ABSTRACT
Osteoblast cell adhesion to the extracellular matrix is established through two main pathways: one is mediated by the binding between integrin and a minimal adhesion sequence (RGD) on the extracellular protein, the other is based on the interactions between transmembrane proteoglycans and heparin-binding sequences found in many matrix proteins. The aim of this study is the evaluation in an in vivo endosseous implant model of the early osteogenic response of the peri-implant bone to a biomimetic titanium surface functionalized with the retro-inverso 2DHVP peptide, an analogue of Vitronectin heparin binding site. The experimental plan is based on a bilateral study design of Control and 2DHVP implants inserted respectively in the right and left femur distal metaphysis of adult male Wistar rats (n=16) weighing about 300grams and evaluated after 15days. Fluorochromic bone vital markers were given in a specific time frame, in order to monitor the dynamic of new bone deposition. The effect inducted by the peptidomimetic coating on the surrounding bone were qualitatively and quantitatively evaluated by means of static and dynamic histomorphometric analyses performed within three concentric and subsequent circular Regions of Interest (ROI) of equivalent thickness (220µm), ROI1 adjacent to the interface, ROI2, the middle, and ROI3 the farthest. The data indicated that these functionalized implants stimulated a higher bone apposition rate (p<0,01) and larger and rapid osteoblast activation in terms of mineralizing surface within ROI1 compared to the control (p<0,01). These higher osteoblast recruitment and activation leads to a greater bone-to-implant contact reached for DHVP samples (p<0,5). This represents an initial stimulus of the osteogenic activity that might results in a faster and better osteointegration process.
Subject(s)
Osteogenesis/drug effects , Peptidomimetics , Prostheses and Implants , Titanium/chemistry , Titanium/pharmacology , Amino Acid Sequence , Animals , Biomimetics , Femur/anatomy & histology , Femur/growth & development , Male , Osseointegration , Peptides/chemistry , Rats , Rats, Wistar , Receptors, Vitronectin/drug effects , Surface PropertiesABSTRACT
OBJECTIVE: Sinus floor elevation with lateral approach is probably the most frequently performed reconstructive procedure to rehabilitate posterior maxilla when a bone deficiency is present. Different graft materials have been proposed and tested, often with high clinical performances and predictable results. Histological analysis is required when evaluating new materials. We investigated human biopsies retrieved after sinus floor elevation procedure by histomorphometric evaluation to test the performance of an equine-derived bone grafting material. STUDY DESIGN: Seventeen consecutive patients were enrolled and sinus lift surgeries were performed using an equine bone graft. Six months after surgery, at implant placement, bone samples were collected. Histomorphometry analysis was carried out on decalcified samples. RESULTS: All surgeries were uneventful and no additional grafting was required prior to implant insertion. Forty percent of new bone formation was detected, which represented the most abundant tissue retrieved, followed by the residual graft material (33%) and fibrous tissue (27%). A significant reduction in particles size demonstrates a remodeling activity of the graft material. CONCLUSION: Within the limitations of this study, this equine-derived bone graft proved to be an effective material to induce new bone formation in the sinus floor elevation procedure.
Subject(s)
Bone Transplantation/methods , Dental Implantation, Endosseous/methods , Maxillary Sinus/surgery , Sinus Floor Augmentation/methods , Adolescent , Adult , Animals , Female , Horses/anatomy & histology , Humans , Male , Maxilla/physiopathology , Maxilla/surgery , Maxillary Sinus/physiopathology , Osteogenesis/physiology , Transplants/growth & development , Transplants/transplantation , Young AdultABSTRACT
Rough titanium surfaces enhance cell response to activation of Wnt canonical signalling, a pathway required for osteoblast differentiation. The present study investigated the effects of GSK3ß-inhibitors SB216763 and SB415286 on osteoblastic differentiation on titanium surfaces with different topography and wettability. Osteoblastic MC3T3 cells were plated on smooth (Pickled), sand-blasted/acid-etched (SLA) or hyper hydrophilic SLA (modSLA) titanium discs and transfected with a reporter vector sys-tem for Wnt canonical signalling. Cells were also seeded in the presence or in the absence of GSK3b-inhibitors SB216763 or SB415286 and their viability, morphology and the expression of Wnt target and osteoblast specific genes was assessed by Real Time PCR. Inhibitors altered cell morphology and mostly reduced cell viability at high concentration. SB415286 markedly increased the expression of ALP in MC3T3 cells on rough surfaces at the concentration of 100 nM before decreasing its expression at higher concentrations. OCN expression was unaffected. Increasing concentrations of SB216763 increased the expression of ALP in MC3T3 cells on rough surfaces but OCN expression was not changed at any con-centration. SB216763 and SB415286 inhibitors should be further investigated as potential tools to improve cell differentiation on titanium surfaces for endosseous implants.
