Comparative analysis of the skim milk and milk fat globule membrane proteomes produced by Jersey cows grazing pastures with different plant species diversity.

The objective of this experiment was to identify and characterize the bovine milk proteome within the skim milk fraction and milk fat globule membrane (MFGM)-associated fraction from 16 organically certified lactating Jersey cows after a short term of grazing pastures with or without annual forage crops (AFC). Cows were offered a partial mixed ration (∼60% of dry matter intake) and approximately 40% of their total dry matter intake as herbage. Eight cows were offered a cool-season grass-legume herbage (GLH), which included orchardgrass (Dactylis glomerata), timothy (Phleum pratense), Kentucky bluegrass (Poa pratensis), and white clover (Trifolium repens). The other 8 cows were offered the same GLH strip-tilled with the AFC, including oat (Avena sativa), millet (Pennisetum glaucum), teff (Eragrostis tef), buckwheat (Fagopyrum esculentum), and chickling vetch (Lathyrus sativus). Milk samples were collected from each cow during a.m. and p.m. milkings on d 19 to 21 of grazing, and composite milk samples per cow were analyzed for (1) the high-abundance milk protein profile, (2) the skim milk low-abundance protein-enriched proteome, and (3) the MFGM proteome. Of the 443 proteins identified in the skim and MFGM proteomes, 433 were included in statistical analysis, including 68 proteins identified in the skim milk fraction and 365 in the MFGM-associated fraction. Analysis of the skim and MFGM proteomes encompassed unique gene ontology profiles and proportions of functional classifications. In response to diet, αS1-casein as well as 8 low-abundance proteins were present in higher concentration or abundance in milk from cows grazing the GLH strip-tilled with the AFC compared with milk from cows grazing GLH, suggesting that even short-term grazing of pastures including some AFC may affect the milk proteome.

Exploration of the bovine colostrum proteome and effects of heat treatment time on colostrum protein profile.

Heat treatment of colostrum is performed on modern dairy farms to reduce pathogenic contamination before hand-feeding the colostrum to newborn calves; however, limited data are available concerning effects of heat treatment on biologically active proteins in colostrum. The objective of this exploratory study was to investigate effects of heat treatment and length of heat treatment on colostrum protein profile. Colostrum samples were collected from Holstein cows within 12 h after parturition and assigned to the following groups: heat treatment at 60°C for 0 (untreated control), 30, 60, or 90 min. Samples were fractionated using acid precipitation, followed by ultracentrifugation and ProteoMiner (Bio-Rad Laboratories, Hercules, CA) treatment, and tandem-mass tagging was used to comparatively assess the low abundance protein profile. A total of 162 proteins were identified with more than 2 peptides in the low abundance protein enriched fraction. Of these, 62 differed in abundance by more than 2-fold in heat-treated samples compared with the unheated control. The majority of proteins affected by heat treatment were involved in immunity, enzyme function, and transport-related processes; affected proteins included lactadherin, chitinase-3-like protein 1, and complement component C9. These results provide a foundation for further research to determine optimum heat treatment practices to ensure newborn calves are fed colostrum-containing proteins with the highest nutritional and biological value.

Multiverse analysis and the Bradley-Terry model: A proposed approach for evaluating palatability and preference.

The palatability of feed for dairy cows is an important consideration but is difficult to measure, particularly when considering more than 2 feeds. We outline how a combination of multiverse analysis and Bradley-Terry modeling, 2 methodological tools that have rarely been applied in dairy science, can be adapted to address this problem. Specifically, we propose to apply multiverse analysis as a way to consider a range of thresholds for how much of a mixed grass-legume (MGL) silage had to be consumed (as a percent of the total DMI) to be designated as preferred. Each threshold gives rise to a separate dataset and a corresponding fitted Bradley-Terry model. Bradley-Terry models attribute to each feed what is commonly referred to as an "ability" in the context of sports or other competitions but can be interpreted as palatability when applied to feeds. This combined approach is a way of estimating palatabilities that appropriately reflect the degree of preference cows express through their feeding behavior. It has the advantages of being transparent and relatively easy to implement. A possible disadvantage is that this method is limited to a paired comparison approach and has difficulties with main-effects statistical inference. We demonstrate the use of this methodology on an example dataset comparing MGL silages under different ensiling conditions and exposed to oxygen for different durations.

