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PubMedDate not supplied

Lasker's coefficient of isonymy between and within 16 Sardinian villages in the periods 1825-1849, 1875-1899, and 1925-1949.

Coefficients of relationship by isonymy, based on the frequency of common surnames, were used to estimate the biological affinity between (R(ib)) and within (R(iw)) 16 Sardinian villages in the periods 1825-1849, 1875-1899, and 1925-1949. The database for the surname analysis consisted of 13,515 marriages registered in the parishes of villages, all belonging to the Sardinian linguistic area. The values of R(ib) and R(iw) tend to decrease in time. In all three periods, the R(ib) values generally decrease as the geographical distance between villages increases. The negative values of the Bravais-Pearson coefficient of correlation between the matrix of R(ib) values and the matrices of geographical distances (calculated both in a straight line and from road distances) are significant for each period (modified Mantel test), with slightly higher correlation coefficients for road distances. Moreover, the plots of isonymic relationships obtained by nonmetric multidimensional scaling for 1825-1849 and 1925-1949 show that the biological relationships between neighboring villages increase in the latter period. The positive values of the Bravais-Pearson coefficients of correlation between R(iw) and altitude are significant in all three periods, while the correlation between R(iw) and population size is not significant, even when altitude is controlled for. Since altitude is considered an indicator of isolation, the results obtained with the coefficient of relationship by isonymy within villages also support the isolation-by-distance model. R(ib) values provided a reliable picture of the biological relationships between 16 Sardinian villages, while R(iw) values illustrated the biological affinity within the communities for a period of 100 years, from 1825-1849 to 1925-1949.

Consanguinity↗
PubMed1989

Analytical evaluation of twenty assays for determination of thyrotropin (TSH).

This paper summarises the results of the evaluation of several assays of thyrotropin in serum. The authors assessed 8 radioimmunoassays (IRMA techniques) and 10 enzyme immunoassays, including ELISA, luminescent immunometry and fluoroimmunometry, also covering a broad spectrum of actual assays. An unique protocol based on identical human samples for all assays was used in order to compare precision, linearity, detection limit, matrix effects and correlation. The authors found a wide spread in precision results between the evaluated assays; as distinct from enzyme immunoassays, discrepant analytical results of the radioimmunoassays are mainly due to differences in standardization procedures. All assays were linear in a smaller or larger range. Sensitivity was generally in agreement with the manufacturer's specification. One assay was not acceptable for clinical use because the practical detection limit only reached 1.5 mU/l. Three other assays (from the same origin) demonstrated particular problems with the stability of the chromogenic reagent. The study of matrix effects was performed using different artificial matrices and the results varied from one assay to another; in the test conditions, some assays were not sensible to matrix manipulations, others were extremely influenced by ion strength or protein concentration. The correlation study confirmed the results of the precision study: assays with a good intraassay precision had also the best correlations. In a general appreciation of the evaluated methods the authors notice that most of the evaluated "classical" enzyme immunoassays operate around the sensitivity limits of spectrophotometry, explaining also the inferior results. Some radioimmunoassays and the enhanced luminescent technique gave the best results in this evaluation study.

Evaluation Studies as Topic↗
PubMed1997

The tomato RBCS3A promoter requires integration into the chromatin for correct organ-specific regulation.

In tomato, the RBCS1, RBCS2 and RBCS3A genes, encoding the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase, are expressed in leaves and light-grown seedlings, but only RBCS1 and RBCS2 are expressed in developing tomato fruits. The activities of the three promoters have been compared in transgenic plants and after transient transformation. Fruit-specific repression of the RBCS3A promoter was observed in transgenic plants, but not after ballistic transient transformation, indicating that chromatin integration is necessary for its correct organ-specific regulation. In addition, matrix attachment regions have been identified in the RBCS1, RBCS2 and RBCS3A promoters. This is the second case in plants of absence of correct regulation of a plasmid-borne plant promoter and correlating potential nuclear matrix attachment of the gene.

Antigens, Nuclear↗
PubMed1985

Development of cell surface linkage complexes in cultured fibroblasts.

