Primary Growth Phase Research Articles

Histological characteristics of oocyte differentiation in the captive longnose seahorse Hippocampus trimaculatus (Leach, 1814).

This study aimed to explore the reproductive histology and oocyte differentiation of the longnose seahorse Hippocampus trimaculatus (Leach, 1814) in captivity. Five mature healthy females were histologically observed. The reproductive systems of the five specimens exhibited similar morphological characteristics with a pair of saccular creamy white ovaries merging caudally into a single gonoduct. There were two germinal ridges lined with a layer of germinal epithelium (GE). The ovarian maturation of this species was considered asynchronous. The oogenic cells were classified into oogonia and oocytes at several developmental phases based on their size and characteristics. Oogonia were identified among the connective tissue in the middle area of the GE. The stromal compartment contained oocytes that were classified into four distinct phases: the primary growth (PG) phase having two steps (perinucleolar and oil droplets-cortical alveolar steps) and the secondary growth (SG) phase with three oocyte types, including early SG oocytes, late SG oocytes, and fully grown oocytes. The atretic oocytes (AO) were observed in all stages of oogenesis. Postovulatory follicles were also seen among the ovarian connective tissue. The occurrence of postovulatory follicles suggested that the specimens analysed in this study were in the spawning period. This research provides new insights into the identification of the reproductive cycles and morphological characteristics of the ovary of H. trimaculatus.

Proteomic analysis of zebrafish folliculogenesis identifies YB-1 (Ybx1/ybx1) as a potential gatekeeping molecule controlling early ovarian folliculogenesis.

As in mammals, ovarian folliculogenesis in teleosts also consists of two phases: the primary growth (PG) and secondary growth (SG) phases, which are analogous to the preantral and antral phases respectively in mammals. In this study, we performed a proteomic analysis on zebrafish follicles undergoing the PG-SG transition aiming to identify factors involved in the event. Numerous proteins showed significant changes, and the most prominent one was Y-box binding protein 1 (YB-1; Ybx1/ybx1), a transcription factor and mRNA-binding protein. YB-1 belongs to the Y-box binding protein family, which also includes the gonad-specific YB-2. Interestingly, phylogenetic analysis showed no YB-2 homolog in zebrafish. Although ybx1 mRNA was expressed in various tissues, its protein Ybx1 was primarily produced in the gonads, similar to YB-2 in other species. In the ovary, Ybx1 protein started to appear in early follicles newly emerged from the germ cell cysts, reached the highest level in late PG oocytes, but decreased precipitously when the follicles entered the SG phase. In PG follicles, Ybx1 might function as a key component of the messenger ribonucleoprotein particles (mRNPs) in association with other RNA-binding proteins. Similar to mammalian YB-1, zebrafish Ybx1 also contains functional signals that determine its intracellular localization. In conclusion, Ybx1 may play dual roles of YB-1 and YB-2 in zebrafish. In the ovary, Ybx1 binds mRNAs to stabilize them while preventing their translation. At PG-SG transition, Ybx1 is removed to release the masked mRNAs for translation into functional proteins, leading to follicle activation.

Sex steroid profile during oocyte development and maturation in the intertidal worm Marphysa madrasi (Polychaeta: Eunicidae) from the east coast of India

Marphysa madrasi is a commercially valuable maturation diet in crustacean aquaculture. This study presents the first detailed investigation of oogenesis in the intertidal polychaete worm M. madrasi and reports the steroid profile during oocyte growth and development. Oogenesis is extraovarian type I, originating from coelomic epithelial cells, with four stages of development - primary growth, early vitellogenic, late vitellogenic, and maturation. The primary growth phase contains oogonial cells and previtellogenic oocyte clusters in the early, mid, and late stages of development form a dispersed ovary attached to blood vessels. The late previtellogenic oocytes detach from the ovary at the onset of vitellogenesis. The detached oocytes complete vitellogenesis and final maturation in the coelomic fluid as solitary free-floating cells without any connection with follicle cells. The worms display asynchronous reproduction with a heterogeneous population of developing oocytes. Steroid extracts from the polychaete homogenates in different stages of oogenesis were identified by HPLC and confirmed by LC-MS/MS. In M. madrasi, two vertebrate-type steroids, pregnenolone (P5) and 17α-hydroxyprogesterone (17-OHP) were detected and quantified. The P5 levels were low in immature worms but increased significantly by ∼ 8.3-fold in the previtellogenic stage and peaked during oocyte maturation. 17-OHP levels were low in immature worms but gradually increase as the oogenesis progress to the primary growth and early vitellogenic phase, with a significant increase (p < 0.001) during the late vitellogenic phase. Although an increase in the concentration of P5 and 17-OHP during vitellogenesis and maturation of oocytes points to a possible role in reproduction, the absence of other vertebrate-type steroids in the investigated polychaete signifies a plausible uptake of P5 and 17-OHP from the environment.

