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Biomedical subjects

M Montana

Publications and source records attributed to M Montana.

At least 37 records · Page 2Linked to original sources
PubMed1999

Diabetes mellitus: polymorphonuclear leukocyte (PMN) filtration parameters and PMN membrane fluidity after chemotactic activation.

The goal of this research was to determine leukocyte rheology at baseline and after chemotactic activation in type I and type II diabetics. In 19 normal subjects, 21 type I diabetics, and 16 type II diabetics at baseline and after in vitro chemotactic activation (prolonged for 5 and 15 minutes) with two stimulating agents (4-phorbol 12-myristate 13-acetate [PMA] and N-formyl-methionyl-leucyl-phenylalanine [fMLP]), we evaluated polymorphonuclear (PMN) filtration parameters (using a St. George filtrometer [Carri-Med, Dorking, UK] and considering the initial relative flow rate [IRFR] and the concentration of clogging particles [CP]) and PMN membrane fluidity (obtained by marking PMNs with the fluorescent probe 1-(4-[trimethylamino]phenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH). At baseline, there was a difference between normals and type I and II diabetics for PMN membrane fluidity only. After activation in normals and diabetics of both types, a significant variation was present in PMN filtration parameters (IRFR and CP) at both 5 and 15 minutes. In normals, no variation was present in PMN membrane fluidity after activation with PMA or fMLP. After PMN activation, only in type I diabetics was a significant decrease in PMN membrane fluidity present at both 5 and 15 minutes. After PMN activation with either PMA or fMLP in comparison to basal values, only the mean variation (delta%) of the IRFR was significantly different between normals, type I diabetics, and type II diabetics at both 5 and 15 minutes. From the data obtained, it is evident that after activation, the PMN filtration pattern shows a specific behavior in diabetics of both types, while PMN membrane fluidity changes only in type I diabetics. The latter finding may be the basis of a metabolic pattern present in PMNs of this type, revealed after in vitro activation.

Adolescent↗
PubMed1999

Leukocyte rheology before and after chemotactic activation in some venous diseases.

OBJECTIVE: to evaluate leukocyte rheology, polymorphonuclear leukocyte (PMN) membrane fluidity and cytosolic Ca2+ concentration in subjects with post-phlebitic leg syndrome (PPS) and acute deep-venous leg thrombosis (DVT). SUBJECTS: twenty-two subjects with leg PPS and 14 subjects with leg DVT. METHODS: we evaluated the leukocyte filtration (unfractionated, mononuclear cells (MN) and PMN), the PMN membrane fluidity and the PMN cytosolic Ca2+ concentration. Subsequently, we evaluated the same PMN variables after in vitro chemotactic activation with 4-phorbol 12-myristate 13-acetate (PMA) and N -formyl-methionyl-leucyl-phenylalanine (fMLP). RESULTS: at baseline we observed a significant difference in the filtration variables of unfractionated and MN cells and in PMN cytosolic Ca2+ concentration. After activation, in normal subjects and subjects with PPS and DVT, a significant variation in PMN filtration at 5 and 15 minutes was evident. In normal subjects, no variation was present in PMN membrane fluidity or cytosolic Ca2+ concentration after activation. In subjects with PPS and DVT, we found a decrease in PMN membrane fluidity and an increase in PMN cytosolic Ca2+ concentration. After PMN activation (at 5 and 15 min) Delta% of IRFR distinguished normal subjects from subjects with PPS and DVT, while no difference was found in Delta% of membrane fluidity or cytosolic Ca2+ concentration. CONCLUSIONS: there is a functional alteration of leukocytes in these patients whose mechanisms are not yet clear.

Calcium↗
PubMed1999

Hydrocortisone levels in the urine and blood of horses treated with ACTH.