The possible role of a 140K membrane-associated protein complex (140K) in fibronectin-cytoskeleton associations has been examined. The 140K was identified by the monoclonal antibody JG22E. Monoclonal and polyclonal antibodies to the 140K showed identical patterns of binding to the cell membranes of fixed and permeabilized chicken embryonic fibroblasts; localization was diffuse, but with marked concentration in cell-to-extracellular matrix contact sites. Correlative localization with interference reflection microscopy and double-label or triple-label immunofluorescence showed that 140K co-distributed with extracellular fibronectin fibrils and intracellular alpha-actinin in microfilament bundles at extracellular matrix contact sites but tended not to co-localize with tropomyosin present in bundles at sites farther from adhesion sites. In addition, binding of antibodies to 140K, alpha-actinin, and fibronectin was excluded from vinculin-rich focal adhesion sites at the cellular periphery. A progressive development of cell surface alpha-actinin-140K-fibronectin associations was observed in early spreading cells. The anti-140K monoclonal antibody JG22E inhibited the attachment and spreading of both normal and Rous sarcoma virus-transformed chicken embryonic fibroblasts to a fibronectin substratum. However, the anti-140K monoclonal antibody became a positive mediator of cell attachment and spreading if it was adsorbed or cross-linked to the substratum. Our results provide the first description of a membrane-associated protein complex that co-localizes with fibronectin and microfilament bundles, and they suggest that the 140K complex may be part of a cell surface linkage between fibronectin and the cytoskeleton.

Animals↗
PubMed2005

Serum fluctuations of total and free tryptophan levels during the menstrual cycle are related to gonadotrophins and reflect brain serotonin utilization.

BACKGROUND: Serotoninergic (5-HT) neurons are suggested to regulate estrous cycle in animal models. In the present study we evaluated whether a relationship exists between the serotoninergic precursors in the central nervous system and the gonadotrophin-ovarian cyclic function. METHODS: We measured 5-HT precursors [free (FT) and total (TT) tryptophan] and LH, FSH and 17beta-estradiol (E2) levels in the serum of 15 fertile women with normal menstrual cycles during the follicular (cycle days 1-5, 7-11), mid-cycle (cycle days 14-16) and luteal (cycle days 17-19, 22-24) phases. RESULTS: TT and FT were significantly increased in the 7-11 and 17-19 cycle days and were decreased at mid-cycle (P < 0.01), with a cyclic and opposite behaviour when compared to that of FSH and LH. Indeed, correlation analysis through the matrix of mean values showed that LH was negatively correlated to TT (r = -0.636) and FT (r = -0.574), as well as FSH (TT, r = -0.655; FT, r = -0.663), and that TT and FT were positively correlated to each other (r = 0.801; P < 0.001). Furthermore, whilst the two FT peaks reached approximately the same levels in the follicular and luteal phase, TT levels were approximately 30% higher in the luteal than in the follicular phase of the cycle: thus in the first (follicular) phase FT peak was relatively higher than that of TT, whereas the contrary occurred in the second (luteal) phase of the cycle. CONCLUSIONS: Both TT and FT levels have cyclic variations throughout the menstrual cycle, being lowest at mid-cycle (14-16 cycle days), concomitant with the highest LH and FSH concentrations, and higher before and after mid-cycle phase, coinciding with the lowest circulating LH/FSH levels. Since TT and FT levels in the plasma have cyclic changes, our study: (i) suggests that a consumption of serum serotonin precursors takes place concomitant with gonadotrophin release during menstrual cycle; (ii) may represent an in vivo model to investigate this relationship in women in different physiopathological conditions.

Adult↗
PubMedDate not supplied

Effect of cell migration on the maintenance of tension on a collagen matrix.

Although it is known that cells promote structural reorganization of the collagen architecture, how individual cells exert mechanical tension on the matrix is not clearly understood. In the present study we have investigated the mechanical interaction of individual corneal fibroblasts with a collagen matrix using an improved version of our previously described in vitro force-measurement system (Roy, P. et al. Exp. Cell Res. 232:106-117, 1997). The elastic distortion of the collagen matrix exerted by cells was temporally recorded and analyzed using a two-dimensional finite-element model to quantify the forces exerted on the matrix. Time-lapse videomicroscopy of serum-cultured cells on the matrix for up to 6 h revealed that individual fibroblasts generated measurable tension on the matrix during pseudopodial extension and slow retraction. Fast retraction, an event observed during active cell migration, was associated with dramatic release of tension on the matrix. An apparent inverse correlation was observed between cell translocation and maintenance of matrix tension. Additional experiments with cells under serum-free conditions revealed that these cells fail to generate any detectable tension on the matrix despite undergoing filopodial extension and retraction. Since serum-free cells do not form focal adhesions or stress fibers, these experimental data suggest that contractility of nonmotile cells, coupled with strong cell-matrix adhesion, is the most favorable mechanism of generating and maintaining tension on the extracellular matrix.

