Cortical Parenchyma Research Articles

Ultrastructural and Molecular Characterization of Platelet-derived growth factor Beta-Positive Leptomeningeal Cells in the Adult Rat Brain.

The leptomeninges, referring to the arachnoid and pia mater and their projections into the perivascular compartments in the central nervous system, actively participate in diverse biological processes including fluid homeostasis, immune cell infiltrations, and neurogenesis, yet their detailed cellular and molecular identities remain elusive. This study aimed to characterize platelet-derived growth factor beta (PDGFR-β)-expressing cells in the leptomeninges in the adult rat brain using light and electron microscopy. PDGFR-β+ cells were observed in the inner arachnoid, arachnoid trabeculae, pia mater, and leptomeningeal sheath of the subarachnoid vessels, thereby forming a cellular network throughout the leptomeninges. Leptomeningeal PDGFR-β+ cells were commonly characterized by large euchromatic nuclei, thin branching processes forming web-like network, and the expression of the intermediate filaments nestin and vimentin. These cells were typical of active fibroblasts with a well-developed rough endoplasmic reticulum and close spatial correlation with collagen fibrils. Leptomeningeal PDGFR-β+ cells ensheathing the vasculature in the subarachnoid space joined with pial PDGFR-β+ cells upon entering the cortical parenchyma, yet perivascular PDGFR-β+ cells in these penetrating vessels underwent abrupt changes in their morphological and molecular characteristics: they became more flattened with loss of immunoreactivity for nestin and vimentin and deficient collagen deposition, which was indicative of inactive fibroblasts termed fibrocytes. In the cortical parenchyma, PDGFR-β immunoreactivity was almost exclusively localized to larger caliber vessels, and significantly decreased in capillary-like microvessels. Collectively, our data identify PDGFR-β as a novel cellular marker for leptomeningeal fibroblasts comprising the leptomeninges and perivascular adventitial cells of the subarachnoid and penetrating large-sized cortical vasculatures.

Pharmacognostic and Preliminary Phytochemical studies on leaf and fruit of Adansonia digitata L.

Background: Adansonia digitata, the baobab, is the well-known tree species of the genus Adansonia. Aim and Objectives: Review reveals that the leaves and fruit of the plant are used traditionally in the treatment of fever, diarrhea, dysentery, ophthalmic, insect bites, inflammation, and pain. Although it is used traditionally, scientifically the plant is yet to be evaluated for its pharmacognostical characters. Hence, the plant parts were subjected to macro-microscopic, photomicrographic, physicochemical, and preliminary phytochemical tests to fix the quality standards for this drug. Method: Plant authentication, pharmacognostical study, physicochemical and phytochemical study was performed by following standard procedures as per Ayurvedic Pharmacopeia of India. Results: Microscopy of T. S. of rachis shows circular to orbicular in shape, in the cortical region parenchyma cells heavily loaded by tannin content, cluster and rosette crystals. Schematic outline of T. S. of the leaflet is bulged in the center and consist of the outer epidermis, mesophyll cells differentiated into upper palisade and lower spongy parenchyma while lower epidermis interrupted by anisocytic type of stomata. T.S. of fruit shows the presence of epidermis followed by layers of stone cells and sclerids and T.S. of seed shows outer hard testa, spool cells layer, inner large endosperm. Both samples show Loss on drying less than 6% and very few amounts of acid insoluble ash which directly indicate the amount of inorganic residue. Fruits are slightly acidic in nature. Qualitative tests showed the presence of carbohydrate, tannin, flavonoid, and steroids in leaves and fruits. Conclusion: This study would be useful in the identification and authentication of the raw drug.

Neuroinflammatory Response in Chronic Hydrocephalus in Juvenile Rats

Hydrocephalus is especially prevalent in countries with limited resources, where its treatment is still a challenge. However, long-term neuropathological changes in untreated hydrocephalus remain largely unexplored. The present study looks at cortical parenchyma and neuroinflammation in acquired, chronic hydrocephalus. Intracisternal kaolin injections were performed in 3-week-old rats, followed by -1, 4- and 8-week survival; matched control rats received saline injections. Ventriculomegaly has been previously reported to stabilize by the third week in this model. Single and triple immunocytochemical approaches were used to highlight neurones, astrocytes, microglia, and the pro-inflammatory cytokine interleukin (IL)-1β in the parietal cortex, utilizing cell counts and densitometry. Microglial protein ionized calcium binding adaptor molecule 1 (Iba1) and IL-1β expressions were monitored with Western blotting in the parietal cortex and hippocampus. In the parietal cortex, which showed progressive disruption of cytoarchitecture, neuronal density was significantly increased at 8weeks post-induction but not at earlier time points, indicating on-going cortical damage in chronic hydrocephalus. Astrocyte and microglia hypertrophy, and Iba1 expression indicated glial cell activation which peaked at 4weeks. IL-1β expression also peaked at 4weeks and was then down-regulated. Overall the findings indicate that neuroinflammatory features build up in the first month after hydrocephalus induction implicating marked IL-1β upregulation. The data also show that astrocytes are the main source of IL-1β in this disorder.

