Bovine Follicular Dynamics, Oocyte Recovery,and Development of Oocytes Microinjected with a Green Fluorescent Protein Construct

The present study was carried out to 1) evaluate the viability of in vitro fertilized zygotes after microinjection of DNA, 2) assess the influence of oocyte quality upon the development rate of embryos when injected with DNA, and 3) determine the integration frequency of green fluorescent protein DNA into microinjected embryos. Oocytes were aspirated from ovaries of nine nonlactating Holsteins and were categorized into grades A, B, C, and D. At 16 h after in vitro fertilization, approximately half of the pronuclear stage presumptive zygotes were classified as having 1 pronucleus or 2 pronuclei, and they were microinjected with DNA constructs. A potential predictor of DNA integration frequency at d 10 was assessment of the incidence of green fluorescing embryos. The proportion of cleaved embryos that developed to morulae or blastocysts was not different between groups with 1 pronucleus injected (45%), 1 pronucleus uninjected (64%), or 2 pronuclei injected (49%). However, the development of morulae or blastocysts was higher in the group with 2 pronuclei uninjected (69%). The overall developmental score of green fluorescent protein-positive embryos was higher for grade A oocytes (1.3 &#;&#;0.1) than for grade B (0.8 &#; 0.1), C (0.6 &#;&#;0.1), or D (0.3 &#;&#;0.1) oocytes. The results show that production of transgenic bovine blastocysts can occur from the microinjection of a presumptive zygote having only one visible pronucleus. Initial oocyte quality is an important factor in selection of oocytes suitable for microinjection of DNA and for preimplantation development to produce bovine transgenic embryos

Sex influence on genetic expressions of early growth in Afshari lambs

Very little is known about the genetic aspects of sexual dimorphism of body weight in domestic sheep, and therefore this study was conducted to quantify the genetic basis of sexual dimorphism for early-growth-related traits in Afshari lambs. Traits evaluated included birth weight (BW), weaning weight (WW), and growth rate (GR) in male and female lambs. Male lambs were 6.6 % heavier at birth, had 14.4 % higher preweaning growth rates and were 16.0 % heavier at weaning compared to female lambs. Levels of sexual-size dimorphism (SSD), expressed as the ratio of male to female means, for BW, WW and GR were 1.07, 1.14 and 1.15, respectively, which indicated low levels of SSD in the traits studied. Fixed effects of year of birth and type of birth interacted with sex effects, with greater variability in birth and weaning weights among years and birth types in male lambs, suggesting greater environmental sensitivity in the males. Bivariate animal models and restricted maximum likelihood (REML) procedures were used to estimate phenotypic variances and their genetic and non-genetic components in male and female lambs. Estimates of the direct heritability (h2) and additive coefficient of variation (CVA) for BW were higher in males. However, for WW and GR, heritability estimates were higher in females. In contrast, whereas the contribution of maternal permanent environmental effects (c2) to variation of BW was higher in females, for WW and GR higher estimates of c2 were observed in males. Respective genetic and maternal permanent environmental correlations between records on males and females were 0.986 and 0.723 for BW, 0.995 and 0.983 for WW, and 0.995 and 0.966 for GR, indicating possible sexual dimorphism only for maternal effects on BW. Based on an approximate 95 % confidence interval, none of the observed differences in variance components between sexes differed from zero and none of the observed genetic or maternal correlations differed from 1.0, indicating no need or opportunity for sex-specific selection strategies

Intraocular retinal transplants

Embryonic rat retinae transplanted into the anterior chamber of adult rat eyes of the same or different strain survive and grow. Light and electron microscopic studies show that the transplants undergo histogenetic differentiation, resulting in the development of mature inner and outer layer neurons and Muller glial cells. Vascular connections develop between the host iris and the retinal transplant. These initial observations indicate that retinal transplantation to a recipient eye is a procedure which offers ample opportunities for the study of problems related to neural development, retinal plasticity and repair

