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Biomedical subjects

M Pellecchia

Publications and source records attributed to M Pellecchia.

At least 19 recordsLinked to original sources
PubMed2006

Feminizing Wolbachia in Zyginidia pullula (Insecta, Hemiptera), a leafhopper with an XX/X0 sex-determination system.

Zyginidia pullula is a grass-dwelling leafhopper characterized by a bisexual reproduction mode. In this species, some females collected in Northern Italy, when mated with males, gave origin to an exclusively female brood. Here, we demonstrated that in these lineages an infection by a new strain of Wolbachia pipientis-designated as wZygpul-was detected by amplifying and sequencing the wsp and 16S rRNA genes. About half of the female progeny were characterized by intersexual phenotypes, i.e. showing upper pygofer appendages, a typical male secondary sexual feature. The karyological analysis proved that while phenotypically normal females had a female genotype, those with upper pygofer appendages had a male genotype and were thus feminized males. The complete removal of W. pipientis after tetracycline treatment of morphologically normal females, and the consequent re-appearance of males in the brood, permitted us to connect the feminizing effect with the presence of the bacterium. This is the first case of feminization by W. pipientis in an XX/X0 sex-determination system, and is the second case reported in insects.

Animals↗
PubMed2006

Quantitative determination of apogossypol, a pro-apoptotic analog of gossypol, in mouse plasma using LC/MS/MS.

A simple and selective liquid chromatography/tandem mass spectrometry (LC/MS/MS) method based on internal standard quantitation using apigenin as the internal standard has been developed and validated for the analysis of the gossypol analog apogossypol, a pro-apoptotic compound, in mouse plasma. The methodology involves protein precipitation of plasma samples followed by LC/MS/MS analysis. Ascorbic acid was added to the spiking solutions and plasma samples to stabilize the easily oxidized compound. Separation of apogossypol and the internal standard from the plasma matrix was achieved using a C18 column with a gradient elution profile consisting of 5mM ammonium acetate and methanol. The validated range of the method extended from 10 to 2000 ng/mL with accuracies of 85-115% and precision of <15%. The average recovery of apogossypol at three concentrations (50, 200 and 1000 ng/mL) assayed in triplicate using this methodology was determined to be 90.8+/-12.9%. Recovery for the internal standard (apigenin) at a concentration of 500 ng/mL was found to be 99.9+/-6.41%. Apogossypol concentrations of 50 ng/mL and above were found to be stable in extracted plasma for 24h when stored at 25 degrees C. This method has been applied to the determination of apogossypol concentrations in plasma collected from mice given an IV dose of apogossypol.

Animals↗
PubMed2006

Tuscany autochthonous cattle breeds: an original genetic resource investigated by AFLP markers.

The aim of this study was to assess the genetic diversity of four autochthonous cattle breeds of Tuscany and their relationships in comparison with Italian Friesian and Italian Brown, using amplified fragment length polymorphism markers. A total of 212 individuals were genotyped with three primer combinations generating 102 polymorphic markers. Average expected heterozygosity ranged from 0.23 in Mucca Pisana to 0.26 in Chianina, Italian Friesian, Italian Brown and Maremmana. The differences resulted not significant (Kruskall-Wallis test, p = 0.416). Gst-B index revealed that 86% of the total genetic variance is retained within population and only 14% is accounted by the between populations component. Multivariate analysis at individual and population level indicated that: (i) Calvana and Chianina are quite separate from the other breeds as an effect of the bottleneck experienced or as a signature of different origin; (ii) Podolian, Maremmana and Italian Brown clustered with the double purpose Mucca Pisana, revealing their contribution to its admixed genetic make up; (iii) Italian Friesian behaved always as out group. The 'analysis of molecular variance' recovered a significant subdivision clustering the six populations into three groups: Italian Friesian and Italian Brown versus Maremmana and Mucca Pisana versus Chianina and Calvana (6% of the total variance).

Animal Husbandry↗
PubMed2005

Traditional Chinese medicines with caspase-inhibitory activity.

