Development of a quantitative Real-Time PCR for micrometastasis detection using CEA in peripheral blood and bone marrow specimens of gastric cancer patients

"n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Gastric adenocarsinoma is the first leading fatal malignancy in Iran. Despite advances in novel therapeutics approaches for gastric cancer (GC) patient, tumor dissemination via blood stream to distant organ is still the major cause of death. Therefore, there is urgent need to establish sensitive methods for early detection of disseminated tumor cells in peripheral blood (PB) and bone marrow (BM) specimens of gastric cancer patients. "n"nMethods: In the present study, we use Carcinoma Embryonic Antigen (CEA) as a tumor marker and Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) as an internal control to detection and quantification of disseminated tumor cells in PB and BM specimens of affected individuals. Total RNA was extracted from AGS (gastric cancer) cell line and CEA and GAPDH fragments were generated by reverse transcription. The amplified fragments were cloned into pTZ57R/T vector separately. Double cloning of these genes has done into one pTZ57R/T vector. Serial dilution of this recombinant plasmid is used to construct standard curve, each containing a known amount of input copy number. Total RNA was extracted from BP and BM specimens of 35 GC patients. cDNA of the specimens were synthesized by reverse transcription and subjected to Quantitative Real-Time PCR (QRT-PCR)."n"nResults: We developed a highly sensitive and specific quantitative PCR for CEA and GAPDH using Real-Time PCR based on TaqMan technology. CEA mRNA was detected in 23% of PB and 20% of BM specimens. There was no CEA mRNA detecting in control group."n"nConclusions: The QRT-PCR for CEA can be a useful technique for detection of micrometastases in the PB and BM specimens of gastric cancer patients."

Modulation of neuronal activity in cortical organoids with bioelectronic delivery of ions and neurotransmitters

Precise modulation of brain activity is fundamental for the proper establishment and maturation of the cerebral cortex. To this end, cortical organoids are promising tools to study circuit formation and the underpinnings of neurodevelopmental disease. However, the ability to manipulate neuronal activity with high temporal resolution in brain organoids remains limited. To overcome this challenge, we introduce a bioelectronic approach to control cortical organoid activity with the selective delivery of ions and neurotransmitters. Using this approach, we sequentially increased and decreased neuronal activity in brain organoids with the bioelectronic delivery of potassium ions (K+) and γ-aminobutyric acid (GABA), respectively, while simultaneously monitoring network activity. This works highlights bioelectronic ion pumps as tools for high-resolution temporal control of brain organoid activity toward precise pharmacological studies that can improve our understanding of neuronal function

Evaluation of the effect of Saccharomyces cerevisiae on fermentation characteristics and volatile compounds of sourdough

The objective of this study was to unveil insights into the effects of Saccharomyces cerevisiae on the development of volatile compounds and metabolites during the dough fermentation in making Chinese steamed bread. Changes in gluten structure under the influence of baker’s yeast were studied using scanning electron micrographs (SEM). A unique aroma profile was found comprising some previously reported aromatic compounds and some unreported aromatic aldehydes ((E)-2-Decenal and 2-Undecenal) and ketones (2-Heptanone and 2-Nonanone) in the baker’s yeast fermentation. Among metabolites, the most preferred sugar for this yeast (glucose) showed a significant decrease in contents during the initial few hours of the fermentation and at last an increase was observed. However, most of the amino acids increased either slightly or decreased by the fermentation time. SEM of fermented dough showed that the yeast had a very little effect on starch stability. This study provided some fermentation features of the bakers’ yeast which could be used for the tailored production of steamed bread