Subject(s)
Aminophenols/pharmacology , Cell Differentiation/drug effects , Glycogen Synthase Kinase 3 beta/antagonists & inhibitors , Indoles/pharmacology , Maleimides/pharmacology , Osteoblasts/enzymology , Titanium/chemistry , Animals , Cell Line , Dose-Response Relationship, Drug , Glycogen Synthase Kinase 3 beta/metabolism , Mice , Osteoblasts/cytology , Surface PropertiesABSTRACT
The aim of the present study was to investigate how the enrichment of chitosan films with anti-fibronectin aptamers could enhance scaffold colonization by osteoblasts, by improving their adhesion and accelerating their proliferation. Chitosan discs were enriched with excess of anti-fibronectin aptamer. Aptamer adsorption on chitosan was monitored by measuring aptamer concentration in the supernatant by spectrophotometry, as well as its release, while functionalization was confirmed by labelling aptamers with a DNA intercalating dye. Chitosan samples were then characterized morphologically with atomic force microscopy and physically with contact angle measurement. Chitosan enrichment with fibronectin was then investigated by immunofluorescence and Bradford assay. 2% chitosan discs were then enriched with increasing doses of aptamers and used as culture substrates for MC3T3-E1 cells. Cell growth was monitored by optical microscopy, while cell viability and metabolic activity were assessed by chemiluminescence and by Resazurin Sodium Salt assay. Cell morphology was investigated by cytofluorescence and by scanning electron microscopy. Chitosan films efficiently bound and retained aptamers. Aptamers did not affect the amount of adsorbed fibronectin, but affected osteoblasts behavior. Cell growth was proportional to the amount of aptamer used for the functionalization, as well as aptamers influenced cell morphology and their adhesion to the substrate. Our results demonstrate that the enrichment of chitosan films with aptamers could selectively improve osteoblasts behavior. Furthermore, our results support further investigation of this type of functionalization as a suitable modification to ameliorate the biocompatibility of biomaterial for hard tissue engineering applications.
Subject(s)
Aptamers, Nucleotide/pharmacology , Chitosan/chemistry , Membranes, Artificial , Osteoblasts/physiology , Raffinose/chemistry , 3T3 Cells , Animals , Mice , Raffinose/metabolism , Tissue ScaffoldsABSTRACT
Androgen hormones play a significant role in regulating bone morphogenesis and in maintaining bone homeostasis throughout life. This study aimed to investigate the local effects of the non-aromatizable androgen stanozolol (ST) on bone regeneration in rats. Bilateral critical-size defects were created in the parietal bone of 26 male Wistar rats: the defect on one side was filled with a deproteinized bovine bone scaffold (DBB) soaked in ST solution (test) and the contralateral with DBB alone (control). Samples were collected at one month and three months. Histomorphometry revealed a significantly higher new bone formation (NB) (24.41% ± 4.14% versus 15.01% ± 2.43%, p < 0.05) and mineral apposition rate (MAR) (9.20 µm/day ± 0.37 versus 6.50 µm/day ± 1.09, p < 0.05) in the test versus control group at one month. Accordingly, real time-polymerase chain reaction revealed a consistently higher Runx2 expression in test samples (fold change test/control: 4.50 ± 1.17, p ≤ 0.05). No morphometrical differences between groups were detected at three months (p > 0.05). However, test samples were characterized by an increase in blood capillary density from one month (11.43 n mm-2 ± 2.01) to three months (28.26 n mm-2 ± 5.62), providing evidence of a vital remodeling tissue. Control samples presented a decrease of anti-Osterix (SP7)/anti-osteocalcin (BGLAP) (3.9 n mm-2 ± 0.32 versus 1.01 n mm-2 ± 0.20) and alkaline phosphatase (ALP) (12.14 n mm-2 ± 6.29 versus 6.29 n mm-2 ± 2.73) immunohistochemical-positive elements, which was suggestive of a stabilized healing phase. Based on these observations, local ST administration boosted bone regeneration in rat calvarial critical-size defects at one month. This study showed the potential of local steroid delivery in bone regeneration.
