderly men.

Human herpes virus-8 (HHV-8)-associated primary effusion lymphoma (PEL) is an unusual lymphoma confined to the body cavities, which primarily affects human immunodeficiency virus (HIV)-positive men at high risk for Kaposi's sarcoma (KS). We describe two HIV-negative elderly Italian men, who developed pleural HHV-8-positive PEL in association with other diseases (systemic hypertension, colonic carcinoma, chronic obstructive airways disease, dilated cardiomyopathy), but without KS. Thoracic computed tomography revealed unilateral pleural effusion and pleural thickening. Thoracentesis disclosed large lymphoma cells, with no T- or B-cell associated antigens, clonal rearrangement of the immunoglobulin heavy chain gene and the presence of HHV-8 but not Epstein-Barr virus deoxyribonucleic acid sequences. Our cases differ from most pleural effusion lymphomas, in that they are non-acquired immunodeficiency syndrome-related. This highlights the possible human herpes virus-8-associated primary effusion lymphoma risk among elderly human immunodeficiency virus-negative patients, particularly Italians, in whom human herpes virus-8 seroprevalence rates and incidence of classic Kaposi's sarcoma are high.

The relationship between glycated hemoglobin and polymorphonuclear leukocyte leukotriene B4 release in people with diabetes mellitus.

In order to evaluate polymorphonuclear leukocyte (PMN) activity in diabetes mellitus, leukotriene B4 (LTB4) levels were measured in sixty patients, 31 affected with Type 1 diabetes mellitus and 29 affected with Type 2 diabetes mellitus. The LTB4 levels (12.1+/-0.2 pg/100 microl) in diabetic patients were higher compared to those of the control group (7.9+/-0.1 pg/100 microl) (P < 0.001), and remained significantly higher (P < 0.001) (12.8+/-0.2 pg/100 microl) than in the control group (11.0+/-0.2 pg/100 microl) after stimulation with calcium ionophore. A significant and positive correlation between glycated hemoglobin and LTB4 was demonstrated (P < 0.001, r = 0.80). This study demonstrates that in diabetic patients there is a PMN activation and that this activation is correlated to glycated hemoglobin level.

The chaotic component of human heart rate variability shows a circadian periodicity as documented by the correlation dimension of the time-qualified sinusal R-R intervals.

OBJECTIVE: This study investigates the hypothesis that the nonlinear component of human heart rate (HR) variability might show a periodic structure over the 24-h span. Such a postulate could explain how the chaotic component might coexist with the deterministic periodic variability of instantaneous HR in beat per minute. MATERIALS AND METHODS: The sinusal R-R intervals (sRRi) of the Holter EKG of 10 clinically healthy subjects (5 M, 5 F, 23-30 years) were analyzed per each hour of the day-night span according to two methods for the nonlinear chaotic variability, i.e., the correlation dimension method, and the linear periodic variability, i.e., periodic regression analysis. RESULTS: The hourly-qualified correlation integrals were found to show a significant circadian rhythm, with an acrophase located during the night in coincidence with the longest duration of the sRRi and the lowest rate of cardiac pulse. CONCLUSIONS: The rhythmic structure of the chaotic component of the human HR variability let us to think that a deterministic periodic chaos of fractal type regulates the nonlinear cardiac dynamics. Such a periodic structure allows the chaos to be compatible with the deterministic linear periodicity of circadian type which characterizes the within-day variability of human HR.

ET-1 plasma levels during cold stress test in sclerodermic patients.

Endothelial cell injury in blood vessels of small arteries and capillaries is considered the primary event in the pathogenesis of systemic sclerosis (SSc). Because endothelin-1 (ET-1) is mainly released in the site of endothelial cell damage, thereby inducing a potent vasoconstriction, it was our intention to study ET-1 release in a group of SSc patients during a cold pressor test (CPT). Twelve SSc patients and a control group of 10 healthy subjects underwent CPT. Blood samples for ET-1 assay were collected at 90 and 180 seconds of exposure to cold stress. Heart rate and blood pressure were recorded at the same intervals. A capillaroscopic examination was performed in both groups before and after CPT. We observed significantly higher levels of plasma ET-1 in SSc patients compared with those of the control group at baseline (P=0.007) and at 90 (P=0.006) and 180 seconds (P=0.03) of CPT. During the test, the capillaroscopic examination showed a dramatic worsening of the vascular picture that was parallel to the increase in ET-1 plasma levels. This suggests that higher ET-1 plasma levels can have a part in the acute vascular reactivity of SSc patients undergoing CPT.

[Lipoprotein (a) and plasminogen in atherosclerosis]. / Lipoproteina (a) e plasminogeno nella malattia aterosclerotica.

The aim of this study was to evaluate plasma levels of lipoprotein (a) [LP(a)] and plasminogen in patients affected with atherosclerotic disease and to understand the mutual relationships. Eighty-four patients affected with atherosclerosis were examined and divided as follows: group I, 24 patients with peripheral arteriopathy; group II, 40 patients with ischemic heart disease (myocardial infarction and/or angina pectoris); group III, 20 patients with multi-infarct dementia; group IV (control group) with 20 healthy young subjects. The results show that Lp(a) plasma levels, in atherosclerotic patients, are higher than 30 mg/dl, while the plasminogen levels are lower than 80 mg/dl. There is an inverse correlation between these two data. Moreover, a different behaviour of Lp(a) and plasminogen rate related to age of patients, to number of atherosclerotic lesions or to acuteness of ischemic heart disease, was observed.