PTEN Deletion in Adult Mice Induces Hypoinsulinemia With Concomitant Low Glucose Levels.

The PI3K/AKT pathway, negatively regulated by PTEN, plays a paramount role in glucose metabolism regulation due to its activation by the insulin receptor signaling pathway. We generated a PTEN-KO mouse to evaluate the systemic effect of the overactivation of the PI3K/AKT pathway in insulin signaling and glucose homeostasis. Our results demonstrate that PTEN-KO mice show very low glucose levels in the fasted state, which poorly respond to glucose and pyruvate administration. Insulinemia decreased without alterations in pancreatic islets. Among the possible reasons, we uncover the deregulation of the expression of proximal tubule glucose transporter and consequent glycosuria. Moreover, we evidence an altered activation of hepatic gluconeogenesis-related genes. In addition, the expression of several genes related to ß-oxidation showed a delayed or even absent response to fasting, suggesting that the lack of PTEN not only impairs glucose metabolism but also slows down the use of lipids as a metabolic fuel. We conclude that the inducible full PTEN-KO mice could be a good model to study the metabolic interactions between glycidic and lipidic metabolism in hypoinsulinemic hypoglycemia and that PTEN could be an important mediator in the disease and/or a potential drug target.

Elimination of Vitamin D Signaling Causes Increased Mortality in a Model of Overactivation of the Insulin Receptor: Role of Lipid Metabolism.

Vitamin D (VD) deficiency has been associated with cancer and diabetes. Insulin signaling through the insulin receptor (IR) stimulates cellular responses by activating the PI3K/AKT pathway. PTEN is a tumor suppressor and a negative regulator of the pathway. Its absence enhances insulin signaling leading to hypoglycemia, a dangerous complication found after insulin overdose. We analyzed the effect of VD signaling in a model of overactivation of the IR. We generated inducible double KO (DKO) mice for the VD receptor (VDR) and PTEN. DKO mice showed severe hypoglycemia, lower total cholesterol and increased mortality. No macroscopic tumors were detected. Analysis of the glucose metabolism did not show clear differences that would explain the increased mortality. Glucose supplementation, either systemically or directly into the brain, did not enhance DKO survival. Lipidic liver metabolism was altered as there was a delay in the activation of genes related to ß-oxidation and a decrease in lipogenesis in DKO mice. High-fat diet administration in DKO significantly improved its life span. Lack of vitamin D signaling increases mortality in a model of overactivation of the IR by impairing lipid metabolism. Clinically, these results reveal the importance of adequate Vitamin D levels in T1D patients.

Influence of background on precision of 3D depth judgment tasks in a real environment.

Presently, little is known about the effect of curved backgrounds against which the target stimulus is presented on precision in stereoacuity. The experiment analyzed the influence of stimulus orientation and 3D background configuration on stereoscopic vision. Participants were instructed to perform 3D visual alignment tasks on a modified version of the Howard-Dolman apparatus, whereupon precision in depth perception for different curved backgrounds (flat, black, concave, and convex) was evaluated. In addition, the influence of stimulus orientation (0 degrees, 45 degrees, and 90 degrees) on precision was examined. The findings revealed an underestimation in the perceived depth in all background configurations, indicating highest and lowest precision outcomes for convex and concave backgrounds, respectively. In addition, a significant interaction of background and orientation was found. It was concluded that, in a real environment, background local depth cues are integrated with target stimuli to contribute to depth perception.

[Acute toxicity by methotrexate used for abortion purpose. Case report]. / Intoxicación aguda por metotrexato utilizado con fines abortivos. Reporte de caso.

We report the case of a 16 years old female patient, with a pregnancy history of 11.4 weeks by ultrasound and intrauterine fetal death. In a private clinic were prescribed methotrexate 500 mg intramuscular single dose, and vaginal misoprostol. She had a clinical feature of five days of evolution characterized by fever of 39 degrees C, nausea, general attack and vomiting. The initial diagnosis was severe sepsis secondary to septic abortion, oral candidiasis and acute poisoning by methotrexate. After that, she was referred to the Instituto Nacional de Perinatologia, where stayed with fever for four days, and was managed with hydration, antibiotics, folinic acid and alkalizing. Her recovery was gradual. She was discharged after 12 days with significant clinical improvement. The literature review describes that the use of methotrexate for abortion purpose with therapeutic-dose presents a similar adverse effects to those found in our patient, however there are no case reports that describe the use of this drug in macrodosis for the same purpose, and their cytotoxic effects. We present this case because the patient used a macrodosis of this antimetabolite and due to the premature and empirical management with folinic acid, joined with alkalinization of urine, is the ideal treatment and as it is illustrated in our case.

