scholarly journals Preliminary Evaluation of Nanobacteria on Crystal Retention, CaSR, and Claudin-14 Expression in HK-2 Cells

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Gang Xu ◽  
Biao Qian ◽  
Liying Zheng
Keyword(s):  
Protein Expression ◽  
Cell Morphology ◽  
Cell Damage ◽  
Urinary Calculi ◽  
Urinary Stone ◽  
Negative Control ◽  
Preliminary Evaluation ◽  
Significant Difference ◽  
Crystal Adhesion ◽  
Claudin 14

To evaluate cell morphology, crystal adhesion, cell damage, Calcium sensitive receptor (CaSR), and Claudin protein-14 (Claudin-14) expression at different time intervals and explore the role of nanobacteria in the formation of urinary calculi. In this experiment, HK-2 cells were cocultured with nanobacteria (NB) in the absence or presence of tetracycline (Tet). Cells treated with calcium oxalate monohydrate (COM) crystals were used as a positive control of urinary stone-induced cell damage. After which, cell morphology was evaluated by hematoxylin-eosin staining in comparison to untreated HK-2 cells (negative control). Use different methods to assess cell damage, crystal adhesion, and protein expression. (The degree of cell damage, crystal adhesion, and protein expression were evaluated by various methods). It was found that the degree of cell damage observed in Tet + NB-treated cells was significantly lower than that in NB-treated cells. Lactate dehydrogenase (LDH) leakage was higher in COM-exposed than in control cells ( P < 0.05 ). However, LDH release from both NB- and Tet + NB-treated cells was significantly lower than from COM-treated cells ( P < 0.05 ). The relative expression of CaSR and Claudin-14 proteins was higher in NB, COM, and TET + NB cells than in control cells ( P < 0.001 ) and was lower in Tet + NB than in NB cells ( P < 0.01 ). And P < 0.05 means that the difference was statistically significant, and P < 0.001 means that there was a significant difference between the both things. From the cell morphology, the cell damage in the COM group was greater than that in the NB group, and the cell damage markers in the COM group and the NB group were elevated. NB caused damage to HK-2 cells by inducing lipid peroxidation, and the degree of damage was increased in processing time. The adhesion of HK-2 cells to COM crystals increased after injury and was proportional to the duration of NB coculture. NB upregulated the expression of CaSR and Claudin-14 in HK-2 cells.

OCT-1 Protein Expression Detected by Flow Cytometry Provided Valuable Information in CML Patients Treated with Imatinib MesylateÃ, FHigher hOCT-1 Expression Predict Better Outcome.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1117-1117
Author(s):  
Huanling Zhu ◽  
Qiurong Zhang ◽  
Nenggang Jiang ◽  
Ting Liu
Keyword(s):  
Flow Cytometry ◽  
Protein Expression ◽  
Conflicts Of Interest ◽  
Organic Cation Transporter ◽  
Negative Control ◽  
Expression Level ◽  
Molecular Response ◽  
Fluorescent Intensity ◽  
Healthy Donors ◽  
Significant Difference

Abstract Abstract 1117 Poster Board I-139 Objective The human organic cation transporter-1(hOCT-1) is the major active influx protein responsible for the transport of imatinib into cells. The functional activity of the hOCT-1 protein using 14-C detected by others demonstrated a link between CML molecular response and hOCT-1 activity. However, 14-C labeled detection is not convenient in routine clinical practice. Hence, we use flow cytometry to detect hOCT-1 protein expression level in CML patient and try to find some relation between hOCT-1 expression and imatinib response. Subjects and methods In this study, 64 CML CP patients and 31 healthy donors were enrolled. Totally, there are 78 patient' peripheral blood (PB) or bone marrow (BM) samples and 31 donor PB samples were measured. The hOCT-1 protein expression levels were detected by indirect immunofluorescent flow cytometry. The hOCT-1 levels were expressed as mean fluorescent intensity (MFI). In avoided to systematic error, lymphocytes which had little hOCT-1 expression were used as internal negative control. Results ‡@ Assessing PB hOCT-1 expressing in patients with donors, hOCT-1 level is higher in healthy donors than in CML patient (mean±standard deviation 9.11±6.04,5.60±3.74,P=0.005). ‡AThe hOCT-1 level was compared with molecular response in patients. Of 39 patients achieved optimal molecular response, the hOCT-1 level was 6.49±3.83, versus 3.86±2.91 in 20 patients with non-optimal response(P=0.009). Comparing 39 optimal responders with 16 sub-optimal responders, hOCT-1 level were 6.49±3.83, versus 3.98±1.23(P=0.025. ‡B Assessing CML stages with hOCT-1 expression, there is no significant difference in chronic stage and advanced stage(5.93±3.87, 3.49±1.64, P=0.085). Conclusions hOCT-1 expression level measured by flow cytometry is very convenient and clinically available. The hOCT-1 expression level can be an important predictor in CML patients treated with imatinib mesylate. Disclosures No relevant conflicts of interest to declare.

scholarly journals Ultrasound microbubble-mediated CRISPR/Cas9 knockout of C-erbB-2 in HEC-1A cells

