地钱卵发育超微结构和细胞化学的研究

采用电镜和细胞化学技术对地钱(Marchantia polymorpha)卵发生过程进行了研究,根据卵发生过程中细胞化学和超微结构特征可将卵发育过程分为幼卵、中期卵和成熟卵3个阶段.幼卵阶段,卵细胞、腹沟细胞及颈沟细胞间有发达的胞间连丝,但卵与腹沟细胞间的胞间连丝很快退化,幼卵细胞内具大量透明的囊泡,均匀分布于细胞质中;卵发育中期,突出特征是卵细胞质内产生嗜锇性的脂滴,位于囊泡中,与此同时,腹沟细胞退化,其细胞质内产生大型囊泡,囊泡内分泌物与卵细胞外的物质类似,呈PAS反应阳性,表明该物质应为多糖类;卵成熟时,腹沟细胞和颈沟细胞完全退化,卵细胞外包被大量粘性多糖类物质,卵细胞核表面不规则,产生明显的核外突,众多的小泡围绕着细胞核,脂滴聚集成簇,卵细胞内其他细胞器不易区分.卵发育过程中,质体不含淀粉粒,线粒体退化,高尔基体相对发达.地钱卵发育的这些特征显著区分于蕨类植物.     

地钱、拳卷地钱和粗裂地钱的紫外谱线组法鉴别

本文建立了紫外谱线组法鉴别地钱、拳卷地钱和粗裂地钱的方法。通过这三种药材的石油醚、氯仿、乙醇和水提取液的紫外谱线比较,发现地钱、拳卷地钱和粗裂地钱的紫外谱线图、最大吸收峰数目及峰位值具有明显差异。该法简单、准确,可用来鉴别地钱、拳卷地钱和粗裂地钱的原药材。     

地钱生殖托形态结构新观察及其对有性生殖的适应

用显微观察技术对苔类植物地钱（Marchantia polymorpha）生殖托的形态结构进行了研究,本研究的新观察包括：（1）生殖托具有明显的背腹分化,被认为是叶状体为适应有性生殖而高度特化的直立枝;（2）雌托盘的9～11个指状裂瓣中边缘两个稍不同,除了两个边缘裂瓣外的其他裂瓣间具雌苞。托柄具有背腹之分,雌托柄背面具光合组织,并向两侧扩展形成纵沟,雌托柄的腹面具2条被鳞片重叠覆盖相互平行的纵沟,内具假根;（3）与雌托柄不同,雄托柄外观平滑,背面无光合组织及纵沟。观察发现,伞状的雌托能滞留水分,并沿雌托柄的纵沟缓慢释放连续的水流,游动精子沿着纵沟内的水流到达雌托下面的颈卵器。上述观察表明地钱生殖托的结构是对有性生殖的一种适应,这有助于我们理解地钱受精作用的机理。     

地钱颈卵器发育和卵发生的显微观察及细胞化学研究

对地钱（Marchantia polymorpha）颈卵器发育和卵发生过程进行了显微观察和细胞化学的研究,颈卵器起始于原始细胞,该细胞呈乳突状,经横分裂产生基细胞和顶细胞,顶细胞经3次纵斜向分裂和1次横分裂产生初生细胞,初生细胞是颈卵器内的第一个细胞,经横分裂产生中央细胞和颈沟母细胞,前者产生1个腹沟细胞和1个卵细胞,后者最终产生4个颈沟细胞。颈卵器的成熟表现为颈部显著伸长和腹部膨大,卵细胞成熟时具有不规则的核,细胞质内含有丰富的囊泡和颗粒物,卵细胞周围充满粘性物质,细胞化学研究表明,该粘性物质为多糖,卵细胞质中深染色的颗粒可能为脂类物质,腹沟细胞自产生后就逐渐退化,颈沟细胞的退化迟于腹沟细胞,其数量通常为4个,偶尔可见5个颈沟细胞或具有双核的现象。     