Subject(s)
Bone Regeneration/drug effects , Bone Substitutes/pharmacology , Bone and Bones/drug effects , Stanozolol/chemistry , Animals , Biocompatible Materials , Bone Matrix/transplantation , Bone Transplantation , Gene Expression Profiling , Guided Tissue Regeneration , Male , Osteocalcin/metabolism , Osteogenesis , Powders , Rats , Rats, Wistar , Transcription Factors/metabolism , Wound HealingABSTRACT
This review focuses on methods used to predict complex traits. Main characteristics of prediction approaches are given: the deterministic or stochastic nature of prediction, the objects of prediction, the sources of information and the main statistical methods. Sources of information discussed are the traditional genealogies and phenotypes, nucleotide sequences, expression data and epigenetics marks. Statistical methods are presented as successive degrees of generalization from the definition of the conditional expectation as the prediction rule, to best linear unbiased prediction, then Bayesian and, recently, machine learning methods, including meta-methods. We highlight the contributions of Daniel Gianola to this methodological evolution.
Subject(s)
Bayes Theorem , Models, Genetic , Models, Statistical , Quantitative Trait Loci , Quantitative Trait, Heritable , Animals , Genetics, Population , Humans , PhenotypeABSTRACT
UNLABELLED: Protein adsorption is the first and decisive step to define cell-biomaterial interaction. Guiding the adsorption of desired protein species may represent a viable approach to promote cell activities conducive to tissue regeneration. The aim of the present study was to investigate whether immobilized anti-Fibronectin aptamers could promote the attachment and growth of osteoblastic cells. Polyethyleneglycole diacrylate/thiolated Hyaluronic Acid hydrogels (PEGDA/tHA) were coated with anti-Fibronectin aptamers. Hydrogel loading and Fibronectin bonding were investigated, through spectrophotometry and Bradford assay. Subsequently, human osteoblasts (hOBs) were cultured on hydrogels for 10days in 2D and 3D cultures. Cells were monitored through microscopy and stained for focal adhesions, microfilaments and nuclei using fluorescence microscopy. Samples were also included in paraffin and stained with Hematoxylin-Eosin. Cell number on hydrogels was quantitated over time. Cell migration into the hydrogels was also studied through Calcein AM staining. Aptamers increased the number of adherent hOBs and their cytoplasm appeared more spread and richer in adhesion complexes than on control hydrogels. Viability assays confirmed that significantly more cells were present on hydrogels in the presence of aptamers, already after 48h of culture. When hOBs were encapsulated into hydrogels, cells were more numerous on aptamer-containing PEGDA-tHA. Cells migrated deeper in the gel in the presence of DNA aptamers, appearing on different focus planes. Our data demonstrate that anti-Fibronectin aptamers promote scaffold enrichment for this protein, thus improving cell adhesion and scaffold colonization. STATEMENT OF SIGNIFICANCE: We believe aptamer coating of biomaterials is a useful and viable approach to improve the performance of scaffold materials for both research and possibly clinical purposes, because different medical devices could be envisaged able to capture bioactive mediators from the patients' blood and concentrate them where they are needed, on the biomaterial itself. At the same time, this technology could be used to confer 3D cell culture scaffold with the ability to store proteins, such as Fibronectin, taking it from the medium and capture what is produced by cells. This is an improvement of traditional biomaterials that can be enriched with exogenous molecules but are not able to selectively capture a desired molecule.
Subject(s)
Aptamers, Peptide/pharmacology , Fibronectins/antagonists & inhibitors , Materials Testing/methods , Tissue Scaffolds/chemistry , Animals , Cattle , Cell Adhesion/drug effects , Cell Movement/drug effects , Cell Proliferation/drug effects , Coated Materials, Biocompatible/pharmacology , Fibronectins/metabolism , Humans , Hydrogels/pharmacology , Osteoblasts/cytology , Protein Binding/drug effects , Reproducibility of ResultsABSTRACT
The aim of the present work was to investigate the morphology and activity of the murine osteoblastic cell line MC3T3 on control smooth (Machined), commercially available rough (ZT) titanium discs, and on titanium samples obtained by modifying the ZT treatment protocol, and herein labelled as ZTF, ZTM and ZTFM. Cells were evaluated at SEM and immunofluorescence for morphology and cell-to-cell interactions and by MTT assay and real time PCR for cell growth and function. Microscopy showed that ZT modified protocols could differently affect cell shape and distribution. All the tested surfaces showed good biocompatibility by viability assay. However, cells on smoother surfaces appeared to express higher levels of transcript for Collagen 1a1, the main component of extracellular matrix, by real time PCR. Expression of the early differentiation marker Alkaline Phosphatase was higher on ZTF surfaces and ZTM enhanced the expression of later osteoblastic markers Osteoprotegerin and Osteocalcin. Noteworthy, the expression of Connexin 43, a component of cell-to-cell contacts and hemichannels, followed a similar pattern to differentiation marker genes and was higher in cells on ZTM surfaces, consistently with the microscopic observation of cell clusters. Taken together, this data showed that ZTF and ZTM treatment protocols appeared to improve the basal sand-blasting/acid-etching ZT procedure with ZTM surfaces promoting the most mature stage of differentiation.