2019 ◽  
Vol 47 (5) ◽  
pp. 2199-2206 ◽  
Author(s):  
Junhong Cai ◽  
Sizhe Huang ◽  
Yuping Yi ◽  
Shan Bao
Keyword(s):  
Protein Expression ◽  
Mrna Expression ◽  
Western Blotting ◽  
Growth Factor Receptor ◽  
Negative Control ◽  
Rt Pcr ◽  
Control Groups ◽  
Transfected Cells ◽  
Significant Difference ◽  
Ultrasound Microbubbles

Objective Epidermal growth factor receptor 2 (C-erbB-2) is one of the most frequently mutated oncogenes in human tumors. We aimed to evaluate the knockout efficiency of clustered regularly interspaced short palindromic repeat (CRISPR) technology using ultrasound microbubble transfection to target C-erbB-2 in human endometrial cancer (HEC)-1A cells. Methods Three single guide RNAs (sgRNAs) targeting C-erbB-2 were designed and used to construct CRISPR/CRISPR-associated (Cas)9-C-erbB-2 plasmids. The constructed plasmids were transfected into HEC-1A cells using ultrasound microbubbles. C-erbB-2 knockout cloned cells were identified by green fluorescence. C-erbB-2 mRNA and protein expression was measured by reverse transcription (RT)-PCR and western blotting, respectively. Results RT-PCR showed that C-erbB-2 mRNA expression was significantly lower in sgRNA1-transfected cells (0.57 ± 0.06) than in blank (1.00 ± 0.09) and negative-control groups (1.02 ± 0.12). Western blotting revealed C-erbB-2 protein expression to be significantly lower in sgRNA1-transfected cells (0.269 ± 0.033) than in blank (0.495 ± 0.059) and negative-control groups (1.243 ± 0.281). However, there was no significant difference in C-erbB-2 protein and mRNA expression in sgRNA2- and sgRNA3-transfected cells compared with controls. Conclusion Ultrasound microbubbles can mediate plasmid transfer into HEC-1A cells to interfere with gene expression and knockout C-erbB-2.

scholarly journals The Effect of Active Gambir (Uncaria gambir) Fraction on TNF-α Protein Expression and Lesion Size in Rats Induced Gastritis

2018 ◽  
Vol 2 (3) ◽  
pp. 1-7
Author(s):  
Ni Wayan Lisa Suasti ◽  
Lunsi Oktafitria ◽  
Ellya Aidia ◽  
Rahmad Dhani
Keyword(s):  
Protein Expression ◽  
Lesion Size ◽  
Negative Control ◽  
Mucosal Lesion ◽  
Control Group ◽  
Gastric Mucosal Lesion ◽  
Control Group Design ◽  
Water Fraction ◽  
White Rats ◽  
Significant Difference

Background: Gambir (Uncaria gambir) is empirically used to treat abdominal pain and vomittus caused by gastritis because of its anti-inflammatory effects, especially flavonoid. This study aims to determine the effect of active gambir fraction on TNF-α protein expression and wound size in white rats gastritis model. Methods: The research method used experimental study design in vivo with post test with control group design. Rats were divided randomly in 11 groups and were induced to be gastritis for 1 day. Group 1 (negative control) was given aquadest of 5 mL, group 2 (positive control) was administered ranitidine 10 mg/kgBW, groups 3, 4, and 5 were given n-hexane fraction, groups 6, 7 and 8 were given a water fraction, and groups of 9, 10, and 11 were given ethyl fractions with each group receives dose of 20, 40, and 80 mg/kgBW and all groups were treated for 3 days. Rats were dissected on 5th day for examination of gastric mucosal lesion size and performed ELISA expression of TNF-α expression of gastric mucosal tissue. The results of this study were assayed by SPSS 18. Results: The result of the research using Kruskal-Wallis test showed that there were significant differences (p <0.05) of the lesions size between the sample groups where  control positive, ethyl fraction 20, 40, 80 mg/kgBW, and water fraction 20, 40 mg/kgBB had the gastric ​​mucosal lesion size differed significantly with the negative control group, while the TNF-α protein expression test  using Kruskal-Wallis showed that there was a significant difference (p <0.05) TNF-α levels of all groups against the negative control. Conclusion: Active gambir fraction had a potention to reduce size of mucose gaster lesion and reduce expression of TNF-α protein.   Keywords: Gambir – TNF-α – antiinflamatory – gastiritis

scholarly journals Preliminary Evaluation of a NitrAdine-Based Brushing Solution for Patients Suffering from Gingivitis: A Prospective Clinical Case–Control Study

2021 ◽  
Author(s):  
Michele Perelli ◽  
Roberto Abunto ◽  
Mario Semenza ◽  
Mauro Centracchio ◽  
Stefano Di Chiara ◽  
...  
Keyword(s):  
Oral Hygiene ◽  
Test Group ◽  
Negative Control ◽  
Preliminary Evaluation ◽  
Meaningful Improvement ◽  
Plaque Score ◽  
Biofilm Removal ◽  
Significant Difference ◽  
Index Value ◽  
Reference Parameters

