Nanoscale Dynamics of Streptococcal Adhesion to AGE-Modified Collagen.

The adhesion of initial colonizers such as Streptococcus mutans to collagen is critical for dentinal and root caries progression. One of the most described pathological and aging-associated changes in collagen-including dentinal collagen-is the generation of advanced glycation end-products (AGEs) such as methylglyoxal (MGO)-derived AGEs. Despite previous reports suggesting that AGEs alter bacterial adhesion to collagen, the biophysics driving oral streptococcal attachment to MGO-modified collagen remains largely understudied. Thus, the aim of this work was to unravel the dynamics of the initial adhesion of S. mutans to type I collagen in the presence and absence of MGO-derived AGEs by employing bacterial cell force spectroscopy with atomic force microscopy (AFM). Type I collagen gels were treated with 10 mM MGO to induce AGE formation, which was characterized with microscopy and enzyme-linked immunosorbent assay. Subsequently, AFM cantilevers were functionalized with living S. mutans UA 159 or Streptococcus sanguinis SK 36 cells and probed against collagen surfaces to obtain force curves displaying bacterial attachment in real time, from which the adhesion force, number of events, Poisson analysis, and contour and rupture lengths for each individual detachment event were computed. Furthermore, in silico computer simulation docking studies between the relevant S. mutans UA 159 collagen-binding protein SpaP and collagen were computed, in the presence and absence of MGO. Overall, results showed that MGO modification increased both the number and adhesion force of single-unbinding events between S. mutans and collagen, without altering the contour or rupture lengths. Both experimental and in silico simulations suggest that this effect is due to increased specific and nonspecific forces and interactions between S. mutans UA 159 and MGO-modified collagen substrates. In summary, these results suggest that collagen alterations due to aging and glycation may play a role in early bacterial adherence to oral tissues, associated with conditions such as aging or chronic hyperglycemia, among others.

Nanomechanical and Molecular Characterization of Aging in Dentinal Collagen.

Methylglyoxal (MGO) is an important molecule derived from glucose metabolism with the capacity of attaching to collagen and generating advanced glycation end products (AGEs), which accumulate in tissues over time and are associated with aging and diseases. However, the accumulation of MGO-derived AGEs in dentin and their effect on the nanomechanical properties of dentinal collagen remain unknown. Thus, the aim of the present study was to quantify MGO-based AGEs in the organic matrix of human dentin as a function of age and associate these changes with alterations in the nanomechanical and ultrastructural properties of dentinal collagen. For this, 12 healthy teeth from <26-y-old and >50-y-old patients were collected and prepared to obtain crown and root dentin discs. Following demineralization, MGO-derived AGEs were quantified with a competitive ELISA. In addition, atomic force microscopy nanoindentation was utilized to measure changes in elastic modulus in peritubular and intertubular collagen fibrils. Finally, principal component analysis was carried out to determine aging profiles for crown and root dentin. Results showed an increased presence of MGO AGEs in the organic matrix of dentin in the >50-y-old specimens as compared with the <26-y-old specimens in crown and root. Furthermore, an increase in peritubular and intertubular collagen elasticity was observed in the >50-y-old group associated with ultrastructural changes in the organic matrix as determined by atomic force microscopy analysis. Furthermore, principal component analysis loading plots suggested different "aging profiles" in crown and root dentin, which could have important therapeutic implications in restorative and adhesive dentistry approaches. Overall, these results demonstrate that the organic matrix of human dentin undergoes aging-related changes due to MGO-derived AGEs with important changes in the nanomechanical behavior of collagen that may affect diagnostic and restorative procedures in older people.

Modulatory Effect of Glycated Collagen on Oral Streptococcal Nanoadhesion.

