Impression Cytology In Patients With Keratoconjunctivitis Sicca

Impression cytology is a simple and painless procedure that allows the collection of the superficial layers of the conjunctival epithelium. Each sample is assigned a grade of epithelial metaplasia, and goblet cell density is calculated in each one. We have studied the superior and temporal bulbar conjuctiva of dry eyes and have compared it with that of normal controls. In normal and dry eyes we find a statistically significant difference both in goblet cell density and grade of metaplasia, between superior and temporal bulbar conjunctiva. the differences in grade of metaplasia and goblet cell density between normal and dry eyes are significant in the superior conjunctiva, but in the temporal conjunctiva we only find significant differences in grade of metaplasia.     

Subretinal Injection of Gene Therapy Vectors and Stem Cells in the Perinatal Mouse Eye

The loss of sight affects approximately 3.4 million people in the United States and is expected to increase in the upcoming years.¹ Recently, gene therapy and stem cell transplantations have become key therapeutic tools for treating blindness resulting from retinal degenerative diseases. Several forms of autologous transplantation for age-related macular degeneration (AMD), such as iris pigment epithelial cell transplantation, have generated encouraging results, and human clinical trials have begun for other forms of gene and stem cell therapies.² These include RPE65 gene replacement therapy in patients with Leber''s congenital amaurosis and an RPE cell transplantation using human embryonic stem (ES) cells in Stargardt''s disease.^3-4 Now that there are gene therapy vectors and stem cells available for treating patients with retinal diseases, it is important to verify these potential therapies in animal models before applying them in human studies. The mouse has become an important scientific model for testing the therapeutic efficacy of gene therapy vectors and stem cell transplantation in the eye.^5-8 In this video article, we present a technique to inject gene therapy vectors or stem cells into the subretinal space of the mouse eye while minimizing damage to the surrounding tissue.     

Maternal effects in vulnerability to eye‐parasites and correlations between behavior and parasitism in juvenile Arctic charr

Hatchery‐reared fish show high mortalities after release to the wild environment. Explanations for this include potentially predetermined genetics, behavioral, and physiological acclimation to fish farm environments, and increased vulnerability to predation and parasitism in the wild. We studied vulnerability to Diplostomum spp. parasites (load of eye flukes in the lenses), immune defense (relative spleen size) and antipredator behaviors (approaches toward predator odor, freezing, and swimming activity) in hatchery‐reared juvenile Arctic charr (Salvelinus alpinus) using a nested mating design. Fish were exposed to eye‐fluke larvae via the incoming water at the hatchery. Fish size was positively associated with parasite load, but we did not find any relationship between relative spleen size and parasitism. The offspring of different females showed significant variation in their parasite load within sires, implying a dam effect in the vulnerability to parasites. However, the family background did not have any effect on spleen size. In the mean sire level over dams, the fish from the bolder (actively swimming) families in the predator trials suffered higher loads of eye flukes than those from more cautiously behaving families. Thus, the results indicate potentially maternally inherited differences in vulnerability to eye‐fluke parasites, and that the vulnerability to parasites and behavioral activity are positively associated with each other at the sire level. This could lead to artificial and unintentional selection for increased vulnerability to both parasitism and predation if these traits are favored in fish farm environments.     

The molecular mechanism of NELL2 movement and secretion in hippocampal progenitor HiB5 cells

Neural epidermal growth factor-like protein-like 2 (NELL2) is a secreted glycoprotein that is predominantly expressed in the nervous system, but little is known about the intracellular movement and secretion mechanism of this protein. By monitoring the localization and movements of enhanced green fluorescent protein (EGFP)-labeled NELL2 in living cultured hippocampal neuroprogenitor HiB5 cells, we determined the subcellular localization of NELL2 and its intracellular movement and secretion mechanism. Cterminal EGFP-fused NELL2 showed a typical expression pattern of secreted proteins, especially with respect to its localization in the endoplasmic reticulum, Golgi apparatus, and punctate structures. Vesicles containing NELL2 exhibited bidirectional movement in HiB5 cells. The majority of the vesicles (70.1%) moved in an anterograde direction with an average velocity of 0.454 μm/s, whereas some vesicles (28.7%) showed retrograde movement with an average velocity of 0.302 μm/s. The movement patterns of NELL2 vesicles were dependent upon the presence of microtubules in HiB5 cells. Anterograde movement of NELL2 did not lead to a detectable accumulation of NELL2 in the peripheral region of the cell, indicating that it was secreted into the culture medium. We also showed that the N-terminal 29 amino acids of NELL2 were important for secretion of this protein. Taken together, these results strongly suggest that the N-terminal region of NELL2 determines both the pattern of its intracellular expression and transport of NELL2 vesicles by high-velocity movement. Therefore, NELL2 may affect the cellular activity of cells in a paracrine or autocrine manner.     