Infiltrating CTLs in human glioblastoma establish immunological synapses with tumorigenic cells.

The immunological synapse between T cells and tumor cells is believed to be important for effective tumor clearance. However, the immunological synapse has never been imaged or analyzed in detail in human tissue. In this work, intercellular interactions between T cells and tumor cells were analyzed in detail in human glioblastoma. After characterization of the population of infiltrating T cells by multiple immunofluorescence staining and stereological quantification, the microanatomy of T cell-tumor cell intercellular communication was analyzed in detail using confocal microscopy and three-dimensional rendering. Cytotoxic T lymphocytes that infiltrated human glioblastoma underwent rearrangement when in contact with tumor cells, to form a three-dimensional structure in the intercellular contact area; this was characterized by microclusters of the CD3/TCR complex, re-arrangement of the cytoskeleton, and granzyme B polarization. In addition, such T cell-targeted cells show fragmentation of the microtubular system and increased expression levels of cleaved caspase 3, which suggests that cytotoxic T lymphocytes likely provoke changes in tumor cells and subsequently induce cell death. These results show that the formation of the cytotoxic T lymphocyte immunological synapse occurs in human tissue and may be relevant for the effective immune-mediated clearance of tumorigenic cells, therefore opening up new avenues for glioblastoma immunotherapy.

CD20, CD3, and CD40 ligand microclusters segregate three-dimensionally in vivo at B-cell-T-cell immunological synapses after viral immunity in primate brain.

The clearance of virally infected cells from the brain is mediated by T cells that engage antigen-presenting cells to form supramolecular activation clusters at the immunological synapse. However, after clearance, the T cells persist at the infection site and remain activated locally. In the present work the long-term interactions of immune cells in brains of monkeys were imaged in situ 9 months after the viral inoculation. After viral immunity, the persistent infiltration of T cells and B cells was observed at the infection sites. T cells showed evidence of T-cell receptor signaling as a result of contacts with B cells. Three-dimensional analysis of B-cell-T-cell synapses showed clusters of CD3 in T cells and the segregation of CD20 in B cells, involving the recruitment of CD40 ligand at the interface. These results demonstrate that immunological synapses between B cells and T cells forming three-dimensional microclusters occur in vivo in the central nervous system and suggest that these interactions may be involved in the lymphocyte activation after viral immunity at the original infection site.

Inflammatory response in Parkinsonism.

Inflammatory responses have been proposed as important factors in dopaminergic neuro-degeneration in Parkinsonism. Increasing evidence suggests that the alteration of the glial microenvironment induced by neuronal degeneration could be deleterious to the remaining neurons. The activation of microglia/macrophages and reactive astrocytes may have a negative effect on the surrounding parenchyma, perpetuating the neurodegenerative process. However, this alteration may also go beyond the brain parenchyma and stimulate other inflammatory changes in other systems, inducing the release of proinflammatory cytokines and probably Acute Phase Proteins (APP) and Glucocorticoids (GC). In this work we review the latest advances in the field to provide a picture of the state of the art of studies of inflammatory responses and Parkinsonism, hopefully opening up new therapeutic perspectives for patients with Parkinson's disease.

Increase of secondary processes of microglial and astroglial cells after MPTP-induced degeneration in substantia nigra pars compacta of non human primates.

Nigral dopaminergic areas from Parkinsonian patients show an increase of reactive astrocytes and active microglia. The reaction of these two cell types is a clear evidence of inflammatory response associated with dopaminergic cell loss. However, the function of this glial reaction remains unclear. This histological hallmark is also reproduced in induced Parkinsonian animals such as MPTP-treated monkeys. In this work, we analyze with confocal microscopy the number of processes of microglial cells and astrocytes in the SNpc of MPTP-treated monkeys and compare with control animals. We observe that secondary branches from microglia and astrocytes increase in MPTP-treated animals, while the scaffold of primary branches does not change. These results demonstrate that glial reaction in MPTP-treated monkeys is characterized by the emission of new filaments after the dopaminergic degeneration, suggesting that glial cells may increase their scanning progress and modify their microanatomy after dopaminergic injury.

A randomized controlled trial comparing isosorbide dinitrate-oxytocin versus misoprostol-oxytocin at management of foetal intrauterine death.

