Biomedical subjects

F Martelli

Publications and source records attributed to F Martelli.

At least 19 recordsLinked to original sources

PubMed2006

Comparative assessment of the consistency and quality of a highly purified FSH extracted from human urine (urofollitropin) and a recombinant human FSH (follitropin alpha).

The revolutionary development of biotechnology-derived therapeutic proteins has provided the expected improvements in quality, purity and consistency, as demonstrated in recombinant human FSH (rhFSH). However, the development of urine-derived gonadotrophins has not always shown comparable improvements. More recently, highly purified urine-derived FSH (uFSH-HP) products have become widely available. The relative purity, level of urine-derived contaminants, and consistency of one such highly purified human uFSH (uhFSH) (urofollitropin) has been assessed and directly compared with rhFSH (follitropin alpha). It has been demonstrated that the highly purified urofollitropin contains variable levels of urine-derived contaminant proteins and demonstrates a variable level of FSH purity, FSH isoforms, and delivered dose. These variable factors may contribute to the control of ovarian stimulation. The relative purity, variable consistency and the presence of contaminants indicates that the urofollitropin is, at best, a partially purified uFSH that is not able to meet the quality attributes of follitropin alpha (rhFSH).

Blotting, Western↗

PubMed2006

Detection of swine torque teno virus in Italian pig herds.

Anellovirus is a recently created, floating genus of viruses. Torque teno virus (TTV), the type species in the genus, was first discovered in a human patient with a post-transfusion hepatitis of unknown aetiology. Recently, TTV genetically related to but distinct from those discovered in humans have also been found in animals, including pigs. The aims of this study were to estimate the prevalence of swine TTV in Italian pig herds and some risk factors possibly associated with this infection. Serum samples from 179 healthy pigs from 10 farms located in north-central Italy were tested by polymerase chain reaction for the presence of swine TTV DNA. Viral DNA was found in the sera of 43 pigs (24.0%), coming from eight of the 10 farms examined. Prevalence was significantly higher in finishing herds (40.1%) than in farrow-to-finish herds (11.0%) and did not depend on the size of the herd. Within the finishing herds the prevalence was significantly higher in weaners (57.4%) than in fatteners (22.9%), but this difference was not observed in farrow-to-finish herds. No relationship was observed between the prevalence of swine TTV and the implementation of some general hygiene practices and biosecurity procedures within the herds.

Animal Husbandry↗

PubMed2006

Concentrations of carnosine, anserine, L-histidine and 3-methyl histidine in boar spermatozoa and sheep milk by a modified HPLC method.

The present study deals with the application of high-performance-liquid-chromatography (HPLC) method for a quantitative detection of carnosine, anserine, L-histidine and 3-methyl-L-histidine in biological material with o-phthaldialdehyde (OPA) post column derivatisation at the constant temperature of 50 degrees C. For this purpose, some mobile-phases were prepared with scalar acetonitrile concentrations. A complete separation of all molecules, particularly for carnosine and 3-methyl-L-histidine, was obtained with a solution of acetonitrile and 6mM hydrochloric acid with 0.48 M sodium chloride (5%:95% v/v). Post column derivatisation reaction at temperature of 50'C permitted to obtain an increase in sensibility of all molecules. This method has been utilised for detection of histidine dipeptides in boar spermatozoa and in sheep milk. Concentrations (mean +/- S.E. nmol/10(9) spermatozoa) of carnosine (0.96 +/- 0.14) and anserine (0.83 +/- 0.18) in boar spermatozoa were significantly lower than those of L-histidine (52.85 +/- 4.86) and 3-methyl-L-histidine (83.07 +/- 7.1). Positive correlation was found between carnosine and anserine contents (r = 0.740; p < 0.01) and between L-histidine and 3-methyl-L-histidine (r = 0.657; p < 0.01). All histidine dipeptides studied were also present in 40 samples of sheep milk. In a case of samples without unit-forming colonies (UFC) of Staphylococcus coagulase-positive, carnosine concentrations (9.17 +/- 0.89 nmol/ml) were higher than anserine (0.51 +/- 0.02 nmol/ml) and both were significantly lower in respect to L-histidine (49.51 +/- 6.48 nmol/ml) and 3-metyl-L-histidine (81.21 +/- 6.82 nmol/ml). A negative correlation was observed between carnosine milk levels (r = -0.773; p < 0.01) and UFC/ml of Staphylococcus coagulase-positive. In conclusion this very simple and fast method can be used to detect histidine dipeptides in biological compartments where their concentrations are very low.