Nutrient utilization and requirement under photoheterotrophic growth of Marchantia polymorpha: Improvement of the culture medium

Previously we reported on a suspension culture of chlorophyllous cells of a liverwort, Marchantia polymorpha L., under photoheterotrophic conditions. The chemically defined medium was a modified Murashige and Skoog's medium, which contained, besides glucose, inorganic salts, a growth regulator (2,4-D), and twenty-four organic compounds as micro constituents. Because of this complexity, we undertook a simplification of the medium. Having examined the utilization of the major nutrients and the requirements for the micro constituents, we have succeeded in improving the medium. The new medium contains phosphate at 3.13 mM and only eight out of the twenty-four original micro organic constituents. In this new medium, the cells grow under a well-balanced nutritional condition, with richer chlorophyll and at a higher rate during the exponential phase than in the original medium.     

地钱在离体条件下的无性繁殖及脱分化与再分化的研究

地钱(Marchantia polymorpha L.)的胞芽和配子体先在 MS 培养基上补加1mg/l 2,4-D和3%蔗糖,经过启动部分脱分化后,再移入1/2KNOP 培养基补加4—8mg/l 2,4-D,0.25～0.5mg/l BA 与 MS 的铁盐,20g 蔗糖,此后愈伤组织肉眼可见,但仍伴有假根。最后移入 White 培养基添加丙酮酸、延胡索酸与柠檬酸三者混合物(5mmol/l)及1mg/l 2,4-D与4%葡萄糖后,始呈现彻底的脱分化状态,愈伤组织才能正常生长。整个脱分化时间长达10个月。而再分化成配子体却比高等植物容易,甚至移入不含激素的 MS 基本培养基即可形成正常的配子体。     

<Emphasis Type=Italic>Marchantia polymorpha</Emphasis> L.: A Bioaccumulator

Mussoorie in the western Himalayas of India, a popular tourist destination, was selected for air pollution monitoring. The study was carried out applying thalloid liverwort species Marchantia polymorpha L. as a tool. Moss bags containing M. polymorpha was transplanted at residential, highly and less polluted areas to obtain comprehensive and comparative data. Lead content, along with some essential micronutrients viz. zinc, manganese, and copper, as well as some physiological parameters total chlorophyll, sugar, protein, catalase and peroxidase activity were analyzed. The highest accumulation of lead (Pb) in M. polymorpha plants was highest in summer season , i.e., 2276 μg/g dry weight. The correlation of metals and physiological parameters has been made to get clear view of the effects of lead on physiology and essential micronutrients.     

Antifungal and antibacterial potential of methanol and chloroform extracts of Marchantia polymorpha L.

The methanol and chloroform extracts of Marchantia polymorpha were tested against three Gram-negative bacterial strains, viz. Xanthomonas oryzae pv. oryzae, Salmonella enterica and Pasturella multocida and four fungal strains, viz. Tilletia indica Mitra, Fusarium oxysporum f. sp. lini, Sclerotium rolfsii Sacc. and Rhizoctonia solani Kuhn. by using disc diffusion and micro broth dilution techniques. Both the extracts showed unique activity against X. oryzae and P.multocida [per cent inhibition (PI) 11.58 and 12.55, minimum inhibitory concentration (MIC) 2.50 and 1.25 μg/mL, minimum bactericidal concentration (MBC) 2.75 and 1.25 μg/mL, respectively] but for fungi, it was shown against S.rolfsii and F. oxysporum [PI 32.65 and 33.44, MIC 2.50 and 0.65 μg/mL, minimum fungicidal concentration (MFC) 4.50 and 0.65 μg/mL, respectively].The extracts possessed antimicrobial activity with different potency against variety of micro-organisms pathogenic to plants as well as animals. Some extracts were fungistatic and bacteriostatic (methanol extract against R. solani and chloroform extract against F. oxysporum and methanol extract for bacteria, respectively), while rest showed fungicidal/bactericidal potential. The results suggest the potential of M. polymorpha for developing a broad spectrum antimicrobial formulation in future.     