Biofilm-mediated oral diseases such as dental caries and periodontal disease remain highly prevalent in populations worldwide. Biofilm formation initiates with the attachment of primary colonizers onto surfaces, and in the context of caries, the adhesion of oral streptococci to dentinal collagen is crucial for biofilm progression. It is known that dentinal collagen suffers from glucose-associated crosslinking as a function of aging or disease; however, the effect of collagen crosslinking on the early adhesion and subsequent biofilm formation of relevant oral streptococci remains unknown. Therefore, the aim of this work was to determine the impact of collagen glycation on the initial adhesion of primary colonizers such as Streptococcus mutans UA159 and Streptococcus sanguinis SK 36, as well as its effect on the early stages of streptococcal biofilm formation in vitro. Type I collagen matrices were crosslinked with either glucose or methylglyoxal. Atomic force microscopy nanocharacterization revealed morphologic and mechanical changes within the collagen matrix as a function of crosslinking, such as a significantly increased elastic modulus in crosslinked fibrils. Increased nanoadhesion forces were observed for S. mutans on crosslinked collagen surfaces as compared with the control, and retraction curves obtained for both streptococcal strains demonstrated nanoscale unbinding behavior consistent with bacterial adhesin-substrate coupling. Overall, glucose-crosslinked substrates specifically promoted the initial adhesion, biofilm formation, and insoluble extracellular polysaccharide production of S. mutans, while methylglyoxal treatment reduced biofilm formation for both strains. Changes in the adhesion behavior and biofilm formation of oral streptococci as a function of collagen glycation could help explain the biofilm dysbiosis seen in older people and patients with diabetes. Further studies are necessary to determine the influence of collagen crosslinking on the balance between acidogenic and nonacidogenic streptococci to aid in the development of novel preventive and therapeutic treatment against dental caries in these patients.

Local influence of the corpus luteum on the ipsilateral oviduct and early embryo development in the ewe.

The objective of this study was to evaluate the local effect of the corpus luteum (CL) on ipsilateral oviduct-uterus functionality and early embryo development in ewes. A total of 499 embryos were transferred on Day 1 after in vitro fertilization into the ipsilateral (n = 250) and contralateral oviducts (n = 249) of 13 ewes on Day 1 after ovulation (18-20 embryos per oviduct). On Day 6, their reproductive tracts were collected and their uterine horns were flushed for embryo recovery. More recovered embryos, a higher proportion of blastocysts, and more viable embryos were collected when the embryos were transferred into the ipsilateral oviducts (P < 0.05). In addition, almost five times higher P4 concentrations and significantly lower E2 concentrations, with higher P4:E2 ratio, were found in the ipsilateral than contralateral oviductal tissue (P < 0.05). Furthermore, a higher concentration of adiponectin was found in the ipsilateral uterine tissue macerates than in the contralateral side to the CL. The ipsilateral oviductal tissue had a lower expression of PGR and IGFBP5, but the transcript expression of ADIPOR1 was higher in the ipsilateral oviductal tissue. In the uterus, the mRNA expression of ESR1, IGFBP3, IGFBP5, and LEPR was higher or tended to be higher in the ipsilateral than contralateral uterine tissue. Uterine flushing fluid collected from the ipsilateral uterine horn had lower insulin concentrations than the contralateral horn, while no differences were found in the P4 and E2 concentrations. In conclusion, on Day 6 post-ovulation, P4 was elevated in the ipsilateral oviductal tissue, embryo development was advanced, and differential gene expression of PGR, ESR1, IGFBP3, IGFBP5, LEPR, and ADIPOR1 in the oviductal or uterine tissue was found between the ipsilateral and contralateral side. This study demonstrates local regulation of the ovary on the ipsilateral oviduct/uterine horn in the ewe.

Cumulus cells during in vitro fertilization and oocyte vitrification in sheep: Remove, maintain or add?

