Morphological Signs of Dystrophy, Regeneration and Hypertrophy of Neurons

Results: Dystrophic changes constitute an extensive group of neuronal disorders and are manifested at the morphological level by deformation of the perikarions and neuropil, wrinkling or swelling of the cell, and changes in the chromatophilia of the cytoplasm. At the electron microscopic level, disorganization of organelles is observed, reflecting gross violations of the vital processes of the neuron. There are several ways to regenerate neurons: intracellular regeneration, restoration of the neuropil, the formation of new neurons (in some parts of the nervous system - the hippocampus, the subventricular layer of the lateral ventricles and olfactory bulbs) and the formation of heterokaryons (fusion of a neuron with an oligodendrocyte). Hypertrophy of neurons may indicate both compensation and the development of a pathological process. To clarify the nature of this phenomenon, it is necessary to conduct an ultramicroscopic study of the organelles of the nerve cell.

Linking Fibrotic Remodeling and Ultrastructural Alterations of Alveolar Epithelial Cells after Deletion of Nedd4-2

Our previous study showed that in adult mice, conditional Nedd4-2-deficiency in club and alveolar epithelial type II (AE2) cells results in impaired mucociliary clearance, accumulation of Muc5b and progressive, terminal pulmonary fibrosis within 16 weeks. In the present study, we investigated ultrastructural alterations of the alveolar epithelium in relation to interstitial remodeling in alveolar septa as a function of disease progression. Two, eight and twelve weeks after induction of Nedd4-2 knockout, lungs were fixed and subjected to design-based stereological investigation at the light and electron microscopic level. Quantitative data did not show any abnormalities until 8 weeks compared to controls. At 12 weeks, however, volume of septal wall tissue increased while volume of acinar airspace and alveolar surface area significantly decreased. Volume and surface area of alveolar epithelial type I cells were reduced, which could not be compensated by a corresponding increase of AE2 cells. The volume of collagen fibrils in septal walls increased and was linked with an increase in blood–gas barrier thickness. A high correlation between parameters reflecting interstitial remodeling and abnormal AE2 cell ultrastructure could be established. Taken together, abnormal regeneration of the alveolar epithelium is correlated with interstitial septal wall remodeling.

Anatomy of the Liver of Mizoram Local Pig (Zovawk)

Background: The present study was aimed for the promotion and advancement of the anatomical knowledge at the gross, light microscopic and electron microscopic level in Zovawk (mizo local pig). Methods: The current investigation was done at the Instructional livestock farm, Department of Veterinary Anatomy and Histology, College of Veterinary Sciences and Animal Husbandry, Central Agricultural University, Selesih, Aizawl, Mizoram and Sophisticated Analytical Instrument Facility (SAIF), North Eastern Hill University (NEHU), Shillong, Meghalaya. Six liver samples were collected from six apparently healthy Zovawk animal of either sex and gross morphological observations were done directly after collection. Thereafter tissue samples were collected as such and were preserved in neutral buffer formalin (NBF) and in Karnovsky’s fixative for routine histology and transmission electron microscopic examination, respectively.Result: Zovawk liver shows four distinct borders, i.e. medial, lateral, dorsal and ventral border, two surfaces, i.e. parietal surface or diaphragmatic surface and caudal or visceral surface and six distinct lobes. The average weight of Zovawk liver was 1.402 kg. Weight of the liver was highly correlated with body weight of animal. Histologically, Zovawk liver was characterized with thick Glisson’s capsule and thick connective tissue septa emerging from it, which gives the hepatic lobules its hexagonal shape. Sinusoids of adjacent hepatocytes were lined by stellate shaped Kupffer cells. The ultra-structural examination of liver shows that, the hepatocytes were rich in mitochondria, endoplasmic reticulum and glycogen granules. Free ribosome and well developed rER and dense lysosomal granules were common in those hepatocyte.