Inactivation of MrSir2 in Monascus ruber Influenced the Developmental Process and the Production of Monascus Azaphilone Pigments.

Monascus species are the producers of Monascus azaphilone pigments (MonAzPs) and lipid-lowering component Monacolin K, which have been widely used as food colorant and health products. In this study, silent information regulator 2 (Sir2) homolog (MrSir2) was characterized, and its impacts on the development and MonAzPs production of Monascus ruber were evaluated. Enzyme activity test in vitro showed that MrSir2 was an NAD+-dependent histone deacetylase. Compared to WT, Δmrsir2 strain accumulated more acetylated lysine residues of histone H3 subunit during its vegetative growth phase, and it exhibited accelerated mycelial aging, more spores, increased resistance to oxidative stress, and more MonAzPs production. RNA-Seq-based transcriptome analysis revealed that MrSir2 mainly regulated the gene expression in macromolecular metabolism such as carbohydrates, proteins, and nucleotides, as well as genes encoding cell wall synthesis and cell membrane component, indicating that MrSir2 probably facilitates the metabolic transition from the primary growth phase to the mycelial aging. Taken together, MrSir2 mainly targets H3 subunit at the vegetative growth phase and affects the development of M. ruber and MonAzPs production.

Successful experience with axenic isolation of Leishmania infantum in culture medium enriched with equine blood

The aim of the study was to demonstrate a successful experience in the axenic isolation of Leishmania infantum (L. infantum) promastigotes in culture using base agar media enriched with defibrinated equine blood (SDE). The study used lymph node aspirate, spleen, liver and lymph node fragments from dogs infected with the disease. The culture media used for the isolation of the parasite were biphasic (solid and liquid) and monophasic (liquid). For a biphasic phase, Blood Agar Base (BAB) and Neal Novy Nicolle (NNN) medium were used, both enriched with SDE, which after solidification was added to Brain Heart Infusion (BHI) broth. For the monophasic phase, Medium 199 (M199) supplemented with 10% fetal bovine serum (SFB) was used. The promastigotes were seeded in duplicate, kept in an oven by biochemical oxygen demand at 25º C for 15 days. Microscopic readings were taken daily to monitor the primary growth phase of the parasite. After primary isolation, they were serialized and diluted with a final concentration of 1x106 parasites/mL, for evaluation of the growth curve. The continuous study showed that in tubes containing SDE enriched NNN and BAB biphasic media, primary parasite growth was 72 hours, whereas in tubes supplemented with SFB it was 96 hours. Regarding the growth curve, it was observed that during an exponential phase, as SDE enriched spleen, liver and lymph node fragments obtained greater promastigotes, when used with SFB supplemented ones, being 189 x 106 e 128 x 106 parasites/mL, respectively. There was a significant difference (P &lt; 0.05) in the percentage of the number of promastigotes in a medium enriched with SDE, when compared to the medium with SFB. The primary tubes remained viable throughout the study period, even after replenishing used to maintain the parasite. The traditional technique of isolation of the parasite in culture medium most commonly used is in base agar enriched with rabbit blood; however, operational difficulties in the acquisition of this enricher have driven the search for other sources of protein supplementation for L. infantum cultivation. The study reveals that the use of SDE as an alternative enrichment medium for the primary isolation of L. infantum was feasible to be used in laboratory routine.

Balbiani body formation and cytoplasmic zonation during early oocyte development in two Clupeiform fishes.

