In Vivo Mapping of Myocardial Injury Outside the Infarct Zone: Tissue at an Intermediate Pathological State.

BACKGROUND: The goal was to determine the feasibility of mapping the injured-but-not-infarcted myocardium using 99mTc-duramycin in the postischemic heart, with spatial information for its characterization as a pathophysiologically intermediate tissue, which is neither normal nor infarcted. METHODS AND RESULTS: Coronary occlusion was conducted in Sprague Dawley rats with preconditioning and 30-minute ligation. In vivo single-photon emission computed tomography was acquired after 3 hours (n=6) using 99mTc-duramycin, a phosphatidylethanolamine-specific radiopharmaceutical. The 99mTc-duramycin+ areas were compared with infarct and area-at-risk (n=8). Cardiomyocytes and endothelial cells were isolated for gene expression profiling. Cardiac function was measured with echocardiography (n=6) at 4 weeks. In vivo imaging with 99mTc-duramycin identified the infarct (3.9±2.4% of the left ventricle and an extensive area 23.7±2.2% of the left ventricle) with diffuse signal outside the infarct, which is pathologically between normal and infarcted (apoptosis 1.8±1.6, 8.9±4.2, 13.6±3.8%; VCAM-1 [vascular cell adhesion molecule 1] 3.2±0.8, 9.8±4.1, 15.9±4.2/mm2; tyrosine hydroxylase 14.9±2.8, 8.6±4.4, 5.6±2.2/mm2), with heterogeneous changes including scattered micronecrosis, wavy myofibrils, hydropic change, and glycogen accumulation. The 99mTc-duramycin+ tissue is quantitatively smaller than the area-at-risk (26.7% versus 34.4% of the left ventricle, P=0.008). Compared with infarct, gene expression in the 99mTc-duramycin+-noninfarct tissue indicated a greater prosurvival ratio (BCL2/BAX [B-cell lymphoma 2/BCL2-associated X] 7.8 versus 5.7 [cardiomyocytes], 3.7 versus 3.2 [endothelial]), and an upregulation of ion channels in electrophysiology. There was decreased contractility at 4 weeks (regional fractional shortening -8.6%, P<0.05; circumferential strain -52.9%, P<0.05). CONCLUSIONS: The injured-but-not-infarcted tissue, being an intermediate zone between normal and infarct, is mapped in vivo using phosphatidylethanolamine-based imaging. The intermediate zone contributes significantly to cardiac dysfunction.

Trocas na selva: uma experiência com grupo de fotoexpressão em meio ao distúrbio psicossomático / Exchanges in the jungle: a photoexpression group experience amid psychosomatic disorder / Cambios en la selva: una experiencia con grupo de fotoexpresión en medio al disturbio psicosomático

Este trabalho é mobilizado pela experiência institucional em um serviço de saúde publica que acolhe pacientes cuja problemática da somatização forma essencialmente seu quadro clínico. No sofrimento que circula no distúrbio psicossomático a incerteza do alojamento da psique no soma é a questão central. Todavia, esta clínica apresenta especificidades que desafiam as constantes de um enquadre psicanalítico tradicional. A técnica da Fotoexpressão como objeto mediador, utilizada em grupos, é observada como uma estratégia bastante apropriada no manejo destes pacientes. A situação clínica que este recurso suscita busca restabelecer a área de jogo e mediante uma zona intermediária, tenta abrir caminhos para processos elementares da experiência. Por meio de um recorte clínico buscamos problematizar a dimensão corporal e o elemento da violência diante do recurso da Fotoexpressão, analisando seus efeitos e possibilidades de deslocamentos.