Cellular and Subcellular Localisation of Kv4-Associated KChIP Proteins in the Rat Cerebellum

The K+ channel interacting proteins (KChIPs) are a family of cytosolic proteins that interact with Kv4 channels, leading to higher current density, modulation of channel inactivation and faster recovery from inactivation. Using immunohistochemical techniques at the light and electron microscopic level combined with quantitative analysis, we investigated the cellular and subcellular localisation of KChIP3 and KChIP4 to compare their distribution patterns with those for Kv4.2 and Kv4.3 in the cerebellar cortex. Immunohistochemistry at the light microscopic level demonstrated that KChIP3, KChIP4, Kv4.2 and Kv4.3 proteins were widely expressed in the cerebellum, with mostly overlapping patterns. Immunoelectron microscopic techniques showed that KChIP3, KChIP4, Kv4.2 and Kv4.3 shared virtually the same somato-dendritic domains of Purkinje cells and granule cells. Application of quantitative approaches showed that KChIP3 and KChIP4 were mainly membrane-associated, but also present at cytoplasmic sites close to the plasma membrane, in dendritic spines and shafts of Purkinje cells (PCs) and dendrites of granule cells (GCs). Similarly, immunoparticles for Kv4.2 and Kv4.3 were observed along the plasma membrane and at intracellular sites in the same neuron populations. In addition to the preferential postsynaptic distribution, KChIPs and Kv4 were also distributed presynaptically in parallel fibres and mossy fibres. Immunoparticles for KChIP3, KChIP4 and Kv4.3 were detected in parallel fibres, and KChIP3, KChIP4, Kv4.2 and Kv4.3 were found in parallel fibres, indicating that composition of KChIP and Kv4 seems to be input-dependent. Together, our findings unravelled previously uncharacterised KChIP and Kv4 subcellular localisation patterns in neurons, revealed that KChIP have additional Kv4-unrelated functions in the cerebellum and support the formation of macromolecular complexes between KChIP3 and KChIP4 with heterotetrameric Kv4.2/Kv4.3 channels.

Subcellular Localization of the TFF Peptides xP1 and xP4 in the Xenopus laevis Gastric/Esophageal Mucosa: Different Secretion Modes Reflecting Diverse Protective Functions

The TFF peptides xP1 and xP4 from Xenopus laevis are orthologs of TFF1 and TFF2, respectively. xP1 is secreted as a monomer from gastric surface mucous cells and is generally not associated with mucins, whereas xP4 is a typical secretory peptide from esophageal goblet cells, and gastric mucous neck and antral gland cells tightly associated as a lectin with the ortholog of mucin MUC6. Both TFF peptides have diverse protective functions, xP1 as a scavenger for reactive oxygen species preventing oxidative damage and xP4 as a constituent of the water-insoluble adherent inner mucus barrier. Here, we present localization studies using immunofluorescence and immunoelectron microscopy. xP1 is concentrated in dense cores of secretory granules of surface mucous cells, whereas xP4 mixes with MUC6 in esophageal goblet cells. Of note, we observe two different types of goblet cells, which differ in their xP4 synthesis, and this is even visible morphologically at the electron microscopic level. xP4-negative granules are recognized by their halo, which is probably the result of shrinkage during the processing of samples for electron microscopy. Probably, the tight lectin binding of xP4 and MUC6 creates a crosslinked mucous network forming a stabile granule matrix, which prevents shrinkage.