Clinical evidence indicates that traditional Chinese medicine (TCM) drugs can reduce stroke-inflicted brain damage. To date, the molecular basis of the apparent neuroprotective effects of these TCM drugs remains largely obscure. Several lines of evidence indicate that the activation of cell death programs leads to the loss of neurons during the reperfusion phase of ischemic stroke. In particular, activation of caspases (cysteinyl aspartate-specific proteinases) is a critical step in neuronal apoptosis. Using nuclear magnetic resonance (NMR) and fluorescence assays, we screened a collection of 58 TCM drugs that are commonly used in stroke therapy for caspase inhibitory activity. We found that aqueous extracts of Lianqiao (Fructus Forsythiae) and Shouwuteng (Caulis Polygoni multiflori) blocked the activity of the initiator caspase-8 as well as the effector caspase-3 and caspase-7 in a dose-dependent manner with an IC(50)10 microg/ml. Identification of caspase inhibitory activity of these TCM drugs, allows the formulation of testable hypotheses and design of further investigations aimed at the elucidation of the molecular basis of TCM stroke therapy.

Caspase 3↗
PubMed2005

Comparative analysis of deformation of two implant/abutment connection systems during implant insertion. An in vitro study.

AIM: The implant/abutment interface represents a crucial element to maintain the structural stability of the implant restoration and to prevent mechanical problems that can compromise the long-term success of the implant treatment. METHODS: In this study deformation of the external hexagonal and internal octagonal extension of 2 implants after applying different values of insertion torque has been tested. The implants were secured in a customized resin baseplate, with the implant neck exposed, then different values of insertion torque were applied by an electronic torque controller. RESULTS: Five internal octagonal implants were tested for 60 seconds to different torque magnitudes (12 Ncm, 21 Ncm, 27 Ncm, 36 Ncm, 42 Ncm). Two external hexagonal implants were tested for 60 seconds to a torque value of 27 Ncm and of 36 Ncm. CONCLUSIONS: After torquing tests, each implant was examined by a scanning electron microscope (SEM) medium and high magnification to evaluate the deformation of the 2 different antirotational systems. The SEM analysis of the internal octagonal and external hexagonal antirotational systems of the TBR implant showed a deformation proportional to the magnitude of the applied torque during implant insertion. The geometric deformation of the antirotational extension is capable of compromising its function after a torque magnitude of 36 Ncm or superior to this value. Moreover it was demonstrated a more evident alteration of the external hexagonal extension in comparison with the octagonal system after applying a torque value of 27 Ncm.

Composite Resins↗
PubMed2005

Comparative analysis of direct and indirect implant impression techniques an in vitro study. An in vitro study.

Implant impression and transfer of the implant position to a working cast for rehabilitative purposes not always represent a predictable reconstructive step. The variable results depend on the transfer technique utilized and on the accuracy of the impression materials. The aim of this study was to compare the accuracy of the direct and indirect impression techniques and to analyze the distortion of different impression materials. An edentulous acrylic resin mandible was used as reference model: in the cuspid position 2 laboratory analogs were inserted perpendicularly to the occlusal plane and fastened with resin. Two impressions for each indirect and direct recording techniques with 4 different impression materials were made. Polyvinyl siloxanes were used for the repositioning implant level method. Polyethers were used for the ''pick up'' impression technique. Analysis of cast distortion was measured computing the inter-implant position on the developed models compared to that of the master model. Measurements were recorded by a digital caliper. The results of this in vitro study demonstrate that the direct impression technique is preferable to the indirect one and that the Impregum polyether impression material is the most precise when used in association with a direct technique.

Dental Impression Technique↗
PubMed2003

[Influence of biphosphonates on the integration process of endosseous implants evaluated using single photon emission computerized tomography (SPECT)].

Biphosphonates are a large class of drugs which inhibits the resorbing activity of osteoclasts. The aim of this study was to evaluate the influence of topical application of the biphosphonate alendronate on the integration process of endosseus implants placed in a partially edentulous patient. After incision and elevation of a mucoperiosteal flap and carrying out the osteotomy, 2 SLA implants 10 mm long and 3.5 mm wide were inserted in the area of # 4.5 and 4.6. Before placement of # 4.5, a paste made of grinded tablets of alendronate and Rifocin was applied in the osteotomy. Fifteen days after stage I, an evaluation of the osteoblastic peri-implant activity was carried out using single photon emission computerized tomography. Healing was within normal limits. Four months after stage I, the 2 implants were uncovered and considered clinically and radiographically integrated. The osteoblastic activity was judged similar bet-ween the 2 implants in terms of quality of biological response. Osseous scintigraphy and the achievement of implant osseointegration demonstrate the lack of toxicity of the topical application of the biphosphonate alendronate during the initial osseous healing. However, new experimental studies and clinical trials are necessary to confirm the results obtained.