Chemical composition of cell walls in velamentous roots of epiphytic Orchidaceae.

The chemical composition of the cell walls strongly affects water permeability and storage in root tissues. Since epiphytic orchids live in a habitat with a highly fluctuating water supply, the root cell walls are functionally important. In the present study, we used histochemistry and immunocytochemistry techniques in order to determine the composition of the cell walls of root tissues of 18 epiphytic species belonging to seven subtribes across the Orchidaceae. The impregnation of lignin in the velamen cells reinforces its function as mechanical support and can facilitate apoplastic flow. Pectins, as well cellulose and lignins, are also essential for the stability and mechanical support of velamen cells. The exodermis and endodermis possess a suberinized lamella and often lignified walls that function as selective barriers to apoplastic flow. Various cortical parenchyma secondary wall thickenings, including phi, reticulated, and uniform, prevent the cortex from collapsing during periods of desiccation. The presence of highly methyl-esterified pectins in the cortical parenchyma facilitates the formation of gels, causing wall loosening and increased porosity, which contributes to water storage and solute transport between cells. Finally, cells with lipid or lignin impregnation in the cortical parenchyma could increase the water flow towards the stele.

Effect of pruning strategy on 'Syrah' bud necrosis and fruitfulness in Brazilian subtropical Southeast

The change of wine grape harvest from wet season (summer) to dry season (winter) by changing the pruning management has improved quality of wines produced in the Brazilian Southeast. However, the vines need to be spur pruned twice a year, i.e. with a 1st pruning in August (winter pruning) for a vegetative cycle during the hot and wet summer, and a 2nd pruning in January (summer pruning) for a productive cycle during the cold and dry season. This double pruning strategy is made necessary by the fact that latent buds developed during the dry season cycle are not fruitful to support a productive cycle in the following year. This histological study, performed in the South of Minas Gerais State (Brazil), showed that annual single pruning done in the wet season (in January) displayed a high rate of necrosis on primary and secondary buds (bud necrosis – BN). In April, 99 days after summer pruning (DASP), the rates of BN were 40 % and 50 % at basal and apical node positions, respectively, reaching 80 % of BN in December (322 DASP). As a consequence of BN, bud potential fertility was drastically reduced from 0.5 inflorescence primordial (IP) per bud (in July) to 0.06 (in December) and bud burst in the next cycle from secondary and tertiary bud axes. Vines managed by double pruning system (submitted to summer and winter pruning) displayed a much higher fruitfulness potential, i.e. 1.46 IP per bud in December (112 days after winter pruning) and limited BN occurrence (20 %). On single pruned vines, we also observed a significant decrease of starch content in canes, trunks and roots. Internal bud anatomy showed that a random cell breakdown started 70 days DASP. At 211 DASP, all buds showed a large starch granule concentration, raphides and crystals of calcium oxalate inside idioblasts of leaf primordia and also in cortical parenchyma of the vegetative axis. The bud starch content was increased and a positive correlation between necrosis and starch accumulation was observed. The impact of carbohydrate availability on bud necrosis development was discussed. This study showed that the necrosis development towards secondary and tertiary axis of the dry season buds is the main reason of unfruitfulness in the vineyards managed by single pruning in the wet season, making the double pruning compulsory.