Subject(s)
Biocompatible Materials , Osteoblasts/metabolism , Titanium/chemistry , 3T3 Cells , Animals , Cell Survival , Gene Expression Regulation/physiology , Hydrophobic and Hydrophilic Interactions , Mice , Microscopy, Electron, Scanning , Real-Time Polymerase Chain Reaction , Surface PropertiesABSTRACT
The aim of the present study was to investigate whether chitosan-based scaffolds modified with D-(+) raffinose and enriched with thiol-modified gelatin could selectively improve osteoblast adhesion and proliferation. 2, 3 and 4.5% chitosan films were prepared. Chitosan suitability for tissue engineering was confirmed by protein adsorption assay. Scaffolds were incubated with a 2.5 mg ml(-1) BSA solution and the decrease of protein content in the supernatants was measured by spectrophotometry. Chitosan films were then enriched with thiol-modified gelatin and their ability to bind BSA was also measured. Then, 2% chitosan discs with or without thiol-modified gelatin were used as culture substrates for MC3T3-E1 cells. After 72 h cells were stained with trypan blue or with calcein AM and propidium iodide for morphology, viability and proliferation assays. Moreover, cell viability was measured at 48, 72, 96 and 168 h to obtain a growth curve. Chitosan films efficiently bound and retained BSA proportionally to the concentration of chitosan discs. The amount of protein retained was higher on chitosan enriched with thiol-modified gelatin. Moreover, chitosan discs allowed the adhesion and the viability of cells, but inhibited their proliferation. The functionalization of chitosan with thiol-modified gelatin enhanced cell spreading and proliferation. Our data confirm that chitosan is a suitable material for tissue engineering. Moreover, our data show that the enrichment of chitosan with thiol-modified gelatin enhances its biological properties.
Subject(s)
Chitosan/chemistry , Gelatin/chemistry , Osteoblasts/physiology , Raffinose/pharmacology , Tissue Engineering/instrumentation , Tissue Scaffolds , Animals , BALB 3T3 Cells , Cell Adhesion/drug effects , Cell Adhesion/physiology , Cell Proliferation/drug effects , Cell Proliferation/physiology , Coated Materials, Biocompatible/chemical synthesis , Equipment Design , Equipment Failure Analysis , Materials Testing , Mice , Osteoblasts/cytology , Osteoblasts/drug effects , Raffinose/chemistry , Sulfhydryl Compounds/chemistry , Tissue Engineering/methodsABSTRACT
Oral movement disorders may lead to prosthesis and implant failure due to excessive loading. We report on an edentulous patient suffering from drug-induced tardive dyskinesia (TD) and oral parafunction (OP) rehabilitated with implant-supported screw-retained prostheses. The frequency and intensity of the movements were high, and no pharmacological intervention was possible. Moreover, the patient refused night-time splint therapy. A series of implant and prosthetic failures were experienced. Implant failures were all in the maxilla and stopped when a rigid titanium structure was placed to connect implants. Ad hoc designed studies are desirable to elucidate the mutual influence between oral movement disorders and implant-supported rehabilitation.