This work is mobilized by an institutional experience in a public health service that welcomes patients whose somatization problem essentially forms their clinical picture. In the suffering that circulates in the psychosomatic disturbance the uncertainty of the psyche's accommodation in the soma is the central issue. However, this clinic has specificities that defy the constants of a traditional psychoanalytic framework. The technique of Photoexpression as a mediator object, used in groups, is observed as a very appropriate strategy in the management of these patients. The clinical situation that this resource raises seeks to reestablish the playing area and through an intermediate zone, tries to open the way for elementary processes of experience. Through a clinical outline we seek to problematize the body dimension and the element of violence in the face of the Photoexpression resource, analyzing its effects and possibilities of displacement.

Este trabajo es movilizado por la experiencia institucional en un servicio de salud pública que acoge pacientes cuya problemática de la somatización forma esencialmente su cuadro clínico. En el sufrimiento que circula en el disturbio psicosomático la incertidumbre del alojamiento de la psique en suma es la cuestión central. Todavía, esta clínica presenta especificidades que desafian las constantes de un encuadre psicoanalítico tradicional. La técnica de la Fotoexpresión como objeto mediador, utilizada en grupos, es observada como una estrategia bastante apropiada en el manejo de estos pacientes. La situación clínica que este recurso suscita busca restablecer la área del juego y mediante una zona intermediaria, intenta abrir caminos para procesos elementares de experiencia. Por medio de un recorte clínico buscamos problematizar la dimensión corporal y el elemento de la violencia ante al recurso de la Fotoexpresión analizando sus efectos y posibilidades de desplazamientos.

Effect of prenatal exposure of green tea extract on the developing central nervous system of rat fetuses; histological, immune-histochemical and ultrastructural studies.

Although, several health benefits were associated with green tea, these effects may be beneficial up to a certain dose. Higher doses of green tea may cause several adverse effects. So, there is a need to test the potential negative effects of green tea during pregnancy. This study was designated to evaluate the effect of prenatal exposure of green tea extract on the development of the central nervous system of 20-day old rat fetuses. The pregnant rats were divided into 4 groups; the control group (received distal water) and the other 3 groups received green tea extract at different doses (200, 600 & 1000 mg/kg/day, respectively) from the 6th to 15th day of gestation i.e., during the organogenesis phase of development. Cerebral cortex, cerebellum and spinal cord specimens were subjected to histological, immunohistochemical and ultrastructure investigations. The body weight of both mothers and fetuses was significantly decreased in the groups that received 600 and 1000 mg green tea extract. Also, the neuronal tissues displayed various signs of degeneration which were evident with the 600 and 1000 mg doses. Green tea extract also increases the glial fibrillary acidic protein (GFAP) and decreases the proliferating cell nuclear antigen (PCNA) which were directly proportional with increasing the dose. Administration of green tea extract during rat organogenesis period induced various histological, immunohistochemical and ultrastructural degenerative changes in the cerebral cortex, cerebellum and spinal cord of 20-day old rat fetuses. These deleterious changes were directly proportional to increasing the green tea extract dose. Thus, it should be stressed that the effect of green tea is dose-dependent and therefore it can be either beneficial or adverse.

Developmental Differences Between the Limbic and Neocortical Telencephalic Wall: An Intrasubject Slice-Matched 3 T MRI-Histological Correlative Study in Humans.

The purpose of the study was to investigate the interrelation of the signal intensities and thicknesses of the transient developmental zones in the cingulate and neocortical telencephalic wall, using T2-weighted 3 T-magnetic resonance imaging (MRI) and histological scans from the same brain hemisphere. The study encompassed 24 postmortem fetal brains (15-35 postconceptional weeks, PCW). The measurements were performed using Fiji and NDP.view2. We found that T2w MR signal-intensity curves show a specific regional and developmental stage profile already at 15 PCW. The MRI-histological correlation reveals that the subventricular-intermediate zone (SVZ-IZ) contributes the most to the regional differences in the MRI-profile and zone thicknesses, growing by a factor of 2.01 in the cingulate, and 1.78 in the neocortical wall. The interrelations of zone or wall thicknesses, obtained by both methods, disclose a different rate and extent of shrinkage per region (highest in neocortical subplate and SVZ-IZ) and stage (highest in the early second half of fetal development), distorting the zones' proportion in histological sections. This intrasubject, slice-matched, 3 T correlative MRI-histological study provides important information about regional development of the cortical wall, critical for the design of MRI criteria for prenatal brain monitoring and early detection of cortical or other brain pathologies in human fetuses.

