soheila soheili

In order to study the early Aptian Oceanic Anoxic Event 1a (OAE 1a), calcareous nannofossils are investigated at the late Barremian ‒ early Aptian sediments of the Garau Formation at south west of the Kabir-Kuh anticline, Qaleh-Darreh section. Ninety species of calcareous nannofossils from 43 genus and 15 families are identified along with marker species like Hayesites irregularis and Eprolithus floralis. Based on index calcareous nannofossil taxa, the studied interval is located between the uppermost part of the CC6/NC5 and the early part of CC7/NC7A. The first occurrence of H. irregularis, the marker of the Barremian ‒ Aptian boundary, used as an index species between CC6/NC5E and CC7/NC6. Statistical analysis of the calcareous nannofossil assemblages at the studied interval indicate the presence of nannoconid decline at the Barremian ‒ Aptian boundary and early Aptian and nannoconid crisis at CC7a/NC6B biozone. Nannoconid crisis is one of the main markers of the early Aptian OAE 1a that is recorded from different parts of the world at the Tethys and Boreal realms, Atlantic and Pacific oceans at the early Aptian (NC6 biozone). At the current study the early Aptian OAE 1a is recorded from the Garau Formation based on calcareous nannofossil assemblages.

Retinal pigment epithelium is responsible for maintaining the structural integrity of the retina by an efficient defense against free radicals, photo-oxidative exposure and light energy. For this purpose the main RPE line of defense is melanosomes. Melanin content of retinal pigment epithelium cells in adults and neonates reveals remarkable variations. In the current study we compared melanogenic factors gene expression levels in human adult and neonate RPE cells in culture. RPE cells from adult and neonate human eye globes were isolated and then cultured in DMEM: F12 (1:1) containing 10% FBS. Expression of RPE65 and Cytokeratin 8/18 markers in isolated cells was confirmed by immunocytochemistry. To evaluate the responsible factors in the pathway of melanin biosynthesis, gene expression of orthodenticle homeobox 2, microphthalmia-associated transcription factor A and H as prominent transcription factors, tyrosinase and dopachrome tautomerase as effective enzymes in the melanin biosynthetic pathway were examined in human neonate derived RPE cells compared to human adult derived RPE cells in culture. According to the Real-Time RT-PCR data, gene expression of MITF-H, OTX2, TYR and DCT in RPE cells of the neonates showed a significant increase compared to the adults. With increasing passage number, gene expression of MITF-H, OTX2, TYR and DCT showed remarkable decline. According to the role of OTX2 and MITF-H as the main transcription factor effectors on the TYR and DCT, restoration of OTX2 and MITF-H gene expression may be retain melanin content in RPE cells.

A cell line spontaneously derived from human retinal pigment epithelium (hRPE) was cultured on alginate film gelatinized with different concentrations of neurobasal cell culture medium (NCCM) to assess its growth and morphological behavior on this naturally occurring polysaccharide. Neonatal human globes were used to isolate hRPE cells. They were cultured in Dulbecco’s modified Eagle’s‑medium‑and‑Ham’s‑F12‑medium‑(DMEM/F12) supplemented with 10% fetal bovine serum (FBS). Cultures were continuously studied using phase contrast microscopy. After the nineth passage, cells were characterized through immunocytochemical analysis for Oct4, Chx10, and Pax6 and Ki67 markers. In each well of a 6‑well microplate, 1 and 2% weight/volume (w/v) alginate in deionized water was added and gelatinized using 1× and 10× NCCM. hRPE cells were cultured at a density of 2 × 105 cells/well in alginate‑coated microplates. After 5 days, hRPE colonies were harvested and re‑plated on polystyrene substrates. Morphology and growth of hRPE cultures were determined during the next 2 weeks. The first few passages of the cultures were purely hRPE cells that revealed typical morphological features of the pigmented epithelium. They made spaces, devoid of cells, between hRPE cell monolayer and fill in the unoccupied spaces. They grew faster than native RPE cells and rapidly overgrew. Immunocytochemical test revealed that the founded cells expressed Chx10, Pax6, Ki67 and Oct4. The hRPE cells survived unlimitedly on alginate film and formed giant adjoining colonies. After re‑plating, hRPE colonies adhered quickly on polystyrene and displayed native hRPE morphological features. Alginate film can support the survival and growth of hRPE cells and induce the cells to re‑organize in tissue‑like structures.