Animals↗

PubMed2005

Comparative analytical characterization of two commercial human follicle-stimulating hormones extracted from human urine.

OBJECTIVE: Pharmaceutical preparations of human menopausal gonadotrophin (hMG), urine-derived follicle-stimulating hormone (u-FSH) and highly purified u-FSH (u-FSH-HP) have been available since the early 1960s and the mid 1980s and 1990s, respectively. Another commercial preparation of u-FSH-HP, Folyrmon P, was launched in Japan in 1999. The aim of this study is to assess the purity of Folyrmon P and to compare it with Fertinorm-P, another commercial preparation of u-FSH-HP that has been available since 1993. METHODS: Folyrmon P and Fertinorm-P were assessed for total protein content, biological activity, immunological activity, specific activity, purity and levels of protein contamination. RESULTS: Folyrmon P, which is extracted from the urine of post-menopausal women, has a specific activity of between 4000 and 5000 IU/mg, while Fertinorm-P, which is also manufactured from the urine of post-menopausal women, has a specific activity of at least 10,000 IU/mg. It has been well documented that commercially available hMG and u-FSH preparations can contain a number of urine-derived protein contaminants. This also proves to be the case for Folyrmon P, in which contaminant proteins other than FSH were shown to be present. It was also demonstrated that both preparations, Folyrmon P and Fertinorm-P, contained high levels of oxidized FSH. CONCLUSIONS: The low specific activity and high level of contaminants in Folyrmon P indicate that this u-FSH is not highly purified. Overall, Fertinorm-P, with higher specific activity and lower levels of contaminant proteins, appears to be of higher quality compared with Folyrmon P.

Female↗

PubMed2004

Assessment of neutrophil components as markers of lung injury in the course of bovine respiratory tract infections.

To define the role of activated neutrophils in lung injury during bovine respiratory tract infections (BRTI) their in vitro function was investigated. As a means to achieve this goal the comparison of secretory action between neutrophils from the BRTI group and control was made on the basis of elastase, myeloperoxidase (MPO), alkaline phosphatase (ALK-P) release, and nitric oxide production. We noted that there is an interdependence between secretory response of neutrophils and clinical severity of BRTI. The release of elastase was greater in the BRTI group than in the control group (49.17+/-4.41 versus 46.43+/-4.95% of the total content). Neutrophils from infected heifers exhibited a significantly (p<0.05) higher value of MPO release than from healthy heifers and reached 39.23+/-10.18 versus 25.54+/-8.41% of the total content. ALK-P containing granules released significantly (p<0.001) more enzyme in the group with BRTI than in the control group (22.42+/-6.27 versus 13.74+/-2.01% of the total enzyme content). The level of nitrite accumulation rose in the culture of cells isolated from heifers with BRTI from 4+/-0.53 microM after 0.5h to 6.9+/-0.52 microM after 72 h. Our data suggest that during BRTI the increase of neutrophil secretory action results in augmentation of enzyme release including elastase, MPO and ALK-P, and the nitrite production. During an excessive secretory response of neutrophils all these factors contribute to lung injury and worsen the course of a disease and might be recognised as markers of lung injury. Moreover, such a destructive action of neutrophils must be taken into account during the introduction of new methods of BRTI treatment.

