RESUMO
In different species, embryonic aneuploidies and genome-wide errors are a major cause of developmental failure. The increasing number of equine embryos being produced worldwide provides the opportunity to characterize and rank or select embryos based on their genetic profile prior to transfer. Here, we explored the possibility of generic, genome-wide preimplantation genetic testing concurrently for aneuploidies (PGT-A) and monogenic (PGT-M) traits and diseases in the horse, meanwhile assessing the incidence and spectrum of chromosomal and genome-wide errors in in vitro-produced equine embryos. To this end, over 70,000 single nucleotide polymorphism (SNP) positions were genotyped in 14 trophectoderm biopsies and corresponding biopsied blastocysts, and in 26 individual blastomeres from six arrested cleavage-stage embryos. Subsequently, concurrent genome-wide copy number detection and haplotyping by haplarithmisis was performed and the presence of aneuploidies and genome-wide errors and the inherited parental haplotypes for four common disease-associated genes with high carrier frequency in different horse breeds (GBE1, PLOD1, B3GALNT2, MUTYH), and for one color coat-associated gene (STX17) were compared in biopsy-blastocyst combinations. The euploid (n = 12) or fully aneuploid (n = 2) state and the inherited parental haplotypes for 42/45 loci of interest of the biopsied blastocysts were predicted by the biopsy samples in all successfully analyzed biopsy-blastocyst combinations (n = 9). Two biopsies showed a loss of maternal chromosome 28 and 31, respectively, which were confirmed in the corresponding blastocysts. In one of those biopsies, additional complex aneuploidies not present in the blastocyst were found. Five out of six arrested embryos contained chromosomal and/or genome-wide errors in most of their blastomeres, demonstrating their contribution to equine embryonic arrest in vitro. The application of the described PGT strategy would allow to select equine embryos devoid of genetic errors and pathogenetic variants, and with the variants of interest, which will improve foaling rate and horse quality. We believe this approach will be a gamechanger in horse breeding.
Assuntos
Variações do Número de Cópias de DNA , Parada Cardíaca , Cavalos , Animais , Haplótipos , Genótipo , Testes Genéticos , AneuploidiaRESUMO
In addition to fulfilling many breeders' curiosity, equine embryonic sex determination can have a profound commercial impact. However, the application of currently described assays for equine embryonic sexing has rendered variable diagnosis and validation rates, with sensitivity being the main problem. In addition, while pregnancy results of in vivo-flushed equine embryos following a needle aspiration biopsy equal those of non-biopsied embryos, the effect on in vitro-produced embryos is unknown. Here, we aimed to develop a highly sensitive and specific assay for equine sex determination that can be directly performed on few embryonic cells, and to test the effect of a needle aspiration biopsy on the viability of the in vitro-produced embryo. To this end, a multiplex quantitative real-time PCR (qPCR) assay with dual-labelled probes was designed to allow the simultaneous generation of both male-specific and control fragments in a single closed-tube reaction, avoiding potential sample loss or contamination. To improve sensitivity, multicopy and polymeric genes were chosen to be specifically amplified, i.e., eight copies of Y-chromosomal ETSTY5 as male-specific and four autosomal UBC monomers as control fragment. Specificity was enhanced by the equine-specific character of ETSTY5 and by using dual-labelled probes. The assay was optimised with equine male and female genomic DNA and demonstrated a 100% accuracy and a >95% qPCR efficiency down to 10 pg of DNA. The assay was subsequently applied to determine the sex of 44 in vitro-produced embryos, collecting trophectoderm biopsies by means of a needle aspiration biopsy and herniating cells. Of all trophectoderm biopsies and herniating cell samples (n = 54), 87% could be diagnosed. Assay results were validated on a second sample obtained from the biopsied embryo (n = 18) or, by ultrasound-based sex determination of the foetus (n = 7) following the transfer of the biopsied embryo to a recipient mare, with about half of the embryos being fillies and colts. The needle aspiration biopsy procedure did not impair initial pregnancy rate or early pregnancy losses as compared to non-biopsied embryos. In conclusion, we report a safe, reliable, fast, and cost-effective assay for equine sex determination which was validated for the sex determination of in vitro-produced embryos based on few embryonic cells, and needle aspiration biopsy did not impair the embryo's viability. The assay and safe biopsy strategy hold potential for other applications.