There exists an association between PI3K pathway licentious activity and the considerable feature of high metastatic potential of the genitourinary cancer cells. Although DU 145 and 5637 have functional phosphatase and tensin homolog (PTEN) tumor suppressor gene, which antagonizes PI3K function, PC-3 is null for PTEN gene. In pursuit to explain why PTEN bearing cell lines display high metastatic behavior, we searched for any discrepancy in PI3K isoforms expression pattern between these cell lines. Gathering gene bank data files, specific primers were designed, for all the genes of 12 studied isoforms from 3 different classes of PI3K. Total RNA was extracted and examined by Real- Time PCR to compare the cells for the type and amount of the isoforms which expressed. Cα and R2 isoforms are indicative of an equal expression for PC3 and DU145, R3 transcripts revealed 80% decrease in DU145 and Cβ, R1 and C2α demonstrated an increased expression in DU145. When a comparison is made between 5637 and PC3, it can be seen that although a little decrease in the level of R3 transcripts was demonstrated, the amount of Cα, Cβ, R2, R1 and C2α increased. In conclusion in this study it is proposed that R1, R2, Cα, Cβ, C2α and R1, Cβ, C2α are candidate genes for silencing via RNAi in 5637 and DU145, respectively, to evaluate their roles in metastatic behavior of the both studied PTEN bearing cell lines.

A detailed study of the Lower Cretaceous Dasyclad algae of the Shahkuh Formation was done in the Chahpalang stratigraphic section (its name comes from the mountain in studied area). According to the study of the 101 thin sections, for the first time, we recognized 7 genus and 8 species of dasyclad algae from Shah Kuh Formation in Chah Palang section including Angioporella cf. fouryae, Griphoporella aff. cretacea, Kopetdagaria sphaerica, Montiella? elitzae, Neomeris sp., Pseudoactinoporella? iranica, Salpingoporella sp., Salpingoporella aff. Muehlbergi and Salpingoporella cf. pygmaea. Based on identified dasyclad algae, and the age of index Orbitolinid fossil that have been recognized in this formation, such as Dictyoconus pachymarginalis, Montseciella arabica, Praeorbitolina sp., Palorbirolina sp., the age of Shahkuh Formation is Barremian to Aptian.

Human retinal pigment epithelium (hRPE) is a cell monolayer located in the outer part of the retina that is in contact with photoreceptors. In many diseases RPE cells damage. One way for treating this disease is the implantation of intact instead of damaged cells. For this reason different types of substrates have been used for cell cultivation. This study has used alginate and a blend of alginate/gelatin (A/G) to study RPE cell growth.

We prepared alginate solutions in concentrations of 1% and 2% (w/v) in water and DMEM/F12. The solutions were infused into each well of 6-well micro plates until a uniform culture substrate that had a 1 mm thickness was generated. Passage-4 hRPE cells were cultivated on the substrate and the cell characteristics studied. hRPE cells did not adhere to alginate in DMEM/F12 and did not exhibit interaction with alginate substrate. For this reason A/G solutions at concentrations of 1% and 2% (w/v) in water were prepared. We prepared A/G blends at weight ratios of: 20:80، 30:70، 40:60، 50:50، 60:40، 70:30، and 80:20. These blends were infused into each well of 6-well plates until the appropriate 1 mm thickness of A/G was achieved. Isolated hRPE cells were cultured on synthetic substrate after which we studied the cells'' characteristics.

hRPE cell generated adhesive colonies on the A/G substrate. In all studied combinations of A/G، the diffused hRPE cells formed a monolayer under the substrate sheets. However the A/G 20:80 ratio had cell growth in the upper face of the substrate. hRPE survived indefinitely on A/G substrate. After the cells were re-cultured on polystyrene، they showed general morphological features of normal hRPE cells.

The A/G blend at a 20:80 ratio was chosen to be used for future studies.

To evaluate the effect of placental growth factor (PlGF) gene knockdown in a murine model of laser-induced choroidal neovascularization. Choroidal neovascularization was induced in the left eyes of 11 mice by infrared laser. Small interfering RNA (siRNA, 20 picomoles/10? l) corresponding to PlGF mRNA was administered intravitreally by Hamilton syringe in all subjects. One month later, fluorescein angiography and histolologic examination were performed. No leakage was apparent in the 11 eyes treated with siRNA cognate to PlGF. The results of histological evaluation were consistent with angiographic findings showing absence of choroidal neovascularization. Knockdown of the PlGF gene can inhibit the growth of laser-induced choroidal neovascularization in mice.

Autism results from developmental factors that affect many or all functional brain systems. Brain is one of tissues which are crucially in need of adenosine triphosphate (ATP). Autism is noticeably affected by mitochondrial dysfunction which impairs energy metabolism. Considering mutations within ATPase 6, ATPase 8 and tRNALys genes, associated with different neural diseases, and the main role of ATPase 6/8 in energy generation, we decided to investigate mutations on these mtDNA-encoded genes to reveal their roles in autism pathogenesis. In this experimental study, mutation analysis for the mentioned genes were performed in a cohort of 24 unrelated patients with idiopathic autism by employing amplicon sequencing of mtDNA fragments. In this study, 12 patients (50%) showed point mutations that represent a significant correlation between autism and mtDNA variations. Most of the identified substitutions (55.55%) were observed on MT-ATP6, altering some conserved amino acids to other ones which could potentially affect ATPase 6 function. Mutations causing amino acid replacement denote involvement of mtDNA genes, especially ATPase 6 in autism pathogenesis. MtDNA mutations in relation with autism could be remarkable to realize an understandable mechanism of pathogenesis in order to achieve therapeutic solutions.