Adhesiveness↗
PubMedDate not supplied

Evaluation of HER-2/neu in serum and tissue of primary and metastatic breast cancer patients using an automated enzyme immunoassay.

Serum HER-2/neu concentrations were evaluated in 172 healthy subjects, 176 primary and 55 metastatic breast cancer patients, employing a new automated assay (Bayer Immuno 1 serum HER-2/neu). Using 13 ng/mL as the cutoff, abnormal HER-2/neu serum levels were found in 8% (14/176) of primary and 50.9% (28/55) of metastatic breast cancer patients. Both in primary and metastatic breast cancer a significant relationship was found with the stage of the disease when serum HER-2/neu was considered as a categorized variable (p=0.0003 and p=0.02, respectively), but not when it was taken as a continuous variable (p=0.247 and p=0.146, respectively). Moreover, we evaluated the correlation between Immuno 1 HER-2/neu and Oncogene Research Products ELISA assay in 53 normal subjects, 46 primary and 34 metastatic breast cancer patients. The correlation was relatively good (p<0.0001), although substantial differences could be found in single cases. The Immuno 1 assay was also evaluated for the first time in breast cancer tissue. The method, which showed good performance both in terms of imprecision and linearity, was used to measure HER-2/neu protein in 140 cytosol samples from primary breast cancer tissue and in homogenates from 40 matched cases. The correlation between the two matrixes was very close (p<0.0001). By contrast, no correlation was found between serum and matched cytosol (p=0.101) or homogenate samples (p=0.511).

Adult↗
PubMed1993

51Cr-release assay adapted to a 96-well format sample reading.

The application of the Matrix 96, a direct beta counter, to monitor cell-mediated lympholysis assays (CML) was investigated. Until now, the gamma rays emitted from 51Cr, released in the supernatant of the sample following lysis of targets by effector cells, were read in gamma counters using individual tubes for each sample. The Matrix 96 has been designed to count 96 wells simultaneously for assays performed in 96-well microplates. Aliquots of supernatants were spotted on a 96-well disposable metal spotting plate and dried prior to reading in the Matrix 96. A tight correlation was observed between the counts obtained in the Matrix 96 and a gamma counter, which indicates that the detection of electron capture beta particles emitted from 51Cr was as accurate as reading gamma rays from the same isotope. Both methods confirmed the minimal level of cytotoxicity mediated by unstimulated lymphocytes and the high levels of nonspecific cytolytic activity mediated by lymphokine-activated killer (LAK) cells against several tumor cell lines.

Beta Particles↗
PubMed2003

Autosomal microsatellite and mtDNA genetic analysis in Sicily (Italy).

DNA samples from 465 blood donors living in 7 towns of Sicily, the largest island of Italy, have been collected according to well defined criteria, and their genetic heterogeneity tested on the basis of 9 autosomal microsatellite and mitochondrial DNA polymorphisms for a total of 85 microsatellite allele and 10 mtDNA haplogroup frequencies. A preliminary account of the results shows that: a) the samples are genetically heterogeneous; b) the first principal coordinates of the samples are correlated more with their longitude than with their latitude, and this result is even more remarkable when one outlier sample (Butera) is not considered; c) distances among samples calculated from allele and haplogroup frequencies and from the isonymy matrix are weakly correlated (r = 0.43, P = 0.06) but such correlation disappears (r = 0.16) if the mtDNA haplogroups alone are taken into account; d) mtDNA haplogroups and microsatellite distances suggest settlements of people occurred at different times: divergence times inferred from microsatellite data seem to describe a genetic composition of the town of Sciacca mainly derived from settlements after the Roman conquest of Sicily (First Punic war, 246 BC), while all other divergence times take root from the second to the first millennium BC, and therefore seem to backdate to the pre-Hellenistic period. A more reliable association of these diachronic genetic strata to different historical populations (e.g. Sicani, Elymi, Siculi), if possible, must be postponed to the analysis of more samples and hopefully more informative uniparental DNA markers such as the recently available DHPLC-SNP polymorphisms of the Y chromosome.