Recovery Processes in the Adrenal Glands of Dogs

The aim of this research was a comparative study of cyclic changes in secretory, destructive and proliferative processes in the adrenal cortex in dogs. Material and methods . Cyclic changes of morphofunctional indices of adrenal cortex in dogs (Canis familiaris) were studied. Healthy mature males were used. The material was collected in the period from December to April with a monthly interval. For light microscopy the fragments of the adrenal glands were fixed in Steve, Bouin and Zenker liquids, 10% neutral formalin solution. PAS-reaction was used. RNA was detected by the Brachet reaction and gallocyanine according to Einarsson. The material fixed in a 10% solution of neutral formalin was used to study lipids. Results. In January, in the adrenal cortex clearly expressed glomerular, fasciculate and reticulae zones. In the glomerular zone elongated cells with oval nuclei are situated, in the fascicular - polygonal cells with rounded nuclei. In February, a decrease in the functional activity of the cortical parenchyma and the development of extensive destructive changes in glandular cells, especially in the glomerular zone, were found in the adrenal cortex. In March, the development of recovery processes and increased secretory activity of the adrenal cortex were noted. Proliferative processes are accompanied by loosening of the capsule, the formation of connective tissue strands going into the fascicular zone. Among the fibers of connective tissue fibroblasts, fibrocytes and small poorly differentiated cells with rounded nuclei are disposed. These cells are converted into elongated cells of the glomerular zone and polygonal cells of the fascicular zone.

In situ histochemical localization of alkaloids in leaves and pods of Prosopis ruscifolia

The tree Prosopis ruscifolia produces indolizidine alkaloids which function in plant defense as antimicrobials. Here, we propose the use of Drangendorff´s reagent, picric acid and lugol for the in situ histochemical localization of indolizidine alkaloids in the leaf parts (petiole, rachis and leaflet) and pods of P. ruscifolia. The main surface and internal anatomical features of the aerial plant parts were visualized with cresyl violet and astra blue safranin. Extracts from leaflets and pods were separated by TLC and revealed with the Dragendorff´s reagent, picric acid and lugol. Fresh cross sections of leaflets, petioles, rachis and pods were also stained with these chromogenic reagents. Glandular trichomes were absent from the surface of mature leaflets, petioles, rachis and pods. Large idioblasts were distributed in the palisade mesophylls of the leaflets and the cortical and medullar parenchyma of the petiole and the rachis. TLC of the extracts revealed with Dragendorff´s reagent, picric acid and lugol gave orange, yellow and light brown spots for the indolizidine alkaloids, respectively. Spraying with picric acid also revealed some non-alkaloid constituents. The three chromogenic reagents indicated the presence of indolizidine alkaloids in the large idioblasts of the leaflets, petioles, rachis and pods. Dragendorff´s reagent and picric acid also showed the alkaloids in the cuticles of all leaf parts and pods. The first was the most sensitive towards alkaloids and the only one that visualizes them in the phloem of the petioles and pods. The indolizidine alkaloids of P. ruscifolia seemed to be transported from green tissues to the pods, and should exert their protective function after the breakdown of leaves and pods.

Sex Differences in Acidosis‐Stimulated Renal Ammonia Metabolism

Renal ammonia excretion is a critical component in maintaining acid‐base homoeostasis and changes in ammonia excretion are the predominant component of increased net acid excretion in response to metabolic acidosis. We reported recently substantial sex‐dependent differences in basal ammonia metabolism that correlate with sex‐dependent structural differences and differences in the expression of key proteins involved in ammonia metabolism. This study's objective was to determine the effect of sex on the renal ammonia response to an exogenous acid load. We compared 4‐month old C57BL/6 male (M) and female (F) mice. Mice were acid‐loaded by adding HCl to powdered chow for 7 days, and control mice had equal amounts of deionized water added. Basal ammonia excretion, factored for body weight, was ~2‐fold greater in female than male mice, but the response to acid‐loading did not differ significantly. There was no significant sex difference in serum Na+, K+, and HCO3− after an acid load. The basal fundamental structural differences between the female and male kidney were maintained after an acid load. After acid‐loading, in the female kidney, proximal tubules account for a lower percentage of the cortical parenchyma than in the male kidney (F, 43±2; M, 61±3%; P < 0.001), whereas the collecting ducts account for a greater percentage of the renal parenchyma (F, 16±1; M, 11±1%; P < 0.01) which are similar to basal conditions. Cortical PCT and PST cell height increased in response to acid‐loading, and the increase was greater in M than F kidney. In the outer medulla (OM), PST cell height was similar in M and F, and sex did not alter the increase with acid‐loading. Similarly, intercalated cell (IC) size in the collecting duct in the OM was greater in F than in M kidney, and increased with acid‐loading in both sexes; sex did not alter the acid‐loading increase. Cortical expression of phosphoenol pyruvate, a major proximal tubule (PT) ammonia generating protein, was greater under control conditions in F, but the relative increase with acid‐loading was greater in M. Expression of glutamine synthetase, which recycles ammonia, significantly decreased in response to an acid load, with no significant sex difference. Expression of total cortical NBCe1, but not NBCe1‐A, increased in response to acid‐loading. Expression of NHE3, believed to be the major mechanism of PT ammonia secretion, increased in response to an acid load in M, but not in F. Cortical NKCC2, which mediates thick ascending limb ammonia reabsorption, was significantly greater in F than M after an acid load. The collecting duct secretes the majority of urinary ammonia and the Rhesus glycoproteins, Rhbg and Rhcg, are the primary collecting duct ammonia transporting proteins. Acid‐loading increased cortical Rhbg expression in M, but not in F in the cortex, whereas in the OM, Rhbg expression increased similarly in both M and F. Rhcg expression in intercalated cells in the inner stripe of the outer medulla was significantly greater in F than in M mice after an acid load. We conclude that there are substantial sex‐dependent differences in the mechanism of renal ammonia response to acid‐loading that enable a similar maximum ammonia excretion response.Support or Funding InformationNIH R01DK045788, R01DK107798 and 5T32DK104721This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Transcriptome analysis of rubber biosynthesis in guayule (Parthenium argentatum gray)