Adult↗
PubMed2002

[Vertical ridge augmentation using a microplate in conjunction with a Gore-Tex membrane. A case report].

Oral implantology is considered an acceptable treatment modality to replace missing teeth when a recipient site with a sufficient horizontal and vertical bone volume exists. Guided bone regeneration is based on the principle of selective exclusion of non osteogenic cells from the bony defect by a barrier membrane. Several supporting systems of the barrier membrane have been successfully used to treat non space-mantaining de-fects. A 35-year-old female affected by a partial edentulism in the molar area of the right mandible showed 8 mm of bone height of the residual ridge above the alveolar canal. A vertical ridge augmentation was performed using a Gore-Tex membrane in conjunction with a microplate and an irradiated freeze-dried cortico-cancellous allograft. Six months after the surgical procedure a 11.5 mm by 5 mm threaded implant was placed into the regenerated bone. After three months of healing, the implant was uncovered and considered integrated. Before the implant placement, a biopsy of the regenerated bone was arried out by a triphine. The histological examination of the bony core showed the attainment of biological goals of guided bone regeneration. The surgical technique followes in this case report has been effective in regenerating a vertical bone volume sufficient for implant treatment.

Adult↗
PubMed2001

NMR in the acceleration of drug discovery.

The field of NMR spectroscopy has grown beyond expectations from the first historic observation of nuclear magnetic resonance in 1946 by Bloch and Purcell, to the first generation of protein structures 20 years ago, to the present where NMR structures now represent 15 to 20% of those submitted to the Protein Data Bank (PDB). However, the applications of NMR go far beyond structure determination. Since NMR is a non-invasive technique that provides a wealth of information about structure and dynamics, it is an ideal analytical method that is now being used in every step of the drug discovery and development process. Indeed, NMR is undergoing a renaissance in the pharmaceutical industry as a plethora of new applications have been developed, ranging from in vitro screening methods in early stage discovery to in vivo magnetic resonance imaging techniques used to assess drug efficacy in clinical trials. In this review, we will present an overview of the role of NMR in these areas, with a special focus on how NMR is poised to revolutionize the high-throughput structural characterization of protein-ligand interactions.

Animals↗
PubMed2000

Magnetization transfer via residual dipolar couplings: application to proton-proton correlations in partially aligned proteins.

A novel three-dimensional NMR experiment is reported that allows the observation of correlations between amide and other protons via residual dipolar couplings in partially oriented proteins. The experiment is designed to permit quantitative measurement of the magnitude of proton-proton residual dipolar couplings in larger molecules and at higher degree of alignments. The observed couplings contain data valuable for protein resonance assignment, local protein structure refinement, and determination of low-resolution protein folds.

Algorithms↗
PubMed2000

Structural insights into substrate binding by the molecular chaperone DnaK.

How substrate affinity is modulated by nucleotide binding remains a fundamental, unanswered question in the study of 70 kDa heat shock protein (Hsp70) molecular chaperones. We find here that the Escherichia coli Hsp70, DnaK, lacking the entire alpha-helical domain, DnaK(1-507), retains the ability to support lambda phage replication in vivo and to pass information from the nucleotide binding domain to the substrate binding domain, and vice versa, in vitro. We determined the NMR solution structure of the corresponding substrate binding domain, DnaK(393-507), without substrate, and assessed the impact of substrate binding. Without bound substrate, loop L3,4 and strand beta3 are in significantly different conformations than observed in previous structures of the bound DnaK substrate binding domain, leading to occlusion of the substrate binding site. Upon substrate binding, the beta-domain shifts towards the structure seen in earlier X-ray and NMR structures. Taken together, our results suggest that conformational changes in the beta-domain itself contribute to the mechanism by which nucleotide binding modulates substrate binding affinity.