ABSTRACT
AIMS: Promoting bone formation at the tissue interface is an important step to improve implant success. This study investigated whether stimulation of Wnt signaling by GSK3b inhibitor lithium chloride (LiCl) could affect the response of mesenchymal or osteoblastic cells growing on titanium surfaces with different topography and wettability, and improve their differentiation along the osteoblastic lineage. MATERIAL AND METHODS: Murine mesenchymal C2C12 cells were plated on Pickled, acid-etched/sand-blasted (SLA), and hydrophilic SLA titanium disks (modSLA) and stimulated with increasing doses of LiCl. Cell viability was measured using chemiluminescence-based ATP quantitation and activation of Wnt canonical signaling was measured using a Luciferase-based reporter assay. Gene expression was measured using real time PCR in C2C12 cells, murine osteoblastic MC3T3 cells or murine primary bone marrow cells. RESULTS: LiCl stimulated Wnt activation and expression of Wnt markers in C2C12 cells on modSLA. Addition of 1 mM LiCl increased levels for bone marker Osteocalcin in MC3T3 cells on modSLA surfaces. Similarly, LiCl potently enhanced Osteoprotegetrin levels in MC3T3 cells on modSLA. When primary bone marrow cells were stimulated with LiCl, the expression of Wnttarget genes and osteoblastic differentiation markers was increased on modSLA surfaces. CONCLUSIONS: Stimulation of the canonical Wnt pathway promoted osteoblast differentiation on hydrophilic modSLA surfaces. Taken together, these results demonstrate that Wnt activators such as LiCl should be further tested as a possible approach to improve implant osseointegration.
Subject(s)
Dental Materials/chemistry , Glycogen Synthase Kinase 3/antagonists & inhibitors , Lithium Chloride/pharmacology , Osteoblasts/drug effects , Titanium/chemistry , Wnt Signaling Pathway/drug effects , 3T3 Cells , Acid Etching, Dental/methods , Alkaline Phosphatase/analysis , Animals , Bone Marrow Cells/drug effects , Cell Culture Techniques , Cell Differentiation/drug effects , Cell Line , Cell Lineage/drug effects , Cell Survival/drug effects , Dental Etching/methods , Glycogen Synthase Kinase 3 beta , Hydrophobic and Hydrophilic Interactions , Luciferases , Luminescence , Luminescent Agents , Mice , Muscle Cells/drug effects , Osteocalcin/drug effects , Osteoprotegerin/drug effects , Surface Properties , WettabilityABSTRACT
Endosseous implants are important tools to replace missing teeth or damaged tissue segments. Their clinical success depends on their integration in bone and, thus, on the response of bone cells to material and surface characteristics. Recent evidence has shown that surface topography and chemistry affect WNT signalling, a pivotal pathway for the commitment of mesenchymal progenitors to the osteoblast lineage and for bone homeostasis. WNT signalling comprises several cascades that, acting through different effectors, modulate several aspects of cell behaviour. It has been shown that cells growing on rough titanium surfaces display a different expression profile for WNT factors, and that surface features can alter the response of bone cells to WNT factors. Although the underlying mechanisms to this regulation are still poorly understood, the present review reports intriguing evidence that that cell cytoskeletal signalling is involved in activating WNT signalling in cells growing on rough implant surfaces.
Subject(s)
Bone and Bones/metabolism , Mesenchymal Stem Cells/metabolism , Osseointegration/physiology , Osteoblasts/metabolism , Wnt Signaling Pathway/physiology , Cell Adhesion/drug effects , Cell Differentiation/drug effects , Dental Implantation, Endosseous/methods , Humans , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/drug effects , Osseointegration/drug effects , Osteoblasts/cytology , Osteoblasts/drug effects , Surface Properties , Titanium/chemistry , Titanium/pharmacology , Wnt Proteins/metabolism , beta Catenin/metabolismABSTRACT
Genomic selection aims to increase accuracy and to decrease generation intervals, thus increasing genetic gains in animal breeding. Using real data of the French Lacaune dairy sheep breed, the purpose of this study was to compare the observed accuracies of genomic estimated breeding values using different models (infinitesimal only, markers only, and joint estimation of infinitesimal and marker effects) and methods [BLUP, Bayes Cπ, partial least squares (PLS), and sparse PLS]. The training data set included results of progeny tests of 1,886 rams born from 1998 to 2006, whereas the validation set had results of 681 rams born in 2007 and 2008. The 3 lactation traits studied (milk yield, fat content, and somatic cell scores) had heritabilities varying from 0.14 to 0.41. The inclusion of molecular information, as compared with traditional schemes, increased accuracies of estimated breeding values of young males at birth from 18 up to 25%, according to the trait. Accuracies of genomic methods varied from 0.4 to 0.6, according to the traits, with minor differences among genomic approaches. In Bayes Cπ, the joint estimation of marker and infinitesimal effects had a slightly favorable effect on the accuracies of genomic estimated breeding values, and were especially beneficial for somatic cell counts, the less heritable trait. Inclusion of infinitesimal effects also improved slopes of predictive regression equations. Methods that select markers implicitly (Bayes Cπ and sparse PLS) were advantageous for some models and traits, and are of interest for further quantitative trait loci studies.