Exploration of endoscopic findings and risk factors of early gastric cancer after eradication of Helicobacter pylori.

AIM: To explore the endoscopic features and risk factors of early gastric cancer (EGC) after eradication of Helicobacter pylori (H. pylori). METHODS: A total of 1961 patients who underwent esophago-gastro-duodenoscopy (EGD) with a history of successful H. pylori eradication were enrolled in this multicenter research. Among them, 162 EGC lesions of 132 patients were detected. The endoscopic features and risk factors of post-eradication EGC were explored. RESULTS: Severe atrophy (75.3% vs. 16.7%, p value <.01), intestinal metaplasia (96.3% vs. 77.1%, p value <.01), map-like redness (89.5% vs. 65.4%, p value <.01), distinct intermediate zone (IZ) (68.5% vs. 23.4%, p value <.01) and xanthoma (58.0% vs. 17.9%, p value <.01) were significantly more frequent in the CA group (patients with newly detected EGC after eradication of H. pylori) than in the NC group (patients without gastric cancer after eradication of H. pylori). In multivariate analysis, severe atrophy (odds ratio (OR) = 8.08; 95% confidence interval (CI), 3.43-20.0; p value<.01), map-like redness (OR = 1.75; 95% CI, 0.11-5.25; p value = .04), distinct IZ (OR = 2.87; 95% CI, 1.20-6.93; p value = .02) and xanthoma (OR = 2.84; 95% CI, 1.20-7.03; p value=.02) were proved to be risk factors for detection of EGC after eradication of H. pylori. CONCLUSIONS: Severe atrophy and map-like redness and distinct IZ and xanthoma are risk factors of EGC after eradication of H. pylori.

Development of an L-type Ca2+ channel-dependent Ca2+ transient during the radial migration of cortical excitatory neurons.

Increasing evidence has shown that voltage-gated L-type Ca2+ channels (LTCCs) are crucial for neurodevelopmental events, including neuronal differentiation/migration and neurite morphogenesis/extension. However, the time course of their functional maturation during the development of excitatory neurons remains unknown. Using a combination of fluorescence in situ hybridization and in utero electroporation-based labeling, we found that the transcripts of Cacna1c and Cacna1d, which encode the LTCC pore-forming subunits, were upregulated in the intermediate zone (IZ) during radial migration. Ca2+ imaging using GCaMP6s in acute brain slices showed spontaneous Ca2+ transients in migrating neurons throughout the IZ. Neurons in the IZ upper layer, especially in the multipolar-to-bipolar transition layer (TL), exhibited more frequent Ca2+ transients than adjacent layers and responded to FPL64176, a potent activator of LTCC. Consistently, nimodipine, an LTCC blocker, inhibited spontaneous Ca2+ transients in neurons in the TL. Collectively, we showed a hitherto unknown increased prevalence of LTCC-dependent Ca2+ transients in the TL of the IZ upper layer during the radial migration of excitatory neurons, which could be essential for the regulation of Ca2+-dependent neurodevelopmental processes.

Basal ganglia involvement in ARX patients: The reason for ARX patients very specific grasping?