An investigation was undertaken to demonstrate whether therapeutic treatment with ACTH raises hydrocortisone (cortisol) levels in horse urine above the limit (1000 ng/ml) established by the International Conference of Racing Authorities with the aim of controlling the abuse of cortisol and ACTH in equine sports. ACTH (200 iu) was administered i.m. to 3 Thoroughbred horses; urine and blood samples were collected at intervals afterwards and analysed by an immunoenzymatic system (ELISA) and HPLC-MS. To ascertain post exercise cortisol levels in untreated horses, 101 urine and 103 serum samples were taken from horses immediately after racing and analysed by ELISA. The peak urine level of cortisol, detected 8 h after ACTH administration, was around 600 ng/ml using either ELISA or HPLC-MS. The peak serum cortisol concentration was found to be around 250 ng/ml by ELISA, but consistently less by HPLC-MS. Mean cortisol levels in post race horses were 135.1+/-72.1 ng/ml in urine and 90.1+/-41.7 ng/ml in serum. High levels of the metabolite 20beta-dihydrocortisol in urine and the cortisol precursor 11beta-desoxycortisol in serum were found. The latter showed high cross-reactivity with cortisol on ELISA. In our experiment, treatment with ACTH 200 iu i.m. did not raise urinary cortisol levels above the 1000 ng/ml threshold proposed by the ICRA.

Adrenocorticotropic Hormone↗
PubMed1999

Chronic renal failure: leukocyte rheology and polymorphonuclear cytosolic Ca2+ concentration.

In a group of subjects with chronic renal failure (CRF), we evaluated the leukocyte rheological activity, expressed as the filtration rate, the polymorphonuclear (PMN) membrane fluidity and the PMN cytosolic Ca2+ content. Using the St. George's Filtrometer, we determined the filtration rate of unfractionated, mononuclear and PMN leukocytes. Using the fluorescent probe 1.4-(trimethylamino)-phenyl-4-phenyl-hexatriene (TMA-DPH), we examined the PMN membrane fluidity and, using the Fura 2-AM, the PMN cytosolic Ca2+ content. From the results obtained, it is evident that only the initial relative flow rate of unfractionated leukocytes was significantly reduced in subjects with CRF, while the filtration parameters of mononuclear and PMN cells did not distinguish normals from CRF subjects. No variation was evident for the PMN membrane fluidity, while the PMN cytosolic Ca2+ content was significantly increased in CRF subjects. In these subjects no correlation was found between leukocyte filtration parameters, PMN membrane fluidity, PMN cytosolic Ca2+ content and plasma parameters (blood urea nitrogen and serum creatinine), reflecting the degree of the CRF. In conclusion, in CRF subjects the abnormality of the leukocyte flow properties was restricted to the initial flow rate of unfractionated leukocytes, whereas the increase of PMN cytosolic Ca2+ content might be attributed to PMN activation.

Calcium↗
PubMed1999

Determination of substances of abuse in biological fluids by GC/MS using chemical ionization with acetonitrile.

GC/ms is an important method to determine drugs in biological fluids. For each of the most commonly used drugs a methanolic solution (100 micrograms/ml) was prepared; 30 microliters of each solution was evaporated using nitrogen flow and than diluted in 100 microliters of a derivatizing mixture constituted by MSTFA and toluene (30/70). After a short reaction time at room temperature 1 ml of each solution was injected into a SE-52 column (15 m x 0.25 mm) using chemical ionization with acetonitrile. This method has a good signal/noise ratio and is very useful in screening techniques.

Acetonitriles↗
PubMed1998

Polymorphonuclear leukocyte membrane fluidity and cytosolic Ca2+ concentration in diabetes mellitus.

We evaluated polymorphonuclear membrane (PMN) fluidity in 32 subjects with type 1 diabetes mellitus, 38 subjects with type 2 diabetes mellitus and 38 normal control subjects, by marking intact and unstimulated PMN cells with the fluorescent probe 1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH). We also evaluated PMN cytosolic Ca2+ content by marking intact and unstimulated PMN cells with the fluorescent probe Fura 2-AM. PMN membrane fluidity differentiated normal subjects from type 1 and 2 diabetic subjects. The PMN cytosolic Ca2+ concentration did not discriminate type 1 and 2 diabetic subjects from normal control subjects. No statistical correlation was found between PMN membrane fluidity and PMN cytosolic Ca2+ concentration in any of the groups of subjects, nor were significant correlations found between PMN membrane fluidity and cytosolic Ca2+ concentration in several plasma parameters (serum glucose, cholesterol and triglycerides). In conclusion, in type 1 and 2 diabetic patients we found a decrease in PMN membrane fluidity and this decrease, which was greater in type 2 diabetic patients, may be a marker of PMN dysfunction.