Alkaline Phosphatase↗

PubMed2003

Mitochondrial alterations induced by serum amine oxidase and spermine on human multidrug resistant tumor cells.

Multidrug resistance (MDR) has been studied extensively because it is one of major problems in cancer chemotherapy. The MDR phenotype is often due to overexpression of P-glycoprotein (P-gp), that acting as an energy-dependent drug efflux pump exports various anticancer drugs out of cells. The major goal of our investigation is to establish whether bovine serum amine oxidase (BSAO), which generates the products H(2)O(2) and aldehyde(s), from the polyamine spermine, is able to overcome MDR of human cancer cells. The cytotoxicity of the products was evaluated in both drug-sensitive (LoVo WT) and drug-resistant (LoVo DX) colon adenocarcinoma cells. A clonogenic cell survival assay demonstrated that LoVo DX cells were more sensitive than LoVo WT cells. Exogenous catalase protected cells against cytotoxicity mainly due to the formation of H(2)O(2). However, spermine-derived aldehyde(s) still induced some cytotoxicity. The cytotoxic effect was totally inhibited in the presence of both enzymes, catalase and NAD-dependent aldehyde dehydrogenase (ALDH). Transmission electron microscopy investigations showed that BSAO and spermine induced evident mitochondria alterations, more pronounced in MDR than in LoVo WT cells. The mitochondrial activity was checked by flow cytometry studies, labelling cells with the probe JC1, that displayed a basal hyperpolarized status of the mitochondria in multidrug-resistant cells. After treatment with amine oxidase in the presence of polyamine-spermine, the cells showed a marked increase in mitochondrial membrane depolarization higher in LoVo DX than in LoVo WT cells. Our findings suggest that toxic oxidation products formed from spermine and BSAO could be a powerful tool in the development of new anticancer treatments, mainly against MDR tumor cells.

Adenocarcinoma↗

PubMed2003

The response of bile secretion and ubiquinone Q10 to hyperglycaemia in sheep.

The aim of these investigations was to establish the secretion of ubiquinone Q10 (UQ10) in bile of sheep under glucose-induced cholestasis. Experiments were performed on 9 cannulated sheep divided into three groups: I-infused with sodium taurocholate, II-with Na-taurocholate plus glucose, III-with Na-taurocholate and glucose plus propranolol, phentolamine and atropine. Infusion of glucose increased plasma glucose concentration from 3.89 +/- 0.593 mM/l to 12.69 +/- 0.852 mM/l in 90 min and produced elevation of plasma insulin from 124.68 +/- 1.984 to 839.54 +/- 29.212 pM/l. Employment of blocking agents reduced insulin release to maximum 685.71 +/- 50.087 pM/l in 90 min. Under infusion of Na-taurocholate, bile flow averaged 14.016 +/- 0.706 microl/min/kg b wt. In the second group, bile flow decreased to 7.08 +/- 0.59 microl/min/kg b wt. in 90 min, and reached 11.25 +/- 0.25 microl/min/kg b wt in 240 min. Addition of the blocking agents in the third group, resulted in a significant (p < 0.05) decrease in bile flow to 3.733 +/- 0.680 microl/min/kg b wt in 105 min. This reduction of bile flow occurred with significant (p < 0.05) reduction of bile acids secretion that averaged 0.032 +/- 0.087 mM/min/kg in the first hour after glucose infusion and was maintained to the end of the experiment. Marked (p < 0.05) increase in UQ10 secretion was observed in both experimental groups. Maximum values of UQ10 secretion were obtained during the second hour of the experiment and averaged 0.449 +/- 0.196ng/min/kg b wt in the second, and 0.338 +/- 0.184ng/min/kg b wt in the third group of animals. Because at the end of the experiment UQ10 secretion gradually decreased we have concluded that free radicals generated during cholestasis lead to reduction of endogenous antioxidant capacity.

Animals↗

PubMed2002

Coenzyme Q10 levels in pigeon (Columba livia) spermatozoa.