PREGNANCY, COMPLICATED BY PREECLAMPSIA: FETOPLACENTAL COMPLEX IMMUNE DEADAPTATION AND HISTOSTRUCTURAL FEATURES

The aim: to study and compare the features of the interleukins levels and morphological changes of placenta at various stages of preeclampsia. Materials and methods: 109 pregnant women with preeclampsia of varying severity (study group) and 30 pregnant women with uncomplicated pregnancy (control group) were examined. Immunohistochemical method, proinflammatory interleukins levels, morphological and morphometric analysis of peripheral and central placental areas biopsies on the optical and electron-microscopic level have been used. Results: Morphofunctional changes in the placenta in case of preeclampsia and the increase in the expression level of the transforming growth factor have a series of regular stages from the formation, strain and disruption of adaptive mechanisms with more pronounced signs of morphological immaturity of parenchymal and stromal elements of the placenta, especially in the area of syncytiotrophoblast and spiral vessels. The degree of clinical manifestation of preeclampsia has a correlation relationship with IL-10 deficiency and with the increase in TNF-α, stimulation of macrophage-protein production that contributes to the change in the ratio of Thl / Th2, which are antagonists and inhibit each other’s development. Conclusions: The severity of the preeclampsia course correlates with the state of morphofunctional changes in the placenta and changes in the ratio of the pro- and anti-inflammatory interleukins.

VITAMIN D DEFICIENCY IN PATIENTS WITH CHRONIC HEART FAILURE AND MORPHO-FUNCTIONAL CONDITION OF PERIPHERAL BLOOD ERYTHROCYTES

The aim: To assess the vitamin D level in blood plasma of patients with chronic heart failure and to identify the effect of its deficiency on the state of peripheral blood erythrocytes during physical exertion. Materials and methods: A total of 25 patients with CHF grade IIA stages II and III functional class were examined. The control group consisted of 25 relatively healthy people. All patients were offered to complete a 6 minutes walking test. The level of 25 (OH) D total in plasma was determined by enzyme immunoanalysis. Morphological studies of erythrocytes were performed on the light-optical (Leica CME) and electron-microscopic level («JEOL-25M-T220»). Results: Patients with chronic heart failure experienced 22.9% decrease in their vitamin D level (17.2±0.04 ng/ml) compared to the control group (38.4±0.05 ng/ml). Correlation analysis showed a direct proportional relationship between vitamin D deficiency and the number of erythrocytes of a modified form (r = 0.58; p <0.05) and erythrocytes with low osmotic resistance (r = 0.87; p <0.05). During the timed physical evaluation patients who experienced chronic heart failure accompanied by vitamin D deficiency developed an increase in the number of their reversibly and irreversibly deformed erythrocytes and a decrease in the cellular osmotic tability. Conclusions: During physical exertion, patients who experienced chronic heart falure accompanied by with vitamin D deficiency experienced morpho-biochemical changes in their red blood cells. These changes indicated structural disturbances in the membranes of their erythrocytes and could potentially have negative consequences for the somatic health of these patients.

Osteogenic Cell Behavior on Titanium Surfaces in Hard Tissue

It is challenging to remove dental implants once they have been inserted into the bone because it is hard to visualize the actual process of bone formation after implant installation, not to mention the cellular events that occur therein. During bone formation, contact osteogenesis occurs on roughened implant surfaces, while distance osteogenesis occurs on smooth implant surfaces. In the literature, there have been many in vitro model studies of bone formation on simulated dental implants using flattened titanium (Ti) discs; however, the purpose of this study was to identify the in vivo cell responses to the implant surfaces on actual, three-dimensional (3D) dental Ti implants and the surrounding bone in contact with such implants at the electron microscopic level using two different types of implant surfaces. In particular, the different parts of the implant structures were scrutinized. In this study, dental implants were installed in rabbit tibiae. The implants and bone were removed on day 10 and, subsequently, assessed using scanning electron microscopy (SEM), immunofluorescence microscopy (IF), transmission electron microscopy (TEM), focused ion-beam (FIB) system with Cs-corrected TEM (Cs-STEM), and confocal laser scanning microscopy (CLSM)—which were used to determine the implant surface characteristics and to identify the cells according to the different structural parts of the turned and roughened implants. The cell attachment pattern was revealed according to the different structural components of each implant surface and bone. Different cell responses to the implant surfaces and the surrounding bone were attained at an electron microscopic level in an in vivo model. These results shed light on cell behavioral patterns that occur during bone regeneration and could be a guide in the use of electron microscopy for 3D dental implants in an in vivo model.