Primary Vesicles Research Articles

Study of the Effect of Fresh Green Asparagus officinalis L. Stems Extract and Metformin Drug on Induced Polycystic Ovary Syndrome (PCOS) in Female Albino Rats

During this research, Polycystic Ovary Syndrome (PCOS) was studied and the effect of Metformin Drug and an extract of fresh Asparagus officinalis L. stems on it was studied. The study involved 20 adult female albino rats divided into four groups, each containing five female rats. The first group received a physiological saline solution, while the second group was induced with Polycystic Ovary Syndrome (PCOS) by being dosed with Letrozole. The third group, after the induction of PCOS, was treated with Metformin, and the fourth group was dosed with Fresh Asparagus officinalis L. stems extract. The study focused on hormonal and Histological changes. The results showed that the group of rats with PCOS induced by Letrozole had a significant decrease in the levels of Progesterone (Prog.) and Oestradiol (E2) hormones, and a significant increase in Testosterone (Testo.) compared to the control group. Dosing rats with Fresh Asparagus officinalis L. stems extract led to a significant decrease in (E2) hormone level and a significant increase in Progesterone (Prog.) hormone level compared with the control group. Dosing rats with Metformin led to a significant decrease in (E2) hormone level and a significant increase in Progesterone (Prog.) and Testosterone (Testo.) hormone levels compared to the control group. The results also indicated significant changes in hormone levels between Treatment groups and PCOS induction group. The histological study revealed changes in the shape of the ovary, the appearance of cystic structures, congestion of blood vessels, and increased ovarian cellularity. Metformin showed fewer Histological changes, while Asparagus officinalis L. stems extract had a therapeutic effect with moderate severity, leading to the disappearance of cystic vesicles and the appearance of corpus luteum and primary vesicles. In conclusion, we conclude that both the therapeutic agents Metformin and the extract of Fresh Asparagus officinalis L. stems showed a therapeutic effect on Polycystic Ovary Syndrome, as Metformin drug showed a treatment for this Syndrome, but it requires a long period to show this result, meaning we need more than 28 days, which is the duration of the study, but Asparagus officinalis L. stems extract had Faster healing effect within just 28 days. Fresh Green, tender Asparagus officinalis L. stems is one of the significant medicinal plants that is used to treat or prevent polycystic ovarian syndrome. Research has shown that it is an effective treatment for the condition. Future researchers studying polycystic ovarian syndrome will find this article useful.

Formation Mechanism and Prediction Method for the Permian Fused Breccia Tuff Reservoir, Wuxia Region, Junggar Basin

Fused breccia tuff occurs globally, but its formation mechanism is very controversial. Volcanic reservoirs have developed at the bottom of the Permian Fengcheng Formation in the Wuxia region of the Junggar Basin, and here, the lithology is fused breccia tuff. The reservoir porosity is mainly vesicles, but the development and relative filling of the vesicles vary spatially, resulting in strong reservoir heterogeneity. Through core and thin section observations and structural analysis, and combined with reconstructions of the paleosedimentary environment, we discussed in detail the formation mechanism of the fused breccia tuff reservoir. Our conclusions are as follows. In the high-temperature and high-pressure environment of the deep crust, intermediate acidic lava containing volatile components rapidly rose to the earth’s surface along a fault. The volatile components in the lava foamed strongly and then exploded due to the sharp decline of pressure and temperature. A small part of the volcanic dust and pyroclastic material was erupted into the upper atmosphere. Most of the magma became magmatic pyroclast, vitric pyroclast, rock debris, dust, and other matter. This material was in a semimolten state and overflowed into a nearby low-lying lake. The extremely high-temperature pyroclastic flow quickly vaporized the water into high-pressure water vapor, which was squeezed into the pyroclastic flow and became mixed with other volatiles in the foam. On cooling, the pyroclastic material solidified into rock, and the vesicles were preserved. In a later period, due to strong tectonic movement, faults and fractures developed, surface water penetrated into the vesicles along the faults and fractures, and silica and other substances were deposited, filling the primary vesicles. To quantify the development and relative filling of vesicles, drilling parameters were used to establish different geologic models, and wave equation forward modeling was used to obtain a relationship between the development and filling of vesicles, and the seismic amplitude. The 3D seismic amplitude attributes were then extracted to predict the extent of the reservoir, yielding prediction results consistent with the drilling observations.