Assuntos
Blastocisto , Embrião de Mamíferos , Gravidez , Animais , Cavalos , Feminino , Masculino , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Biópsia/veterinária , DNARESUMO
The combined health and economic impact of Taenia solium urges for control and, if possible, elimination of this neglected parasitic zoonosis. Up till now there is still no consensus about the most cost-effective and feasible approaches for control. The objective of this systematic review is to identify and summarize the evidence in English scientific literature on the control and elimination of T. solium since 2014, based on the rapidly evolving field of evidence on control and elimination of T. solium. The search resulted in the identification of 458 records of which 31 were included, covering 13 field trials and 18 articles containing experimental data, mathematical models, and other information directly relevant the control of T. solium. Recent field studies confirm that combinations of interventions or multiple rounds are more successful in obtaining rapid reductions in transmission and parasite occurrence, with the quick impact of the combination of human and pig treatment confirmed in a South Asian and Peruvian context. Moreover, elimination of transmission through a one-year intensive program, combining human and pig treatment/vaccination was described in a Peruvian study. Recent studies also provide more data on the positive impact of specific health education, as well as newly developed electronic educational tools, providing opportunities for area specific community-engaged participatory interventions. Once control has been achieved, monitoring of migration of both potentially infected people and pigs from outside the control area is important for sustained disease control.
RESUMO
This paper evaluates maxillary expansion by synthesizing the scientific literature, focusing its role in treatment of maxillary insufficiency, buccal cross bites, and deviations in mandibular closure as well as its effectiveness in correcting a lack of harmony between tooth and jaw size. By its creation of maxillary bone "capital" it facilitates treatment of arch length discrepancies in an environment of periodontal health. And by restoring the maxilla to normal size it improves the orthopedic effectiveness of Class II treatment of mandibular advancement appliances. In a similar fashion rapid palatal expansion, by relaxing circum-maxillary sutures, prepares the way for maxillary advancement in Class III cases with masks of Delaire or Petit. And, by broadening the floor of the nose, palate splitting improves nasal airflow and helps mouth breathers learn how to breathe through their noses. Most authors emphasize the benefits of early intervention through truly orthopedic movement that eliminates tipping of posterior teeth and reduces the risk of buccal fenestration.
Assuntos
Má Oclusão Classe II de Angle/terapia , Maxila/crescimento & desenvolvimento , Ortodontia Interceptora/métodos , Técnica de Expansão Palatina , Fatores Etários , Humanos , Respiração Bucal/terapia , Obstrução Nasal/terapia , Odontometria , Aparelhos Ortodônticos Funcionais , Ortodontia Interceptora/instrumentação , Prevenção SecundáriaRESUMO
The objectives of this experimental study were to evaluate histological responses following cross-transplantations of the contrastive metopic and internasal sutures and to determine the nature of their integration and metamorphosis in the respective new functional environments. Twenty-two Hartly guinea pigs were utilized. They were divided into three groups: a nonsurgical control group, a surgical control group, and an experimental group. Vital dyes were injected weekly to each animal. Two histological procedures were used: paraffin embedded decalcified sections for observations of fine cellular structures, ground sections for UV visualisation of the vital dye labelling and new bone formation. Amalgam implants markers were also used around the metopic and internasal sutures for measurements of the amount of expansion taking place at the levels of these sutures. The results demonstrate that expansion and growth activity at these sutural junctions are minimal, that the sutural characteristics relate to the nature of the functional environment, and that, following transplantation to exchanged sites each suture will adopt the characteristics and come to conform with the functional requirements of the new site. It is concluded that sutural tissues are appropriate for early manipulations in craniofacial surgery as transplantable sites for subsequent growth. Nevertheless, a great deal of experimental study will be required to determine possible donor sites and appropriate timings for surgical transplants.
Assuntos
Suturas Cranianas/cirurgia , Osso Frontal/cirurgia , Osso Nasal/cirurgia , Animais , Suturas Cranianas/anatomia & histologia , Suturas Cranianas/crescimento & desenvolvimento , Osso Frontal/crescimento & desenvolvimento , Cobaias , Métodos , Osso Nasal/crescimento & desenvolvimento , Osteogênese , ReimplanteRESUMO
The objectives of this study is to challenge the soft tissue profile standards of esthetics and to compare them with the feeling and the sense of beauty of the large public. The facial profile of one hundred fifty-two cinema stars and top models (seventy-one men and eighty-one women) were analysed. Twenty three measurements were taken among the most used and widely accepted. These references come from the works of very reknown orthodontists and researchers: BURSTONE, HOLDAWAY, MERRIFIELD, PECK and PECK, RICKETTS, STEINER, WORMS (as described by MOSHIRI). The results demonstrate, after statistical analysis that: some values are very different compared with the references; those discrepancies neither concern the same values nor for the same amount, or to the same direction for the male or the female samples. The mean male soft tissue profile is more concave, prognathic type with more retruded upper and lower lips and more reduced lower face height than the standards described by the authors. The mean women soft tissue profile tends to be more convex with a relatively more protruded upper lip and a more retrognathic mandible than the "standards". The whole face height is significantly reduced with a more marked discrepancy for the nasal height and the lower lip length. The nasal prominence is also reduced compared with the norms. It appears then that we cannot trust anymore references that do not take into account differences between men and women ideal facial profiles for the analysis of soft tissue profile in orthodontics and maxillo-facial surgery. The values described by the authors, which are our daily standards, do not seem to match the large public sense of esthetics and therefore our patients feeling of beauty.