Lactate induces tumour necrosis factor-alpha, interleukin-6 and interleukin-1beta release in microglial- and astroglial-enriched primary cultures

Hyperammonaemia has deleterious effects on the CNS in patients with liver dysfunction. Cellular mechanisms underlying the effects of hyperammonaemia are largely unknown, although astrocytes have been the main target of interest. This study investigated how treatment with NH4Cl and lactate, which increase in the brain as a consequence of hyperammonaemia, affects cells in primary rat cultures enriched in either astrocytes or microglia. Morphological changes were studied over time using light microscopy. Release of the proinflammatory cytokines tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-6 and IL-1beta was measured using ELISA. NH4Cl was found to induce vacuole formation in both culture systems. Lactate treatment altered astrocytic appearance, resulting in increased space between individual cells. Microglia adopted a round morphology with either NH4Cl or lactate treatment. Lactate, but not NH4Cl, induced release of TNF-alpha and IL-6 in both astroglial- and microglial-enriched cultures, while IL-1beta was released only in microglial cultures. Cytokine release was higher in the microglial- than in the astroglial-enriched cultures. Additionally, the astroglial-enriched cultures containing approximately 10% microglial cells released more cytokines than cultures containing about 5% microglial cells. Taken together, our data suggest that most TNF-alpha, IL-6 and IL-1beta release comes from microglia. Thus, microglia could play an important role in the pathological process of hyperammonaemia.     

Additional morphological and physiological heterogeneity within the midgut of larval Aedes aegypti (Diptera: Culicidae) revealed by histology, electrophysiology, and effects of Bacillus thuringiensis endotoxin

Analysis of larval Aedes aegypti midgut using scanning electron microscopy, nuclear and mitochondrial dyes, response to Bacillus thuringiensis israelensis CryIVB toxin, and electrophysiology is described. The anterior ventriculus ("stomach") region is found to have much lower mitochondrial densities than other midgut regions. The transitional region is distinguished by apical surface architecture, and by region-specific effects of CryIVB endotoxin. In this region CryIVB causes holes ranging from 1.0 to 7.0 microm in diameter (mean 3.3+/-0.53 microm, N=12), blisters 16.9+/-1.54 microm in diameter (N=10), and separation of adjacent cells. The holes are not consistent with damage due to the colloid osmotic lysis model of delta-endotoxin activity. The posterior ventriculus possesses a distinctive cellular architecture consisting of hemispherical, domed apical membranes surrounded by deep clefts. Functional and morphological heterogeneity is revealed within the posterior ventriculus, with the anterior end dominating the electrical profile of isolated, perfused preparations and showing the greatest response to serotonin. Hyperpolarization of the transepithelial potential by serotonin occurred in conjunction with a decrease in the space constant lambda, ruling out closure of ion channels as the mechanism of action of serotonin.     

The use of whole genome amplification in the study of human disease

The availability of large amounts of genomic DNA is of critical importance for many of the molecular biology assays used in the analysis of human disease. However, since the amount of patient tissue available is often limited and as particular foci of interest may consist of only a few hundred cells, the yield of DNA is often insufficient for extensive analysis. To address this problem, several whole genome amplification (WGA) methodologies have been developed. Initial WGA approaches were based on the polymerase chain reaction (PCR). However, recent reports have described the use of non-PCR-based linear amplification protocols for WGA. Using these methods, it is possible to generate microgram quantities of DNA starting with as little as 1mg of genomic DNA. This review will provide an overview of WGA approaches and summarize some of the uses for amplified DNA in various high-throughput genetic applications.     