Interaction between Gemini Dodecyl O-Glucosides-Based Multilayer Vesicles and β-Lactoglobulin: The Dominant Role of Surface Charge.

Novel Gemini dodecyl O-glucoside-based primary, secondary, and tertiary vesicles were developed in this work utilizing layer-by-layer deposition of polysaccharides (e.g., sodium carboxymethyl cellulose and chitosan), and their interaction with β-lactoglobulin (BLG) was carefully investigated. The increase of polysaccharide layers on primary vesicles led to a monotonic increase in size and consecutive reversal of surface charge. Polysaccharide deposition significantly retarded the vesicle aggregation and degradation of entrapped catechin laurate during storage. Steady-state fluorescence, isothermal titration calorimetry, and protein precipitation analyses revealed the surface charge dependence of the interactions between vesicles and a model milk protein BLG, which were much stronger when they were charged oppositely than when they presented the same type of surface charge. It was highlighted that the surface charge of vesicles could be tuned by differently charged coatings to accommodate to that of the milk proteins in the food matrix. This work will contribute to the practical application of niosomal vesicles loaded with bioactive compounds to fortify dairy products.

Reservoir characteristics and forming controls of intrusive-metamorphic reservoir complex: A case study on the diabase-metamudstone rocks in the Gaoyou sag, eastern China

Hydrocarbon reservoirs associated with magmatic intrusion have significance in expanding hydrocarbon exploration. However, this type of reservoir is rarely documented in previous studies. Taking the diabases and their contact metamudstones from the Funing Formation, northern slope of the Gaoyou sag, as study objectives, the current work mainly investigated lithofacies, reservoir porosities and properties, and C, O isotopes of intrusive diabases and/or contact metamudstones, aiming to unravel reservoir characteristics and forming controls of diabase and metamudstones. After diabase intrusion, four lithofacies, i.e., diabase center, diabase margins, slates, and hornfels, were formed. Meanwhile, various porosity types were developed, including primary vesicles, grain-dissolved micropores, contractive micropores, cooling fractures, and structural fractures in the diabases, and recrystallizational and matrix-dissolved micropores, hydrothermal and contractive microfractures, and structural fractures in the contact metamudstones. Controls on diabase reservoir development include uneven cooling, dissolution, authigenic minerals filling, and structural movements. For contact metamudstone reservoirs, the forming controls include physical fracturing, hydrothermal baking, and authigenic carbonate dissolution. The controlling factors exerted diverse influences on reservoir properties with respect to different parts in contact metamudstones or diabase, thus causing obvious heterogeneity regarding to reservoir quality, i.e., hornfelses and diabase margins exhibiting preferable reservoir properties than slates and diabase center, respectively. Finally, an evolutional model depicting the forming process of diabase-metamudstone reservoir complex was tentatively proposed.

Abstract A006: Skp2 associates with Slug and androgen receptor in aggressive prostate cancer