A geoarchaeological review of Balzi Rossi, Italy: A crossroad of Palaeolithic populations in the northwest Mediterranean

The Balzi Rossi archaeological complex (comprised of caves, rock shelters, and open-air sites) is a globally significant site for Palaeolithic culture and understanding the transition from Neanderthal to Anatomically Modern Human populations in Europe. It also retains some of the earliest evidence of human interactions with their coastal environment. Balzi Rossi has been subject to excavation for over 150 years - traditionally as individual site locations - with most deposits removed when the discipline of archaeology was nascent, and the science not yet developed. The consequence was the unfortunate loss of materials and critically important stratigraphic context. However, valuable information regarding the Palaeolithic population, their coastal environment, and earlier sea-level change, remains in the literature and in museum repositories. In this work we have compiled and reviewed the extensive resources, available largely in French and Italian, to provide a summary and catalogue for each individual site. These "Site Summaries" are available as appendices to this review, which provides a comprehensive synopsis of the history of excavations conducted at Balzi Rossi, a reconstruction of stratigraphy where possible, the evidence of Palaeolithic occupations, the evidence of Pleistocene sea-level fluctuations, and an assessment of the chronological constraint available for both the Palaeolithic populations and sea level. Finally, this synopsis identifies gaps in knowledge and provides comments on pathways for future research, suggesting a consilient approach that can be applied in other archaeological contexts

Platelet-Rich Fibrin (PRF) Analyzed for Cytokine Profiles - A Misguided Hope for Osteogenesis in Jawbone Defects? Research and Clinical Observational Study

Johann Lechner,1 Volker vonBaehr,2 Cornelia Doebis,3 Florian Notter,4 Fabian Schick4 1Department of Osteoimmunology, Clinic for Integrative Dentistry, Munich, Germany; 2Department of Immunology and Allergology, Institute for Medical Diagnostics, Berlin, Germany; 3Department of Analysis, Institute for Medical Diagnostics, Berlin, Germany; 4Department of Implantology, Clinic for Integrative Dentistry, Munich, GermanyCorrespondence: Johann Lechner, Department of Osteoimmunology, Clinic for Integrative Dentistry, Gruenwalder Str. 10A, Munich, 81547, Germany, Tel +49 89 697 0129, Fax +49 89 692 5830, Email [email protected]: Platelet-rich fibrin (PRF) blood concentrates are used in oral implantology and defect surgery to promote osteoneogenesis in Bone Marrow Defects in Jawbone (BMDJ), according to the morphology of fatty-degenerative osteonecrosis also called FDOJ.Question: Can the benefit of PRF on alveolar osteoneogenesis be confirmed by cytokine analysis?.Methods: The cytokine expressions of the PRF samples in 26 patients undergoing BMDJ/FDOJ surgery in the same session were analysed for seven cytokines (RANTES/CCL5; FGF-2; IL-1RA; Il-6; IL-8; MCP-1; TNF-a) by multiplex (Luminex). The FDOJ samples of these 26 BMDJ/FDOJ patients were analysed for the RANTES/CCL5 expression only.Results: Cytokine expression in PRF is compared to reference values for healthy medullary bone of the jaw and BMDJ/FDOJ and shows that the cytokine expressions of the PRF samples do not compensate or counteract prima vista for the cytokine dysregulations present in the BMDJ/FDOJ areas.Discussion: To define the aid of cytokines studied in PRF in the restoration of the immunological dysregulation in areas of BMDJ/FDOJ, literature is reviewed comparing RANTES/CCL5, IL-1ra, TNF-α and MCP-1/CCL2 expression in PRF and BMDJ/FDOJ. Immunoregulatory properties of PRF in alveolar bone restoration are evaluated.Summary: PRF was mistakenly thought to be a cure for bone healing, which is here shown to be incorrect. Enoral Ultrasound Sonography of bone density is available for the clinical measurement of individually developed osteoneogenesis by PRF.Conclusion: The multiplex analysis of PRF shows a dynamic and cytokine-based interaction with osteoneogenesis that is not yet fully clarified. Keywords: platelet-rich fibrin, multiplex analysis, cytokines, bone marrow defects of the jaw, osteoneogenesis, enoral transalveolar ultrasonograp