AIM: To assess the CoQ10 levels in pigeon spermatozoa and to verify their possible correlation with spermatic kinetic parameters. METHODS: In pigeons the sperm motility percentage (MOT%), mean linear velocity (VCM) and morphology were determined in ejaculated semen. In addition intracellular CoQ10 concentrations were also detected with a HPLC method. RESULTS: Intracellular CoQ10 levels demonstrated wide individual variations, averaging 4.85 +/- 2.31 (SD) ng/106 spermatozoa. Statistical analysis showed a positive correlation of the substance with the sperm concentration (r= 0.63; P<0.05) and with the VCM (r=0.66; P<0.05), and a negative correlation with the MOT% (r = -0.78; P=0.01). No correlation was found between the CoQ10 concentration and the percentage of normal spermatozoa. CONCLUSION: Our results suggest a possible role for CoQ10 as a "fertility marker" in pigeons, which may be employed to monitor the pharmacological effects of cytostatic substances often used to reduce the pigeon fertility in urban environment.

Animals↗

PubMed2001

p19ARF targets certain E2F species for degradation.

p19ARF suppresses the growth of cells lacking p53 through an unknown mechanism. p19ARF was found to complex with transcription factors E2F1, -2, and -3. Levels of endogenous or ectopically expressed E2F1, -2, and -3, but not E2F6, were reduced after synthesis of p19ARF, through a mechanism involving increased turnover. p19ARF-induced degradation of E2F1 depended on a functional proteasome, and E2F1 was relocalized to nucleoli when coexpressed with p19ARF. Consistent with reduced levels of E2F1 and E2F3, the proliferation of cells defective for p53 function was suppressed by p19ARF, and the effect was partially reversed by ectopic overexpression of E2F1. These results suggest a broader role for p19ARF as a tumor suppressor, in which targeting of certain E2F species may cooperate with stimulation of the p53 pathway to counteract oncogenic growth signals.

Blotting, Northern↗

PubMed2001

Assessment of trans, trans-muconic acid in human seminal plasma.

Trans, trans-muconic acid (tt-MA) is one of the most important metabolites of benzene, a pollutant ubiquitously distributed in ambient air and classified in 1982 as a group I carcinogen. For its sensitivity and specificity, tt-MA excreted in urine is considered a good biological marker of benzene exposure. In this study, seminal tt-MA levels in occupationally nonexposed subjects (n = 32) have been determined. The seminal fluid of normozoospermic subjects contained an average tt-MA concentration (170 +/- 100 ng ml-1) significantly lower than that of teratozoospermic (310 +/- 180 ng ml-1; P < 0.01), oligozoospermic (400 +/- 180 ng ml-1; P < 0.001), and oligoasthenozoospermic (430 +/- 230 ng ml-1; P < 0.01) subjects. A negative correlation existed between tt-MA levels and sperm concentration (r = - 0.62; P < 0.001), percentage of normal spermatozoa (r = - 0.41; P < 0.05), and percentage of vital spermatozoa (r = - 0.89; P < 0.001). Average tt-MA levels detected in seminal plasma were higher in smokers (350 +/- 160 ng ml-1) than in nonsmokers (280 +/- 210 ng ml-1). These results show that seminal plasma tt-MA content could be an important biological indicator for evaluating the negative effects of benzene on spermatogenesis.

Adult↗

PubMed2001

Unsteady flow modelling in turbine stage.

The paper deals with the problems of unsteady flow modelling in turbine stage. The impact of unsteadiness in the design procedure as well as in the real flow phenomena are briefly addressed. After a discussion of the physical aspects of the real flow in turbine stage which differ from the unsteady flow in a single row environment, the paper reports a survey of the actual and more recent approaches to the numerical modelling on the unsteady flow in multiple rows environment. The classical steady models, used for long time in the design procedure, are addressed and their limits are focused. The more complex procedures ranging from quasi-unsteady methods to the fully unsteady methods are reported and investigated in details. The discussion on the specific features of the different approaches is pointed to assess the accuracy, the feasibility, the robustness and the usability (i.e. the computer time and storage requirements). Sample results from the author research activity are reported and presented. Discussion of crucial and open questions to improve the prediction capability of the methods are finally reported as well as some comments on the possibility to joint classical (steady methods) and advanced approaches (fully unsteady methods) to improve the actual design procedure are addressed.