Abstract Introduction: The Skp2 F-box protein is the substrate recruiting component of the SCF (Skp1-Cullin 1-F-box) type of E3 ubiquitin-ligase complexes. Skp2 plays an important role in prostate tumorigenesis (e.g., via recently reported stabilization of EZH2 or Twist1), which needs further elucidation. Patients and Methods: Prostate cancer patient cohort (N=101) was analyzed by immunohistochemistry for the following proteins: Skp2, AR, Ki-67, Slug, E-cadherin, MdmX, Mdm2, and p53. The indicated proteins were also analyzed in seminal vesicles and lymph node metastases (SV=35 and LN=18 patients, respectively), as well as in benign prostatic hyperplasia (BPH=34). Colocalization analysis was performed using Perkin Elmer Opal Multiplex kit, Vectra 3.0 imaging system, and confocal microscope Carl Zeiss LSM 780. Independent prostate cancer cohort (N=30) was evaluated for TMPRSS2-ERG fusion by a triple-color deletion assay and assessed for Skp2, ERG, and NQO1 protein expression. Cox regression analysis and Kaplan-Meier curves were used to assess importance of Skp2, Slug, EZH2, and Twist1 for biochemical relapse in the TCGA prostate cancer dataset (N=488, RNA seq). Results: Expression of E-cadherin was lower in primary prostate cancer and seminal vesicles compared to BPH and LN, which may be linked to epithelial-mesenchymal transition. Opposite expression profile was observed for Skp2, MdmX, and Slug. High Gleason score (≥8, N=30) was significantly associated with higher Skp2 and lower E-cadherin expression (p<0.001 and 0.011, respectively). Skp2 slightly correlated with AR and Slug in the whole patient cohort (Rs 0.37 and 0.322, respectively). These correlations were stronger in patients with high Gleason score (Rs 0.53 and 0.56, respectively). In patients with metastasis into lymph nodes, Skp2 similarly correlated with Slug and AR (Rs 0.56 and 0.37, respectively) while androgen receptor further correlated with Ki-67 (Rs 0.50). Confocal microscopy revealed colocalization of Skp2 and Slug in prostate cancer cells. In an independent cohort, Skp2 correlated with expression of ERG, NQO1, and percentage of cancer cells positive for TMPRSS2-ERG (Rs 0.503, 0.387 and 0.615, respectively). In the TCGA dataset, high mRNA levels of Skp2 and EZH2 (but not Slug or Twist1) associated with shorter time to biochemical relapse, compared to low mRNA levels [HR 3.62 (2.1 – 6.23), p<0.001]. Conclusion: Immunohistochemistry and colocalization studies revealed association between Skp2 and Slug in aggressive prostate cancer. Other factors such as TMPRSS2-ERG fusion or oxidative stress may contribute to prostate cancer progression. Citation Format: Jan Bouchal, Gvantsa Kharaishvili, Alena Mickova, Mariam Gachechiladze, Alena Burdova, Martin Mistrik, Milan Kral, Balazs Gyorffy, Zdenek Kolar. Skp2 associates with Slug and androgen receptor in aggressive prostate cancer [abstract]. In: Proceedings of the AACR Special Conference: Prostate Cancer: Advances in Basic, Translational, and Clinical Research; 2017 Dec 2-5; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(16 Suppl):Abstract nr A006.

Abstract 5649: Multi-omics data indicate that primary and lymph node oral cancer cells-derived extracellular vesicles carry cargo molecules with a specific aggressive pattern

Abstract Oral squamous cell carcinoma (OSCC) exhibits high incidence and morbidity, representing 300,000 new cases and 145,000 deaths per year. Moreover, the 5-year survival rate is 50% and this is directly associated with regional lymph node metastases. To explore how the extracellular vesicles (EVs) play a significant role on OSCC lymph node metastasis, we characterized the EVs released by tumor primary site cells and their paired lymph node metastasis cells using nanoparticle tracking analysis, high-resolution microscopy (Cryo-EM), EVs uptake by HUVEC, BJ-5ta and gingival primary fibroblasts recipient cells and the proteins, lipids, metabolites and miRNAs contents. Both cells-derived EVs showed a similar size, morphology, and the presence of annexin-2 and flotillin-1 surface markers. Interesting, only gingival primary recipient cells were more likely to uptake primary OSCC-derived EVs compared with lymph node-derived EVs. This effect is probably associated with the differential abundance of 490 proteins, 316 miRNAs, 63 lipids and 17 metabolites between the oral cancer cells-derived EVs. Markedly, multilevel omic data integration indicates that metabolic processes were over-represented mainly in primary OSCC-derived EVs, suggesting that these EVs carry specific cargo molecules associated with a more aggressive pattern compared with lymph node OSCC-derived EVs. Financial support: FAPESP (Process number 2016/50005-7). Citation Format: Ariane F. Busso-Lopes, Carolina M. Carnielli, Flávia Winck, Diego M. Riaño-Pachón, Ana K. Oliveira, Rute A. Costa, César Rivera, Camila Caldana, Jay W. Fox, Adriana F. Paes Leme. Multi-omics data indicate that primary and lymph node oral cancer cells-derived extracellular vesicles carry cargo molecules with a specific aggressive pattern [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5649.