BackgroundNatural rubber is currently produced nearly exclusively from latex of the Para rubber tree, Hevea brasiliensis. The desire to reduce the environmental cost of rubber production, fears of pathogen susceptibility in clonal Hevea plantations, volatility in the price of natural rubber, and increasing labor costs have motivated efforts to diversify the supply of natural rubber by developing alternative rubber crops such as guayule (Parthenium argentatum Gray). In Hevea, latex is produced as an exudate following wounding while in guayule, rubber is deposited within the cortical parenchyma and its production is strongly influenced by environmental conditions.ResultsTo better understand the enzymology and regulation of guayule rubber biosynthesis and to identify genes with potential uses in the improvement of rubber yields, we conducted de novo transcriptome assembly and differential gene expression analyses of this process in guayule. This analysis supports a role for rubber in the defense against pathogens, identified new enzymes potentially involved in the biosynthesis of rubber as well as transcription factors specifically expressed in rubber-producing tissues.ConclusionsData presented here will be useful in the improvement of guayule as an alternative source of natural rubber and in better understanding the biosynthesis of this critical polymer. In particular, some of the candidate transcription factors are likely to control the rubber biosynthesis pathway and are good targets for molecular breeding or engineering of guayule plants with higher and more consistent production of rubber.

Root structural changes of two remediator plants as the first defective barrier against industrial pollution, and their hyperaccumulation ability.

In the present day, plants are increasingly being utilized to safeguard the environment. In this study, we used Salsola crassa M. B. and Suaeda maritima L. Dumort for phytoremediation of water contaminated with heavy metals and simultaneous examination of the effect of industrial pollution on their root structures. After irrigation of a treatment group with wastewater and a control group with fresh water for 3months, we fixed the root parts in the FAA fixator for developmental study, and measured the concentrations of Co, Ni, Zn, As, Cu, and Pb in the roots, shoots, soil, and irrigating water. The plants irrigated with wastewater showed significant accumulation of heavy metals in the roots and some translocation of heavy metals from the roots to the shoots. We also performed an experiment with two 0.3m3 pools to more closely study the feasibility of these plants for filtering water of contaminants, including mineral compounds, and altering its chemical characteristics. In our anatomical studies, the cells of the treatment roots showed irregularities and abnormal appearances in all tissue layers. The diameter and area of the xylem and the size of the cortical parenchyma have increased in the treatment plants of both species, confirmed by Stereolite software. Phytoremediation studies indicated that S. crassa accumulated As, Cu, Zn, Pb, Co, and Ni, and S. maritima accumulated As, Co, Zn, and Cu. S. crassa accumulated more heavy metals in its roots, whereas S. maritima accumulated more in its shoots. The biological oxygen demand and chemical oxygen demand were also significantly reduced in the wastewater passed through pools with S. crassa. Our results indicate that both genera are hyperaccumulators of heavy metals and therefore hold promise for industrial wastewater treatment, especially the absorption of As.