国产爵床科芦莉花族植物的花粉形态

报道了国产爵床科Acanthaceae芦莉花族Ruellieae芦莉花亚族Ruelliinae 2属7种、假杜鹃亚族Barlerinae 1属3种和马蓝亚族Strobilanthinae 16属34种植物扫描电镜下的花粉形态.芦莉花亚族的地皮消属Pararuellia和喜花草属Eranthemum的花粉均为圆球形,具3孔或3孔沟,外壁为不同的网状结构; 假杜鹃亚族的假杜鹃属Barleria的花粉为长球形,具3孔沟,外壁亦为网状结构;马蓝亚族植物(包含广义的马蓝属Strobilanthes s.l.)花粉形态多样,结构复杂.依据花粉萌发孔和外壁纹饰特征,可将马蓝亚族16属植物和上述两亚族3属植物的花粉形态归纳成3大类型: 1. 具3孔类型.其中又有(1)外壁具网状纹饰者,见于地皮消属; (2)外壁具芽胞状纹饰者,见于黄猄草属Championella; (3)外壁具刺状(棒状)纹饰者,见于南一笼鸡属Paragutzlaffia、叉花草属Diflugossa和假蓝属Pteroptychia.2. 具3孔沟及具3孔沟与假沟类型(肋条带型).其中又有(1)具3孔沟和网状纹饰者,见于喜花草属和假杜鹃属; (2)具刺状(棒状)纹饰者,见于南一笼鸡属、叉花草属和假蓝属; (3)具3孔沟与假沟,外壁纹饰具节隔、肋条带状或网状,网眼纵向排列成行,网眼内有细网纹者,见于耳叶马蓝属Perilepta、马蓝属Pteracanthus(大部分)、金足草属Goldfussia、紫云菜属Strobilanthes(部分)和合页草属Sympagis; (4)具3孔沟与假沟类型,肋条带状,但不具节隔,外壁纹饰网状,网眼不成行或不明显纵向排列,网内无细网纹者,见于尖蕊花属Aechmanthera、板蓝属Baphicacanthus、马蓝属(部分)和糯米香属Semnostachya; (5)具双脊及细网状纹饰者,见于环毛紫云菜Strobilanthes cycla.3. 具(4-)5孔沟及假沟类型(肋条带型),外壁具网状或拟网状纹饰,见于腺背蓝属Adenacanthus.另外兰嵌马蓝属Parachampionella、山一笼鸡属Gutzlaffia和肖笼鸡属Tarphochlamys的花粉有无萌发孔尚不清楚,有待进一步研究.综上所述,芦莉花族植物的花粉形态具有较高的多样性,是重要的分类性状.利用花粉形态特征能较好地区分高级分类群如亚科、族以及亚族,有时也有助于阐明类群之间的相互关系,甚至也能用于区分属、种和阐明其关系.     

Do “glanduliferous” larvae of Galerucinae (Coleoptera: Chrysomelidae) possess defensive glands? A scanning electron microscopic study

Larvae of the chrysomelid taxon Galerucinae, Sermylini, are known to release a fluid from dorsolateral segmental openings when disturbed. The release of this fluid resembles the discharge of secretion from well-studied exocrine defensive glands in larvae of Chrysomelinae, the putative sister group of Galerucinae. Thus, Sermylini larvae have been named “glanduliferous” ones. However, no comparative analyses of the internal structures of the segmental openings in Sermylini larvae have been available prior to this study. Therefore, segmental larval openings in 10 Sermylini species were investigated by scanning and light microscopy. Two types of segmental openings were detected: (1) the opening is visible as an integumental slit which can be opened and closed by muscles. No specific glandular structures are associated with this slit. (2) The opening is covered internally by a cuticular sac which can be everted to the outside. A cuticular duct of a gland is leading into this sac. Haemolymph is discharged from both types of openings. These structures are discussed with respect to reflex bleeding in other taxa. Furthermore, a common origin of the segmental openings in larvae of Sermylini and the segmental exocrine glands in chrysomeline larvae is critically questioned.     

Comparative genomics of hemiascomycete yeasts: genes involved in DNA replication, repair, and recombination

Among genes conserved from bacteria to mammals are those involved in replicating and repairing DNA. Following the complete sequencing of four hemiascomycetous yeast species during the course of the Genolevures 2 project, we have studied the conservation of 106 genes involved in replication, repair, and recombination in Candida glabrata, Kluyveromyces lactis, Debaryomyces hansenii, and Yarrowia lipolytica and compared them with their Saccharomyces cerevisiae orthologues. We found that proteins belonging to the replication fork and to the nucleotide excision repair pathway were-on the average-more conserved than proteins involved in the checkpoint response to DNA damage or in meiotic recombination. The meiotic recombination proteins Spo11p and Mre11p-Rad50p, involved in making meiotic double-strand breaks (DSBs), are conserved as is Mus81p, involved in resolving meiotic recombination intermediates. Interestingly, genes found in organisms in which DSB-repair is required for proper synapsis during meiosis are also found in C. glabrata, K. lactis, and D. hansenii but not in Y. lipolytica, suggesting that two modes of meiotic recombination have been selected during evolution of the hemiascomycetous yeasts. In addition, we found that SGS1 and TOP1, respectively, a DEAD/DEAH helicase and a type I topoisomerase, are duplicated in C. glabrata and that SRS2, a helicase involved in homologous recombination, is tandemly duplicated in K. lactis. Phylogenetic analyses show that the duplicated SGS1 gene evolved faster than the original gene, probably leading to a specialization of function of the duplicated copy.     

Application of Image Analysis Techniques in Activated Sludge Wastewater Treatment Processes

Image analytical techniques have been extensively developed to evaluate complex microbial aggregates such as sludge flocs and biofilms. This review covers the latest contributions concerning the application of image analysis to the activated sludge systems with respect to the most frequently used morphological parameters and relations between them and traditional wastewater treatment parameters. Recent developments have indicated that image analysis can be successfully used for the quantification of flocs and filamentous bacteria in the operating wastewater treatment plants, which enables prediction of bulking events and pinpoint flocs formation.