Schistosoma mansoni: Mechanisms in regulation of glycolysis

Adult pairs of Schistosoma mansoni convert glucose to lactate rapidly and almost quantitatively under aerobic and anaerobic conditions E. Bueding, 1950, Journal of General Physiology33, 475–495). Glycolysis is the principal source of energy of schistosomes and its inhibition by trivalent organic antimonials, at the phosphofructokinase step [EC 2.7.1.11], may be the basis for the chemotherapeutic effects of these agents E. Bueding and J. M. Mansour, 1957, British Journal of Pharmacology and Chemotherapy12, 159–165). We have developed standardized conditions for the comparison of rates of glucose consumption and lactate production by intact schistosomes in vitro and by centrifuged homogenates of worms. The rates of glycolysis of homogenates prepared from freshly isolated worms, and from worms that have been lyophilized immediately after harvesting and stored for prolonged periods at ?80 C were identical, when measured in media containing appropriate concentrations of glucose, NAD, ATP, MgCl₂, KCl, and phosphate. The specific activities of the 11 glycolytic enzymes and of 3 related enzymes (fructose-biphosphatase [EC 3.1.3.11], glycerol-3-phosphate dehydrogenase [EC 1.1.1.8], and malate dehydrogenase [EC 1.1.1.37]) were measured in homogenates under optimal conditions. The profile of the relative activities of glycolytic enzymes of S. mansoni resembles closely that of Ehrlich ascites tumor cells, and differs markedly from that observed in erythrocytes or skeletal muscle. As is the case in many animal tissues, hexokinase [EC 2.7.1.1] was the enzyme of lowest specific activity, and the rate of glycolysis of homogenates was almost the same as the hexokinase activity. Several other lines of evidence support the view that the hexokinase reaction is the rate-limiting step in the glycolysis of worm homogenates. Hexokinase activity was not particulate in schistosome homogenates, and there was no detectable high K_m glucokinase-like activity. The rate of glycolysis by homogenates exceeded that of intact worms by a factor of nearly 5. The contributions of glucose transport, availability of ADP and inorganic phosphate, regulatory enzymes, and a substrate cycle catalyzed by fructose-bisphosphatase are considered as possible mechanisms for the restraint of glycolysis in intact worms. The mechanisms contributing to the rapid rates of glycolysis of adult S. mansoni have not been identified, although several can be excluded (unusually high capacity of the glycolytic enzymes, the presence of mitochondrial hexokinase, the occurrence of glycosomes, and the operation of defective mitochondrial shuttles). In view of the regulatory role of hexokinase in the glycolysis of S. mansoni, inhibition of this enzyme is a potentially important target for the development of new antischistosomal drugs.     

Soybean Recombination Sites are Present as Dispersed Segments in Arabidopsis and Liverwort Mitochondrial DNA

In previous studies we have shown that recombination across a 299-bp interspersed sequence accounts for the diversity of the mitochondrial genome in wild and cultivated soybeans. In this study, a computer-assisted survey of databases was performed using sequences of the repeat and its neighboring regions as query sequences. The sequences of soybean were found to be present as many short segments that include repeated sequences in the mitochondrial genomes of Arabidopsis and liverwort. Taken together with the results of a DNA gel-blot analysis, this suggests that the soybean sequences were found to have originated during land plant evolution and are present as small-interspersed segments in many taxa of land plants.     

Asymmetric transformation of enol acetates with esterases from Marchantia polymorpha

Two esterases catalyzing the asymmetric hydrolysis of enol acetates to give optically active -alkylated ketones were isolated from cultured cells of Marchantia polymorpha by a three-step procedure: hydrophobic chromatography, anion exchange chromatography and gel-filtration chromatography. These esterases had opposite stereoselectivities on protonation of the enol intermediate in the hydrolysis and one of them obviously reversed the stereoselectivity when the chain length and the bulkiness of substituents at the β-position to the acetoxyl group were increased. The internal amino acid sequences of peptide fragments obtained by the proteolysis of the esterases with lysyl endopeptidase had no similarity to those of other hydrolytic enzymes.     

