Depth-targeted intracortical microstroke by two-photon photothrombosis in rodent brain

Significance: Photothrombosis is a widely used model of ischemic stroke in rodent experiments. In the photothromboris model, the photosensitizer Rose Bengal is systemically introduced to the blood stream and activated by green light to induce aggregation of platelets that eventually cause vessel occlusion. Since the activation of Rose Bengal is a one-photon phenomenon and the molecules in the illuminated area (light path) are subject to excitation, targeting of thrombosis is unspecific especially in the depth dimension. We have developed a photothrombosis protocol that can target a single vessel in the cortical parenchyma by two-photon excitation. Aim: We aim to induce a thrombotic stroke in the cortical parenchyma by two-photon activation of Rose Bengal so that we confine photothrombosis within a vessel of a target depth. Approach: FITC-dextran is injected into the blood stream to visualize the cerebral blood flow in anesthetized adult mice with a cranial window. After a target vessel is chosen by two-photon imaging (950 nm), Rose Bengal is injected into the blood stream. The scanning wavelength is changed to 720 nm and photothrombosis was induced by scanning the target vessel. Results: Two-photon depth-targeted single vessel photothrombosis was achieved with a success rate of 84.9+/-1.7% within 80 s. Attempts without Rose Bengal (i.e., only with FITC) did not result in photothrombosis at the excitation wavelength of 720 nm. Conclusions: We described a protocol that achieves depth-targeted single vessel photothrombosis by two-photon excitation. Simultaneous imaging of blood flow in the targeted vessel using FITC dextran enabled the confirmation of vessel occlusion and prevention of excess irradiation that possibly induces unintended photodamage.

Ultrastructural Study of the Outer Cortex, Deep Cortex and Medulla of Mesenteric Lymph Node in Adult Goats

Background: A mesenteric lymph node in the sub pyloric region which is the longest among all mesenteric lymph nodes in adult goats was identified for study under the scanning electron microscope. Methods: The outer cortical, deep cortical and medullary parenchyma of the collected lymph nodes was thoroughly screened under VEGA3 TESCAN. The outer cortical parenchyma revealed T lymphocytes, B lymphocytes, Sinus Lining Cells (SLC) and Sinus Reticular Cells (SRC) at a magnification of 9600x. The deep cortical parenchyma revealed Reticular cells, T lymphocytes and Interdigitating cells (IDC) at 8000x magnification.Result: The medullary parenchyma revealed T lymphocytes, B lymphocytes, Sinus Lining Cells (SLC), Fibrous Reticular Cells (FRC) and medullary sinuses at a magnification of 8500x. Plasma cells and lymphocytes also remained in the sinus. The morphology and relations of the cells observed were recorded.

Densitometric analysis of brain computed tomography as a new prognostic factor in patients with acute subdural hematoma

OBJECTIVEAcute subdural hematoma (ASDH) is a major cause of mortality and morbidity after traumatic brain injury (TBI). Surgical evacuation is the mainstay of treatment in patients with altered neurological status or significant mass effect. Nevertheless, concerns regarding surgical indication still persist. Given that clinicians often make therapeutic decisions on the basis of their prognosis assessment, to accurately evaluate the prognosis is of great significance. Unfortunately, there is a lack of specific and reliable prognostic models. In addition, the interdependence of certain well-known predictive variables usually employed to guide surgical decision-making in ASDH has been proven. Because gray matter and white matter are highly susceptible to secondary insults during the early phase after TBI, the authors aimed to assess the extent of these secondary insults with a brain parenchyma densitometric quantitative CT analysis and to evaluate its prognostic capacity.METHODSThe authors performed a retrospective analysis among their prospectively collected cohort of patients with moderate to severe TBI. Patients with surgically evacuated, isolated, unilateral ASDH admitted between 2010 and 2017 were selected. Thirty-nine patients were included. For each patient, brain parenchyma density in Hounsfield units (HUs) was measured in 10 selected slices from the supratentorial region. In each slice, different regions of interest (ROIs), including and excluding the cortical parenchyma, were defined. The injured hemisphere, the contralateral hemisphere, and the absolute differences between them were analyzed. The outcome was evaluated using the Glasgow Outcome Scale–Extended at 1 year after TBI.RESULTSFifteen patients (38.5%) had a favorable outcome. Collected demographic, clinical, and radiographic data did not show significant differences between favorable and unfavorable outcomes. In contrast, the densitometric analysis demonstrated that greater absolute differences between both hemispheres were associated with poor outcome. These differences were detected along the supratentorial region, but were greater at the high convexity level. Moreover, these HU differences were far more marked at the cortical parenchyma. It was also detected that these differences were more prone to ischemic and/or edematous insults than to hyperemic changes. Age was significantly correlated with the side-to-side HU differences in patients with unfavorable outcome.CONCLUSIONSThe densitometric analysis is a promising prognostic tool in patients diagnosed with ASDH. The supplementary prognostic information provided by the densitometric analysis should be evaluated in future studies.