The Balbiani body (Bb) was examined in primary growth phase oocytes for the first time in two clupeoid fish species, the Mediterranean sardine, Sardina pilchardus, and the European anchovy, Engraulis encrasicolus, which belong to different families, Clupeidae and Engraulidae, respectively. Cytoplasmic morphological changes of early secondary growth oocytes were also investigated using confocal laser scanning microscopy, light and transmission electron microscopy. The ultrastructural observations showed that the two species develop a distinct spherical Bb. However, differences in the cytoplasm, mainly in the perinuclear area, were observed. Briefly, in sardine the Bb coexists with a thick perinuclear ring containing mitochondria, nuage, endoplasmic reticulum and small vesicles, while in anchovy this perinuclear ring is thinner, consisting of complexes of nuage and mitochondria. After the disassembly of the Bb, a prominent cytoplasmic zonation develops in the secondary growth oocytes of sardine and anchovy, although with different organelle distribution between the two species. Sardine oocytes exhibit a thick zone of endoplasmic reticulum around the nucleus, whereas in those of anchovy, a thick mitochondria-rich ring surrounding the nucleus was observed. The cytoplasmic characteristics, such as the perinuclear ring in primary oocytes in sardine and the mitochondria-rich ring of early secondary oocytes in anchovy, are also discernible in histological sections by standard procedures and could thus be used as indicators of maturity or imminent spawning period in routine light microscopy observations, providing a valuable tool for applied fisheries biology.

Histological observations on the oocyte development in the Picnic seabream, Acanthopagrus berda (Forsskål 1775)

Histological analysis was carried out on ovarian development in the picnic seabream, Acanthopagrus berda (Forsskål 1775) caught in the estuarine waters of Calicut, Kerala, India from January to December 2016. Four developmental phases of oocyte development were identified viz., oogonia proliferation and transformation into the primary oocyte, primary growth phase, secondary growth phase and maturation phase and these are similar to those described for other species. The reproductive development process begins in a very young ovary with the formation of oogonia, the cells originally derived from special primordial germ cells formed early in the embryonic development; their multiplication and differentiation are followed by the release of mature oocytes. Oocyte development stages observed in A. berda were chromatin nucleolus, early perinucleolar, late perinucleolar, lipid vesicle stage I oocyte, lipid vesicle stage II oocyte, primary yolk granule stage, secondary yolk granule stage, tertiary yolk granule stage, hydrated oocyte and atretic oocyte. The oocytes entered the maturation phase when they attained the maximum size of 459.6±71.83 µm. Histological examination of the mature ovaries showed the presence of all the different oocyte stages in the ovary of A. berda with an asynchronous mode of ovarian development shortly before spawning, indicating that the species is a multiple spawner.

Histological and Ultrastructural Characterization of the Gonads of the Grunting Toadfish Allenbatrachus grunniens (Linnaeus, 1758) from the Pranburi River Estuary, Thailand

Information on the reproductive biology of toadfish remains limited. In this study, we examined the structure and development of gonads in the grunting toadfish Allenbatrachus grunniens (Linnaeus, 1758) using morphological, histological and ultrastructural methods. The fish were collected from the Pranburi River Estuary, Thailand, during the annual reproductive period for this species (January to December 2018). The ovary of this species was paired and had elongated sac-like structures parallel to the kidneys and the digestive tract. In females, we identified 4 oocyte differentiation phases in the ovary including oogonia proliferation phase, a primary growth phase that was further classified into 2 steps (perinucleolar and oil droplets-cortical alveolar steps), and a secondary growth phase that also contained 2 steps (secondary growth and full-grown oocyte steps) and post-ovulatory phases, indicating an asynchronous pattern in ovarian development for this species. Transmission electron microscopy showed the 4 layers including the zona pellucida, basement membrane, granulosa cells and theca cells, all of which initially appeared in the oil droplets-cortical alveolar stage. The zona pellucida and the granulosa cells were highly developed during the secondary growth stage. The granulosa cells contained abundant smooth endoplasmic reticulum near the mitochondria. In males, the spermatogenesis was classified into spermatogonium to spermatozoa. Finally, we associated the morphological gonadal developments (stage I - IV) and the gonadosomatic index (GSI value) with the cellular developmental processes in both sexes. These results help integrate various levels of reproductive observations, which will be applied to understanding the reproductive cycle and development for aquaculture.