Osteoimmune Interaction and TH-1/TH-2 Ratio in Jawbone Marrow Defects: An Underestimated Association – Original Research

Johann Lechner,1 Volker von Baehr,2 Florian Notter,1 Fabian Schick1 1Clinic for Integrative Dentistry, Munich, Germany; 2Department of Immunology and Allergology, Institute for Medical Diagnostics, Berlin, GermanyCorrespondence: Johann Lechner, Gruenwalder Str. 10A, Munich, 81547, Germany, Tel +49 89 697 0129, Fax +49 89 692 5830, Email [email protected]: Osteoimmunology recognizes the relationship between bone cells and immune cells. Chronic osteoimmune dysregulation is present in bone marrow defects of the jaw (BMDJ) as fatty-degenerative osteonecrosis (FDOJ). In comparison to samples from healthy jaw bone, the cytokine analysis of samples of BMDJ/FDOJ from 128 patients showed downregulated TNF-α and IL-6 expression and the singular overexpression of the chemokine RANTES/CCL5.Aim and Objectives: This paper raises the question of whether the osteoimmune defects due to incomplete wound healing in BMDJ/FDOJ in 128 patients are related to dysregulation of the Th1/Th2 ratio and regulatory T cell (T-reg) expression in a control group of 197 BMDJ/FDOJ patients, each presenting with BMDJ/FJOD and one of seven different immune disorders.Material and Methods: In the control group, serum concentrations of the cytokines IFN-y and IL-4 were determined after stimulated cytokine release and displayed as Th1/Th2 ratios.Results: Data show a shift in Th2 in more than 80% (n = 167) of the control cohort of 197 chronically ill patients with concomitant BMDJ/FDOJ. In these 167 subjects, the Th1/Th2 ratio was 1.900 pg/mL, indicating strongly downregulated immune activity.Discussion: BMDJ/FDOJ is characterized by a lack of Th1 cytokines and an excessive expression of RANTES/CCL5 and IL-1ra and, thus, the inversion of an acute inflammatory cytokine pattern. In contrast, abdominal fat contains a very high proportion of regulatory Th1 cells and produces an inflammatory immune response through the high overexpression of TNF-α and IL-6. The lack of Th1 activation in BMDJ/FDOJ areas inhibits normal wound healing and supports the persistence of BMDJ/FDOJ.Conclusion: The Th1/Th2 ratio requires greater consideration, especially with respect to wound healing following dental surgical interventions, such as jaw surgery, implantation and augmentation, to avoid the emergence of the osteoimmune situation that is characteristic of BMDJ/FDOJ. Keywords: osteoimmunology, T-cells, Th1/Th2 shift, jawbone, osteoclastogenesis, RANTES/CCL

Dental Ultrasonography for Visualizing Osteoimmune Conditions and Assessing Jaw Bone Density: A Narrative Review

Robert Huber,1 Elisa Choukroun,2 Harald Fahrenholz,3 Florian Notter4 1Department of Ultrasonography, Digital Dental & Healthcare Technology, Gruenwald, 82031, Germany; 2Department of Immunology, Private Practice, Saint Laurent du Var, 06700, France; 3Department of Integrative Dentistry, Zahnaesthetik-Wien, Vienna, 1010, Austria; 4Department of Implantology, Clinic for Integrative Dentistry, Munich, 81547, GermanyCorrespondence: Robert Huber, Email [email protected]: Despite the widespread use of ultrasonography (US) in medical diagnostics, there is no similar US device available for visualizing jawbone density. This study is a narrative review of the possible applications of US in dentistry. This review is divided as follows: (a) Pulse–echo ultrasonography: the applications offer new perspectives for periodontal and peri-implant assessment. (b) Through-transmission alveolar US (TTAU): this technique was a novel imaging modality until 2004, when TTAU devices were last available. Quantitative US scaling made the device useful for diagnosing chronic inflammatory conditions in the jaw. (c) Ultrasound transmission velocity (UTV): in 2008, this technique was introduced in German university dental clinics to analyze the mechanical properties of the jawbone without translating the scientific findings into a practical device. (d) Trans-alveolar US device (TAU): the growing importance of “osteoimmune focal bone marrow defects” has led practitioners to develop a new TAU device. The attenuation of US was used for imaging of jawbone density. (e) Patients who benefit from TAU-guided jawbone surgery: research has shown remarkable results in specific disease cases. This review concludes that US has been undervalued as a diagnostic tool in dentistry. The new TAU-n unit offers the opportunity to change this in the future. Keywords: ultrasonography, dentistry, jawbone density, osteonecrosis, trans-alveolar ultrasound, radiation protectio