Subject(s)
Breeding/methods , Sheep/genetics , Animals , Bayes Theorem , Dairying/methods , Female , France , Genotype , Lactation/genetics , Least-Squares Analysis , Male , Pedigree , Phenotype , Quantitative Trait, HeritableABSTRACT
Goat milk somatic cell counts have been collected for several years in France by the national milk recording organization. Information is used for health management, because repeatedly elevated somatic cell counts are a good indirect predictor of intramammary infection. Genetic parameters were estimated for 67,882 and 49,709 primiparous goats of the dairy Alpine and Saanen breeds, respectively, with complete information for milk somatic cell counts and milk production traits. About 40% of the goats had additional information for 11 udder type traits scored by official classifiers of the breeders' association CAPGENES. Estimates were obtained by REML with an animal model. The studied trait was lactation somatic cell score (LSCS), the weighted mean of somatic cell score (log-transformed SCC) adjusted for lactation stage. Heritability of LSCS was 0.20 and 0.24 in the Alpine and Saanen breeds, respectively. Relationships with milk production and udder type traits were additionally estimated by using multitrait analyses. Heritability estimates in first lactation ranged from 0.30 to 0.35 for lactation milk, fat, and protein yields; from 0.60 to 0.67 for fat and protein contents; and from 0.22 to 0.50 for udder type traits. Genetic correlations of somatic cell score with milk production traits were generally low, ranging from -0.13 to 0.12. Slightly more negative correlations were estimated for fat content: -0.18 and -0.20 in Saanen and Alpine breeds, respectively. Lactation somatic cell score was genetically correlated with udder floor position (r(g)=-0.24 and -0.19 in the Alpine and Saanen breeds, respectively), and, in Saanen, teat length, teat width, and teat form (r(g)=0.29, 0.34 and -0.27, respectively). These results suggest that a reduction in somatic cell count can be achieved by selection while still improving milk production and udder type and teat traits.
Subject(s)
Goats/genetics , Lactation/genetics , Mammary Glands, Animal/anatomy & histology , Milk/cytology , Animals , Cell Count/veterinary , Female , France , Goat Diseases/microbiology , Mastitis/microbiology , Mastitis/veterinary , Parity , PregnancyABSTRACT
Multiplex PCR for the detection of sea, seb, sec, sed and see genes of Staphylococcus aureus. Characterization of isolates from food. The presence of Staphylococcus aureus in food represents a potential risk to public health, being its enterotoxins the major virulence factor. Enterotoxin detection can be determined by ELISA, but only for the pool of enterotoxins SEA, SEB, SEC, SED and SEE. The main aims of this study were to optimize two PCR techniques for detection of S. aureus sea, seb, sec, sed and see, and to characterize Staphylococcus spp. isolates associated with food intoxication. Two PCR techniques were optimized and 115 Staphylococcus spp. isolates from Ciudad Aut6noma de Buenos Aires, and Buenos Aires, Córdoba, and Neuquén provinces were characterized. The characterization was performed by biochemical tests, ELISA and PCR. Sixty-eight isolates (59.1%) were positive by ELISA, while 61 (53%) were positive by PCR. Out of the positive PCR isolates, 34 (55.7%) carried the sea gene, 9 (14.8%) the seb gene, 5 (8.1%) the see gene, 4 (6.5%) the sec gene, 6 (9.9%) were positive for sea and seb genes, 2 (3.3%) for sea and sec genes, and 1 (1.7%) for sea and sedgenes. This is the first study of genotypic characterization of S. aureus isolates associated with food intoxication from different provinces of Argentina.