The ARX (Aristaless Related homeoboX) gene was identified in 2002 as responsible for XLAG syndrome, a lissencephaly characterized by an almost complete absence of cortical GABAergic interneurons, and for milder forms of X-linked Intellectual Disability (ID) without apparent brain abnormalities. The most frequent mutation found in the ARX gene, a duplication of 24 base pairs (c.429_452dup24) in exon 2, results in a recognizable syndrome in which patients present ID without primary motor impairment, but with a very specific upper limb distal motor apraxia associated with a pathognomonic hand-grip, described as developmental Limb Kinetic Apraxia (LKA). In this study, we first present ARX expression during human fetal brain development showing that it is strongly expressed in GABAergic neuronal progenitors during the second and third trimester of pregnancy. We show that although ARX expression strongly decreases towards the end of gestation, it is still present after birth in some neurons of the basal ganglia, thalamus and cerebral cortex, suggesting that ARX also plays a role in more mature neuron functioning. Then, using morphometric brain MRI in 13 ARX patients carrying c.429_452dup24 mutation and in 13 sex- and age-matched healthy controls, we show that ARX patients have a significantly decreased volume of several brain structures including the striatum (and more specifically the caudate nucleus), hippocampus and thalamus as well as decreased precentral gyrus cortical thickness. We observe a significant correlation between caudate nucleus volume reduction and motor impairment severity quantified by kinematic parameter of precision grip. As basal ganglia are known to regulate sensorimotor processing and are involved in the control of precision gripping, the combined decrease in cortical thickness of primary motor cortex and basal ganglia volume in ARX dup24 patients is very likely the anatomical substrate of this developmental form of LKA.

Expanded alleles of the FMR1 gene are related to unexplained recurrent miscarriages.

Up to 50% of recurrent miscarriage cases in women occur without an underlying etiology. In the current prospective case-control study, we determined the impact of CGG trinucleotide expansions of the fragile-X mental retardation 1 (FMR1) gene in 49 women with unexplained recurrent miscarriages. Case group consisted of women with two or more unexplained consecutive miscarriages. Blood samples were obtained and checked for the presence of expanded alleles of the FMR1 gene using PCR. Patients harboring the expanded allele, with a threshold set to 40 repeats, were further evaluated by sequencing. The number of abortions each woman had, was not associated with her respective CGG repeat number (P=0.255). The repeat sizes of CGG expansion in the FMR1 gene were significantly different in the two population groups (P=0.027). All the positive cases involved intermediate zone carriers. Hence, the CGG expanded allele of the FMR1 gene might be associated with unexplained multiple miscarriages; whether such an association is coincidental or causal can be confirmed by future studies using a larger patient cohort.

Validation of Polymerase Chain Reaction-Based Assay to Detect Actual Number of CGG Repeats in FMR1 Gene in Indian Fragile X Syndrome Patients.

Molecular genetic testing for fragile X (FX) is complicated due to the large variation in the size of CGG expansion. The aim of this study was to apply this new technique using AmplideX FMR1 PCR assay, which is considered a better diagnostic tool for detecting expanded alleles in Indian population. The primary objective was to identify the carrier status of females and to correlate the instability of premutation alleles in females with the repeat sizes. 24 children with FX based on rapid PCR and 29 female relatives of these patients were included. Out of the 29 females screened, those whose child (or children) was affected by FX, were all premutation carriers confirming their role in transmission. The smallest PM allele that expanded into FM in the next generation was 78 repeats and the smallest PM allele detected was 63 repeats, and when transmitted from mother to offspring remained in the premutation range. In 4 families, the repeat size of the allele reduced from PM to normal repeat numbers in their daughters and in 1 case to borderline PM range. Thus, apart from the reduced turnaround time, this PCR based assay offers advantage by its sensitivity to detect CGG repeats in the intermediate region and lower range of premutation alleles. It also provides added information of AGG interruptions, which may have an impact on the counseling of women with intermediate and PM alleles.

In and out from the cortex: development of major forebrain connections.

In this review we discuss recent advances in the understanding of the development of forebrain projections attending to their origin, fate determination, and axon guidance. Major forebrain connections include callosal, corticospinal, corticothalamic and thalamocortical projections. Although distinct transcriptional programs specify these subpopulations of projecting neurons, the mechanisms involved in their axonal development are similar. Guidance by short- and long-range molecular cues, interaction with intermediate target populations and activity-dependent mechanisms contribute to their development. Moreover, some of these connections interact with each other showing that the development of these axonal tracts is a well-orchestrated event. Finally, we will recapitulate recent discoveries that challenge the field of neural wiring that show that these forebrain connections can be changed once formed. The field of reprogramming has arrived to postmitotic cortical neurons and has showed us that forebrain connectivity is not immutable and might be changed by manipulations in the transcriptional program of matured cells.