Intraretinal Transplantation for Rod-Cell Replacement in Light-Damaged Retinas

Blindness from retinal disease is often the consequence of extensive damage to the photoreceptor cell population, while other cell types which form the neural retina are relatively spared. In this setting, transplantation of photoreceptor cells could offer hope for the restoration of some degree of visual function. We testd the feasibility of this approach by transplanting immature retinal cells into the eyes of adult rats affected by late stage phototoxic retinopathy, which are almost totally devoid of photoreceptor cells

Administration of intrapulmonary sodium polyacrylate to induce lung injury for the development of a porcine model of early acute respiratory distress syndrome.

BACKGROUND: The loss of alveolar epithelial and endothelial integrity is a central component in acute respiratory distress syndrome (ARDS); however, experimental models investigating the mechanisms of epithelial injury are lacking. The purpose of the present study was to design and develop an experimental porcine model of ARDS by inducing lung injury with intrapulmonary administration of sodium polyacrylate (SPA). METHODS: The present study was performed at the Centre for Comparative Medicine, University of British Columbia, Vancouver, British Columbia. Human alveolar epithelial cells were cultured with several different concentrations of SPA; a bioluminescence technique was used to assess cell death associated with each concentration. In the anesthetized pig model (female Yorkshire X pigs (n = 14)), lung injury was caused in 11 animals (SPA group) by injecting sequential aliquots (5 mL) of 1% SPA gel in aqueous solution into the distal airway via a rubber catheter through an endotracheal tube. The SPA was dispersed throughout the lungs by manual bag ventilation. Three control animals (CON group) underwent all experimental procedures and measurements with the exception of SPA administration. RESULTS: The mean (± SD) ATP concentration after incubation of human alveolar epithelial cells with 0.1% SPA (0.92 ± 0.27 μM/well) was approximately 15% of the value found for the background control (6.30 ± 0.37 μM/well; p < 0.001). Elastance of the respiratory system (E RS) and the lung (E L) increased in SPA-treated animals after injury (p = 0.003 and p < 0.001, respectively). Chest wall elastance (E CW) did not change in SPA-treated animals. There were no differences in E RS, E L, or E CW in the CON group when pre- and post-injury values were compared. Analysis of bronchoalveolar lavage fluid showed a significant shift toward neutrophil predominance from before to after injury in SPA-treated animals (p < 0.001) but not in the CON group (p = 0.38). Necropsy revealed marked consolidation and congestion of the dorsal lung lobes in SPA-treated animals, with light-microscopy evidence of bronchiolar and alveolar spaces filled with neutrophilic infiltrate, proteinaceous debris, and fibrin deposition. These findings were absent in animals in the CON group. Electron microscopy of lung tissue from SPA-treated animals revealed injury to the alveolar epithelium and basement membranes, including intra-alveolar neutrophils and fibrin on the alveolar surface and intravascular fibrin (microthrombosis). CONCLUSIONS: In this particular porcine model, the nonimmunogenic polymer SPA caused a rapid exudative lung injury. This model may be useful to study ARDS caused by epithelial injury and inflammation