Oocyte Differentiation and Reproductive Health of Solitary Tunicate (Styela plicata) from Eastern Coast of Thailand

Histopathological examination is a widely acknowledged technique to assess the reproductive health of aquatic organisms, but it has never been applied to the tunicate Styela plicata, a potential indicator species of water quality. In this study, we examined the oocyte differentiation of S. plicata obtained from the eastern coast of the Gulf of Thailand in order to provide basic information for future assessment of its reproductive health. The mature gonad of S. plicata comprised several ovo-testicular convoluted tubes, in which each tube was divided into apical and terminal portions. The ovarian tissue is located in the apical part of the tunicate body and contained oocytes of various differentiation stages (asynchronous development type) consisting of the four phases namely oogonial proliferation phase, primary growth phase, secondary growth phase (secondary growth and full-growth stages), and post-ovulatory phase. Changes in the morphology of oocytes and follicular cells were described for each differentiation stage. In addition, we unexpectedly observed a high prevalence of atretic follicles (24.5%), which might indicate the oocyte damage by environmental stresses. These findings would be useful for future assessment of reproductive histology of S. plicata and other tunicate species from environmental perspectives.

Crushed maize seeds enhance soil biological activity and salt tolerance in caper (Capparis spinosa L.)

Caper (Capparis spinosa L.) is an ancient plant with ecological, industrial, medicinal and economical benefits which at its primary growth phase is sensitive to salt stress. This study was conducted to investigate the biostimulant effect of supplementing crushed maize (Zea mays L.) seeds (CMS) (0, 10 and 20 %) into growing media on caper salt tolerance and soil biological activity under different salinity level (0, 5, 10 and 15 dS/m). Applying 20 % biostimulant increased the number of leaves and roots by 200.0 and 56.0 %, respectively, compared with untreated seedlings at the same salinity level. The application of 20 % biostimulant also enhanced the proline content (by 40.5 %), total phenolic contents (34.0 %), rutin (356.4 %) and leaf K content (29.8 %) of treated capers compared with untreated seedlings. Applying 20 % biostimulant also significantly increased the soil enzymes activity including urease, dehydrogenase, and phosphatase as well as soil bacteria population. Whereas, using biostimulant significantly decreased the content of Na in different caper’s organs. However, the total flavonoid content, soil fungi population and most of the morphological parameters of caper were not significantly affected by the application of CMS. In total, the soil application of CMS had a substantial effect on physiological and biochemical parameters of salinized capers and soil biological activity based on principal component analysis (PCA). In conclusion, using CMS as a soil supplement can be recommended as a safe and natural biostimulant to alleviate the negative impacts of salt stress in caper seedlings.

Comparative stages of atretic oocytes between Episesarma versicolor and E. singaporense from Thailand: Implication on the reproductive success

Informative reviews on the oocyte atresia referring to degeneration and resorptive mechanism are warranted in common animals, but it is rarely still found in carbs. In this current study, we focused on characteristics and stage of the atretic oocyte from two important female sesarmid carbs (violet vinegar crab, Episesarma versicolor, and Singapore vinegar crab, E. singaporense) during ovarian maturation. A total of 30 female samples in each carb species were collected from the Palian mangrove area of Trang province, Thailand. The results shared among those carb samples that both primary growth (PGs) and secondary growth (SGs) phases underwent a degenerative process of atresia. The latter of atresia during SGs was also divided into five stages (I, II, III, IV, and V). The degeneration of follicular cell and yolk architecture identifying in the SGs were highlighted. The number of atresias was slightly compared between E. versicolor and E. singaporense; therefore, there were no significant differences. Our observation suggested that the low number of atresia might be associated with the increased reproductive success of two female sesarmid carb living in natural habitats.

Reproductive development of the priapium fish Neosthetus lankesteri Regan, 1916 (Atheriniformes: Phallosthethidae) from Pranburi river estuary, Thailand using the histological approach