DNA damage response in microcephaly development of MCPH1 mouse model.

MCPH1 encodes BRCT-containing protein MCPH1/Microcephalin/BRIT1, mutations of which in humans cause autosomal recessive disorder primary microcephaly type 1 (MCPH1), characterized by a congenital reduction of brain size particularly in the cerebral cortex. We have shown previously that a deletion of Mcph1 in mice results in microcephaly because of a premature switch from symmetric to asymmetric division of the neuroprogenitors, which is regulated by MCPH1's function in the centrosome. Because MCPH1 has been implicated in ATM and ATR-mediated DNA damage response (DDR) and defective DDR is often associated with neurodevelopmental diseases, we wonder whether the DDR-related function of MCPH1 prevents microcephaly. Here, we show that a deletion of Mcph1 results in a specific reduction of the cerebral cortex at birth, which is persistent through life. Due to an effect on premature neurogenic production, Mcph1-deficient progenitors give rise to a high level of early-born neurons that form deep layers (IV-VI), while generate less late-born neurons that form a thinner outer layer (II-III) of the cortex. However, neuronal migration seems to be unaffected by Mcph1 deletion. Ionizing radiation (IR) induces a massive apoptosis in the Mcph1-null neocortex and also embryonic lethality. Finally, Mcph1 deletion compromises homologous recombination repair and increases genomic instability. Altogether, our data suggest that MCPH1 ensures proper neuroprogenitor expansion and differentiation not only through its function in the centrosome, but also in the DDR.

Growth defects in the dorsal pallium after genetically targeted ablation of principal preplate neurons and neuroblasts: a morphometric analysis.

The present study delineates the large-scale, organic responses of growth in the dorsal pallium to targeted genetic ablations of the principal PP (preplate) neurons of the neocortex. Ganciclovir treatment during prenatal development [from E11 (embryonic age 11) to E13] of mice selectively killed cells with shared S-phase vulnerability and targeted expression of a GPT [golli promoter transgene; GPT linked to HSV-TK (herpes simplex virus-thymidine kinase), τ-eGFP and lacZ reporters] localized in PP neurons and their intermediate progenitor neuroblasts. The volume, area and thickness of the pallium were measured in an E12-P4 (postnatal age 4) longitudinal study with comparisons between ablated (HSV-TK(+/0)) and control (HSV-TK(0/0)) littermates. The extent of ablations was also systematically varied, and the effect on physical growth was assessed in an E18 cross-sectional study. The morphological evidence obtained in the present study supports the conclusion that genetically targeted ablations delay the settlement of the principal PP neurons of the dorsal pallium. This leads to progressive and substantial reductions of growth, despite compensatory responses that rapidly replace the ablated cells. These growth defects originate from inductive cellular interactions in the proliferative matrix of the ventricular zone of the pallium, but are amplified by subsequent morphogenic and trophic cellular interactions. The defects persist during the course of prenatal and postnatal development to demonstrate a constrained dose-response relationship with the extent of specific killing of GPT neurons. The defects propagate simultaneously in both the horizontal and vertical cytoarchitectural dimensions of the developing pallium, an outcome that produces a localized shortfall of volume in the telencephalic vesicles.

Molecules and mechanisms involved in the generation and migration of cortical interneurons.

The GABA (γ-aminobutyric acid)-containing interneurons of the neocortex are largely derived from the ganglionic eminences in the subpallium. Numerous studies have previously defined the migratory paths travelled by these neurons from their origins to their destinations in the cortex. We review here results of studies that have identified many of the genes expressed in the subpallium that are involved in the specification of the subtypes of cortical interneurons, and the numerous transcription factors, motogenic factors and guidance molecules that are involved in their migration.