Journal Article↗

PubMed2001

Application of a constant blood withdrawal method in equine exercise physiology studies.

The aim of the present study was to test a constant blood withdrawal method (CBWM) to collect blood samples from horses during treadmill exercise. CBWM was performed in 4 Standardbreds and 5 Haflinger horses. A peristaltic pump was used to control blood aspiration from an i.v. catheter via an extension line. Blood was collected using an automatic fractions collector, with a constant delay time between the drawing of blood and sample collection. Blood withdrawal using CBWM was made during a treadmill standardised exercise test (SET). A blood flow of 12 m/min was used and samples collected every 60 s during the entire period of exercise. The volume of blood collected in each sample tube was 12.1+/-0.2 ml, with a delay time of mean +/- s.d. 25.3+/-0.8 s. Plasma lactate kinetics based on measurement of lactate in each fraction showed an exponential increase during the first 13 min of exercise (10.5 min of SET and 2.5 min recovery). The peak plasma lactate concentration was observed between 2.5 and 5.5 min after the end of SET. CBWM permits the kinetics of lactate and other blood-borne variables to be studied over time. This method could be a valuable aid for use in studying equine exercise physiology.

Animals↗

PubMed2000

Potential involvement of ubiquinone in myotonic dystrophy pathophysiology: new diagnostic approaches for new rationale therapeutics.

An impairment of mitochondrial function may contribute to the pathophysiology of myotonic dystrophy (MyD). Coenzyme Q10 (CoQ10) deficiency has been previously observed, even if in a restricted sample of patients. The aim of this investigation was to obtain more information about coenzyme Q10 and its relationships to the aerobic metabolism in a group of MyD patients. Serum CoQ10 appeared significantly reduced with respect to normal controls: 0.93 +/- 0.22 vs. 1.58 +/- 0.28 micrograms/ml (p < 0.05). Moreover, the results demonstrated an inverse tendency between CoQ10 levels and the CTG expansion degree. Basal blood lactate levels were significantly higher than controls (p < 0.05). A borderline inverse correlation between CoQ10 and lactate, corresponding to lactate threshold, was found. These data suggest a possible role of CoQ10 in the pathogenesis of MyD, which may be mediated by mechanisms of cellular damage common to the oxidative pathway. Therapeutic strategies may be devised by virtue of this rationale.

Coenzymes↗

PubMed2000

Accuracy of the diffusion equation to describe photon migration through an infinite medium: numerical and experimental investigation.

The accuracy of results obtained from the diffusion equation (DE) has been investigated for the case of an isotropic point source in a homogeneous, weakly absorbing, infinite medium. The results from the DE have been compared both with numerical solutions of the radiative transfer equation obtained with Monte Carlo (MC) simulations and with cw experimental results. Comparisons showed that for the cw fluence rate, discrepancies are of the same order as statistical fluctuations on MC results (within 1%) when the distance r from the source is > 2/mu(s)', (mu(s)' is the reduced scattering coefficient). For these values of r, discrepancies for the time-resolved fluence rate are of the same order of statistical fluctuations (within 5%) when the time of flight is t > 4t0 with to time of flight for unscattered photons. For shorter times the DE overestimates the fluence discrepancies are larger for larger values of the asymmetry factor. As to the specific intensity, for small values of r the MC results are more forward peaked than expected from the DE, and the forward peak is stronger for photons arriving at short times. We assumed r > 2/mu(s)' and t > 4t0 for the domain of validity of the DE and we determined the requirements for which the simplifying assumptions necessary to obtain the DE, expressed by two inequalities, are fulfilled. Comparisons with cw experimental results showed a good agreement with MC results both at high and at small values of r mu(s)', while the comparison with the DE showed significant discrepancies for small values of r mu(s)'. Using MC results we also investigated the error made on the optical properties of the medium when they are retrieved using the solution of the DE. To obtain accuracy better than 1% from fitting procedures on time-resolved fluence rate data it is necessary to disregard photons with time of flight < 4t0. Also from cw data it is possible to retrieve the optical properties with good accuracy: by using the added absorber technique discrepancies are < 1%, both on mu(s)' and on mu(a), if the absorption coefficient is small (mu(a)/mu(s)' < 0.005).