Paracrine activity of heparin-binding EGF-like growth factor in atherogenesis: an immunohistochemical study.

AIM: Cells involved in atherogenesis produce growth factors crucial for the progression of the atherosclerotic lesions. One of them is the heparin-binding EGF-like growth factor (HB-EGF), a member of the epidermal growth factor (EGF) family, synthesized as a transmembrane precursor (proHB-EGF). This anchored insoluble juxtacrine growth factor can be converted into a soluble molecule with paracrine activity and mature HB-EGF is released in the extracellular matrix from the cell surface. HB-EGF is a potent stimulator of cell proliferation, migration and cell motility and several studies show that HB-EGF is associated with pathologies of hyperplasia of smooth muscle cells including atherosclerosis. METHODS: We localized HB-EGF by immunohistochemistry within the atherosclerotic lesions collected from right or left internal carotid artery of 20 patients with evident clinical symptoms. RESULTS: In the 20 samples we tested, the proportion of positive samples was significant. Considering the only positive samples the proportion difference related to the gender of patients was highly significant. CONCLUSION: The aim of our investigation was to better understand if this growth factor exerts its role through a juxtacrine or paracrine mechanism, or both in the process of atherogenesis. According to the results, the paracrine role of HB-EGF was clear.

Carotid intima-media thickness and liver histology in hemodialysis patients with nonalcoholic Fatty liver disease.

The prevalence of atherosclerotic cardiovascular disease in chronic hemodialysis (HD) patients has been demonstrated to be higher than in healthy people. Severe liver fibrosis is strongly associated with early carotid atherosclerosis and it might reduce the survival of patients who undergo both renal replacement therapy and transplantation. We wanted to assess whether nonalcoholic fatty liver disease (NAFLD) was associated with altered intima-media thickness (IMT) in HD patients as an independent marker of subclinical atherosclerosis. We enrolled 42 patients undergoing HD and 48 patients with normal renal function, all of them with high levels of aminotransferases and an ultrasonographic diagnosis of liver steatosis. The control group consisted of 60 healthy subjects. Laboratory tests for inflammatory and oxidative markers, ultrasonographic liver evaluation, carotid IMT measurement, and liver biopsy were performed. Different degrees of fibrosis were detected in our study cohort. Worse liver histopathological scores and higher plasmatic levels of C-reactive protein, reactive oxygen species, and vascular cell adhesion molecule-1 were found in HD patients. Carotid IMT was significantly higher (p < 0.005) in patients with histological steatosis. HD patients may develop active and progressive chronic hepatitis faster than patients with normal renal function and the thickness of their carotid intima-media might be markedly increased. These two conditions seem to be independent on classical risk factors and on metabolic syndrome. They might be related to the high levels of oxidants and to the inflammatory state, which are typical of patients undergoing HD. Independently related with the traditional risk factors for cardiovascular disease, nonspecific inflammation and oxide-reductive imbalance may play an important role in the progression of NAFLD and atherosclerotic disease in HD patients.

[Central diabetes insipidus as a first manifestation of lung adenocarcinoma]. / Diabete insipido centrale come prima manifestazione di adenocarcinoma polmonare.