The objective was to evaluate the developmental competence of immature and matured ovine oocytes after removing, maintaining or adding cumulus cells (CC) associated to vitrification by Cryotop method. Three experiments were performed involving 3,144 oocytes. In Experiment 1, CC were removed from immature, matured or fertilized oocytes subjected to in vitro embryo production. In Experiment 2, oocytes were vitrified either in MI or MII stage with or without CC, while a control group with CC remained unvitrified. In Experiment 3, oocytes partially denuded from CC were vitrified either in MI or MII stage, and a co-culture of fresh CC was added or not soon after warming to complete in vitro maturation (IVM) and in vitro fertilization (IVF), or IVF, respectively, while a control group remained unvitrified. In Experiment 1, the cleavage rate, development rate on Day 6 and blastocyst rate on Day 8 were improved when CC were maintained until the end of IVF (P < 0.05). In Experiment 2, vitrification of oocytes with enclosed CC showed a tendency to increase cleavage (P = 0.06) and improved blastocyst rate (P < 0.05). In Experiment 3, adding CC as co-culture after vitrification-warming tended to improve cleavage rate (P = 0.06) and increased hatching rate (P < 0.05). Regarding oocyte stage, vitrification of in vitro matured oocytes resulted in greater developmental competence than immature stages (P < 0.05). In conclusion, CC seems to have a relevant role for in vitro embryo development in either fresh or vitrified oocytes.

Serum progesterone concentrations during FSH superstimulation of the first follicular wave affect embryo production in sheep.

The aim of the present study was to evaluate the effect of serum progesterone concentrations during the superstimulatory treatment of the first follicular wave on fertilization rate and embryo development in sheep. A total of 71 Merino ewes received a superstimulatory FSH treatment during Wave 1 of ovarian follicular development (Day 0 Protocol), which was administrated under low progesterone concentrations typical of the early luteal phase (control group, n = 33) or under high progesterone concentrations induced by the administration of an intravaginal device from Day 0 to Day 3 containing 0.3 g progesterone (n = 38). Intrauterine insemination after FSH superstimulation was followed by uterine flushing 6 days later. Serum progesterone concentrations from Day 0 to 3 were greater in those ewes treated with progesterone (P < 0.05), while serum estradiol-17ß concentrations were not affected by the treatment. Although the mean number of corpora lutea per donor was not affected by the progesterone treatment, the number of collected ova and embryos was greater in progesterone treated than untreated ewes (6.6 ± 0.7 compared with 4.6 ± 0.9, respectively; P < 0.05). Furthermore, progesterone treatment increased fertilization rate (93.3% compared with 83.3%; P < 0.05) and the proportion of Grade 1 embryos (67.7% compared with 52.7%; P < 0.05) compared with the control group. In conclusion, oocyte fertilization rate and embryo quality are improved by high progesterone concentrations during FSH superstimulation, which suggests an important role of progesterone during preovulatory follicular development.

Oocyte developmental competence is improved by relatively greater circulating progesterone concentrations during preovulatory follicular growth.

This study evaluated the effect of progesterone priming during follicular growth on oocyte competence to undergo oocyte cleavage and embryo development in sheep. Two experiments were performed on a total of 195 females that either received or did not receive a progesterone treatment (CIDR-type device) during the first follicular wave, beginning soon after ovulation (i.e., Day 0 of the experiment). On Day 3, the follicular population and oocyte quality (Experiment 1 and 2) and the competence of oocytes for cleavage and embryo development (Experiment 2) were evaluated after laparoscopic ovum pickup (LOPU) and in vitro fertilization. In Experiment 1, in a 2 × 2 factorial study the progesterone priming treatment (treated or not) was or was not associated with a single dose of FSH in a slow-release hyaluronic acid preparation given on Day 0. The follicular population on Day 3 and the number and morphology of recovered cumulus oocyte complexes (COCs) were not affected by the progesterone treatment (P = NS) but were improved by the FSH administration (P <  0.05). An interaction between both treatments was observed (P <  0.05), with more desirable outcome with the females that received both the progesterone and the FSH treatments. In Experiment 2, half of the females received the exogenous progesterone priming, and all females received FSH on Day 0. After follicular aspiration on Day 3, the cleavage rate and the embryo development rate following in vitro fertilization and culture were greater in those females that received the progesterone treatment (P <  0.05). In conclusion, these studies provide evidence that progesterone treatment during follicular growth affects oocyte competence, with the greater progesterone concentrations enhancing the oocyte's capacity to undergo cleavage and embryo development.

Embryo survival and birth rate after minimum volume vitrification or slow freezing of in vivo and in vitro produced ovine embryos.