Adult↗
PubMed1998

Type II diabetics with macrovascular complications: polymorphonuclear leukocyte (PMN) filtration, PMN membrane fluidity and cytosolic Ca2+ content after activation.

We evaluated polymorphonuclear (PMN) filtration parameters, membrane fluidity and cytosolic Ca2+ content in 21 normal subjects and in 18 type II diabetics with macrovascular complications (MVC). Evaluations were carried out at baseline and after in vitro activation prolonged for 5 and 15 min. PMA (4-phorbol 12-myristate 13-acetate) and fMLP (N-formyl-methionyl-leucyl-phenylalanine) were used as stimulating agents. TMA-DPH (1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene) was used as fluorescent probe for the membrane fluidity tests and Fura 2-AM for the cytosolic Ca2+ content. A significant variation was evident in PMN filtration parameters at 5 and 15 min. No variation was present in PMN membrane fluidity and cytosolic Ca2+ content in normals. In type II diabetics with MVC, we found an increase solely in PMN cytosolic Ca2+ content after PMA activation and an early decrease in PMN membrane fluidity and a late increase in PMN cytosolic Ca2+ content after fMLP activation. After PMA activation alone (at 15 min), PMN filtration distinguishes normals from type II diabetics with MVC. The PMN filtration parameters behave similarly in the two groups, but PMN membrane fluidity and cytosolic Ca2+ content behave differently.

Aged↗
PubMed1998

Essential hypertension: leukocyte rheology and polymorphonuclear cytosolic Ca2+ content at baseline and after activation.

In 24 hypertensives we evaluated, at baseline, the leukocyte filtration parameters (using the St. George's Filtrometer), polymorphonuclear (PMN) membrane fluidity (with the fluorescent probe 1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene [TMA-DPH]) and PMN cytosolic Ca2+ content (with the fluorescent probe Fura 2-AM). In a subgroup of hypertensives (n = 17) the PMN filtration parameters, PMN membrane fluidity and cytosolic Ca2+ content were evaluated after in vitro chemotactic activation (prolonged for 5 and 15 min) with two stimulating agents (4-phorbol 12-myristate 13-acetate [PMA] and N-formyl-methionyl-leucyl-phenylalanine [fMLP]). It was evident, from the baseline data, that there was a significant difference in the mononuclear (MN) initial relative flow rate (IRFR), clogging rate (CR) and clogging particles (CP), and in PMN cytosolic Ca2+ content. There were, however, no differences in the filtration parameters of unfractionated leukocytes and PMNs or in PMN membrane fluidity. After activation, in normals and in hypertensives, a significant variation in PMN filtration parameters was evident. In normals no variation was present in PMN membrane fluidity or cytosolic Ca2+ content after activation. In hypertensives, however, we found an increase solely in PMN cytosolic Ca2+ content after fMLP activation. After PMN activation (at 15 min) one parameter (IRFR) of PMN filtration distinguished normal subjects from hypertensives. No difference between the two groups was found in PMN membrane fluidity or PMN cytosolic Ca2+ content after PMN activation.

Adult↗
PubMed1997

Leukocyte flow properties, polymorphonuclear membrane fluidity and cytosolic Ca2+ content in subjects with vascular atherosclerotic disease.