FcεRI, The high affinity receptor for IgE plays a critical role in triggering the allergic reactions. It is responsible for inducing mast cell degranulation and deliberation of allergy mediators when it is aggregated by allergen and IgE complexes. FcεRI on the mast cells consists of three subunits; α chain directly binds IgE, β chain and dimmer of γ chains together mediate intracellular signaling. Cross-linking of IgE-bound FcεRI on the surface of mast cells and basophils by the multivalent antigen induces release of chemical mediators. The present in vitro study was designed to investigate the effect of synthetic FcεRI-α siRNA on the antigen-induced activation of MC/9 cells. MC/9 cells which are murine mast cells were transfected by FcεRI-α siRNA and negative control siRNA. After 6 h, anti-DNP (Dinitrophenyl) IgE was used for the cells sensitization. Then the cells were challenged with Dinitrophenyl-Human Serum Albumin (DNP-HSA) for mast cell degranulation induction before collection of supernatants. The amount of mRNA and protein expression was measured by Real Time PCR and western blot analysis, respectively. Determination of the expression rate of FcεRI-α on cell surface was achieved by flow cytometry. ELISA and spectrophotometry methods were used subsequently for measuring the effects of FcεRI-α siRNA on antigen-induced histamine and β-hexosaminidase release. FcεRI-α siRNA treated cells showed significant decrease in FcεRI-α mRNA and protein expression in comparison to control cells. FcεRI-mediated mast cell release of β-hexosaminidase and histamine were also inhibited. In this study it was shown that FcεRI-α siRNA could suppress FcεRI-α expression and inhibited degranulation and histamine release in antigen-stimulated MC/9 cells. In conclusion, knock-down of FcεRI-α by siRNA could be a promising method for inhibition of the mast cell-mediated allergic reactions.

To establish human retinal pigment epithelial (RPE) cell culture as a source for cell replacement therapy in ocular diseases. Human cadaver globes were used to isolate RPE cells. Each globe was cut into several pieces of a few millimeters in size. After removing the sclera and choroid, remaining tissues were washed in phosphate buffer saline and RPE cells were isolated using dispase enzyme solution and cultured in Dulbecco''s Modified Eagle''s Medium: Nutrient Mixture F-12 supplemented with 10% fetal calf serum. Primary cultures of RPE cells were established and spheroid colonies related to progenitor/stem cells developed in a number of cultures. The colonies included purely pigmented or mixed pigmented and non-pigmented cells. After multiple cellular passages, several types of photoreceptors and neural-like cells were detected morphologically. Cellular plasticity in RPE cell cultures revealed promising results in terms of generation of stem/progenitor cells from human RPE cells. Whether the spheroids and neural-like retinal cells were directly derived from retinal stem cells or offspring of trans-differentiating or de-differentiating RPE cells remains to be answered.

Interferon producing cells have an important role in controlling HIV. The interferon producing cell count was evaluated in order to study the possibility of immune modulating qualification of HGV co-infection in HIV patients’ safety. This descriptive study was performed on 83 HIV positive patients with CD4+ cell count >400 to evaluate the interferon producing cell count. These 83 patients were categorized in 4 groups based on their current infection status. Patients infected with HCV and HGV (HGV+/HCV+), patients infected with HGV but not with HCV (HGV-C+/HCV-), patients infected with HCV but not with HGV (HGV-/HCV+) and patients not infected by HGV and HCV (HGV-/HCV-). Interferon producing cell analysis was performed by flow cytometric method and the presence of -HGV RNA by using RT-PCR and PCR-Elisa. Interferon producing cell count in HGV co-infected individuals revealed a wider range of numbers however there was no significant difference in the interferon producing cells count between HIV infected patients with and without HGV co-infection. HGV/HCV co-infection doesnt have an effective role on increasing of production or rotation of interferon producing cell in HIV infected patients with relatively intact immune system.

Hepatitis virus infections are common in patients suffering from human immunodeficiency virus (HIV) infection due to similar transmission routes of these viruses. Hepatitis virus infections lead to impaired survival in HIV positive patients. In contrast to the expectations, some studies indicate that HGV infection slows down the progression and eventually death of AIDS patients. The aim of this study was to estimate the frequency of active HGV infection among HIV infected patients in Tehran. Flow cytometric analysis of lymphocyte subpopulations was performed on blood samples of 103 patients with HIV infection referring to the central laboratory of Iranian blood transfusion organization. Their plasma samples were analyzed for the presence of HGV RNA by RT-PCR. HGV Positive cases were confirmed by PCR-ELISA. Of 103 HIV-positive patients, 16 (15.5%) were HGV RNA-positive. It was apparent that the frequency of HGV in patients with CD4 count above 500 cell/μl is higher. Sucha frequency is not high compared with those found in other studies. The high frequency of active HGV infection in patients with asymptomatic HIV infection may be due to the loss of active HGV infection in patients with advanced HIV infection.