The pituitary gland and infundibulum can be involved in a variety of medical conditions, including infiltrative diseases, fungal infections, tuberculosis, primary and metastatic tumors. Metastases to the pituitary gland are absolutely rare, and they are generally secondary to pulmonary carcinoma in men and breast carcinoma in women. Pituitary metastases more commonly affect the posterior lobe and the infundibulum than the anterior lobe. The posterior lobe involvement may explain why patients with pituitary metastases frequently present with diabetes insipidus. We are presenting a case report of a 48-year-old male patient with sudden onset of polyuria and persistent thirst. Laboratory results revealed central diabetes insipidus. Computed tomography (CT) scan of the brain showed a mass located in the sella turcica and suprasellar region. CT scan of the chest showed a mass in the right superior lobe with mediastinal lymphadenopathy. Bronchoscopy and biopsy demonstrated a pulmonary adenocarcinoma. Thus, we made a diagnosis of lung cancer with local and pituitary metastases. The patient received radiotherapy on the pituitary gland and adjuvant chemotherapy. As a result the intrasellar and suprasellar mass decreased in size and urinary output accordingly decreased. In conclusion, in patients presenting with sudden onset of diabetes insipidus pituitary metastases should be taken in account in differential diagnosis.

Acute lymphoblastic leukemias from relapse engraft more rapidly in SCID mice.

It is generally believed that relapse of acute leukemia heralds progression of the disease into a more aggressive stage. The biological behavior of leukemic cells collected from four patients with adult acute lymphoblastic leukemia (ALL) prior to treatment and at relapse was studied after engraftment into 28 unconditioned mice with severe combined immunodeficiency (SCID). Leukemic cells engrafted in all but one mouse, with major differences observed in the growth and aggressiveness of the leukemias. Recipient mice of cells derived from all patients at relapse died more rapidly in overt leukemia than those which were injected with cells obtained prior to induction treatment (P=0.0002). SCID mice that received cells from one patient at the time of diagnosis also died in terminal leukemia. Other SCID mice however, that received cells from the remaining three patients prior to treatment developed occult leukemia that was detectable in the blood or bone marrow with the use of polymerase chain reaction (PCR) or flow cytometry only. Leukemic cells recovered from mice with terminal leukemia exhibited a larger proliferating fraction than cells originally injected (P=0.004). Our results demonstrate, that during the evolution from initial presentation to relapse, ALL cells may acquire biological properties which render them more aggressive in SCID mice.

[Phosphorus: a new cardiovascular risk factor?]. / Il Fosforo: nuovo fattore di rischio cardiovascolare?

Phosphorus is an essential mineral in the regulation of many metabolic processes. However, is known as alterations in serum phosphate levels, compared to the normal range, have clinical relevance: many studies about phosphorus and cardiovascular risk have shown that high serum phosphate levels are associated with clinical and subclinical cardiovascular disease, in CKD and non-CKD patients. In recent years, serum phosphate level within the upper limits of normal range is also identified as a "stealthier killer", and has emerged as a risk factor of cardiovascular mortality and progression of CKD. This mounting evidence suggests the possibility that lowering serum phosphate levels may be a future target of cardiovascular disease management, also through the use of early biomarkers of phosphate overload, such as FGF23, Klotho or the urinary fractional excretion of phosphate. The goal must be an early diagnosis and treatment of disordered phosphorus metabolism, before end-organ damage occurs. Since the western diet is rich in phosphate, a dietary restriction associated with the use of phosphate binders, as well as the use of intervention such as calcitriol supplementation, certainly will have a positive influence on the phosphate-regulatory axis.

Nitric oxide induces apoptosis in NALM-6, a leukaemia cell line with low cyclin E protein levels.

Intracellular nitric oxide levels may differ in resting and stimulated cells and contribute to the regulation of cell survival and proliferation through a variety of mechanisms and effects. We exposed two B-cell lines to a range of S-nitroso-N-acetyl-D,L-penicillamine (SNAP) concentrations in order to examine their susceptibility to exogenous nitric oxide and the participation of nitric oxide as modulator of cell proliferation. Although both FLEB and NALM-6 decreased their levels of thymidine incorporation, only NALM-6 cells were induced to undergo G1 arrest, phosphatidyl serine exposure and DNA fragmentation when cultured in the presence of 250 microm SNAP. This higher sensitivity of NALM-6 coincided with initially low cyclin E protein levels which were increased 7.8-fold after culture for 24 h with 250 microm SNAP. In contrast, there was no difference in cyclins A and D3, Bcl-2 and actin levels, neither at the beginning nor at the end of the 24 h culture. Our study reveals that FLEB and NALM-6 exhibit different response to the same concentration of nitric oxide, that nitric oxide can simultaneously induce cell cycle alterations and apoptosis, and further suggests an association between these two processes, with the involvement of cell cycle regulatory molecules.