Histological description of the larval development of Brycon gouldingi Lima, 2004 (Teleostei, Characidae)

Piabanha, Brycon gouldingi , is an endemic species in the Tocantins-Araguaia basin. It has aroused the interest of both fish farmers, who started its creation in confinement, and riverine people who appreciate it as a food source. In order to provide information about organic systems of B. gouldingi larvae, a histological description was performed after capturing adult specimens in the Rio das Mortes (Mato Grosso, Brazil), adapted to captivity and induced to spawn at Buriti Fisheries (Nova Mutum, MT, Brazil). The collection of samples took place at pre-defined moments after hatching, and the processes relating to morphological differentiation of digestive, excretory, cardiorespiratory, nervous/sensory systems and gas bladder were characterized. At the hatching were found: undifferentiated digestive system; pronephros (primitive kidney), rudimentary heart, central nervous system characterized by primary vesicles, optic vesicle forming the optic cup and crystalline lens. In the course of ontogeny, these organic systems were developed and at the time of the total absorption of the yolk at 55 hours post-hatching were found: the digestive system consisting of the head gut, foregut, midgut and hindgut; two heart chambers and branching of gill arches; three regions of the brain (forebrain, midbrain and hindbrain), neuromasts, olfactory cavity, taste buds; eye consisting of well-defined layers; presence of gas bladder. The results of this study may be useful in providing support for the captive breeding of B. gouldingi during the larval stage.

Morphological characteristics of mule conceptuses during early development.

Hybrids between species are often infertile and extremely rare among mammals. Mules, i.e. crossing between the horse and the donkey, on the other hand are very common in agricultural and leisure practices due to their enhanced post-natal physical characteristics that is believed to occur for outbreeding or hybrid vigor. Since no reports are availableon the effects of hybrid vigor during early development, this study focused on characterizing the intrauterine development of mule conceptuses during critical embryo-to-fetus transition period. Nine embryos and fetuses of early gestation, obtained after artificial insemination and transcervical flushing, were evaluated by means of gross anatomy and histology and compared to data available for the equine. We found that some events, such as C-shape turning, apearence of branchial archs, limb and tail buds, formation of primary and secondary brain vesicles, heart compartmentalization, and development of somites, occurred slightly earlier in the mule. Nonetheless, no major differences were observed in other developmental features, suggesting similarities between the mule and the horse development. In conclusion, these data suggest that the effect of hybrid vigor is present during intrauterine development in the mule, at least with regard to its maternal parent.

Primary vesicles, vesicle-rich segregation structures and recognition of primary and secondary porosities in lava flows from the Paraná igneous province, southern Brazil

This study focuses on a volcanic succession of pāhoehoe to rubbly lavas of the Parana–Etendeka Province exposed in a single road profile in southernmost Brazil. This work provides an integrated approach for examining primary vesicles and vesicle-rich segregation structures at the mesoscopic scale. In addition, this study provides a quantitative analysis of pore types in thin section. We documented distinct distribution patterns of vesicle and vesicle-rich segregation structures according to lava thickness. In compound pāhoehoe lavas, the cooling allows only vesicles (<1 cm size) and pipe vesicles to be frozen into place. In inflated pāhoehoe lavas, vesicles of different sizes are common, including pipe vesicles, and also segregation structures such as proto-cylinders, cylinders, cylinder sheets, vesicle sheets, and pods. In rubbly lavas, only vesicles of varying sizes occur. Gas release from melt caused the formation of primary porosity, while hydrothermal alteration and tectonic fracturing are the main processes that generated secondary porosity. Although several forms of porosity were created in the basaltic lava flows, the precipitation of secondary minerals within the pores has tended to reduce the original porosities. Late-stage fractures could create efficient channel networks for possible hydrocarbon/groundwater migration and entrapment owing to their ability to connect single pores. Quantitative permeability data should be gathered in future studies to confirm the potential of these lavas for store hydrocarbons or groundwater.