Subject(s)
Genes, Bacterial/genetics , Staphylococcus aureus/genetics , Food Microbiology , Polymerase Chain Reaction/methods , Staphylococcus aureus/isolation & purificationABSTRACT
Optimal management of retinal vein occlusion (RVO) is still a matter of debate. Antithrombotic and fibrinolytic drugs have been investigated after demonstration of a role of thrombosis in the complex pathogenesis of the disease. Aim of our study was to systematically summarise best available evidence on the acute treatment and on the secondary prevention of RVO with antithrombotic and fibrinolytic drugs. A computer-assisted search of the MEDLINE and EMBASE electronic databases up to January 2009 was performed. Two review authors selected all published randomised controlled trials (RCTs) from the search, assessed study quality and extracted data. Based on Jadad's score, RCTs were stratified into three quality categories. A total of six RCTs were included. Only one RCT of high quality was identified. A total of 384 patients were investigated, 234 with central retinal vein occlusion and 150 with branch retinal vein occlusion. No study enrolled more than 100 patients. Three studies compared therapeutic doses of low-molecular-weight heparin (LMWH) with low-dose aspirin, one study compared ticlopidine with placebo and two studies compared intravenous fibrinolytic therapy followed by warfarin or aspirin with either haemodilution or no treatment. A partial improvement of visual acuity was reported in every study, independently of the study drug. No long-term secondary prevention study was published. The present systematic review suggests that antithrombotic therapy, in particular LMWH, may be part of the therapeutic armamentarium for patients with recent onset RVO. No firm recommendation can be provided given the limited available evidence.
Subject(s)
Fibrinolytic Agents/therapeutic use , Retinal Vein Occlusion/drug therapy , Aspirin/therapeutic use , Drug Evaluation , Heparin, Low-Molecular-Weight/therapeutic use , Humans , Randomized Controlled Trials as Topic , Ticlopidine/therapeutic use , Treatment Outcome , Visual Acuity/drug effects , Warfarin/therapeutic useABSTRACT
The phenotypic observation of some reproduction traits (e.g., insemination success, interval from lambing to insemination) is the result of environmental and genetic factors acting on 2 individuals: the male and female involved in a mating couple. In animal genetics, the main approach (called additive model) proposed for studying such traits assumes that the phenotype is linked to a purely additive combination, either on the observed scale for continuous traits or on some underlying scale for discrete traits, of environmental and genetic effects affecting the 2 individuals. Statistical models proposed for studying human fecundability generally consider reproduction outcomes as the product of hypothetical unobservable variables. Taking inspiration from these works, we propose a model (product threshold model) for studying a binary reproduction trait that supposes that the observed phenotype is the product of 2 unobserved phenotypes, 1 for each individual. We developed a Gibbs sampling algorithm for fitting a Bayesian product threshold model including additive genetic effects and showed by simulation that it is feasible and that it provides good estimates of the parameters. We showed that fitting an additive threshold model to data that are simulated under a product threshold model provides biased estimates, especially for individuals with high breeding values. A main advantage of the product threshold model is that, in contrast to the additive model, it provides distinct estimates of fixed effects affecting each of the 2 unobserved phenotypes.
Subject(s)
Models, Genetic , Reproduction/genetics , Animals , Female , Male , PhenotypeABSTRACT
En febrero de 2006 ocurrió un brote epidémico de gastroenteritis aguda de origen alimentario, en ocasión de un festejo popular en una pequeña localidad de la provincia de Neuquén, Argentina. Aproximadamente 800 personas participaron de un almuerzo en las instalaciones del Gimnasio Municipal, y unas tres horas después de finalizado, cerca de 150 asistentes consultaron al hospital local, afectados por síndrome gastroentérico agudo. Se realizó una investigación epidemiológica caso-control a través de un muestreo representativo no probabilístico. Los resultados epidemiológicos establecieron un brote de ETA a fuente común, con una relación caso-control de 1:1,8. Los principales síntomas fueron cólicos abdominales (88%), vómitos (73,5%) y diarrea (60%). La torta que se sirvió en ese evento fue identificada como el alimento causal (OR 9,79; IC 95%; 2,66-36,00; valor p = 0,0001), sujeto a condiciones higiénico-sanitarias insatisfactorias en los diferentes procesos de elaboración, conservación y manipulación. De una porción de la torta se aisló una cepa de Staphylococcus aureus subespecie aureus, coagulasa positiva, enterotoxigénica, con un recuento de 2,4x10(6) UFC/g, y también se aisló este microorganismo de tres muestras de manos y narinas de personas involucradas en la preparación y el servicio. Las cepas aisladas de un operador y de la torta portaron el gen sea y presentaron el mismo patrón de SmaI-PFGE. Se atribuyó el brote de ETA a la contaminación durante el proceso de preparación de la torta consumida durante ese almuerzo popular, lo que podría estar relacionado con deficiencias en aspectos higiénicos y con la falta de refrigeración y de mantenimiento de la cadena de frío.