Changes in splenoportal axis calibre and flow in a patient affected by hereditary angioedema.

The authors describe a case of hereditary angioedema characterised by abdominal pain accompanied by ascites. Ultrasound (US) examination performed after acute abdominal attack implied the presence of increased splenoportal axis calibre and reduced blood flow. According to the authors, this may confirm the pathogenic role of C1-inhibitor deficiency induced oedema that is capable of creating major haemodynamic involvement also of abdominal vessels. US findings of transient appearance, especially related to the specific treatment, may help physicians make early diagnosis and avoid dangerous invasive procedures resulting from incorrect diagnosis of acute abdomen.

Evaluation of cardiovascular parameters by continuous 24 hour monitoring in diabetic hypertensive patients with autonomic neuropathies.

Correlations between scores of autonomic neuropathy severity in diabetic hypertensive subjects and behaviour of cardiovascular parameters detected by continuous 24 hour monitoring were investigated in order to reveal any alterations hat indicated onset and progression of disautonomy. R-R and Q-T patterns, circadian pressure rhythm and ECG Holter over 24 hours were analysed in 30 patients divided into various groups (age, duration of disease, treatment) and 10 controls. Three autonomic tests (deep breathing, postural hypotension, lying to standing) were performed and the relative Ewing scores recorded. Data obtained were analysed using Pearson's correlation test and simple linear regression. The results not only confirm circadian rhythm demodulation of arterial pressure, but also show progressive correlation between the scores obtained and modified cardiovascular parameters. According to the authors, detection of these intermediate alterations may be useful in forecasting possible onset or evolution of dysautonomic pathologies.

Ultrasonography in diagnosis and evaluation of fibrocystic liver diseases: two case reports.

This paper describes 2 cases of a rare biliary cystic disease. A patient with Caroli's disease and another with bile duct swelling that were initially diagnosed by ultrasound and power color Doppler. Diagnosis was later confirmed by additional instrumental examinations. The first case was totally asymptomatic and only identified by chance, while the second was symptomatic and determined severe pain and dyspepsia. In both pathologies US B mode imaging and power color Doppler examination revealed the characteristic lesions of the intrahepatic biliary tree diseases and ruled out their vascular nature and communication with the portal and arterial tree.

[Atheroembolism renal disease: diagnosis and etiologic factors]. / La malattia renale ateroembolica: diagnosi ed eziologia.

Atheromatous renal disease is the major cause of renal insufficiency in the elderly, and cholesterol embolism is a manifestation of this disease. Cholesterol embolism occurs in patients suffering from diffuse erosive atherosclerosis, usually after triggering causes, such as aortic surgery, arterial invasive procedures (angiography, left heart catheterization and coronary angioplasty) and anticoagulant or thrombolytic therapy. It is characterized by occlusion of small arteries with cholesterol emboli deriving from eroded atheromatous plaques of the aorta or large feeder arteries. The proximity of the kidneys to the abdominal aorta and the large renal blood supply make the kidney a frequent target organ for cholesterol atheroembolism. The exact incidence of atheroembolic renal disease (AERD) is not known. The reported incidence AERD varied in the literature because of the differences in study design and the different criteria used for making the diagnosis. Retrospective data derived from autopsy or biopsy studies may exaggerate the frequency by including many subclinical cases. Clinical observations that are based on a short duration of follow-up after an invasive vascular procedure and the infrequency of the confirmatory renal biopsies can lead to an underestimation of the true incidence of AERD. The initial signs and symptoms in patients diagnosed with cholesterol embolism were blue toes syndrome, livedo reticularis, gangrene, leg, toe or foot pain, abdominal pain and flank or back pain, gross haematuria, accelerated hypertension and renal failure. Cholesterol embolism may also be associated with fever, increased erythrocyte sedimentation rate and eosinophilia. Thus, in the cases of spontaneous cholesterol embolism, differential diagnosis includes, polyarteritis nodosa, allergic vasculitis and subacute bacterial endocarditis. Skin and renal biopsy specimens are the best sample for histologic diagnosis. There is, at present, no pharmacological treatments shown to be effective in altering the course of the disease. Management is limited to supportive therapy and avoidance of anticoagulation; aortic procedures should be postponed.