The objective was to evaluate pregnancy outcomes and birth rate of in vivo derived vs. in vitro produced ovine embryos submitted to different cryopreservation methods. A total of 197 in vivo and 240 in vitro produced embryos were cryopreserved either by conventional freezing, or by vitrification with Cryotop or Spatula MVD methods on Day 6 after insemination/fertilization. After thawing/warming and transfer, embryo survival rate on Day 30 of gestation was affected by the source of the embryos (in vivo 53.3%, in vitro 20.8%; P < 0.05) and by the method of cryopreservation (conventional freezing 26.5%, Cryotop 52.0%, Spatula MVD 22.2%; P < 0.05). For in vivo derived embryos, survival rate after embryo transfer was 45.6% for conventional freezing, 67.1% for Cryotop, and 40.4% for Spatula MVD. For in vitro produced embryos, survival rate was 7.3% for conventional freezing, 38.7% for Cryotop, and 11.4% for Spatula MVD. Fetal loss from Day 30 to birth showed a tendency to be greater for in vitro (15.0%) rather than for in vivo produced embryos (5.7%), and was not affected by the cryopreservation method. Gestation length, weight at birth and lamb survival rate after birth were not affected by the source of the embryo, the cryopreservation method or stage of development (average: 150.5 ± 1.8 days; 4232.8 ± 102.8 g; 85.4%; respectively). This study demonstrates that embryo survival and birth rate of both in vivo and in vitro produced ovine embryos are improved by vitrification with the minimum volume Cryotop method.

Efficient Generation of Myostatin Knock-Out Sheep Using CRISPR/Cas9 Technology and Microinjection into Zygotes.

While CRISPR/Cas9 technology has proven to be a valuable system to generate gene-targeted modified animals in several species, this tool has been scarcely reported in farm animals. Myostatin is encoded by MSTN gene involved in the inhibition of muscle differentiation and growth. We determined the efficiency of the CRISPR/Cas9 system to edit MSTN in sheep and generate knock-out (KO) animals with the aim to promote muscle development and body growth. We generated CRISPR/Cas9 mRNAs specific for ovine MSTN and microinjected them into the cytoplasm of ovine zygotes. When embryo development of CRISPR/Cas9 microinjected zygotes (n = 216) was compared with buffer injected embryos (n = 183) and non microinjected embryos (n = 173), cleavage rate was lower for both microinjected groups (P<0.05) and neither was affected by CRISPR/Cas9 content in the injected medium. Embryo development to blastocyst was not affected by microinjection and was similar among the experimental groups. From 20 embryos analyzed by Sanger sequencing, ten were mutant (heterozygous or mosaic; 50% efficiency). To obtain live MSTN KO lambs, 53 blastocysts produced after zygote CRISPR/Cas9 microinjection were transferred to 29 recipient females resulting in 65.5% (19/29) of pregnant ewes and 41.5% (22/53) of newborns. From 22 born lambs analyzed by T7EI and Sanger sequencing, ten showed indel mutations at MSTN gene. Eight showed mutations in both alleles and five of them were homozygous for indels generating out-of frame mutations that resulted in premature stop codons. Western blot analysis of homozygous KO founders confirmed the absence of myostatin, showing heavier body weight than wild type counterparts. In conclusion, our results demonstrate that CRISPR/Cas9 system was a very efficient tool to generate gene KO sheep. This technology is quick and easy to perform and less expensive than previous techniques, and can be applied to obtain genetically modified animal models of interest for biomedicine and livestock.

Embryo development, fetal growth and postnatal phenotype of eGFP lambs generated by lentiviral transgenesis.