In the summer of 2006, an epidemic outbreak of acute gastrointestinal illness related to food consumption occurred in a small town in the province of Neuquén, Argentina. During a popular feast, approximately 800 local residents attended lunch held in the facilities of the Municipal Gymnasium. About three hours later, nearly 150 attendees sought medical assistance at the local hospital due to acute gastroenteritis. A case-control epidemiological investigation was conducted using representative non-probability sampling. The epidemiological investigation showed a common-source foodborne disease outbreak with a case-control ratio of 1:1.8. The main symptoms were abdominal cramps (88%), vomiting (73.5%) and diarrhea (60%). The cake was identified as the source of infection (OR 9.79; IC 95%, 2.66-36.00; p = 0.0001), and unsatisfactory hygienic conditions in food production, conservation and handling steps were identified. Coagulase positive, enterotoxigenic Staphylococcus aureus, subspecies aureus was detected in a piece of cake, with a count of 2.4x10(6) CFU/g, and in samples from the hands and nostrils of three people involved in food preparation and service. The strains isolated from both the cake and one of the food handlers carried the sea gene, and presented the same SmaI-PFGE pattern. The foodborne disease outbreak was considered to be due to contamination in the preparation process of the cake consumed at the feast, which was related to inadequate hygienic conditions, lack of refrigeration and cold chain disruption.
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Disease Outbreaks , Food Contamination , Gastroenteritis/epidemiology , Staphylococcal Food Poisoning/epidemiology , Argentina/epidemiology , Bacterial Typing Techniques , Case-Control Studies , Carrier State/metabolism , Food Handling , Gastroenteritis/etiology , Gastroenteritis/microbiology , Refrigeration , Staphylococcal Food Poisoning/microbiology , Staphylococcus aureus/classification , Staphylococcus aureus/genetics , Staphylococcus aureus/isolation & purificationABSTRACT
A total of 416,670 lactations for 189,101 ewes from 3,603 sires and distributed across 1,978 herd-year groups were used to estimate genetic and environmental parameters of standardized milk yield (SMY(T)), fertility in ewe lambs (PR(1)), and fertility in adult ewes (PR(A)). Parameters were estimated with a multiple-trait sire linear model. Heritabilities for SMY(T), PR(1), and PR(A) were 0.27 (0.009), 0.04 (0.004), and 0.05 (0.004), respectively. These results were in accordance with the literature. The genetic correlation between PR(1) and PR(A) was 0.55, indicating that fertility is not the same trait in ewe lambs and adult ewes. The genetic correlation between milk yield and lamb fertility was not significantly different from zero. The genetic correlation between milk yield and fertility in adult ewe (-0.23) was in the range of antagonistic correlations reported in dairy cattle. Consequently, these results show that selection for milk yield can induce an indirect decrease in fertility. Nevertheless, no phenotypic decrease in fertility in artificial insemination matings has been observed in this population. This is the first time that correlation between milk yield and fertility is reported in sheep and further investigations are needed to confirm this result.
Subject(s)
Fertility/genetics , Lactation/genetics , Milk/metabolism , Sheep/genetics , Animals , Female , Insemination, Artificial , Male , PhenotypeABSTRACT
Reproductive seasonality observed in all breeds of goats originating from temperate latitudes and in some breeds from subtropical latitudes can now be controlled by artificial changes in photoperiod. Short days stimulate sexual activity, while long days inhibit it. This knowledge has allowed the development of photoperiodic treatments to control sexual activity in goats, for both the buck and doe. In the French intensive milk production system, goat AI plays an important role to control reproduction and, in conjunction with progeny testing, to improve milk production. Most dairy goats are inseminated out of the breeding season with deep frozen semen, after induction of oestrus and ovulation by hormonal treatments. This protocol provides a kidding rate of approximately 65%. New breeding strategies have been developed, based on the buck effect associated with AI, to reduce the use of hormones. With the development of insemination with frozen semen, a classical selection programme was set up, including planned mating, progeny testing and the diffusion of proved sires by inseminations in herds. Functional traits have become important for efficient breeding schemes in the dairy goat industries. Based on knowledge gained over the past decade, the emphasis in selective breeding has been placed on functional traits related to udder morphology and health. New windows have been opened based on new molecular tools, allowing the detection and mapping of genes of economic importance.