[Immunolocalization of HB-EGF in human atherosclerotic plaques]. / Immunolocalizzazione di HB-EGF in placche aterosclerotiche umane.

AIMS: Heparin-binding epidermal growth factor is a member of the EGF family, it is a potent mitogen for smooth muscle cells and has been implicated in atherosclerosis, angiogenesis. In athererogenesis, HB-EGF has been detected in medial smooth cells and foamy macrofages. In this work, we have investigate about immunohistocemical localization of HB-EGF in atherosclerotic plaques. MATERIALS AND METHODS: Three cases of man affected by atherosclerosis have been examined. We have collected and examined atherosclerotic plaques by immunohistochemical procedure in optical microscopy. Samples have been incubated with primary Ac (anti-human HB-EGF- goat IgG). RESULTS: In the three examined cases, results are partly overlap-ping, but with some difference in relation to location of positivity to HB-EGF. Only in one case, HB-EGF staining is rather weak and located just below endothelium where is a thickened area of tissue rich in fibres and few cells, In another case, positivity to HB-EGF is present in an area of connective tissue of the intima. In the last case, positivity to HB-EGF is evident in the context of a presumed elastic tissue with fusiform cells following fibres orientation, and that could be fibroblasts or smooth muscle cells. CONCLUSIONS: These results indicate that HB-EGF is involved in the development of atherosclerotic plaques and that HB-EGF is a possible target for atherosclerosis therapy.

[Heparin-binding epidermal growth factor (HB-EGF): myth or reality?]. / Heparin-binding epidermal growth factor (HB-EGF): mito o realtà?

Heparin-Binding Epidermal Growth Factor (HB-EGF) is growth factor member of EGF family that stimulate differentiation and growth. HB-EGF was initially identified as a secreted product of human macrophage-like cells, it is sinthetized as a transmembrana protein; proHB-EGF; that is shed by specific metalloproteases, releasing soluble growth factor(sHB-EGF). It exerts biological activity trough activation of the EGFR. sHB-EGF is implicated in diverse biological processes: angiogenesis, shin wound, blastocysts implantation, atherosclerosis, tumor formation moreover it acts as the Diphtheria Toxin (DT) receptor. HB-EGF It's an important molecule because could be a novel targeting for cancer and atherosclerosis therapy.

Ultrasound imaging in diagnosis of superior mesenteric artery syndrome.

OBJECTIVES: We screened a cohort of subjects affected by various degree of dyspepsia to reveal if they presented a reduction of the aorto-mesenteric angle and to diagnose suspected cases of superior mesenteric artery (SMA) syndrome. DESIGN: Controlled, prospective, study. SETTING: Subjects were studied as outpatients. SUBJECTS: The study investigated a total of 3622 subjects referred to our department by their general practitioners for dyspepsia and/or abdominal pain. Interventions. Abdominal ultrasonography with power colour Doppler, gastroduodenoscopy, hypotonic duodenography, contrast-enhanced spiral computerized tomography were performed. MAIN OUTCOME MEASUREMENT AND RESULTS: Color Doppler revealed a significant reduction of the SMA angle in 29 of 950 subjects; gastroscopy showed duodenal compressive pulsation in 14 of 29 patients and X-ray revealed compression of the third segment of the duodenum in 28 of 29 patients. CT confirmed the presence of a reduced angle and various degrees of duodenal compression in all patients. Ultrasonography and CT examinations gave overlapping results (P > 0.05) in diagnosing pathological aorto-mesenteric angle. CONCLUSION: The authors believe that the incidence of reduced aorto-mesenteric angle and SMA syndrome might be underrated. Ultrasound power colour Doppler imaging is useful in epidemiological screening of reduced aorto-mesenteric angle to diagnose suspected cases of SMA syndrome.