Reproduction in the family Phallostethidae is an area that has long been of interest especially in gonadal structure and gametogenesis. However, no published data concerning as mentioned above in Neostethus lankesteri Regan, 1916 from Thailand is noted. This fish is important for rural livelihood in the estuarine fisheries of Pranburi River estuary, Thailand. The construction of the female reproductive system of N. lankesteri composed of the ovary and short oviduct. The mature ovarian morphology is a single organ, which is histologically considered to be an asynchronous development ovary. The differentiating oocytes were observed and classified into oogonia proliferation, primary growth phase, secondary growth phase, post-ovulatory phase and atretic follicle phase. Under the ovarian differentiation, our observation indicated that the ovarian parenchyma of N. lankesteri during 1.8 – 2.4 cm in standard length (SL) was accompanied by the secondary growth phases. Identification of the post-ovulatory phase was detected during our sampling periods, indicating to entire life in the Pranburi River estuary as well as true resident species. In the male reproductive system of N. lankesteri, the mature testicular parenchyma was organized morphologically as a single organ and histologically appeared to be a restricted spermatogonial lobular testis type. The differentiation of the spermatogenetic stage was composed of three phases including spermatogonial, spermatocyte and spermiogenetic phases. It was to identify possible trends that the size at first maturity of the female fish was estimated at 1.8 cm SL. Certainly, results from this study clearly showed on the reproductive pattern of N. lankesteri, which could be applied to further studies (the reproductive cycle, spawning season and histopathology).

Studies on chemotaxonomic properties of tomato (&lt;i&gt;Solanum lycopersicum&lt;/i&gt; Linn.)

This research investigated the chemotaxonomic properties of Solanum lycopersicum Linn belonging to Solanaceae. The annual herbaceous free branching cultivated crop grows up to 60 cm or more in height. It has simple uniseriate trichomes densely dispersed all over the plant with adaxial and abaxial foliar trichome indices of 27.19% and 19.44% respectively. The compound leaves measure up to 6±3 cm long and 4 ± 2cm wide with serrated margin. The inflorescence has a cyme of 3 to 5 flowers or more. The petals are yellowish, up to 2±1 cm long and 1.5±0.7 cm wide with greenish sepals up to 1.5±0.7 cm long and 0.7±0.3 cm wide. Flowers are axile, hexa-pentamerous up to 1 cm in diameter with bilocular 2 celled ovary. The berry fruit is greenish when unripe and reddish when ripe, up to 5 cm in diameter with seed measuring up to 0.3 cm in diameter. The epidermal studies revealed anisocytic stomata with adaxial and abaxial stomatal indices of 19.30% and 19.64% respectively. The anatomy of mid-ribs and petioles showed bicollateral vasculation. The node is unilacunar. The stem anatomical section is made of 5 to 6 vascular bundles, with petioles associated with 2 rib traces at primary growth phase while at secondary growth phase, the mid-rib and petiole revealed vascular arcs and the stem, rings of open vascular system. Alkaloids, saponins, tannins, phlobatannins, flavonoids and combined anthraquinones were present while free anthraquinones and cardiac glycosides were absent.Keywords: Morphological, Anatomical, Cytological, Phytochemical, Studies

Primary dendrite growth kinetics and rapid solidification mechanism of highly undercooled Ti-Al alloys

The rapid solidification processes of undercooled Ti-(47, 50, 54) at.% Al alloys were investigated by electromagnetic levitation (EML) method combined with a high-speed photoelectric detector. The maximum undercoolings of the three liquid alloys were 376 K, 352 K and 316 K, respectively. Recalescence processes corresponding to the primary dendrite growth and subsequent phase transition were recorded at various undercoolings. The primary dendrite growth velocity V meets a double exponential relationship with the undercooling ΔT. Besides, a novel formula with physical meaning is proposed to explain that the more ordered liquid metal atoms accelerate the primary dendrite growth. Three recalescences are found at all undercoolings for Ti-47 at.% Al alloy and at high undercoolings for Ti-50 at.% Al alloy. The microstructures of solidified Ti-47 at.% Al alloys successively appear as coarse lamellar dendrites and finally evolve to refined parallel lamellar dendrites with the increasing undercooling. When ΔT rises, the microstructures of solidified Ti-50 at.% Al alloys appear from coarse primary dendrites and interdendritic dendrites to refined lamellar dendrites. In the process from low undercooling to high undercooling, the primary phase of undercooled Ti-54 at.% Al alloys changes from α-Ti (α) to γ-TiAl (γ) and the microstructures of solidified alloys evolve from spherical primary dendrites and matrix phases to cellular dendrite phases. Meanwhile, for the Ti-(47, 50) at.% Al, the transformation temperature of metastable intermediate α phase decreases with the increase of undercooling. Moreover, the microhardness of the three solidified alloys reaches the maximum when the undercoolings are 185 K, 270 K and 316 K, respectively.