High prevalence of asthma symptoms in Warao Amerindian children in Venezuela is significantly associated with open-fire cooking: a cross-sectional observational study

Background

The International Study on Asthma and Allergies in Childhood (ISAAC) reported a prevalence of asthma symptoms in 17 centers in nine Latin American countries that was similar to prevalence rates reported in non-tropical countries. It has been proposed that the continuous exposure to infectious diseases in rural populations residing in tropical areas leads to a relatively low prevalence of asthma symptoms. As almost a quarter of Latin American people live in rural tropical areas, the encountered high prevalence of asthma symptoms is remarkable. Wood smoke exposure and environmental tobacco smoke have been identified as possible risk factors for having asthma symptoms.

Methods

We performed a cross-sectional observational study from June 1, 2012 to September 30, 2012 in which we interviewed parents and guardians of Warao Amerindian children from Venezuela. Asthma symptoms were defined according to the ISAAC definition as self-reported wheezing in the last 12 months. The associations between wood smoke exposure and environmental tobacco smoke and the prevalence of asthma symptoms were calculated by means of univariate and multivariable logistic regression analyses.

Results

We included 630 children between two and ten years of age. Asthma symptoms were recorded in 164 of these children (26%). The prevalence of asthma symptoms was associated with the cooking method. Children exposed to the smoke produced by cooking on open wood fires were at higher risk of having asthma symptoms compared to children exposed to cooking with gas (AOR 2.12, 95% CI 1.18 - 3.84). Four percent of the children lived in a household where more than ten cigarettes were smoked per day and they had a higher risk of having asthma symptoms compared to children who were not exposed to cigarette smoke (AOR 2.69, 95% CI 1.11 - 6.48).

Conclusion

Our findings suggest that children living in rural settings in a household where wood is used for cooking or where more than ten cigarettes are smoked daily have a higher risk of having asthma symptoms.     

The long periodicity phase (LPP) controversy part I: The influence of a natural-like ratio of the CER[EOS] analogue [EOS]-br in a CER[NP]/[AP] based stratum corneum modelling system: A neutron diffraction study

This study used neutron diffraction to investigate a ceramide-[NP] C24/[AP] C24 /[EOS]-br C30/cholesterol/lignoceric acid (0.6: 0.3: 0.1: 0.7: 1) based stratum corneum modelling system. By adding specifically deuterated ceramides-[NP]-D₃, [AP]-D₃, and [EOS]-br-D₃, detailed information on the lamellar and the nanostructure of the system was obtained. For the short periodicity phase a natural-like lamellar repeat distance of 5.47?±?0.02?nm was observed, similar to the [NP]/[AP] base system without the [EOS]-br. Unlike in this system the ceramides here were slightly tilted, hinting towards a slightly less natural arrangement. Due to the deuteration it was possible to observe that the long ceramide chains were overlapping in the lamellar mid-plane. This is considered to be an important feature for the natural stratum corneum. Despite the presence of a ceramide [EOS] analogue – able to form a long phase arrangement – no distinct long periodicity phase was formed, despite a slightly higher than natural ω-acyl ceramide ratio of 10?mol%. The deuterated variant of this ceramide determined that the very long ceramide was integrated into the short periodicity phase, spanning multiple layers instead. The – compared to the base system – unchanged repeat distance highlights the stability of this structure. Furthermore, the localisation of the very long ceramide in the short periodicity phase indicates the possibility of a crosslinking effect and thus a multilayer stabilizing role for the ceramide [EOS]. It can be concluded, that additionally to the mere presence of ceramide-[EOS] more complex conditions have to be met in order to form this long phase. This has to be further investigated in the future.