Morphological characterisation of vesicular structures in the canine ejaculate

Membrane vesicles (MV) have been identified in seminal plasma from various species and they are thought to have a significant impact on semen quality and fertilisation. Although recently presence of MV has been also described in the canine ejaculate, detailed knowledge on their morphology is missing by now. This is, however, needed to provide a basis for detailed biochemical and functional studies as it is generally assumed that different MV populations are responsible for distinct tasks. MV were prepared for light (LM) and transmission electron microscopy (TEM) analysis using samples from normospermic dogs (n=15), hypokinozoospermic dogs (n=2, h) and one castrated azoospermic dog (a). For TEM, a new preparation protocol was used resulting in a higher MV retrieval rate. Using fractionated semen samples, most MV were identified in the second (sperm-rich) fraction in LM. Using pooled ejaculates, three different MV types could be identified in LM: (1) large MV with a marginal accumulation of opaque, granulated material, (2) medium- to small size MV with dense, opaque content and (3) small MV with no further defined contents. No direct contact between sperm and MV could be visualised. In TEM, 11 different MV types were identified based on diameter, structure, contents and electron density of contents as well as presence, number and size of smaller MV inside the MV itself. In normospermic males, secondary vesicles (type i, H, K1/2) included smaller vesicles and had a weighted mean diameter of 409.46nm; hereof types i, H and K1 were smaller (mean: 287.55nm, range: 51.25–994.86nm) and type K2 was larger (mean: 1746.43nm, range: 1003.66–3289.34nm). Primary vesicles (mean diameter: 135.29nm) – without vesicles inside – were differentiated into larger MV (A, B, C1/2) with a mean diameter of 219.63nm (range: 39.08–1300.13nm) and small primary MV (F, G) with a mean diameter of 66.12nm (range: 24.62–99.84nm). Whereas all mentioned MV were round to oval and mostly double-, rarely multiple-membrane surrounded, one longish primary MV type (L) was identified. In general, small primary vesicles were most common independent of semen quality, but distribution frequency of vesicle types differed between normospermic, pathospermic dogs and the castrated male. Mean weighted diameter of MV was 195.14nm (range: 24.62–3289.34nm) in normospermic males with the maximum diameter being smaller in the other dogs (h: 2096.78nm; a: 1314.06nm). Our results provide new information about ultrastructure and distribution frequency of canine MV in normospermic males and point to possible differences in MVs depending on semen quality. They provide the basis for further detailed functional analysis of MV subpopulations. Furthermore, the presence of MV in the castrated azoospermic male confirms an at least partly prostatic origin of canine MV.

Transvaginal three-dimensional sonographic assessment of the embryonic brain: a pilot study.

AimsA very good knowledge of human embryology is mandatory not only for the correct sonographic assessment of the developing brain, but also for better understanding the origins of congenital anomalies involving the central nervous system. 3D transvaginal sonography may be an effective technique for imaging the developing brain. The aims of this explorative study are to demonstrate the feasibility of imaging the embryonic brain between 7 and 10 weeks of gestation for clinical studies by using a 3D high-frequency vaginal ultrasound transducer and to provide a reference for the morphology of the brain in the embryonic period.Materials and methodsFour embryos of 9 mm, 17 mm, 23 mm and 31 mm crown-rump length respectively were assessed in vivo by transvaginal sonography. We gave a special attention to the embryonic brain. All patients were examined with a Voluson E10, BT 15 ultrasound scanner (GE Healthcare, Zipf, Austria), using a high-frequency 6–12 MHz/ 256-element 3D/4D transvaginal transducer. Three-dimensional sonography was performed routinely as the patients were scanned. The multiplanar display was used after selecting the best volume. The Omni view® software was used for digitally slicing the selected volumes.ResultsWe describe the morphological details of the developing brains of four embryos ranging from 7 to 10 gestational weeks. In the human embryo 9 mm CRL the hypoechogenic cavities of the three primary vesicles (prosencephalon, mesencephalon, rhombencephalon) could be observed on a sagittal section. In the human embryo 17 mm CRL the prosencephalon was divided into the median diencephalon and two telencephalic vesicles, which were partially separated by the falx cerebri. In the human embryo 23 mm CRL the cerebral hemispheres developed and they were completely separated by the falx cerebri. The choroid plexus was evident inside the lateral ventricles and the fourth ventricle. In the human embryo 31 mm CRL the ventral thalamus was evident, and the ganglionic eminence, as the precursor of the basal ganglia, was well seen on the floor of the cerebral hemispheres.ConclusionsStudies of embryology are still needed for a complete understanding of the developing brain. 3D sonography using a high-frequency vaginal ultrasound transducer is feasible for imaging the embryonic brain with an acceptable quality for clinical studies.

First Report of Choanephora Blight Caused by Choanephora infundibulifera on Hibiscus rosa-sinensis in Korea.