Management of Fire Blight Using Pre-bloom Application of Prohexadione-Calcium

Fire blight, a bacterial disease of rosaceous plants caused by Erwinia amylovora, is one of the most important diseases affecting commercial apple production worldwide. Antibiotics, applied at bloom to protect against blossom infection, are the most effective means of management but raise concern due to the potential for antibiotic resistance in both the pathogen population and nontarget organisms. In addition, most fire blight outbreaks in New York State often emerge in late June to July as shoot blight, calling into question the role of blossom infections and the antibiotic applications made to manage them. Prohexadione-calcium (PhCa) is a gibberellic acid inhibitor used post-bloom to control shoot vigor and to manage shoot blight. However, the magnitude of shoot blight management is directly related to the suppression of shoot growth, which is undesirable, especially in young orchards during establishment years. PhCa is believed to control shoot blight by thickening cell walls in cortical parenchyma, preventing invasion of host tissues by E. amylovora. We hypothesize that PhCa applied pre-bloom could similarly prevent invasion of blossom pedicels following infection, leading to reduced disease incidence. We evaluated novel pre-bloom PhCa programs for their effects on disease management (blossom and shoot blight) as well as their impact on shoot growth for three years in a mature ‘Gala’ orchard in New York. In all three years of the study, all PhCa programs resulted in less than 27% incidence (71% control) of blossom blight and less than 13% incidence (77% control) of shoot blight with minimal effect on tree growth. Inclusion of a biopesticide during bloom further reduced the incidence of blossom blight in one year of three. Using light microscopy, we found that cell walls in the cortical parenchyma of fruitlet pedicels on trees receiving pre-bloom PhCa applications were significantly thicker than those of untreated trees 40 days after full bloom and inoculation. Overall, we found that pre-bloom applications of PhCa had utility in reducing blossom blight and shoot blight with minimal impacts on tree growth. These pre-bloom programs would fit with standard production practices and may contribute toward the development of fire blight management programs without the use of antibiotics.

МОРФОЛОГО-АНАТОМІЧНЕ ДОСЛІДЖЕННЯ БЕДРИНЦЮ ЛОМИКАМЕНЕВОГО (PIMPINELLA SAXIFRAGA L.)