Cyclin E overexpression in relapsed adult acute lymphoblastic leukemias of B-cell lineage.

Using Western blot analysis, we examined cyclin E and cyclin A protein levels in 19 patients with acute lymphoblastic leukemia ([ALL] 15 B-ALL and four T-ALL). Whereas normal, nonproliferating peripheral blood mononuclear cells (PBMCs) expressed low levels of the 50-kD cyclin E, ALL blasts in the peripheral blood, although showing low-level or no proliferation as judged by FACS/cell-cycle analysis and cyclin A protein levels, expressed high levels of cyclin E, with a mean value similar to that of the proliferating Burkitt's lymphoma cell line, Akata. The accumulation of a protein shown to shorten the G1 phase of the cell cycle, cyclin E, in growth-delayed leukemic blasts may reflect the malignant status of these cells. Before treatment, B-ALL cells expressed predominantly the 50-kD cyclin E. T-ALL samples displayed the 50-kD cyclin E protein and a smaller, approximately 43-kD cyclin E species. In paired B-ALL samples taken before treatment and at relapse, we found a significant overexpression of the 50-kD protein in relapsed samples (P < .006), plus the presence of up to four additional smaller-molecular-weight species of cyclin E, illustrating clear diagnosis versus relapse differences. Cyclin E expression in ALL blasts may correlate to the relative malignant status of the cells, with higher protein levels reflecting a more advanced stage of the disease and a greater potential to proliferate under permissive conditions.

Phenotype-related differences in the expression of D-type cyclins in human B cell-derived lines.

Exposure of normal resting B lymphocytes to EBV in vitro leads to activation, subsequent immortalization, and the establishment of lymphoblastoid cell lines (LCLs). The endemic form of Burkitt's lymphoma (BL) is associated with EBV. EBV-positive BL lines maintain the original tumor phenotype (group I BL) initially and express only one EBV-encoded protein, EBV nuclear antigen (EBNA)-1. Most of them drift toward a LCL-like phenotype during in vitro culturing and express all nine EBV-encoded growth transformation-associated proteins (group III BL). Cyclin D2 and D3 have been found previously to differ in their mRNA expression in BL and LCL. Cyclin D2 expression has been attributed to EBV gene expression and to EBNA-2 and EBNA-5 in particular. We have studied cyclin D2/D3 expression in larger series of LCLs, BL lines, and freshly EBV-infected peripheral blood B lymphocytes, both at the mRNA and protein levels. The predominant cyclin D2 expression in the LCLs and group III BLs correlated with an activated B-cell phenotype. In contrast, only cyclin D3 was expressed in the group I BL lines. EBV-carrying group II BL lines that retained the BL-associated CD10 did not express cyclin D2. A collection of in vitro EBV-converted sublines of originally EBV-negative BLs that expressed a full set of the growth transformation-associated EBV proteins maintained their CD10 and cyclin D3 expression. Blood B lymphocytes expressed no D2 or D3 as judged by immunostaining. Cyclin D2 could be detected by immunostaining in a proportion of the activated blasts 40 h postinfection. Cyclin D3 that is expressed at a low level in the established LCLs was stained easily in B blasts at this time. The level of cyclin D3 at 72 h postinfection was two to three times higher than in the exponentially growing LCLs, as detected by immunoblotting. It was still 5-10 times lower than in group I BLs. Mitogen stimulation of primary B cells induced cyclin D3 at a similar level as in the LCLs. Our data are consistent with the notion of cell lineage-specific D-type cyclin expression. They also indicate that B cells may switch their D-type cyclin expression during differentiation.