BACKGROUND: The metabolic activity of endogenous nitric oxide (NO) and the medical use of nitrovasodilatory drugs like isosorbide dinitrate have been shown to be potential inducers inducers of cervical ripening prior to surgical evacuation of the uterus. OBJECTIVE: To assess the therapeutic efficacy and safety of combined isosorbide dinitrate-oxytocin in the management of intrauterine foetal death (IUFD). METHODS: Sixty women with IUFD after 20 weeks of gestation requesting uterine evacuation were randomly selected to receive isosorbide dinitrate gel solution (80 mg/1.5 mL; n = 30) or misoprostol gel solution (100 mcg/1.5 mL; n = 30) every 3 h with a maximum of four doses or until a Bishop score >7 was reached. Subsequently, patients received a high dose of intravenous oxytocin until complete uterus evacuation was achieved. Therapeutic efficacy was evaluated by mean the relative risk of the foetal expulsion based on comparison of event rates, and the proportion of women induced to labor at 7, 10 and 15 h after the administration of isosorbide dinitrate or misoprostol. Safety was assessed on the basis of woman´s vital signs and evaluation of adverse effects, including headache, abdominal pain, pelvic pain, lower back pain, nausea, dizziness and vomiting. RESULTS: The foetal expulsion rate using the isosorbide dinitrate-oxytocin combination was approximately 4.4 times, and at least 2.1 times, the foetal expulsion rate with the misoprostol-oxytocin regimen at any given point in time. The proportion of women achieved vaginal delivery at 15 hours was 100% for the isosorbide dinitrate-oxytocin group and 86.7% for the misoprostol-oxytocin group. The average delivery induction interval was significantly lower when isosorbide dinitrate-oxytocin was used (8.7 ± 3.1 h) than when misoprostol-oxytocin (11.9 ± 3.1 h) was used. A total of 20% of patients in the isosorbide dinitrate-oxytocin group recorded headache, and no cases of uterine tachysystole, haemorrhage or coagulopathy were recorded. CONCLUSION: This study indicates that intravaginal isosorbide dinitrate followed by intravenous oxytocin was more effective than the conventional method used to induce labour in the medical management of foetal death in pregnancies after 20 weeks of gestation. TRIAL REGISTRATION: Clinicaltrials.gov NCT02488642.

The acquisition of productive vocabulary in Spanish children with Down syndrome.

BACKGROUND: It is generally assumed that children with Down syndrome (DS) present a deficit in lexical production relative to their cognitive abilities. However, the literature on this topic has recently shown several contradictory results. In addition, most studies only consider vocabulary production in its vocal modality. However it is also necessary to take into account gesture production, since this is a strength in children with DS. Our main purpose in this study, therefore, was to investigate the relationship between cognitive development and vocabulary size in both its vocal and gestural modalities in a broad sample of Spanish children with DS. METHOD: Participants in the study were 66 children with DS and 66 children with typical development (TD), with a mental age (MA) of 14-28 months (divided into five groups with a MA of 14-16, 17-19, 20-22, 23-25 and 26-28 months). Children were matched on the basis of their gender and MA. Productive vocabularies were collected using an adaptation of the MacArthur-Bates CDI for children with DS. RESULTS: In vocal modality, the number of words produced by children with DS and children with TD is similar. As in previous studies, our data confirmed that gestural communication is superior in children with DS. However, when words and gestures are combined, the performance of both groups of children is practically equal. CONCLUSION: Our results do not support a specific dissociation between cognitive and lexical development in children with DS.

Malformations in newborns associated to anticonvulsant consumption during pregnancy. experience in third level hospital of Mexico.

AIM: The purpose of the present study is to determine the relationship between the anticonvulsant drug use during pregnancy and the presence of malformations in the newborns. METHODS: The frequency of malformations in the neonates of epileptic mothers under anticonvulsant treatment was analyzed in two periods, one from 1988 to 1992, which included 76 epileptic mothers, and another from 1996 to 2003 with 170 patients. RESULTS: In the first period, 51 (67.1%) of mothers received monotherapy and 25 (32.9%) received polytherapy of phenytoin with carbamazepine, valproic acid or phenobarbital. In this period, 4 newborns (16%) with congenital malformations were registered. In the second period, 159 (93.5%) of the epileptic mothers received monotherapy and 11 (6.5%) received polytherapy of valproic acid with carbamazepine or phenytoin. During this period only 3 newborns 27.3% with malformations were registered. DISCUSSION: Clinical treatment should consider the risk of using polytherapy, mainly if phenytoin or valproic acid are combined with other anticonvulsants.

Visual fatigue while watching 3D stimuli from different positions.