Ovarian morphology and development of 3 to 8 months old African catfish, Clarias gariepinus

Thirty female African catfish of 3, 4, 5, 6, 7, and 8 months (n = 5 per age) cultured in south-east Nigeria were used to investigate ovarian morphology and development using gross anatomical and histological techniques. The results illustrated that the gonadosomatic indices of the African catfish peaked at 7 months of age (20.05 ± 1.45). Histologically, the ovaries of 3-month-old fish exhibited only pre-vitellogenic oocytes, while 4- to 8-month-old fish showed all stages of oocyte development, with predominance of pre-vitellogenic at 4 and 5 months old, and mature oocytes at 6 to 8 months old. The African catfish demonstrated an asynchronous mode of ovarian development. Oocytes representing the primary, secondary, and maturation growth phases were observed. Based on the gonadosomatic index and the histological evaluation of these ovaries, it was suggested that the African catfish mature for spawning at 7 months of age; moreover, its asynchronous mode of ovarian development may indicate that oocyte development and onset of sexual maturity of the catfish may be influenced by the amount and quality of food available in the culture environment. The study has provided essential information on the oogenesis and ovarian development of C. gariepinus, which may be useful for effective production and management of the fish in captivity.

Sirtuin E is a fungal global transcriptional regulator that determines the transition from the primary growth to the stationary phase

In response to limited nutrients, fungal cells exit the primary growth phase, enter the stationary phase, and cease proliferation. Although fundamental to microbial physiology in many environments, the regulation of this transition is poorly understood but likely involves many transcriptional regulators. These may include the sirtuins, which deacetylate acetyllysine residues of histones and epigenetically regulate global transcription. Therefore, we investigated the role of a nuclear sirtuin, sirtuin E (SirE), from the ascomycete fungus Aspergillus nidulans An A. nidulans strain with a disrupted sirE gene (SirEΔ) accumulated more acetylated histone H3 during the stationary growth phase when sirE was expressed at increased levels in the wild type. SirEΔ exhibited decreased mycelial autolysis, conidiophore development, sterigmatocystin biosynthesis, and production of extracellular hydrolases. Moreover, the transcription of the genes involved in these processes was also decreased, indicating that SirE is a histone deacetylase that up-regulates these activities in the stationary growth phase. Transcriptome analyses indicated that SirE repressed primary carbon and nitrogen metabolism and cell-wall synthesis. Chromatin immunoprecipitation demonstrated that SirE deacetylates acetylated Lys-9 residues in histone H3 at the gene promoters of α-1,3-glucan synthase (agsB), glycolytic phosphofructokinase (pfkA), and glyceraldehyde 3-phosphate (gpdA), indicating that SirE represses the expression of these primary metabolic genes. In summary, these results indicate that SirE facilitates the metabolic transition from the primary growth phase to the stationary phase. Because the observed gene expression profiles in stationary phase matched those resulting from carbon starvation, SirE appears to control this metabolic transition via a mechanism associated with the starvation response.

Effect of Feed and Feeding Strategies on Oocytes Development in Labeo rohita: A Histomorphological Study

The present study was conducted to study the effect of different diets and feeding strategies on the development of oocytes during the pre spawning and post spawning period in Labeo rohita. The study showed the presence of oocytes in different developmental stages viz., primary growth phase, secondary growth phase, maturation phase, post ovulatory phase and atretic oocytes during both pre spawning and post spawning period. The primary growth phase comprised of oogonia stage, chromatin nucleolus stage, early perinucleolar stage and late perinucleolar stage, whereas, the secondary growth phase was differentiated as cortical alveolus stage, early vitellogenic stage and late vitellogenic stage, and early and late types of atresia. The percentage occurrence (92–28%) of these different stages varied during pre spawning and post spawning period respectively The study revealed that (65%) of the oocytes were in the vitellogenic and maturation phases during pre spawning period, while as perinuclear stage and atretic oocytes (27.6%) were seen during post spawning period. All the developmental stages showed more active oocytes in the ovaries of fish fed with vitamins and minerals indicating that diet influences the reproductive efficiency of the fish