The effect of biochar on the physiological, morphological and anatomical characteristics of mung bean roots after exposure to salt stress

To investigate the changes in the anatomical and physiological characteristics of mung bean roots in response to biochar treatment during salt stress, a pot experiment was conducted. Mung bean plants were subjected to three biochar concentrations (0, 50 and 100 g kg-1 soil) and three salinity treatments (0, 5 and 10 dSm-1 NaCl). Salinity decreased root growth, vascular cylinder (VC) and cortical parenchyma (CP) areas, affecting the VC/CP ratio, shoot dry weight, the relative water content (RWC) of roots and leaves, and the root indole-3-acetic acid (IAA) content. It increased specific root length, the shoot/root ratio and root abscisic acid (ABA), and 1-aminocyclopropane-1-carboxylic acid (ACC) contents. Plant growth, RWC, the shoot/root ratio, specific root length, total root area, VC and CP areas, and the IAA/ABA and IAA/ACC ratios were increased by biochar under saline media. Biochar improved xylem structure, the plant growth regulator IAA, and decreased stress hormones, ABA and ACC, which accelerate plant senescence, consequently increasing mung bean growth under salt stress.

Robust kidney perfusion mapping in pediatric chronic kidney disease using single-shot 3D-GRASE ASL with optimized retrospective motion correction.

To develop a robust renal arterial spin labeling (ASL) acquisition and processing strategy for mapping renal blood flow (RBF) in a pediatric cohort with severe kidney disease. A single-shot background-suppressed 3D gradient and spin-echo (GRASE) flow-sensitive alternating inversion recovery (FAIR) ASL acquisition method was used to perform 2 studies. First, an evaluation of the feasibility of single-shot 3D-GRASE and retrospective noise reduction methods was performed in healthy volunteers. Second, a pediatric cohort with severe chronic kidney disease underwent single-shot 3D-GRASE FAIR ASL and RBF was quantified following several retrospective motion correction pipelines, including image registration and threshold-free weighted averaging. The effect of motion correction on the fit errors of saturation recovery (SR) images (required for RBF quantification) and on the perfusion-weighted image (PWI) temporal signal-to-noise ratio (tSNR) was evaluated, as well as the intra- and inter-session repeatability of renal longitudinal relaxation time (T1 ) and RBF. The mean cortical and/or functional renal parenchyma RBF in healthy volunteers and CKD patients was 295 ± 97 and 95 ± 47 mL/100 g/min, respectively. Motion-correction reduced image artefacts in both T1 and RBF maps, significantly reduced SR fit errors, significantly increased the PWI tSNR and improved the improved the repeatability of T1 and RBF in the pediatric patient cohort. Single-shot 3D-GRASE ASL combined with retrospective motion correction enabled repeatable non-invasive RBF mapping in the first pediatric cohort with severe kidney disease undergoing ASL scans.

The proteome microenvironment determines the protective effect of preconditioning in cisplatin-induced acute kidney injury

Acute kidney injury (AKI) leads to significant morbidity and mortality; unfortunately, strategies to prevent or treat AKI are lacking. In recent years, several preconditioning protocols have been shown to be effective in inducing organ protection in rodent models. Here, we characterized two of these interventions-caloric restriction and hypoxic preconditioning-in a mouse model of cisplatin-induced AKI and investigated the underlying mechanisms by acquisition of multi-layered omic data (transcriptome, proteome, N-degradome) and functional parameters in the same animals. Both preconditioning protocols markedly ameliorated cisplatin-induced loss of kidney function, and caloric restriction also induced lipid synthesis. Bioinformatic analysis revealed mRNA-independent proteome alterations affecting the extracellular space, mitochondria, and transporters. Interestingly, our analyses revealed a strong dissociation of protein and RNA expression after cisplatin treatment that showed a strong correlation with the degree of damage. N-degradomic analysis revealed that most posttranscriptional changes were determined by arginine-specific proteolytic processing. This included a characteristic cisplatin-activated complement signature that was prevented by preconditioning. Amyloid and acute-phase proteins within the cortical parenchyma showed a similar response. Extensive analysis of disease-associated molecular patterns suggested that transcription-independent deposition of amyloid P-component serum protein may be a key component in the microenvironmental contribution to kidney damage. This proof-of-principle study provides new insights into the pathogenesis of cisplatin-induced AKI and the molecular mechanisms underlying organ protection by correlating phenotypic and multi-layered omics data.