PURPOSE: When observers focus their stereoscopic visual system for a long time (e.g., watching a 3D movie) they may experience visual discomfort or asthenopia. We tested two types of models for predicting visual fatigue in a task in which subjects were instructed to discriminate between 3D characters. One model was based on viewing distance (focal distance, vergence distance) and another in visual direction (oculomotor imbalance). METHOD: A 3D test was designed to assess binocular visual fatigue while looking at 3D stimuli located in different visual directions and viewed from two distances from the screen. The observers were tested under three conditions: (a) normal vision; (b) wearing a lens (-2 diop.); (c) wearing a base-out prism (2▿) over each eye. Sensitivity and specificity were calculated (as Signal Detection Theory parameters: SDT). RESULTS: An ANOVA and SDT analyses revealed that impaired visual performance were directly related to short distance and larger deviation in visual direction, particularly when the stimuli were located nearer and at more than 24° to the centre of the screen in dextroversion and beyond. CONCLUSION: This results support a mixed model, combining a model based on the visual angle (related to viewing distance) and another based on the oculomotor imbalance (related to visual direction). This mixed model could help to predict the distribution of seats in the cinema room ranging from those that produce greater visual comfort to those that produce more visual discomfort. Also could be a first step to pre-diagnosis of binocular vision disorders.

Pyridinium-1-yl bisphosphonates are potent inhibitors of farnesyl diphosphate synthase and bone resorption.

We report the design, synthesis and testing of a series of novel bisphosphonates, pyridinium-1-yl-hydroxy-bisphosphonates, based on the results of comparative molecular similarity indices analysis and pharmacophore modeling studies of farnesyl diphosphate synthase (FPPS) inhibition, human Vgamma2Vdelta2 T cell activation and bone resorption inhibition. The most potent molecules have high activity against an expressed FPPS from Leishmania major, in Dictyostelium discoideum growth inhibition, in gammadelta T cell activation and in an in vitro bone resorption assay. As such, they represent useful new leads for the discovery of new bone resorption, antiinfective and anticancer drugs.

3-D QSAR investigations of the inhibition of Leishmania major farnesyl pyrophosphate synthase by bisphosphonates.

We report the activities of 62 bisphosphonates as inhibitors of the Leishmania major mevalonate/isoprene biosynthesis pathway enzyme, farnesyl pyrophosphate synthase. The compounds investigated exhibit activities (IC(50) values) ranging from approximately 100 nM to approximately 80 microM (corresponding to K(i) values as low as 10 nM). The most active compounds were found to be zoledronate (whose single-crystal X-ray structure is reported), pyridinyl-ethane-1-hydroxy-1,1-bisphosphonates or picolyl aminomethylene bisphosphonates. However, N-alicyclic aminomethylene bisphosphonates, such as incadronate (N-cycloheptyl aminomethylene bisphosphonate), as well as aliphatic aminomethylene bisphosphonates containing short (n = 4, 5) alkyl chains, were also active, with IC(50) values in the 200-1700 nM range (corresponding to K(i) values of approximately 20-170 nM). Bisphosphonates containing longer or multiple (N,N-) alkyl substitutions were inactive, as were aromatic species lacking an o- or m-nitrogen atom in the ring, or possessing multiple halogen substitutions or a p-amino group. To put these observations on a more quantitative structural basis, we used three-dimensional quantitative structure-activity relationship techniques: comparative molecular field analysis (CoMFA) and comparative molecular similarity index analysis (CoMSIA), to investigate which structural features correlated with high activity. Training set results (N = 62 compounds) yielded good correlations with each technique (R(2) = 0.87 and 0.88, respectively), and were further validated by using a training/test set approach. Test set results (N = 24 compounds) indicated that IC(50) values could be predicted within factors of 2.9 and 2.7 for the CoMFA and CoMSIA methods, respectively. The CoMSIA fields indicated that a positive charge in the bisphosphonate side chain and a hydrophobic feature contributed significantly to activity. Overall, these results are of general interest since they represent the first detailed quantitative structure-activity relationship study of the inhibition of an expressed farnesyl pyrophosphate synthase enzyme by bisphosphonate inhibitors and that the activity of these inhibitors can be predicted within about a factor of 3 by using 3D-QSAR techniques.

[Plasma and milk concentrations of acenocoumarin in breast-feeding women during post partum]. / Concentraciones de acenocumarina en el plasma y la leche de madres anticoaguladas durante el puerperio.