Ultrastructural morphometry of the rat pituitary intermediate zone after stimulation and suppression.

The intermediate zone of the rat pituitary was studied after stimulation for 3 weeks with the adrenostatic drugs metyrapone and aminogfutethimide and after suppression for 3 weeks with the glucocorticoid 6α-methylprednisolone using morphometry by the point-counting method on electron micrographs. After metyrapone, intermediate cells showed a significant (P <.05) increase in volume percentages of the smooth endoplasmic reticulum, the Golgi apparatus, and the lucent immature secretory granules, as well as a significant decrease in the lucent secretory granules, the vacuoles, the lysosomes, and the cell membranes in comparison with the untreated controls. After aminoglutethimide, the volume percentages of the nuclei, the smooth endoplasmic reticulum, and the dense and lucent immature secretory granules were significantly increased in comparison with the controls, whereas the unorganized cytoplasm, the lucent secretory granules, the vacuoles, and the lysosomes were significantly decreased. The morphological changes after both adrenostatic drugs clearly indicate the pathophysiologically expected increase of secretory activity. After 6α-methyfprednisolone, the rough endoplasmic reticulum, the vacuoles, and the lysosomes significantly decreased in comparison with the controls, and the smooth endoplasmic reticulum, the Golgi fields, and the immature secretory granules were increased. These changes cannot be interpreted without certain reservations as the first alterations indicate decreased activity, whereas the increased Golgi fields and immature secretory granules are usually signs of increased secretory activity, which cannot be expected after glucocorticoid treatment. The possible causes are discussed.

DISTRIBUTION OF MONOAMINERGIC AND SOME PEPTIDERGIC AFFERENT FIBERS IN THE INTERMEDIATE ZONE OF THE RAT SPINAL CORD / 解剖学报

This study demonstrated with the immunocytochemical ABC method that thedistribution pattern of TH,5-TH,SP,L-ENK and OXY immunoreactive fiberswere alike in the intermediate zone of the thoracic and upper lumbar spinal cord.These fibers located in the lateral funiculus,lateral horn,intermediate gray and thearea surrounding the central canal,forming a transversal fiber band with most ofthe fibers in the lateral horn.In the lateral horn,TH,5-HT,SP and L-ENKimmunoreactive fibers and terminals aggregated in fiber clusters and numerous fib-ers made up a longitudinal bundle running through the fiber clusters.These immuno-reative fibers and terminals surrounding the central canal and distributed mainly dorsalto it.TH and 5-HT fibers accumulated in midplane where some fibers ran in alongitudinal fiber bundle and bilateral fibers and terminals distributed sparsely.SP and L-ENK fibers and terminals were relatively less in the midplane whiledense laterally.All kinds of the immunoreactive fibers were found close to theependyma.Two small 5-HT fiber bundles ran parallel and lateral to the centralcanal and a small SP fiber bundle ventral to it.Moderate immunoreactive fiberswere found periodically in the intermediate gray and formed distinct bands to linkfiber clusters in the lateral horn and fibers dorsal to the central canal.The imm- unoreactive fibers in lateral funiculus accumulated near the lateral horn and mostdensely in upper thoracic cord.OXY-containing fibers were much less than theothers,thus obvious fiber clusters,longitudinal bundle or transversal bands had notbeen found.In general,the distribution pattern of some monoaminergic and pepti-dergic fibers in the intermediate zone was provided with a laddershaped configura-tion.The two longitudinal immunoreactive fiber bundles——dorsal to the centralcanal and within the lateral horn,located medial and lateral in the spinal grayrespectively,and the transversal bands appeared periodically in the intermediate graybridging between the two longitudinal bundles.This configuration of immunorea-ctive fibers coincided with nuclear regions which contained preganglionic sympatheticneurons as well as their dendrites.