Hibiscus rosa-sinensis L., commonly known as Chinese hibiscus, is an evergreen flowering shrub belonging to the Malvaceae and is widely cultivated throughout Asia including Korea. In August 2013, blight was observed on Chinese hibiscus in a commercial flower nursery in Seoul, Korea. Initial symptoms began as reddish purple spots at the tip of flowers and expanded to encompass entire flowers. Infected lesions appeared water-soaked, reddish brown, and were followed by rapid rotting of infected tissues. Approximately 50% of the plants surveyed were affected. Monosporous sporangiola formed on infected tissue were transferred to potato dextrose agar (PDA). Fungal colonies were obtained that were at first white with abundant aerial mycelium, and then became yellowish with the appearance of sporangiola. Sporangiophores bearing sporangiola were erect to slightly curved, unbranched, and hyaline. Funnel-shaped secondary vesicles formed on the primary vesicles. Sporangiola were indehiscent, ovoid to subglobose, smooth, non-striated, brown to dark brown, 10 to 27.5 × 8.5 to 17 μm, and sometimes germinated in culture. The fungus was identified as Choanephora infundibulifera (Curr.) D.D. Cunn. based on the morphological and cultural characteristics (2). Voucher specimens were housed in the Korea University Herbarium (KUS). An isolate obtained from KUS-F27535 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC47643) and used for a pathogenicity test and molecular analyses. To confirm identity of the fungus, genomic DNA was extracted with DNeasy Plant Mini Kits (Qiagen Inc., Valencia, CA). The internal transcribed spacer (ITS) region of rDNA and the D1/D2 region of the large subunit (LSU) were amplified with the primers ITS1/ITS4 and NL1/LR3, respectively (3), and sequenced. The resulting 635-bp ITS and 680-bp D1/D2 sequences were deposited in GenBank (Accession Nos. KF486539 and KF486538). A GenBank BLAST search revealed that the ITS sequences showed 100% similarity with that of C. infundibulifera (JN943009) and D1/D2 sequences also showed 100% identity with that of C. infundibulifera (JN939193). A sporangiola suspension (2 × 104 cells/ml) was sprayed over three pots of the shrub, kept in a humid chamber for 2 days, and placed in greenhouse (28°C and 80 to 100% RH). Another three potted plants of the same age were sprayed with sterile water and served as controls. After 4 days, typical blossom blight symptoms, identical to the ones observed in the nursery, developed on the inoculated flowers. No symptoms were observed on controls. C. infundibulifera was re-isolated from inoculated plants. Pathogenicity test was conducted twice with the same results, fulfilling Koch's postulates. Choanephora blight caused by C. infundibulifera on H. rosa-sinenesis has been reported in Japan, Myanmar, Nepal, Guinea, and the United States (1). In Korea, there was one record of this fungus on H. syriacus (1). To our knowledge, this is the first report of C. infundibulifera on H. rosa-sinensis in Korea. This pathogen could be a potential threat to the production of this ornamental shrub over a prolonged period of hot and humid weather.

Regional differences in actomyosin contraction shape the primary vesicles in the embryonic chicken brain

In the early embryo, the brain initially forms as a relatively straight, cylindrical epithelial tube composed of neural stem cells. The brain tube then divides into three primary vesicles (forebrain, midbrain, hindbrain), as well as a series of bulges (rhombomeres) in the hindbrain. The boundaries between these subdivisions have been well studied as regions of differential gene expression, but the morphogenetic mechanisms that generate these constrictions are not well understood. Here, we show that regional variations in actomyosin-based contractility play a major role in vesicle formation in the embryonic chicken brain. In particular, boundaries did not form in brains exposed to the nonmuscle myosin II inhibitor blebbistatin, whereas increasing contractile force using calyculin or ATP deepened boundaries considerably. Tissue staining showed that contraction likely occurs at the inner part of the wall, as F-actin and phosphorylated myosin are concentrated at the apical side. However, relatively little actin and myosin was found in rhombomere boundaries. To determine the specific physical mechanisms that drive vesicle formation, we developed a finite-element model for the brain tube. Regional apical contraction was simulated in the model, with contractile anisotropy and strength estimated from contractile protein distributions and measurements of cell shapes. The model shows that a combination of circumferential contraction in the boundary regions and relatively isotropic contraction between boundaries can generate realistic morphologies for the primary vesicles. In contrast, rhombomere formation likely involves longitudinal contraction between boundaries. Further simulations suggest that these different mechanisms are dictated by regional differences in initial morphology and the need to withstand cerebrospinal fluid pressure. This study provides a new understanding of early brain morphogenesis.

The primary brain vesicles revisited: Are universal the three primary vesicles (forebrain/midbrain/hindbrain) in vertebrates?

The primary brain vesicles revisited: Are universal the three primary vesicles (forebrain/midbrain/hindbrain) in vertebrates?