BACKGROUND: Patients are receiving anticoagulants during postpartum. Literature data still is controversy in milk excretion of acenocoumarin; there are conducts in favor and against. Because of the benefits of maternal milk it's necessary to probe if acenocoumarin is excreted by human milk. OBJECTIVE: To determine the milk excretion of acenocoumarin in different periods of the postpartum and the milk excretion index in anticoagulant women. MATERIAL AND METHODS: It's an observational, descriptive and prospective study. The milk and plasma concentrations of acenocoumarin were determined in breast-feeding mothers with anticoagulation during the postpartum. ANOVA was used to determine the differences in pharmacokinetic constants in the different days of study. RESULTS: Two patients required light anticoagulation, nine moderate and five intense. The 37.5% of the new born were full term hypotrophy and the 18.75% were preterm. The highest plasma average concentration of acenocoumarin was found in day 45th postpartum (0.21 microg/mL). Acenocoumarin present in milk was found until day 30th; the average concentrations were low 0.011 microg/mL. The value of the maternal milk excretion index was 0.057 in day 45, what represents that approximately the 5% of acenocoumarin is eliminated by milk. The calculated dose of acenocoumarin that a new born could receive through maternal milk was lower than the recommended doses (1.79 microg/kg/day). CONCLUSION: These results allowed us to recommend breastfeeding in patients who are been anticoagulated with acenocoumarin.

Critical evaluation of the anatomical location of the Barrington nucleus: relevance for deep brain stimulation surgery of pedunculopontine tegmental nucleus.

Deep brain stimulation (DBS) has become the standard surgical procedure for advanced Parkinson's disease (PD). Recently, the pedunculopontine tegmental nucleus (PPN) has emerged as a potential target for DBS in patients whose quality of life is compromised by freezing of gait and falls. To date, only a few groups have published their long-term clinical experience with PPN stimulation. Bearing in mind that the Barrington (Bar) nucleus and some adjacent nuclei (also known as the micturition centre) are close to the PPN and may be affected by DBS, the aim of the present study was to review the anatomical location of this structure in human and other species. To this end, the Bar nucleus area was analysed in mouse, monkey and human tissues, paying particular attention to the anatomical position in humans, where it has been largely overlooked. Results confirm that anatomical location renders the Bar nucleus susceptible to influence by the PPN DBS lead or to diffusion of electrical current. This may have an undesirable impact on the quality of life of patients.

Persistent phagocytic characteristics of microglia in the substantia nigra of long-term Parkinsonian macaques.

Patients with Parkinson's disease show persistent microglial activation in the areas of the brain where the degeneration of dopaminergic neurons takes place. The reason for maintaining this activated state is still unknown, but it is thought that this persistent microglial activation may contribute to the degeneration of dopaminergic neurons. In this study, we report the microanatomical details of microglia and the relationship between microglia and neurons in the substantia nigra pars compacta of Parkinsonian monkeys years after insult with MPTP. We observed that microglial cells appear polarized toward dopaminergic neurons in MPTP-treated macaques compared to untreated animals and present clear phagocytic characteristics, such as engulfing gliaptic contacts, an increase in Golgi apparatus protein machinery and ball-and-chain phagocytic buds. These results demonstrate that activated microglia maintain phagocytic characteristics years after neurotoxin insult, and phagocytosis may be a key contributor to the neurodegenerative process.

Visual mechanisms governing the perception of auto-stereograms.

BACKGROUND: Single image random dot stereograms (SIRDS) have been used to study diverse visual parameters and skills. The aim of the present study was to identify the main optometric factors involved in the perception of SIRDS and to obtain a discriminant model to categorise our participants in terms of their skill in perceiving SIRDS. METHODS: Response time was determined to assess the ability of 69 participants to perceive the hidden three-dimensional shape in an auto-stereogram presented under controlled conditions, whereupon three skill level groups were defined. The same participants were administered a battery of optometric tests to evaluate various aspects of accommodation and convergence, as well as stereopsis and phoria. Linear discriminant analysis, which served to examine the relationship between response times and the evaluated visual parameters and skills, provided a set of discriminant functions (or model), thus allowing for the categorisation of participants according to their skill to perceive SIRDS. RESULTS: Two discriminant functions were obtained, which allowed for an overall predictive accuracy of 66.67 per cent (p = 0.024), with a higher predictive accuracy for groups 1 (minimum time less than 10 seconds, 78.26 per cent) and 2 (minimum time greater than 10 seconds, 75.86 per cent) than for group 3 (SIRDS not perceived, 35.29 per cent). Stereoacuity, negative relative convergence, phoria at near and, to a lesser extent, the accommodative convergence and accommodation ratio were found to be the most relevant discriminant variables, although between-group statistically significant differences were only disclosed for stereoacuity (p = 0.001) and negative relative convergence (p = 0.003). CONCLUSION: The ability to perceive SIRDS was related to many visual parameters and skills, including, but not limited to, stereoacuity and negative relative convergence. It is uncertain whether SIRDS might be considered a useful tool in clinical practice.