In a group of subjects with monovascular atherosclerotic disease (MVAD) and in a group of subjects with polyvascular atherosclerotic disease (PVAD) we evaluated white blood cell (WBC) filtration (unfractionated, mononuclear -MN-, polymorphonuclear -PMN- cells), using the St. George Filtrometer and considering respectively the initial relative flow rate (IRFR) and the clogging rate (CR), the polymorphonuclear leukocyte membrane fluidity, employing the fluorescent probe 1.4-(trimethylamino)-phenyl-4-phenylhexatriene (TMA-DPH) and calculating the fluorescence polarization degree, and the polymorphonuclear leukocyte cytosolic Ca2+ content, adopting the fluorescent probe Fura 2-AM. Only the filtration parameters (IRFR, CR) of unfractionated WBCs discriminate normals from MVAD and PVAD subjects, and also monovascular and polyvascular VAD subjects between them. The filtration parameters of mononuclear and polymorphonuclear leukocytes do not distinguish normals from MVAD and PVAD subjects. PMN membrane fluidity does not differentiate normals from MVAD and PVAD subjects, while PMN cytosolic Ca2+ content discriminates normals from MVAD and PVAD subjects, but does not distinguish the two groups of VAD subjects. In conclusion, in subjects with vascular atherosclerotic disease we noted an alteration of the unfractionated leukocyte flow properties, more evident in PVAD subjects, and an increase of the PMN cytosolic Ca2+ content.

Aged↗
PubMed1996

Leukocyte flow properties, polymorphonuclear membrane fluidity, and cytosolic Ca2+ content in subjects with vascular atherosclerotic disease with and without noninsulin-dependent diabetes mellitus.

The aim of this research was the evaluation of white blood cell (WBC) filtration, reflecting WBC flow properties, polymorphonuclear cell membrane fluidity, and cytosolic Ca2+ content in subjects with vascular atherosclerotic disease (VAD) and in VAD subjects with noninsulin-dependent diabetes mellitus (NIDDM), in good hemodynamic balance. The authors examined WBC filtration (unfractionated, mononuclear [MN], polymorphonuclear [PMN] cells), using the St. George Filtrometer and considering, respectively, the initial relative flow rate (IRFR) and the clogging rate (CR); the PMN membrane fluidity, employing the fluorescent probe TMA-DPH and calculating the fluorescence polarization degree; and the PMN cytosolic Ca2+ content, adopting the fluorescent probe Fura 2-AM and considering the ratio between the Fura 2-Ca2+ complex and the unchelated Fura 2 fluorescence intensity. The obtained data showed that only the filtration parameters (IRFR, CR) of unfractionated WBCs discriminated normal subjects from VAD groups, whereas the filtration parameters of MN and PMN cells did not demonstrate any distinction. PMN membrane fluidity did not distinguish normal subjects from VAD groups, whereas PMN cytosolic Ca2+ content was significantly increased in VAD groups in comparison with normal subjects. No relationship was evident between WBC filtration and plasma metabolic parameters; the correlations obtained between PMN filtration and other PMN parameters need further investigation.

Aged↗
PubMed1995

Detection of beta 2-agonists in milk replacer.

beta 2-Agonist drugs may be illegally used as growth promoters for feedlot calves, when mixed into milk replacer immediately before feeding. To check for the presence of clenbuterol, salbutamol and terbutaline in such food, an analytical system was established using a screening method based on two commercial qualitative competitive ELISA tests, with antibodies raised against the arylamino group and the t-butyl group. The extraction procedure was based on precipitation of the milk samples with acetonitrile followed by filtration. The absence of any significant interference by other substances in the filtrate allowed detection of beta 2-agonist drugs in spiked samples at the lowest concentration having a repartitioning effect (50 ppb for clenbuterol, mabuterol and terbutaline, 500 ppb for salbutamol). In view of a false positive response with tetracycline in milk samples and a cross-reaction between clenbuterol and mabuterol, an HPLC-MS technique was developed which, after extraction and purification of the samples with SPE C18 Polar Plus, was able to confirm the presence of these drugs. The good recovery after extraction (ranging from 84% to 90.2%) and the low detection limit with this method (250 ng/ml for clenbuterol, mabuterol and terbutaline, and 2.5 micrograms/ml for salbutamol) allowed easy confirmation and simultaneous detection of the four beta 2-agonists at the lowest concentrations at which they are used in adulterated milk for calves.

Adrenergic beta-Agonists↗