Lentiviral technology has been recently proposed to generate transgenic farm animals more efficiently and easier than traditional techniques. The objective was to evaluate several parameters of lambs obtained by lentiviral transgenesis in comparison with non-transgenic counterparts. In vitro produced embryos were microinjected (TG group) at two-cell stage with a lentiviral construct containing enhanced green fluorescent protein (eGFP) gene, while embryos produced by in vitro fertilization (IVF group) or intrauterine insemination (IUI group) were not microinjected. Microinjection technique efficiently generated eight-cell transgenic embryos (97.4%; 114/117). Development rate on day 5 after fertilization was similar for TG (39.3%, 46/117) and IVF embryos (39.6%, 44/111). Pregnancy rate was detected in 50.0% (6/12) of recipient ewes with TG embryos, in 46.7% (7/15) with IVF embryos, and in 65.0% (13/20) of IUI ewes (P = NS). Nine lambs were born in TG group, six lambs in IVF group, and 16 lambs in IUI group. All TG lambs (9/9) were GFP positive to real-time PCR and eight (88.9%) showed a strong and evident GFP expression in mucosae, eyes and keratin tissues. Fetal growth monitored every 15 day by ultrasonography did not show significant differences. Transgenic lambs neither differ in morphometric variables in comparison with non transgenic IVF lambs within 3 months after birth. Transmission of the transgene to the progeny was observed in green fluorescent embryos produced by IVF using semen from the TG founder lambs. In conclusion, this study demonstrates the high efficiency of lentiviral technology to produce transgenic sheep, with no clinic differences in comparison with non transgenic lambs.

Cryotolerance of Day 2 or Day 6 in vitro produced ovine embryos after vitrification by Cryotop or Spatula methods.

This study was conducted to evaluate the cryotolerance of in vitro produced ovine embryos submitted to vitrification at different developmental stages using two methods of minimum volume and rapid cooling rate. Embryos were vitrified at early stage (2 to 8-cells) on Day 2 or at advanced stage (morulae and blastocysts) on Day 6 after in vitro fertilization. Vitrification procedure consisted of the Cryotop (Day 2, n=165; Day 6, n=174) or the Spatula method (Day 2, n=165; Day 6, n=175). Non vitrified embryos were maintained in in vitro culture as a control group (n=408). Embryo survival was determined at 3h and 24h after warming, development and hatching rates were evaluated on Day 6 and Day 8 after fertilization, and total cell number was determined on expanded blastocysts. Embryo survival at 24h after warming increased as the developmental stage progressed (P<0.05) and was not affected by the vitrification method. The ability for hatching of survived embryos was not affected by the stage of the embryos at vitrification or by the vitrification method. Thus, the proportion of hatching from vitrified embryos was determined by the survival rate and was lower for Day 2 than Day 6 vitrified embryos. The percentage of blastocysts on Day 8 was lower for the embryos vitrified on Day 2 than Day 6 (P<0.05), and was lower for both days of vitrification than for non-vitrified embryos (P<0.05). No interaction of embryo stage by vitrification method was found (P=NS) and no significant difference was found in the blastocyst cell number among vitrified and non-vitrified embryos. In conclusion, both methods using minimum volume and ultra-rapid cooling rate allow acceptable survival and development rates in Day 2 and Day 6 in vitro produced embryos in sheep. Even though early stage embryos showed lower cryotolerance, those embryos that survive the vitrification-warming process show high development and hatching rates, similar to vitrification of morulae or blastocysts.

Hidrocolpos e hidrometrocolpos neoanatal: a propósito de un caso clínico / Neonatal hydrocolpos and hydrometrocolpos: a clinical case

Hydrocolpos and hydrometrocolpos is a condition characterized by a cystic dilatation od the vagina and/or uterus with accumulation of fluid as a result of congenital vaginal obstructions. It can be of secretory or urinary types, the last one when a vagino-vesical communication develops such as a sinus or chloaca. Other causes are vaginal septum, imperforated hymen. Clinical Case: Ten day old newborn, 46 XX with genital virilization (Prader IV) confirmed as due to Congenital Adrenal Hyperplasia, sonogram showed dilated vagina with fluid content due to neonatal hydrocolpos. Conclusion: It is important to maintain a high index of suspicion when a female newborn shows urogenital sinus, chloaca, genital virilization or imperforated hymen, as well as a female newborn with an abdominal mass. Diagnostic test of choice is a sonogram. Evaluation must be completed by a multidisciplinary team, including urology, endocrinology and pediatric gynecology for optimal patient management.