<p><strong>MORPHOLOGICAL AND ANATOMICAL RESEARCH </strong></p><p><strong>OF </strong><strong>P</strong><strong>IMPINELLA SAXIFRAGA</strong><strong> L.</strong><strong></strong></p><p><strong>S.M.Marchyshyn, T.M. Hontova, E.A. Panasiuk</strong></p><p> </p><p>“I.Ya.HorbachevskyTernopilStateMedicalUniversityof the Ministry of Health ofUkraine” higher educational establishment</p><p>NationalPharmaceuticalUniversity</p><p><strong>Summary.</strong> The morphological and anatomical research of the herb and rhizome of pimpinella saxifrage l. has been made. Basic macro- and microscopic have been established to identify the raw material.</p><p><strong>Key words: </strong>pimpinella saxifrage l., herb, rhizome, macro- and microscopic features.</p><p><strong>Introduction. </strong>Pimpinella saxifraga L. Is a perennial herbaceous glabrous or puberulent plant of the Apiaceae family. It grows scattered on slopes, grass hills, forest glades, among the shrubs and along the roads on the entireterritory ofUkraine. For a long time pimpinella saxifrage l. has been used in folk medicine as an anti-spastic, ant-inflammatory, uretic, expectorant, anti-tussic, photosensitizing drug. The plant inUkraine is considered as non-officinal.</p><p><strong>The purpose</strong> of the research is to make a macro- and microscopic analysis of the herb and rhizome of pimpinella saxifrage l.</p><p><strong>Methods of the research. </strong>The micro-preparations were made of freshly collected raw material in the mixture of 96% ethanol – glycerol – purified water (1:1:1). The diagnostic microscopic features were studied by means of the microscope “Granum” with magnification power of x40, x100, x400. The photographs were taken with a camera Sony DSC-W80.</p><p><strong>Findings of the research. </strong>The macroscopic features of the rhizome of pimpinella saxifrage l. Spindle-shaped, longitudinally-wrinkled rhizomes up to5 cm in length and 2 –5 mm. in thickness. Grey-brown in colour, with uneven bend, yellowish-brown in colour, with yellow-brown spots. Strong, fragrant and acrid odour. Strong, bittersweet flavor.</p><p><strong>Microscopic features of the rhizome. </strong>The rhizome is covered with a thick layer of periderm. The cortical parenchyma is composed of parenchymal cells, small cells (phloem elements) are located lower. The phloem spreads evenly, in wide primary rays. The upper part of the cortical parenchyma contains oblong air vessels. The cortical parenchyma may have small schizogenous spaces; the parenchyma cells surrounding them contain starch. The central cylinder is clearly separated by the layer of cambium. The vessels are lined in oblong chains. The wood parenchyma accumulates starch. The rhizome center has a distinct lacune. </p><p><strong>Microscopic features of the herb. </strong>The stalks are thin-ribbed, branched and puberulent. The leaves are pinnated and macropodous; the margin is <a href="http://www.multitran.ru/c/m.exe?t=3157693_1_2&amp;s1=%EF%E8%EB%FC%F7%E0%F2%EE-%E7%F3%E1%F7%E0%F2%FB%E9">serrate-dentate</a>. The flowers are small, pentapetalous and in compound umbel inflorescence. The fruit are small, egg-shaped mericarps. The blade is green on the upper part and light green on the lower part; the flowers are white. The odour is faint and pleasant. The flavor is spicy and bitterish.</p><p><strong>Microscopic analysis if herb. </strong>The stalk is multi-ribbed. The stalk epiderm is small-celled; the cellular membrane has straight and slightly thickened walls. The stomata are oval, large and rare. The stomatal mechanism is anomocytic and anysocytic. The epiderm is unevenly puberulent. The trichomes are long, bent and many-celled, with a small basilar cell and oblong main cells, cone-shaped, 2-celled, long 3-celled straight, erect with oblong apical cell.</p><p>The leaf is dorsiventral. Palisade mesophile, bilarious, cancellate mesophile – tetrastichous. The cells of the palisade mesophile are small, not distinct, cylinder-shaped or oval; the second-row cells of the inner layer are loose. The cells of the cancellate mesophile are small, round or horizontally oblong. The stomata on the lower epiderm are protruding. The upper epiderm is represented by the parenchymal cells of various shape – starting with multi-angled ones and finishing with isodiametric-shaped ones. The cells of the lower epiderm are parenchymal, thin-walled and with rather sinuated membranes. The stomata are multiple, they are large and oval. The stomatal mechanism is anomocytic and paracytic. The leaf is unevenly puberulent. There are more trichomes on the lower side.</p><p><strong>Conclusions. </strong>The morphological and anatomical features of pimpinella saxifrage l. have been studied, and the main macro- and microscopic diagnostic features of rhizome, stalks, flowers and leaves have been defined to be later used for standardization of the medical raw material and design of the project “Pimpinella herb” and “Pimpinella rhizome”.</p>