Ultracytochemistry of cholera-toxin binding sites in ciliary processes

Summary Cholera toxin reduces the rate of formation of aqueous humor in concentrations (10^–11 M) that do not disturb the morphology of the aqueoushumor forming epithelial cells of the ciliary processes of the rabbit eye. The search for an endogenous mediator of aqueous-humor formation comparable to cholera toxin in its mode of operation prompted us to map the distribution of cell surface receptors for cholera toxin in the ciliary processes of the eyes of rabbits. Cytochemical studies were carried out with the use of conjugates of cholera toxin to fluorescein isothiocyanate (CT-FITC) and to horseradish peroxidase (CT-HRP), and of the B subunit of cholera toxin to horseradish peroxidase (B-HRP). Multiple fluorescent CT-FITC binding sites were observed on the outer nonpigmented epithelial layer near the crests of the processes. Processes incubated with CT-HRP in vitro showed surface staining of 30–40% of the nonpigmented epithelial cells. A prominent reaction product was observed along the basal and lateral plasma membranes of these cells. In vivo studies carried out after arterial infusion of B-HRP showed a reproducible dense reaction product between the apical surfaces of the pigmented epithelium (PE) and of the nonpigmented epithelium (NPE) facing each other. Aggregations of reaction product were observed with the electron microscope in the extracellular space between the apices of PE and NPE. The apical plasma membrane of the endothelium of the blood vessels near the crests of the ciliary processes was stained after either in vivo or in vitro exposure to peroxidase conjugates. These findings indicate that the cell-surface receptors which mediate the action of cholera toxin on aqueous humor formation are very likely localized in the apical plasma membranes of the epithelium of the ciliary processes.Supported in part by USPHS grant # EY-00237, the Connecticut Lions Eye Research Foundation, Inc., and Research to Prevent Blindness, Inc.     

The lateral photoreceptor of the barnacle,Balanus eburneus

Summary The lateral eye of the barnacle, Balanus eburneus, fixed in highly concentrated osmium is a lens-shaped body of approximately 250 m in diameter and about 75 m thick. It contains three photoreceptor cells which occupy about 42% of its volume. The photoreceptor cells are irregularly shaped and extend countless dendritic processes which bear rhabdomeres at their ends. Individual rhabdomeres come into contact with rhabdomeres originating from dendrites of the same or of one of the other visual cells. Thirteen per cent of the volume of the photoreceptor cells is taken up by the rhabdomeres. The membranes of the rhabdomeric microvilli contain globular subunits which suggest a 70 Å spacing of rhodopsin molecules. There are two kinds of glial cells. One kind, type A glial cells, makes contact with the fibrous capsule of the photoreceptor. The other kind, type B glial cells, is associated with the photoreceptor cells and extends countless tiny cytoplasmic extensions which interdigitate with similar extensions of the receptor cells. There are approximately 95 type B glial cells and 130 type A glial cells in the receptor. The cytoplasm of the photoreceptor cells contains countless small Golgi fields, mitochondria, microtubules, multivesicular and multilamellar bodies. The extracellular space of the photoreceptor is less than 0.1% of its total volume.The authors wish to thank Mrs. G. Theisen and Miss D. Hupp for expert technical assistance and Drs. M. Behrens, C. Helrich, and C.C. Krischer for many inspiring discussions. This study was partly supported by the SFB 160 of the Deutsche Forschungsgemeinschaft     