Alleles↗
PubMed2003

Specific alpha v integrin receptors modulate K1735 murine melanoma cell behavior.

Expression of beta 3 integrins is increased in invasive melanoma. In this study we show that K1735 cell proliferation is enhanced by the expression of either beta 3 or a constitutively active Src. We investigated possible modulators of FN matrix assembly and found that matrix metalloproteinase 2 (MMP2) was activated by alpha v beta 3. alpha v beta 3 integrin was localized to focal contacts whereas alpha v beta 5 was peripherally distributed. MMP2 was also activated by expression of CASrc. MMP2 activation inversely correlated with FN matrix assembly, in that it dramatically reduced the organization of a FN matrix. K1735 cell migration on VN and invasion through a reconstituted basement membrane were decreased in the presence of anti-MMP2 antibodies. These results demonstrate that the expression of the alpha v beta 3 complex modulates melanoma cell behavior including activation of Src, organization of the cytoskeleton, assembly of the extracellular matrix, cell motility, and activation of MMP2.

Animals↗
PubMed2005

A Bayesian framework for noise covariance estimation using the facet model.

In image processing literature, thus far, researchers have assumed the perturbation in the data to be white (or uncorrelated) having a covariance matrix sigma2I, i.e., assumption of equal variance for all the data samples and that no correlation exists between the data samples. However, there have been very few attempts to estimate noise characteristics under the assumption that there is a correlation between data samples. In this work, we propose a new and a novel approach for the simultaneous Bayesian estimation of the unknown colored or correlated noise (population) covariance matrix and the hyperparameters of the covariance model using the well-known facet model. We also estimate the facet model coefficients. We use the facet model because of its simple, yet elegant, mathematical formulation. We use the generalized inverted Wishart density as the prior model for the noise covariance matrix. We place a structure on the covariance matrix using the parameters of a correlation filter. These hyperparameters are estimated by a new extension of the expectation-maximization algorithm called the generalized constrained expectation maximization algorithm that we developed.

Algorithms↗
PubMed2000

Expression of integrin alpha(v)beta(3) correlates with activation of membrane-type matrix metalloproteinase-1 (MT1-MMP) and matrix metalloproteinase-2 (MMP-2) in human melanoma cells in vitro and in vivo.

Activation of matrix metalloproteinase-2 (MMP-2) is mediated by binding to the complex of membrane-type matrix metalloproteinase-1 (MT1-MMP) with tissue inhibitor of MMP-2 (TIMP-2) on the cell surface. Binding of MMP-2 to integrin alpha(v)beta(3) has been implicated in presenting activated MMP-2 on the cell surface of invasive cells, but interactions with the MT1-MMP-TIMP-2 system have not been considered. Therefore, we studied the expression and interaction of MT1-MMP, MMP-2 and TIMP-2 in the alpha(v)beta(3)-negative melanoma cell line BLM and in its beta(3)-transfected, alpha(v)beta(3)-expressing counterpart BLM-beta(3), both on cell lines and in xenografts. Total expression levels of MMP-2, MT1-MMP and TIMP-2 did not differ markedly between the alpha(v)beta(3)-negative and alpha(v)beta(3)-positive cells. Remarkable differences, however, exist in the presence of active MMP-2 and MT1-MMP. Zymography on cell lysates revealed that active MMP-2 was restricted to alpha(v)beta(3)-positive cell line and clearly accumulated in xenografts derived from the BLM-beta(3) cells, confirming the relevance of this integrin for MMP-2 function. Western blotting of cell lysates showed that processing of proMT1-MMP to the activated form was enhanced in BLM-beta(3). The ratio of active and inactive MT1-MMP was 3-fold higher in the beta(3)-transfectants. Immunofluorescence double-labeling followed by confocal laser microscopy showed co-localization of MT1-MMP and alpha(v)beta(3) on BLM-beta(3) cells. In xenografts from BLM-beta(3) cells, active MT1-MMP was markedly increased. Our results demonstrate that expression of alpha(v)beta(3) in cell lines and xenografts was accompanied by an accumulation of active MT1-MMP and MMP-2. Furthermore, MT1-MMP and alpha(v)beta(3) are co-localized on the cell membrane of tumor cells. These findings suggest that activated MT1-MMP co-localized with alpha(v)beta(3) may be involved in activation of alpha(v)beta(3)-bound MMP-2.