Metabolomics of mammalian brain reveals regional differences

BackgroundThe mammalian brain is organized into regions with specific biological functions and properties. These regions have distinct transcriptomes, but little is known whether they may also differ in their metabolome. The metabolome, a collection of small molecules or metabolites, is at the intersection of the genetic background of a given cell or tissue and the environmental influences that affect it. Thus, the metabolome directly reflects information about the physiologic state of a biological system under a particular condition. The objective of this study was to investigate whether various brain regions have diverse metabolome profiles, similarly to their genetic diversity. The answer to this question would suggest that not only the genome but also the metabolome may contribute to the functional diversity of brain regions.MethodsWe investigated the metabolome of four regions of the mouse brain that have very distinct functions: frontal cortex, hippocampus, cerebellum, and olfactory bulb. We utilized gas- and liquid- chromatography mass spectrometry platforms and identified 215 metabolites.ResultsPrincipal component analysis, an unsupervised multivariate analysis, clustered each brain region based on its metabolome content, thus providing the unique metabolic profile of each region. A pathway-centric analysis indicated that olfactory bulb and cerebellum had most distinct metabolic profiles, while the cortical parenchyma and hippocampus were more similar in their metabolome content. Among the notable differences were distinct oxidative-anti-oxidative status and region-specific lipid profiles. Finally, a global metabolic connectivity analysis using the weighted correlation network analysis identified five hub metabolites that organized a unique metabolic network architecture within each examined brain region. These data indicate the diversity of global metabolome corresponding to specialized regional brain function and provide a new perspective on the underlying properties of brain regions.ConclusionIn summary, we observed many differences in the metabolome among the various brain regions investigated. All four brain regions in our study had a unique metabolic signature, but the metabolites came from all categories and were not pathway-centric.

The Hypomorphic Gata1low Mutation Induces Fibrosis in Multiple Organs

Mice carrying the hypomorphic mutation which reduces the transcription factor GATA1 in megakaryocytes (Gata1low mice), develop by 8-months myelofibrosis a phenotype resembling primary myelofibrosis, the most severe of myeloproliferative neoplasms (Vannucchi et al Blood 2002;100:1123). The high levels of TGF-β expressed by the abnormal Gata1low megakaryocytes that drive myelofibrosis in these mice alter the bone marrow (BM) expression profiling up-regulating expression of the transcription factor c-Jun (Ciaffoni et al BCD 2015; 54:234). Recently, Dr. Weissman reported that c-Jun over-expression in response to TGF-β activation induces in mice fibrosis in multiple organs (Werning et al PNAS 2017, 114, 4757), suggesting that also Gata1low mice may develop multi-organ fibrosis. To test this hypothesis, we compared morphology (by haematoxylin-eosin staining) and fibrosis (reticulin fibres by trichrome Mallory and collagen fibres by Gomory or Sirius Red Picrate staining) of organs from Gata 1low and wild-type (WT) mice at 1-, 8- and 15-months of age (3 mice/group).With age, the organs from WT mice presented histological abnormalities consistent with the mild one expected for being associated with aging and were seldom positive for fibrosis. By contrast, all the organs from Gata1low mice analysed had profound abnormal morphologies with reticulin or collagen fibers detectable, in addition to BM and spleen, in skin, lung, and kidney in an age-specific fashion (Fig 1). Gata1lowskin was thicker than normal and the connective layer presented numerous reticulin strikes already by 1-month and heavily dense connective regions strongly positive for collagen fibres by 8-15 months, resembling the skin from scleroderma patients. In lung, the alveoli had thickened walls with reticulin fibers detectable by 8-months and collagen bundles by 15-months near the bronchus walls. The abnormal morphology of kidney included reduced numbers of glomeruli and poorly organized cortical parenchyma. Reticulin and collagen fibres were observed in the medullary and nephron region by 8- and 15-months, respectively. In heart, cardiomyocytes presented a strong reduction of intercalary disks by 8-months and reticulin fibers were detectable at 15-months, suggesting that this mild fibrosis is driven by lung insufficiency. The liver presented abnormal localization and morphology of hepatocytes and presence of extramedullary hematopoiesis in perisinusoid areas. Sirius Red Picrate staining revealed few reticulin fibres mostly within erythroid islands.To clarify the mechanisms leading to multi-organ fibrosis in Gata1low mice, the transcription signature of 8-months Gata1low BM was compared with that published for murine liver, lung and kidney fibrosis (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi? token = ijgpwgsyjhwpdoj&acc = GSE89630). Gata1low BM presented abnormal expression of 1724 genes (821 up- and 903 down). Pathway analyses indicated that most of the down genes were in the Gata1 pathway while the most prominent up genes were in the c-Jun, EZH2, SCL and p53 pathways. Some of the gene abnormal in Gata1low BM were also abnormal in liver (64), lung (29) and kidney (432) fibrosis. Only 28 genes were abnormal in more than one organ (25 liver and kidney, 7 lung). These common genes were obvious markers for fibrosis (5 collagen genes, fibronectin, TGF-β) and did not include c-Jun. Among genes up in Gata1low BM, kidney and liver there was lipocalin 2 (LCN2), a growth factor overexpressed in primary myelofibrosis patients (Lu et al Blood 2015;126:972) that exerts positive and negative effects, respectively, on fibrosis in BM and liver. By comparing the plasma levels of LCN2 in Gata1low and WT mice at 8-9 -and 15-17-months (8-29 mice/group), we determined that levels of LNC2 do not change with age in WT mice but increase by 3-fold in old Gata1low mice (p=0.028), explaining why liver was not one of the organs in which the mutants develop fibrosis.In conclusion, in addition to myelofibrosis, Gata1low mice develop fibrosis in skin, lung and kidney but not in heart and liver and represent genetic models for studies on the pathogenesis of fibrosis in multiple organs. Moreover, these results suggest that although the initiation factor(s) for fibrosis in the various organs are likely different, most of them are expressed by Gata1low megakaryocytes highlighting the importance for studies on the secretome profile of these cells. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Karyometric and electron microscopic studies of dark and light cells of the adrenal cortex in mammals