Adenosine Triphosphatases↗
PubMed2000

Distal extension mandibular removable partial denture connected to an anterior fixed implant-supported prosthesis: a clinical report.

This clinical report demonstrates the possibility of achieving positive results with a removable prosthesis connected to an implant-supported fixed prosthesis. Bone quality and quantity at the implant sites are essential requirements for the success of the treatment. The support of the RPD and its connection with the fixed prosthesis creates stability during chewing activity and allows a functional activity similar to that involving an overdenture. The adoption of an occlusal scheme with subocclusion of the fixed prosthesis contacts compared with those of the removable prosthesis, together with the stress-director system of the RPD can be effective factors that decrease the risk of loss of implant integration. Further long-term follow-up studies with a larger patient population are needed to confirm the clinical and biomechanical validity of the prosthetic solution described in this clinical report.

Aged↗
PubMed1999

NMR structure of the human oncofoetal fibronectin ED-B domain, a specific marker for angiogenesis.

BACKGROUND: The process of angiogenesis (i.e. the formation of new blood vessels from pre-existing ones) is fundamental to physiological processes such as reproduction, development and repair, as well as to pathological conditions such as tumor progression, rheumathoid arthritis and ocular disorders. The oncofoetal ED-B domain, a specific marker of angiogenesis, consists of 91 amino acid residues that are inserted by alternative splicing into the fibronectin (FN) molecule. RESULTS: The NMR structure of the ED-B domain is reported and reveals important differences from other FN type III domains. A comparison of the ED-B domain with the crystal structure of a four-domain FN fragment shows the novel features of ED-B to be located in loop regions that are buried at interdomain interfaces, and which therefore largely determine the global shape of the FN molecule. The negatively charged amino acids in this highly acidic protein are uniformly distributed over the molecular surface, with the sole exception of a solvent-exposed hydrophobic patch that represents a potential specific recognition site. Epitope mapping with 82 decapeptides that span the ED-B sequence revealed that three ED-B-specific monoclonal antibodies, which selectively target newly forming blood vessels in tumor-bearing mice, bind to adjacent regions on the ED-B surface. CONCLUSIONS: The NMR structure enables the identification of a large surface area of the ED-B domain that appears to be accessible in vivo, opening up new diagnostic and therapeutic opportunities. Furthermore, the mapping of specific monoclonal antibodies to the three-dimensional structure of the ED-B domain, and their use in angiogenesis inhibition experiments, provides a basis for further investigation of the role of the ED-B domain in the formation of new blood vessels.

Amino Acid Sequence↗
PubMed1999

Line narrowing in spectra of proteins dissolved in a dilute liquid crystalline phase by band-selective adiabatic decoupling: application to 1HN-15N residual dipolar coupling measurements.

Residual heteronuclear dipolar couplings obtained from partially oriented protein samples can provide unique NMR constraints for protein structure determination. However, partial orientation of protein samples also causes severe 1H line broadening resulting from residual 1H-1H dipolar couplings. In this communication we show that band-selective 1H homonuclear decoupling during data acquisition is an efficient way to suppress residual 1H-1H dipolar couplings, resulting in spectra that are still amenable to solution NMR analysis, even with high degrees of alignment. As an example, we present a novel experiment with improved sensitivity for the measurement of one-bond 1HN-15N residual dipolar couplings in a protein sample dissolved in magnetically aligned liquid crystalline bicelles.

Anisotropy↗
PubMed1999

Pilus chaperone FimC-adhesin FimH interactions mapped by TROSY-NMR.

The 23 kDa two-domain periplasmic chaperone FimC from Escherichia coli is required for the assembly of type-1 pili, which are filamentous, highly oligomeric protein complexes anchored to the outer bacterial membrane that mediate adhesion of pathogenic E. coli strains to host cell surfaces. Here we identified the contact sites on the surface of the NMR structure of FimC that are responsible for the binding of the 28 kDa mannose-binding type-1 pilus subunit FimH by 15N and 1H NMR chemical shift mapping, using transverse relaxation-optimized spectroscopy (TROSY). The FimH-binding surface of FimC is formed nearly entirely by the N-terminal domain, and its extent and shape indicate that FimC binds a folded form of the pilus subunits.

Adhesins, Bacterial↗