ABSTRACT
We developed a method of reducing the background fluorescence of samples made from formalin-fixed and paraffin-embedded blocks of the brain of the second-trimester human fetuses. For reducing excess background fluorescence, the samples were subjected to photobleaching using an LED lamp with blue and red emission peaks in the range of visible spectrum in a construction of an original design. The decrease in the background autofluorescence was checked by measuring the intensity of the emitted background fluorescence of the samples and relative abundance of immunopositive structures after immunohistochemical staining. It was found that the proposed method reliably reduced the background fluorescence of the samples, which improved the quality of multicolor immunofluorescence images of the cerebral cortex.
Subject(s)
Brain , Formaldehyde , Fetus , Humans , Paraffin Embedding , PhotobleachingABSTRACT
We studied peculiarities of the structure of human entorhinal cortex at weeks 20-26 of gestation (10 hemispheres). The samples were Nissl-stained and immunohistochemically treated with antibodies to parvalbumin, calretinin, calbindin, and cytoskeleton proteins (MAP2 and N200). 3D-reconstruction of the entorhinal cortex from serial sections was performed, caudomedial and rostrolateral areas were isolated. Parvalbumin+ cells in layer I, discrete distribution of layer II cells with colocalization of MAP2 and calretinin at the border with layer I, and two sublayers Va and Vb with MAP2+ neurons were typical for the caudomedial area. Rostrolateral area was characterized by the homogenous layer II with big amount of cells, high density of MAP2+ neurons only in layer III, and the unique layer V. Reelin+ Cajal-Retzius cells and N200+ fiber plexus in layer I were observed in the caudomedial and rostrolateral areas of the entorhial cortex. Layer IV was represented by a cell-free desiccant.
Subject(s)
Asphyxia/metabolism , Entorhinal Cortex/ultrastructure , Gene Expression Regulation, Developmental , Neurons/metabolism , Asphyxia/pathology , Calbindin 2/genetics , Calbindin 2/metabolism , Calbindins/genetics , Calbindins/metabolism , Entorhinal Cortex/anatomy & histology , Entorhinal Cortex/metabolism , Fetus , Gestational Age , Humans , Image Processing, Computer-Assisted , Immunohistochemistry , Microtomy , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Neurofilament Proteins/genetics , Neurofilament Proteins/metabolism , Neurons/ultrastructure , Parvalbumins/genetics , Parvalbumins/metabolism , Reelin ProteinABSTRACT
The age dynamics of corpus callosum development was studied on magnetic resonance images of the brain in children aged 2-11 years without neurological abnormalities and with infantile cerebral palsy. The areas of the total corpus callosum and its segments are compared in the midsagittal images. Analysis is carried out with the use of an original formula: proportion of areas of the anterior (genu, CC2; and anterior part, CC3) and posterior (isthmus, CC6 and splenium, CC7) segments: kCC=(CC2+CC3)×CC6/CC7. The results characterize age-specific dynamics of the corpus callosum development and can be used for differentiation, with high confidence, of the brain of children without neurological abnormalities from the brain patients with infantile cerebral palsy.
Subject(s)
Cerebral Palsy/pathology , Corpus Callosum/pathology , Age Factors , Case-Control Studies , Cerebral Palsy/diagnosis , Cerebral Palsy/physiopathology , Child , Child, Preschool , Corpus Callosum/physiopathology , Disease Progression , Female , Humans , Magnetic Resonance Imaging , MaleABSTRACT
A comparative morphometric analysis of MRI brain tomograms of term ant preterm born children was conducted. Common morphometric indexes and a novel quantitative parameter, a coefficient of corpus callosum (kCC), developed with regard to prenatal cortex ontogenesis pattern were applied to the study in order to get objective structural characteristics of the brain. All these quantitative indexes display anatomical features of the preterm brain. Reduced values of corpus callosum coefficient at preterm born children are known to associate with altered proportion between its rostral and caudal segments. A threshold coefficient value was established to differentiate preterm brain from the term one and it has proven its statistical significance.
Subject(s)
Cerebral Cortex/anatomy & histology , Corpus Callosum/physiology , Infant, Premature/physiology , Magnetic Resonance Imaging , Brain Mapping , Cerebral Cortex/diagnostic imaging , Child , Corpus Callosum/diagnostic imaging , Female , Gestational Age , Humans , Infant , Infant, Newborn , Male , RadiographyABSTRACT
Prenatal ontogeny of human neocortex have distinctive features that make it unique. Experimental data obtained on animal models could not be easily extrapolated on human corticogenesis of middle and late gestational period. Our research was aimed at features of human cortical pyramidal neurons development within 16-26 gestational weeks. Material was obtained during legal autopsies. Neurons were marked using indirect immunofluorescence with primary antibodies against phosphorylated a nd dephosphorylated microtubule associated protein MAP2.Expression of this protein marks the beginning of dendrogenesis. Morphotype and position of neuron within embryonic cortical layers could be easily identified due to abundance of MAP2 in neuron body and dendrites. It was shown, that MAP2 positive neurons are identifiable in embryonic cortical layer eV as early as 18th gestational week. At 25th gestational week two populations of pyramidal neurons are apparent inside cortical plate. In addition to layer eV neurons that have had differentiated earlier, layer eIII neurons appears. According to fact that differentiating neurons are more vulnerable to damaging factors than neuroblasts and mature neurons, our results suggest that critical periods for cortico-cortical and corticofugal populations of pyramidal cells occurs at different stages of second gestational trimester.
Subject(s)
Dendrites/metabolism , Neocortex/embryology , Neurogenesis/physiology , Pyramidal Cells/metabolism , Female , Gene Expression Regulation/physiology , Humans , Male , Microtubule-Associated Proteins/biosynthesis , Neocortex/cytology , Pregnancy , Pregnancy Trimester, Second/metabolism , Pyramidal Cells/cytologyABSTRACT
A female patient aged 50 is described, in whom candidiasis of the large folds has been erroneously diagnosed for 10 years; the diagnosis has not been confirmed by bacteriologic analysis, the therapy has been ineffective, the recurrences developed every year. The patient has developed symmetrical erythematous infiltrative areas in the armpits, groin, and in the folds under mammary glands, with solitary bullae up to 0.5-1.0 cm in diameter, with transparent contents, with thin flaccid covers and fragments thereof, as well as small pigmented vegetation. Basing on the histologic findings (suprabasal vesicles above the basal layer, acantholysis in the epidermis, and negligible lymphocytic infiltrate in the derma), Gougerot-Hailey-Hailey's disease has been diagnosed. The familial nature of the disease could not be established. Therapy with diphenhydramine, methyluracil, dimocifon, aloe extract, and aniline dyes, administered for 20 days, has resulted in clinical cure.