The purpose of this study is a comparative study of dark and light cells of cortical parenchyma of the adrenal glands in mammals using light and electron microscopy. Material and methods. Cariometric and electron microscopic studies of the adrenal glands of 9 mammalian species have been carried out: Vulpes vulpes (fox), Vulpes lagopus (arctic fox), Canis familiaris (dog), Мartes zibellina (sable), Mustela vison (mink), Enhydra lutris (sea ape); Castor fiber (river beaver); Callorhinus ursinus (fur seal); Ovis aries (sheep). Serial paraffin sections were stained with hematoxylin and eosin, iron hematoxylin, Heidenheim azan, the PAS-reaction was used, the color of the trichrome-PAS and the tetrachromium-PAS. RNA was detected by the reaction of Brashe and gallocyanin by Einarsson. For the detection of lipids, frozen sections were stained with Sudan III + IV, Black Sudan. The number of dark and light cells in the cortex of the adrenal mink in different seasons was determined. On electron micrographs, the amount of lipid droplets and mitochondria in the fascicle of the adrenal cortex was counted. Results. In the cortex of the adrenal glands of mammals, dark and light cells are present. Dark cells, as far as accumulation of sudanophilic substances are converted into light cells, which with further accumulation of lipids undergo destructive changes. Also found are the dark dying cells of the adrenal cortex, characterized by densification of the cytoplasm, pycnosis of nuclei, accumulation of yellow-brown pigment. Dark cells have larger nuclei, give more intense reaction to RNA and ketosteroids and are more active secretory cells compared to light adrenocorticocytes. Electron microscopy in dark cells reveals a large number of mitochondria and a significant amount of lipid droplets are present. Light cells contain many lipid inclusions and few mitochondria.

Morphological and anatomical changes in soybean roots subjected to indole-3-acetic acid and tryptophol: indole compounds present in plant auxin metabolism

The auxin metabolism is practically elucidated, and the compounds that are part of the biosynthesis are well characterized, but the indole-3-ethanol or tryptophol, a molecule that has a regulatory position in the indole-3-acetic acid biosynthesis, still represents a gap in the understanding of this pathway. We examined the hypothesis that tryptophol present the function of plant growth regulation on soybean root development. We evaluated two doses of auxin and two doses of tryptophol (100 e 200 mg L− 1), respectively, beside a control treatment (water), via leaf application, in soybean plants under V1–V2 phonological stages. After 18 days of application, the roots were collected for their volume and area measurement, thereafter small segments (0.5 cm of length), were collected at 1 cm below the root-collar, for anatomical analysis. We observed that the control showed greater area and root volume, but using 200 mg L− 1 auxin and 100 mg L− 1 tryptophol led to a radial increase of roots with significant increases in width radius vascular and cortical parenchyma. These results suggest that the application of both compounds had a potential of modify the vascular and ground tissues in soybean roots, which may be beneficial for the development of plants.

Initial stages of minimal processing of red beets result in significant loss of bioactive compounds

This study aims to detect the main stages in which bioactive compounds are lost during minimal processing of red beet and to comprehend more deeply the influence of each processing stage in the quality of minimally processed beet. Samples of beets were collected during the stages of preparation of this minimally processed produce, analyzed for betalains (betacyanins and betaxanthins) and phenolic compounds content, as well as polyphenoloxidase (PPO) activity, and performed anatomical analysis. Initial stages of minimal processing of red beet result in significant losses of bioactive compounds. Peeling accounts for one-quarter of betacyanins losses. Betaxanthins were expressively reduced at the first sanitation, resulting in almost half decrease. Anatomical sections have shown that cells of the cortical parenchyma are smaller and vascular parenchymal tissue was characterized by larger cells, well visible intercellular spaces and containing single vacuole. In conclusion, initial stages result in bigger losses of betalains and phenolic compounds during minimal processing of red beet. These stages promote PPO leaching, as its location in organelles is compromised by cell decompartmentalization. However, because these stages are essential for minimally processed produce, strategies aiming to reduce losses should focus on other factors.