Chemotaxonomic studies on &lt;i&gt;Schwenckia americana&lt;/i&gt; LINN

This study investigated the chemotaxonomic studies on Schwenckia americana Linn., a member of the family Solanaceae predominantly found mostly in low grass fields, Nigeria. The habit is annual herbaceous weed with slender stem characterized with free branching and growing up to 45cm in height.They are used mainly as medicine. The leaves are simple, entire, elliptic to ovate in shape, smooth, variable, petiolate and larger at the lower region of stem and narrowing to smaller almost sessile and oblanceolate towards the apical regions which are alternately arranged and acrescently structured from the top to the base up to 3.7 ± 1.5cm long and 2.4±0.6cm wide. The inflorescence is a panicle of 15 or more flowers occurring at stem terminal.The flowers are whitish tubular structures measuring up to 1.0±0.2cm in length and 0.1cm in diameter. The petals are whitish up to 0.9 ±0.2cm in length and sepals are greenish up to 0.1cm in length. The berry fruit is greenish when unripe and pale yellowish when ripe up to 0.3±0.1cm in diameter. The seeds are blackish and spherical or triangular shaped with rough edges measuring up to 0.1cm in diameter. The epidermal studies revealanomocytic stomata whereas the trichomes are simple uniseriate forms wth bulge heads. The anatomy of mid-ribs and petioles showed bicollateral vascular systems. There are three vascular traces and the node is unilacunar. The petioles have 2 rib traces at primary growth phase. At secondary growth phase, the mid-rib and petiole revealed vascular arcs and the stem has a ring of open vascular system. The cytological studies showed a diploid chromosome number of 2n = 24 and n = 12 for the haploids. Alkaloids, saponins, tannins, phlobatannins, flavonoids, combined anthraquinones, free anthraquinones and cardiac glycosides are present.© JASEMKeywords: Morphological, Anatomical, Cytological, Phytochemical, Studies.

Molecular characterization and expression profiling of novel laccase isoenzyme genes from Fusarium solani X701, a wood-rotting ascomycete

Fusarium solani X701 is a wood-rotting ascomycete. Four laccase isoenzyme genes, lcc1, lcc2, lcc3 and lcc4, were cloned from this fungus, expression of these genes was profiled, and their relationship with other laccases was explored. Expression profiling of the isoenzymes in different media clearly shown that lcc4 mRNA was significantly induced by lignin, suggesting that lcc4 be the responsible gene for catabolizing lignin. Expression of lcc1 was triggered by the switch from the presence to the absence of glucose, while lcc2 was significantly induced in the glucose medium. Expression levels of lcc3 were not affected by different carbon sources in the primary growth phase, but a short induction by lignin was observed in early secondary growth phase. Bioinformatical analyses revealed unique features in our isoenzymes in comparison with other laccase isoenzymes from ascomycete.

Modeling the pattern of contrast extravasation in acute intracerebral hemorrhage using dynamic contrast-enhanced MR.

Contrast extravasation (CE) in spontaneous intracerebral hemorrhage (ICH), coined the spot sign, predicts hematoma expansion (HE) and poor clinical outcome. The dynamic relationship between CE and the mode of ICH growth are poorly understood. We characterized the in vivo pattern and rate of HE using a novel animal model of acute ICH. Basal ganglia ICH was created in 14 Yorkshire swine utilizing a novel MRI integrated model, permitting real-time CE observation using dynamic contrast-enhanced (DCE) MRI. Computerized planimetry measured CE volume at each time point. Spatial vector analysis along three orthogonal axes determined distance vectors. Maximizing and minimizing the coefficient of determination defined the temporal phases of growth and stability, respectively. CE rate was calculated using a Patlak model. Asymmetric growth and variable rates of expansion characterized HE defining three distinct growth phases and patterns. A primary growth phase (duration 160 s; IQR 50-130) demonstrated rapid linear growth (0.04 mm/s IQR 0.01-0.10) accounting for 85 ± 15 % of total HE. The stationary phase demonstrated stability (duration 145 s; IQR 0-655). A secondary growth phase (duration 300; 130-600 s) accounted for 23 ± 8 % of total HE. In the primary and secondary growth phase, asymmetric growth occurred in the anterior-posterior (AP) planes (0.056 mm/s; p = 0.026 and 0.0112 mm/s; p = 0.03). Monophasic 2 (14 %), biphasic 4 (35 %) (primary followed by secondary growth), and triphasic 8 (56 %) patterns (primary, stationary, and secondary growth phase) were observed. A novel model of ICH provides real-time study of the dynamics and rate of CE. This data facilitates the understanding of pattern and rate of ICH formation.