Animals↗
PubMed1988

Immunohistochemical studies on the tissue localization of collagen types I, III, IV, V and VI in schwannomas. Correlation with ultrastructural features of the extracellular matrix.

The distinctive tissue localization of collagen types in typical schwannomas with Antoni type A and B areas was demonstrated immunohistochemically using affinity-purified antibodies against types I, III, IV, V and VI collagen and comparative ultrastructural studies were made on the extracellular matrix components. Antoni type A tissue, which was composed of tightly packed spindle cells with long cytoplasmic processes surrounded by a continuous basement membrane and a few fibrillar components of the extracellular matrix, was almost exclusively immunoreactive for type IV collagen, presumably representing the basement membrane. Verocay bodies, which are organoid structures of Antoni type A tissue, had a variety of more abundant extracellular fibrous components, such as banded collagen fibrils, fibrous long-spacing fibrils and microfibrils. These were positive for type I and III, as well as type IV collagen. In Antoni type B areas, where two types to tumor cells designated Schwann cell-like and fibroblast-like were scattered in large amounts of amorphous extracellular matrix containing microfibrils and thick banded collagen fibrils, type VI collagen as well as types I, III and IV collagen were consistently detected. Type V collagen was localized in dense fibrous tissue areas and around blood vessels. These findings indicate that the differently organized cellular patterns of schwannomas, identified as Antoni types A and B, are characterized not only by the ultrastructural features of the extracellular matrix, but also by the distinctive collagen types produced by neoplastic Schwann cells.

Adult↗
PubMed2005

Handling of computational in vitro/in vivo correlation problems by Microsoft Excel: IV. Generalized matrix analysis of linear compartment systems.

A linear system comprising n compartments is completely defined by the rate constants between any of the compartments and the initial condition in which compartment(s) the drug is present at the beginning. The generalized solution is the time profiles of drug amount in each compartment, described by polyexponential equations. Based on standard matrix operations, an Excel worksheet computes the rate constants and the coefficients, finally the full time profiles for a specified range of time values.

Linear Models↗
PubMed2005

Lipid nanoparticles for skin penetration enhancement-correlation to drug localization within the particle matrix as determined by fluorescence and parelectric spectroscopy.

With topical treatment of skin diseases, the requirement of a high and reproducible drug uptake often still is not met. Moreover, drug targeting to specific skin strata may improve the use of agents which are prone to cause local unwanted effects. Recent investigations have indicated that improved uptake and skin targeting may become feasible by means of nanoparticular systems such as solid lipid nanoparticles (SLN), nanostructured lipid carriers (NLC) and nanoemulsions (NE). Here we describe techniques to characterize drug loading to carrier systems and skin penetration profiles by using the lipophilic dye nile red as a model agent. Since the mode of drug association with the particle matrix may strongly influence the efficiency of skin targeting, parelectric spectroscopy (PS) was used to differentiate between matrix incorporation and attachment to the particle surface and fluorescence spectroscopy (FS) to solve dye distribution within NLC particles. Nile red was incorporated into the lipid matrix or the covering tensed shell, respectively, of SLN and NLC with all the lipids studied (Compritol, Precirol, oleic acid, Miglyol). In NLC, the dye was enriched in the liquid phase. Next, nile red concentrations were followed by image analysis of vertical sections of pigskin treated with dye-loaded nanoparticular dispersions and an oil-in-water cream for 4 and 8 h in vitro. Following the SLN dispersions, dye penetration increased about fourfold over the uptake obtained following the cream. NLC turned out less potent (<threefold increase) and penetration appeared even reduced when applying a NE. In contrast to previous studies with glucocorticoids attached to the surface of SLN, a targeting effect was not detected here. Therefore, drug targeting appears to be more strictly related to the mode of interaction of drug and particle than penetration enhancement.

Administration, Topical↗