Arterial spin labeling MR imaging aids to identify cortical venous drainage of dural arteriovenous fistulas.

Cortical venous drainage (CVD) increases the probability of intracranial hemorrhage and mortality rate of dural arteriovenous fistulas (DAVF). Although digital subtraction angiography (DSA) is the most accurate method to determine CVD in DAVFs, this modality has limitations due to its invasive nature and radiation issues. The purpose of this study was to evaluate the diagnostic utility of arterial spin-labeling perfusion-weighted images (ASL-PWI) to identify CVD in patients with DAVF.The Institutional Review Board of our hospital approved this retrospective study. ASL-PWI features of 22 patients with DAVF were retrospectively reviewed for the presence of bright signal intensity in cortical veins and brain parenchyma. DAVF with bright signal intensity in cortical veins and/or brain parenchyma was regarded as having CVD. Using DSA as a reference standard, sensitivity, specificity, positive predictive value, and negative predictive value of ASL-PWI for detecting CVD were calculated.Based on DSA features, 11 (11/22, 50%) patients were classified as having “aggressive” pattern with CVD. Eleven (11/22, 50%) patients also showed bright signal intensity in cortical veins (9/22, 41%) and/or brain parenchyma (4/22, 18%) on ASL-PWI. The 11 patients who had “Aggressive” pattern on DSA were the same 11 patients who were classified as having “aggressive” pattern on ASL-PWI. ASL-PWI showed perfect diagnostic performance for identifying CVD with sensitivity, specificity, positive predictive value, and negative predictive value of 100% for all.Thus, ASL-PWI could be used as a noninvasive mean to predict the presence of CVD in patients with DAVFs. It has the potential as a screening tool to evaluate DAVF prior to invasive DSA.

Improving Microcirculatory Reperfusion Reduces Parenchymal Oxygen Radical Formation and Provides Neuroprotection

Reperfusion is the most significant determinant of good outcome after ischemic stroke. However, complete reperfusion often cannot be achieved, despite satisfactory recanalization. We hypothesized that microvascular protection was essential for achieving effective reperfusion and, hence, neuroprotection. To test this hypothesis, we have developed an in vivo model to differentially monitor parenchymal and vascular reactive oxygen species (ROS) formation. By comparing the ROS-suppressing effect of N-tert-butyl-α-phenylnitrone (PBN) with its blood-brain barrier impermeable analog 2-sulfo-phenyl-N-tert-butylnitrone (S-PBN), we assessed the impact of vascular ROS suppression alone on reperfusion and stroke outcome after recanalization. The distal middle cerebral artery was occluded for 1 hour by compressing with a micropipette and then recanalized (n=60 Swiss mice). ROS formation was monitored for 1 hour after recanalization by intravital fluorescence microscopy in pial vasculature and cortical parenchyma with topically applied hydroethidine through a cranial window. PBN (100 mg/kg) or S-PBN (156 mg/kg) was administered shortly before recanalization, and suppression of the vascular and parenchymal hydroethidine fluorescence was examined (n=22). Microcirculatory patency, reperfusion, ischemic tissue size, and neurological outcome were also assessed in a separate group of mice 1 to 72 hours after recanalization (n=30). PBN and S-PBN completely suppressed the reperfusion-induced increase in ROS signal within vasculature. PBN readily suppressed ROS produced in parenchyma by 88%. S-PBN also suppressed the parenchymal ROS by 64% but starting 40 minutes later. Intriguingly, PBN and S-PBN comparably reduced the size of ischemic area by 65% and 48% (P>0.05), respectively. S-PBN restored the microvascular patency and perfusion after recanalization, suggesting that its delayed parenchymal antioxidant effect could be secondary to improved microcirculatory reperfusion. Promoting microvascular reperfusion by protecting vasculature can secondarily reduce parenchymal ROS formation and provide neuroprotection. The model presented can be used to directly assess pharmacological end points postulated in brain parenchyma and vasculature in vivo.