荒漠条件下甘草气孔振荡的水被动证据

生长在中国西北干旱荒漠的甘草（Glycyrrhiza inflata Batalin),当白天大气水蒸汽压差（ＶＰＤ）高于１kPa时,其气孔导度随时间的变化趋势为从稳态转为振荡态。通过茎木质部注射代谢抑制剂（ＮaN3或羰基氰化物－间－氯苯腙（ＣＣＣＰ）使气孔导度有些微降低,但是并不能显改变气孔振荡强度（振幅／平均值）。气孔振荡强度与ＶＰＤ和根阻力显相关,但与呼吸速度无明显相关,在荒漠条件下,当ＶＰＤ大于０．８kPa和至少存在１／４全根阻力的条件下才能出现气孔振荡。结果说明荒漠干旱条件诱发的甘草气孔振荡可能主要是一种水被动过程 。     

Effect of light quality on movement of Pterostichus melanarius (Coleoptera: Carabidae)

Abstract Behaviour of nocturnal insects is routinely observed under red light, but it is unclear how the behaviour under red light compares to behaviour in complete darkness, or under a source of white light. Here, we measure movement behaviour of the nocturnal carabid beetle Pterostichus melanarius Illiger (Coleoptera: Carabidae) using camera recording under a near‐infrared (nir), red or white radiation source. Red light significantly reduced movement speed in females similar to the effect of white light and different from nir. Also movement activity and pause length were affected by radiation source, with a significant difference between nir and white light, and with intermediate values in red light. The results presented here indicate that P. melanarius has different movement behaviour under the three radiation sources and suggest that nir rather than red radiation is most appropriate for measuring behaviour in total darkness. However, in the field total darkness is rare both because of natural light sources such as the moon and stars but increasingly also because of ecological light pollution, and therefore red light may still be of use for observing ecologically and practically relevant natural night‐time behaviour.     

北京西山地区油松林水文过程中营养元素迁移特征

对北京西山地区31a油松林降水化学进行定位监测与分析.结果表明：（1）大气降水流经油松林过程中,其营养元素（除Na^＋外）的浓度出现梯度变化：树干茎流高于穿透雨高于大气降水,并且有明显的月际变化,表明从林冠、树干淋洗大量的营养元素,林冠和树干中养分元素迁移是森林生态系统养分循环重要的组成部分.（2）营养元素变化中,浓度差异变化较大的元素是NO3^--N和K^＋,NO3^--N在穿透雨和树干茎流的浓度分别是大气降水的4.4倍、9.9倍,它们中的K^＋浓度分别为大气降水的4.1倍和8.1倍.（3）降水经过油松林,养分淋溶总量为54.12kg hm^-2,淋溶量较多的元素是Ca和K,淋溶强度最大的元素是K^＋.（4）北京西山地区大气降水输入林地的养分66.38kg hm^-2,较多的元素是Ca、N.降水经过油松林输入林地养分总量为120.50kg hm^-2,Ca元素最高,为61.22kg hm^-2,其次是N元素（NH4^＋-N＋N3^--N）,为31.34kg hm^-2,K元素为16.49kg hm^-2,Mg元素为8.11kg hm^-2,Na元素最少,为3.34kg hm^-2.     

细胞松弛素B对蚕豆气孔运动的影响

以蚕豆叶片下表皮条为材料,研究了微丝在气孔运动中的作用。利用肌动蛋白纤丝专一性抑制剂──细胞松弛素Ｂ（ＣＢ）预处理后,再用诱导气孔运动的因子处理表皮条,在显微镜下观测气孔孔径的变化。结果显示,用ＣＢ处理开放或关闭状态气孔,其开度均不发生变化;ＣＢ处理使微丝解聚,气孔运动被抑制;且ＣＢ处理后气孔的运动是可以恢复的。实验进一步表明,开放气孔经１０ｍｇ／Ｌ的ＣＢ预处理后,ＡＢＡ、Ｃａ２＋及暗诱导气孔关闭的作用均不同程度地受到抑制,推测微丝可能参与ＡＢＡ、Ｃａ２＋及暗诱导的气孔关闭过程;关闭气孔经１０ｍｇ／Ｌ的ＣＢ预处理后,Ｋ＋和（或）光诱导气孔开放的作用受到抑制,推测微丝可能参与光及Ｋ＋诱导的气孔开放过程。