Biophysical Phenomena↗

PubMed1999

Regulation of endogenous E2F1 stability by the retinoblastoma family proteins.

Certain E2F transcription factor species play a pivotal role in regulating cell-cycle progression. The activity of E2F1, a protein with neoplastic transforming activity when unregulated, is tightly controlled at the transcriptional level during G0 exit. In addition, during this interval, the stability of endogenous E2F1 protein increased markedly. E2F1 stability also was dynamically regulated during myogenic differentiation and in response to gamma irradiation. One or more retinoblastoma family proteins likely participate in the stability process, because simian virus 40 T antigen disrupted E2F1 stability regulation during G1 exit in a manner dependent on its ability to bind to pocket proteins. Thus, endogenous E2F1 function is regulated by both transcriptional and posttranscriptional control mechanisms.

Carrier Proteins↗

PubMed1999

Study on the propagation of ultra-short pulse light in cylindrical optical phantoms.

A detailed study about ultra-short pulse light propagation inside cylindrical scattering and absorbing phantoms is presented. Some comparisons between a Monte Carlo code and the analytical solution of diffusion equation for a delta(t) source and point-like detectors placed at different angles of detection are shown. To better quantify the agreement between the temporal profiles obtained from the two different methods of calculation, a fitting procedure based on the Levenberg-Marquardt algorithm has been implemented. Some examples on the retrieval of the optical properties for both the Monte Carlo and experimental data are presented. In the case of inhomogeneous cylindrical phantoms, some comparisons between the results of the time-domain 3D model of finite element method and 3D Monte Carlo are also shown.

Algorithms↗

PubMed1999

Properties of the light emerging from a diffusive medium: angular dependence and flux at the external boundary.

By using the diffusion approximation of the radiative transfer equation and the partial-current boundary condition, an analytical expression for the angular dependence of the specific intensity emerging from a diffusive medium has been obtained. The analytical expression for the angular distribution has been validated by comparisons with the results of Monte Carlo simulations. By using the diffusion equation and the extrapolated boundary condition, an heuristic analytical expression for the diffuse time-resolved reflectance has also been obtained by assuming that the photon flux is simply proportional to the fluence rate. For the case of the semi-infinite medium, comparisons with Monte Carlo results are presented and time-resolved reflectance data are fitted with the simple fluence rate formula. The results obtained show that the simple expression correctly describes the time-resolved reflectance giving an error in the retrieved optical parameters smaller than that of other commonly used expressions.

Biophysical Phenomena↗

PubMed1998

A source of false-negative results in clenbuterol analysis in tissues of veal calves.

The possibility of false-negative results in clenbuterol analysis was investigated in bovine tissues. An extraction procedure currently in use was adapted to process 100 specimens of different tissues each time. Its efficiency and accuracy were investigated radiometrically by means of a series of different molar concentration of the tritiated drug. In samples not submitted to extensive delipidation, unreliability of the analysis was evident. The measurement of tissue clenbuterol content, by a competitive ELISA, gave results numerically similar to those existing in literature, but with an accuracy high enough to minimize the frequency of false-negative results.

Adrenergic beta-Agonists↗