Malformations of Ventral Induction

t o s t p o T f a Ventral induction occurs during 5-10 weeks of gestation. During this phase 3 primary vesicles (prosencephalon, esencephalon, and rhombencephalon) are differentiated to orm forebrain, midbrain, and hindbrain, respectively.1 Norally, early forebrain vesicles, prosencephalon, develop in he rostral end of the neural tube between 22 and 24 days of estation. The prosencephalon forms 2 laterally directed dierticula by 5-6 weeks and is divided into telencephalon future cerebrum) and diencephalon (future thalamus and ypothalamus). Diencephalon and telencephalon differentition occurs from the provesicle before the neural tube loses. Any insult during this phase affects the development f brain vesicles and the formation of the facial skeleton. cular and nasal anomalies are frequently associated with orebrain malformation because the optic placode and forerain develop at the same time, with subsequent formation of he olfactory vesicle 1 week later. The rhombencephalon diides into metencephalon (future pons and cerebellum) and yelencephalon (future medulla). Malformation of the hombencephalon is discussed in detail in this issue under Posterior Fossa Malformations.” The commonly seen forebrain ventral induction malforations are (1) holoprosencephaly, (2) atelencephaly, (3) lfactory aplasia, (4) agenesis of the corpus callosum, (5) genesis of the septum pellucidum (septo-optic dysplasia and avum vergae and pellucidum).

Polymerization-like Multilevel Hierarchical Self-Assembly of Polymer Vesicles into Macroscopic Superstructures with Controlled Complexity

We demonstrate a high-leveled hierarchical self-assembly process into fractal structures. Two hyperbranched multiarm copolymers are first coassembled into binary isotropic vesicles in the primary self-assembly. Then, these primary vesicles are in situ endowed with anisotropic hydrophobic "binding sites" through a pH-induced lateral microphase separation, undergoing an isotropic-anisotropic transition. Subsequently, the anisotropic vesicles further assemble together through the specific self-recognition between the binding sites into linear, branched, cyclization, and network-like vesicle chains. Furthermore, the obtained vesicle chains can transform into linear, branched, ring-like, and network tubes through successive vesicle fusion. Such a hierarchical process is pH-triggered and well-controlled by adjusting the vesicle compositions and is coined as "polymerization-like" self-assembly due to the striking analogy between the vesicle association model and polymerization process.

Lithofacies and reservoir properties of Tertiary igneous rocks in Qikou Depression, East China

SUMMARY Tertiary igneous rocks in Qikou Depression, East China are primarily basic rocks. Two types of igneous reservoirs including the effusive basaltic reservoir and the near-surface intrusive doleritic reservoir have been identified so far and each has different reservoir spaces. In this paper, the lithofacies and reservoir properties of Tertiary igneous rocks are described through a systematic analysis of the characteristics of seismic reflection and well logs and selected cores observation. Both of the two types of igneous rocks present strong and low-frequency reflection events on seismic profiles, however, intrusive diabases exhibit the characteristic of strata penetration with a small intersection angle. Effusive basaltic lavas are divided into upper, middle and lower subfacies. This can be distinguished from the well log curves. Observation of sampled cores finds that primary vesicles and amygdales are abundant in the upper and lower subfacies of basaltic lavas. These vesicles can be favourable reservoir space if connected by cracks formed in the late-stage. Intrusive diabases can also be separated into inner subfacies as well as the upper and lower marginal subfacies, of which the former is well crystallized because of slowly cooling, whereas the latter is vitreous due to fast descending temperature at the contact zone between intrusive rocks and host rocks. Hydrothermal metamorphism left the surrounding mudstones metamorphosed to form shrunk fissures and structural fractures as reservoir space, making mudstones of poor porosity and permeability to be metamorphic and have certain reservoir potential.

Cloning of gene fragment of estrogen receptor-βand its expression in mouse embryo

In order to study the expression and regulation effects of estrogen receptor-beta (ERbeta) in the development of mouse embryo, the primer of ERbeta was designed, the ERbeta fragment was first obtained by RT-PCR and subcloned into plasmids pGEM- 3Z, then the recombinant plasmids were linearized with the restriction enzymes of EcoRand Hind. Using Sp6 and T7 RNA polymerase, the digoxigenin(dig) labeled sense and anti-sense probes were transcriped in vitro, respectively. Then the expression of ERbeta in mouse embryo was examined with the probes by whole-mount in situ hybridization. The results indicated that ERbeta is expressed in the brain, spinal neural tube, genital ridge, pericardium, limb bud and mandibular arch of 10.5 dpc embryo, and is also expressed in the telencephalon, mesencephalon, medulla oblongata, spinal cord and limb bud of 13.5 dpc embryo. These results suggest that ERbeta maybe play a role of regulation in sexual differentiation, primal differentiation of neural tube, further differentiation of three primary cerebral vesicles and spinal cord, generation and differentiation of bone and cartilage of limb bud, development of pericardium and configuration differentiation of mandibular in mouse embryo.