El Hidrocolpos e Hidrometrocolpos es una condición caracterizada por dilatación quística de la vagina y/o del útero, con acumulación de líquido como resultado de obstrucciones vaginales congénitas. Puede ser de tipo secretorio o urinario, este último ocurre cuando existe comunicación vagino-vesical, como en el seno urogenital o anomalía tipo cloaca. Otras causas son septo vaginal, himen imperforado, malformación tipo cloaca y senourogenital. Caso: Recién nacida de 10 días, 46 XX, con virilización de genitales grado IV de Prader, cuyo estudio confirmó una Hiperplasia Suprarrenal Congénita y cuya ecografía demostró una vagina dilatada con contenido liquido correspondiendo a un hidrocolpos neonatal.

Hipertiroidismo en niños y adolescentes / Hyperthyroidism in children and adolescents

Background: Hyperthyroidism (HT) prevalence is 0.1/100,000 children and 1/100,000 adolescents and Graves Disease is the most frequent etiology. Objective: To evaluate the clinical presentation, etiology and treatment in hyperthyroid children. Method: Retrospective review of clinical charts of children under 15 years-old, between June 2004 and August 2005. Hyperthyroidism diagnosis was performed with suppressed TSH and increased thyroid hormones levels. Etiological study was done by TRAb, ATPO, ATG, thyroid echotomography and I131 capture. Results: 26 patients were evaluated; 84.6 percent females and age at diagnosis was 9.8 +/- 3,5 years-old (range: 3,8 - 14,5). Goiter was the most frequent clinical sign (96,2 percent), tachicardy and swelling. Etiology: Graves Disease (73 percent),Hashitoxicosis (15,3 percent) and unknown etiology (11,5 percent). Treatment: 88,4 percent began with anti-thyroid drugs (DAT): 78 percent PTU and 22 percent Tiamazol. 62.5 percent became euthyroid after 6 months and 79.1 percent after 12 months. 31.5 percent of GD presented hypothyroidism at 6.3 +/- 4 months of DAT, requiring LT4 substitution. I131 was applied to 4 children (16.6 percent); 3 due to hepatic compromise pre or post PTU use and 1 girl for missing treatment, developing a thyrotoxic torment. Thyroidectomy was done in 2 patients (8.3 percent), both with GD; 1 for giant goiter without DAT response at 19 months and 1 for persistant hyperthyroidism after 25 months of DAT. 92 percent received (3-blockers (Propanolol) for adrenergic symptoms for 5 +/- 4 months. Conclusions: Goiter was the most frequent pediatric HT symptom and Graves disease the main etiology. DAT treatment control HT in 76.9 percent patients and no adverse reactions with I131 were observed. These resUIts promote DAT treatment as first line in HT management, prefering Tiamazol for its better adherence and less adverse reactions. Radioiodide therapy and thyroidectomy are alternatives if treatment fails...

El hipertiroidismo (HT) tiene una prevalencia de 0,1/100 000 en niños y 1/100 000 en adolescentes, siendo la enfermedad de Graves (EG) la etiología más frecuente. Objetivo: Revisar presentación clínica, etiología y manejo de niños con HT. Método: Estudio retrospectivo de fichas clínicas de niños con HT menores de 15 años, evaluados entre Junio/04 y Agosto/05. El diagnóstico de HT se hizo con TSH suprimida y hormonas tiroideas elevadas. El estudio etiológico se realizó en base a anticuerpos TRAb, ATPO, ATG; Ecotomograña tiroidea, y captación de I131. Resultados: Se evaluaron 26 pacientes; 84,6 por ciento fueron mujeres. Edad promedio al diagnóstico fue 9,8 +/- 3,5 años (rango 3,8 a 14,5). La presentación clínica más frecuente fue bocio (96,2 por ciento), seguidos por taquicardia y sudoración. Etiología: Enfermedad de Graves 73 por ciento, Hashitoxicosis 15,3 por ciento y etiología no precisada 11,5 por ciento. Manejo: 88,4 por ciento inician con drogas antitiroideas (DAT); 78 por ciento PTU y 22 por ciento con Tiamazol. 62,5 por ciento se hizo eutiroideo a los 6 meses y 79,1 por ciento a los 12 meses. El 31,5 por ciento de EG presentó hipotiroidismo a los 6,3 +/- 4 meses de uso de DAT, requiriendo sustitución con LT4. El I131 fue indicado a 4 niños (16,6 por ciento): en 3 casos por compromiso hepático importante pre o post uso de PTU y 1 niña por abandono de tratamiento y reingreso con tormenta tiroidea. Tiroidectomía: se indicó a 2 pacientes (8,3 por ciento), ambos con EG; uno por bocio gigante, sin respuesta a DAT después de 19 meses de uso y el otro por persistir hipertiroideo después de 25 meses de uso de DAT. El 92 por ciento recibió (3 bloqueador (propanolol) para manejo de los síntomas adrenérgicos, (5 +/- 4 meses). Discusión y conclusiones: El bocio es el síntoma principal en pediatría. La etiología más frecuente es la Enf de Graves. Las DAT permitieron controlar el HT en 76,9 por ciento de los pacientes, no observamos complicaciones con el uso de I131...