The effect of some endogenic substances contained in the cortical parenchyma of apple trees on growth in vitro of the fungus Phytophthora cactorum (Leb. of Cohn) Schroeter

Gel filtration of an alkaline extract of the cortical parenchyma yielded four fractions of which two had activity <i>in vitro</i> on the growth of <i>Phytophthora cactorum</i>. Fraction I was stimulatory and fraction IV inhibitory; one of the inhibitors in fraction IV was identified as phloridzin. When the extraction solvent contained sodium sulphite in addition to sodium hydroxide, fraction II showed inhibitory activity.

Sclerification in the bark tissues of common fir (Abies alba Mill.)

The sclerification process in bark tissues of common fir (<em>Abies alba</em> Mill.) has been described. The sclerification begins in 3 years old stems. Sclereids differentiate from cortical parenchyma cells and from secondary phloem parenchyma cells that do not contain phenolic deposits. The first single sclereids are formed at the interface of the cortex and nonfunctional phloem. Hereafter, a continuous layer of them is formed. Later, new sclereid layers are formed successively in nonfunctional secondary phloem and cortex. The consecutive layers are separated tangentially by phloem parenchyma cells, that accumulate large amounts of phenolic substances, and by compressed phloem cells. Laterally they are separated by phloem rays that except of some dislocations are continuous. Structural net of the cortical phloem ray cells and phloem parenchyma delineates the areas where the formations of sclereid layers occurs in nonfunctional secondary phloem. Older cortex contains more sclereid layers and the time period of their formation extends continuously.

Adaptive anatomical structure for nastic movement in Mimosa pudica L.

In order to study the adaptive anatomical structures during nastic movement of Mimosa pudica L., anatomical structures of main pulvinus, common petioles, rachis and leaflets were compared with Albizia julibrissin Durazz. (taken as control). The anatomical structures of main pulvinus and common petiole of M. pudica were different from that of A. julibrissin. Upon stimuliti, the protoplast volume of M. pudica in the lower cortical parenchyma cells become smaller than that in upper ones, a feature seldom found in A. julibrissin. There were found many reticulate lacunas on the two side of adaxial petiole of M. pudica, but nil in A. julibrissin. Similarly some ill developed lacunas were found in the pulvinus of rachis and leaflet of M. pudica, but absent in A. julibrissin. It appears that reticulate lacunas in common petiole of M. pudica are responsible for its strong nastic movement. The main sensitivity position lies at the base of common petiole, where the lower cortex is more sensitive than the upper cortex, and the ordinal sensitivity positions are rachis and leaflets. DOI: http://dx.doi.org/10.3329/bjb.v42i1.15876 Bangladesh J. Bot. 42(1): 131-137, 2013 (June)

Anatomical changes in the stems of highbush blueberry (Vaccinium australe Small), caused by the fungus Godronia cassandrae f. vaccinii (Peck) Groves

Some histological changes were observed in blueberry stems infected by the fungus <i>G.cassandrae</i>. Dead cells of subepidermal collenchyma and cortical parenchyma filled with brown flocculent deposits were seen in the lesion areas. Pycnidia characteristic for the conidial stage of the fungus (<i>Topospora myrtilli</i>) were found in the collenchyma layer. The diseased tissues were found to be separated from the healthy ones by a layer of cork cells which was initiated under the epidermis and ended under the pericycle. Beneath this cork layer lamellar collenchyma and collenchyma-like phelloderma formed. Similar histopathological changes were observed in blueberry stems infected by seven other fungi.