Characterization of sequential exocytosis in a human neuroendocrine cell line using evanescent wave microscopy and “virtual trajectory” analysis

Secretion of hormones and other bioactive substances is a fundamental process for virtually all multicellular organisms. Using total internal reflection fluorescence microscopy (TIRFM), we have studied the calcium-triggered exocytosis of single, fluorescently labeled large, dense core vesicles in the human neuroendocrine BON cell line. Three types of exocytotic events were observed: (1) simple fusions (disappearance of a fluorescent spot by rapid diffusion of the dye released to the extracellular space), (2) "orphan" fusions for which only rapid dye diffusion, but not the parent vesicle, could be detected, and (3) events with incomplete or multi-step disappearance of a fluorescent spot. Although all three types were reported previously, only the first case is clearly understood. Here, thanks to a combination of two-color imaging, variable angle TIRFM, and novel statistical analyses, we show that the latter two types of events are generated by the same basic mechanism, namely shape retention of fused vesicle ghosts which become targets for sequential fusions with deeper lying vesicles. Overall, approximately 25% of all exocytotic events occur via sequential fusion. Secondary vesicles, located 200-300 nm away from the cell membrane are as fusion ready as primary vesicles located very near the cell membrane. These findings call for a fundamental shift in current models of regulated secretion in endocrine cells. Previously, sequential fusion had been studied mainly using two-photon imaging. To the best of our knowledge, this work constitutes the first quantitative report on sequential fusion using TIRFM, despite its long running and widespread use in studies of secretory mechanisms.

Oogenesis in calanoid copepods

Summary An ultrastructural investigation of ovary structure and oogenesis was conducted on ten species of cold and warm water calanoid copepods from six families with broad distributions ranging from coastal to mesopelagic habitats. They include Calanus finmarchicus, C. hyperboreus, C. glacialis, Euchaeta marina, Centropages furcatus, C. velificatus, Labidocera aestiva, Acartia tonsa, Pleurommama xiphias, and Undinula vulgaris. All ten species show similarities with respect to ovarian morphology and oogenesis. Oogenesis is asynchronous with all stages of oocytes present within each sexually mature female. The oocytes differentiate to diplotene of the first meiotic division and undergo a growth phase within the ovary before initiating vitellogenesis within the diverticulae. Vitellogenesis involves both an autosynthetic (endogenous) and a heterosynthetic (exogenous) process resulting in the deposition of two different types of yolk bodies. Endogenous yolk formation is initiated by the proliferation of vesicular endoplasmic reticulum (ER) through the blebbing of the outer lamina of the nuclear envelope. These ER vesicles contain a variable number of granules that arise before the vesicles detach from the nuclear envelope. Parallel arrays of annulate lamellae (AL) also appear in this region and their swollen, distal cisternae release ER vesicles into the surrounding ooplasm that are indistinguishable from those generated by the nuclear envelope. Golgi complexes proliferate and produce secretory products that fuse with both the ER cisternae and newly formed yolk bodies. Simultaneous biosynthetic activity of the ER, AL, and Golgi results in the formation of Type I yolk bodies, and primary and secondary cortical vesicles that likely participate in the formation of the egg envelope. In the later stages of vitellogenesis, extraoocytic yolk precursors originating in the hemocoel pass through the follicle cells that encompass the oocytes in an endocytotic process that releases precursors into the pervitelline space. Oocytes then incorporate these precursors endocytotically resulting in the gradual assembly of Type II yolk bodies. To our knowledge, this is the first ultrastructural study to describe the biosynthesis of specific products (yolk bodies and cortical granules) by annulate lamellae in any metazoan oocyte. Vitellogenesis in calanoid coperpods is significantly different from that described in any other crustacean group. Further investiation of this process in related crustaceans might provide useful phylogenetic insights.