Pesquisa precoz de nefropatía diabética en niños y adolescentes portadores de diabetes mellitus tipo 1 / Early detection of diabetic nephropathy in children and adolescents with type 1 diabetes mellitus

La nefropatía diabética es la principal causa de insuficiencia renal crónica terminal (IRCT) en adultos a nivel mundial, a la vez que la IRCT es la principal causa de mortalidad de los pacientes con diabetes mellitus (DM) tanto tipo 1 como tipo 2.

La intervención en el lenguaje en el contexto del aula. Un estudio de caso / The language intervention at the classroom. A single case study

La finalidad de este artículo es presentar un estudio llevado a cabo con una niña deficiente mental en dos situaciones diferenciadas: la primera, desarrollada en el aula de logopedia y la segunda, en el aula ordinaria. Para la evaluación inicial se recurrió tanto a procedimientos informales (observación, entrevista y sociograma) como formales (inventario de desarrollo de atención temprana). Los resultados indicaron una mejora de las habilidades comunicativas básicas, una mayor participación, grado de compromiso y realización de las tareas, una efectividad del apoyo por pares, y un cambio hacia la colaboración y el desarrollo profesional, entre todos los agentes educativos

The aim of this paper is to show a single case carried out with a mental deficient girl in two different contexts: the speech therapy classroom and the ordiml1Y classroom. The procedures used were the following ones: observation, interview and inventory of the early intervention. The results showed best basic skills communication, bigger participation, an increase on the tasks and compromise, higher peer support intervention, and a change towards collaboration and professional development

Predicting vertical dimension with cephalograms, for edentulous patients.

OBJECTIVES: To provide information on vertical dimension (VD), occlusal plane (OP), and size of future occlusal rims, through lateral Rx and cephalometric tracings that analyse and compare hard and soft tissue. To show that these measures are compatible with methods routinely used to prescribe for full. DESIGN: Ten patients age range 53-81. Lateral Rx, with and without dentures, were taken and traced cephalograms were used to compare hard and soft tissue angles and planes. MAIN OUTCOME MEASURES: Based on three cephalometric analyses: Rickets, and McNamara for skeletal, and Legan-Burstone for soft tissue, to identify the following landmarks: 13 skeletal points: N, ANS, PNS, Me, Xi, Pm, FC, Or, Po, A, B, Co, Gn and four soft tissue points: G, Gv, Sn, Me'; seven skeletal planes: HP, FP, NF, N-ANS-Me, A-B, ANS-Me, Co-Gn, and three soft tissue planes: G-Sn, Sn-Me', Sn-Gv, and six angles: N-FC-A, ANS-Xi-Pm, INF, IMP, 6NF, interincisor angle. RESULTS: Stability in skeletal VD was observed with the proportion of 0.8 +/- 0.2 being present, between the middle third and lower third facial heights, N-ANS/ANS-Me. In addition, soft tissue proportion remained near 1, G-Sn,/Sn-Me. The length and position of first upper molar, upper and lower incisors were used to predict the OP. When compared with cephalograms of the same patient with dentures, similar measurements were observed. CONCLUSIONS: It was possible to provide information on skeletal, facial proportions, VD, OP, and rim size using cephalograms for edentulous patients.