Analysis of growth layers in the teeth of Tursiops truncatus using light microscopy, microradiography, and SEM

Preliminary results show microradiography and scanning electron microscopy (SEM) to be more accurate methods of accessing growth layer groups (GLGs) in the teeth of Tursiops truncatus than transmitted light microscopy. Microradiography shows the rhythmic deposition of mineral as alternating radiopaque and radiolucent layers. It improves the resolution of GLGs near the pulp cavity in older individuals, better than either SEM or light microscopy. SEM of etched sections show GLGs as ridges and grooves which are easily counted from the micrograph. SEM also shows GLGs to be composed of fine incremental layers of uniform size and number which may allow for more precise age determination. Accessory layers are usually hypomineralized layers within the hypermineralized layer of the GLG and are more readily distinguishable as such in SEM of etched sections and microradiographs than in thin sections viewed under transmitted light. The neonatal line is hypomineralized, appearing translucent under transmitted light, radiolucent in a microradiograph, and as a ridge in SEM. (PDF contains 6 pages.)

Eggs of British Meniscus midges (diptera;dixidae) observed by scanning electron microscopy

The egg of Dixella martinii is described for the first time. The eggs of the Dixidae are placed in three morphological groups: bulbous and meshed; streamlined and smooth; streamlined and minutely spiculated. Ten of the fourteen species known from Britain are placed in these groups. After a detailed description of the egg of D. martinii, the three morphological groups are described and scanning electron micrographs are provided.

Cloning and expression of zebra fish ferroportin and investigating its influence on anemia

Iron is required for many microbes and pathogens for their survival and proliferation including Leishmania which cause leishmaniasis. Leishmaniasis is an increasingly serious infectious disease with a wide spectrum of clinical manifestations. These range from localized cutaneous leishmaniasis (CL) lesions to a lethal visceral form. Certain strains such as BALB/c mice fail to control L. major infection and develop progressive lesions and systemic disease. These mice are thought to be a model of non-healing forms of the human disease such as kala-azar or diffuse cutaneous leishmaniasis. Progression of disease in BALB/c mice has been associated with the anemia, in last days of their survival, the progressive anemia is considered to be one of the reasons of their death. Ferroportin (Fpn), a key regulator of iron homeostasis is a conserved membrane protein that exports iron across the duodenal enterocytes as well as macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival and proliferation of many microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immune responses and pathogenesis of micoorganisms. To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP–N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of FPN-EGFP protein in Hek 293T cells. The expression was confirmed by fluorescence microscopy and flow cytometery. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 server and NetNGlyc 3.1 server. Data emphasised that obtained Fpn from indian zebrafish contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 mucin-type glycosylated amino acid. The results indicate that the prepared and characterized recombinant Fpn protein has no membrane topology difference compared to other Fpn described by other researcher. Our next aim was to deliver recombinant plasmid (pEGFP-ZFpn) to entrocyte cells. However, naked therapeutic genes are rapidly degraded by nucleases, showing poor cellular uptake, nonspecificity to the target cells, and low transfection efficiency. The development of safe and efficient gene carriers is one of the prerequisites for the success of gene therapy. Chitosan and alginate 139 polymers were used for oral gene carrier because of their biodegradability, biocompatibility and their mucoadhesive and permeability-enhancing properties in the gut. Nanoparticles comprising Alginate/Chitosan polymers were prepared by pregel preparation method. The resulting nanoparticles had a loading efficiency of 95% and average size of 188 nm as confirmed by PCS method and SEM images had showed spherical particles. BALB/c mice were divided to three groups. The first and second group were fed with chitosan/alginate nanoparticles containing the pEGFP-ZFpn and pEGFP plasmid, respectively (30 μgr/mice) and the third group (control) didn’t get any nanoparticles. The result showed BALB/c mice infected by L.major, resulted in higher hematocryte and iron level in pEGFP-ZFpn fed mice than that in other groups. Consentration of cytokines determined by ELISA showed lower levels of IL-4 and IL-10 and higher levels of IFN-γ/IL-4 and IFN-γ/IL-10 ratios in pEGFP-ZFpn fed mice than that in other groups. Morover more limited increase of footpad thickness and significant reduction of viable parasites in lymph node was seen in pEGFP-ZFpn fed mice. The results showed the first group exhibited a highr hematocryte and iron compared to the other groups. These data strongly suggests the in vivo administration of chitosan/alginate nanoparticles containing pEGFP-ZFpn suppress Th2 response and may be used to control the leishmaniasis .

Survey of plankton blooms and red tides and their interaction with environmental factors through monitoring satellite images

One of the most important marine ecological phenomena is red tide which is created by increasing of phytoplankton population, influenced by different factors such as climate condition changes, utrification hydrological factors and can leave sever and undesired ecological and economical effects behind itself in the case of durability. Coast line of Hormozgan is about 900km from east to west, within the range of geographical coordinates of 56 16 23.8, 26 58 8.8 to 54 34 5.33 and 26 34 32 eastern longitude and northern latitude, seven sampling stations were considered and sampled for a period of one year from October 2008 to October 2009. after the analysis of Satellite images, monthly, during the best time. In several stages, samplings were performed. In each station, three samples were collected for identification and determination of Bloom- creating species abundance. Cochlodinium polykrikoides was the species responsible for the discoloration which occurred at October 2008 in Hormozgan marine water. Environmental parameters such as sea surface temperature, pH, salinity, Dissolved Oxygen concentration, Total Dissolved Solids (T.D.S.), conductivity, nitrate, nitrite and phosphate and also chlorophyll a were measured and calculated. Kruscal Wallis test was used to compare the densities between different months, seasons and the studied stations. Mann-whitney test from Nonparametric Tests was used for couple comparison. Pearson correlation coefficient was used to determine the relationship between physical and chemical data set and the abundance of Cochlodinium polykrikoides. Multivariate Regression and analysis of variance (ANOVA) also were used to obtain the models and equations of red tide occurrence relationship, environmental parameters and nutrient data. The highest density was 26 million cells per liter in Qeshm station. A meaningful difference was observed between sampling months and seasons but there was no between sampling stations which indicates that in favorable conditions, the occurrence of this phenomenon by the studied species is probable. Regarding to β coefficients of nitrate, temperature, phosphate, Total Dissolvable Solutions (T.D.S) and pH these parameters are effective on the abundance of this species and red tide occurrence. Increase in these factors can represent the effects and outcomes of human activities and increase in marine pollution.