Abstract Objectives This article aimed to evaluate the clinical efficacy of a nonantibiotic biofilm-removal formulation based on NitrAdine (PerioTabs), combined with a regular home oral hygiene regimen, in Caucasian patients with gingivitis. Materials and Methods A sample of 60 patients were included in this clinical prospective study. All selected subjects underwent regular prophylaxis and professional oral hygiene at baseline; 30 days later, they were recalled for the measurements of the reference parameters about bleeding on probing (full-mouth bleeding upon probing score [FMBS]) and plaque index (full-mouth plaque score [FMPS]); no other clinical procedure was performed. Consequently, half of the patients (n = 30) were instructed to use PerioTabs for 10 days. The remaining patients (n = 30) were used as the negative control, only instructed to continue with their usual oral hygiene regimen. Fifteen days after, the clinical parameters of FMBS and FMPS were re-evaluated in both groups. Results Changes in the scores of clinical indices FMBS and FMPS were calculated and compared. A significant difference between pre- and post-values, for both FMBS and FMPS, was noticed in the test group; in particular, the bleeding index value demonstrated the more significant changes: 22 participants showed a clinically meaningful improvement, and 5 had a small improvement. Only three patients had no evidence of change. In addition, 50% of patients had a reduction in plaque levels. No side effects were reported. Conclusions The adjunctive use of 10-day PerioTabs treatment in the daily oral hygiene routine seemed to be efficient in reducing gingival bleeding and plaque accumulation, with absence of adverse effects. These results should be confirmed in studies with a larger number of participants following a controlled-blinded design.

Aktivitas Antifungi Air Perasan Bawang Merah (Allium ascalonicum L.) Terhadap Candida albicans Secara In Vitro

2017 ◽  
Vol 9 (2) ◽  
pp. 71
Author(s):  
Nurhasanah Nurhasanah ◽  
Fauzia Andrini ◽  
Yulis Hamidy
Keyword(s):  
Candida Albicans ◽  
Antifungal Activity ◽  
Allium Sativum ◽  
Fungicidal Activity ◽  
Positive Control ◽  
Negative Control ◽  
Randomized Design ◽  
Significant Difference ◽  
Completely Randomized Design

Shallot (Allium ascalonicum L.) has been known as traditional medicine. Shallot which has same genus with garlic(Allium sativum L.) contains allicin that is also found in garlic and has been suspected has fungicidal activity toCandida albicans. It is supported by several researches. Therefore, shallot is suspected has antifungal activity too.The aim of this research was to know antifungal activity of shallot’s water extortion againsts Candida albicans invitro. This was a laboratory experimental research which used completely randomized design, with diffusion method.Shallot’s water extortion was devided into three concentrations, there were 50%, 100% and 200%. Ketoconazole 2%was positive control and aquadest was negative control. The result of this research based on analysis of varians(Anova), there was significant difference between several treatments and was confirmed with Duncan New MultipleRange Test (DNMRT) p<0,05, there was significant difference between 100% shallot’s water extortion with othertreatments, but there was no significant difference between 50% shallot’s water extortion with 200% shallot’s. Theconclusion was shallot’s water extortion had antifungal activity againsts Candida albicans with the best concentration100%, but it was lower than ketoconazole 2%.

scholarly journals Urinary stone composition analysis and clinical characterization of 1520 patients in central China

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Daling Zhang ◽  
Songchao Li ◽  
Zhengguo Zhang ◽  
Ningyang Li ◽  
Xiang Yuan ◽  
...  
Keyword(s):  
Calcium Oxalate ◽  
Urinary Stone ◽  
Geographic Region ◽  
Central China ◽  
Urinary Stones ◽  
Composition Analysis ◽  
Stone Composition ◽  
Calcium Oxalate Stones ◽  
Significant Difference ◽  
Lower Urinary

AbstractA total of 1520 patients with urinary stones from central China were collected and analysed by Fourier transform infrared spectroscopy between October 1, 2016 and December 31, 2019. For all patients, age, sex, comorbidities, stone location, laboratory examination and geographic region were collected. The most common stone component was calcium oxalate (77.5%), followed by calcium phosphate (8.7%), infection stone (7.6%), uric acid (UA) stone (5.3%)and cystine (0.9%). The males had more calcium oxalate stones (p < 0.001), while infection stone and cystine stones occurred more frequently in females (p < 0.001). The prevalence peak occurred at 41–60 years in both men and women. UA stones occurred frequently in patients with lower urinary pH (p < 0.001), while neutral urine or alkaline urine (p < 0.001) and urinary infection (p < 0.001) were more likely to be associated with infection stone stones. Patients with high levels of serum creatinine were more likely to develop UA stones (p < 0.001). The proportion of UA stones in diabetics was higher (p < 0.001), and the incidence of hypertension was higher in patients with UA stones (p < 0.001). Compared to the other types, more calcium oxalate stones were detected in the kidneys and ureters (p < 0.001), whereas struvite stones were more frequently observed in the lower urinary tract (p = 0.001). There was no significant difference in stone composition across the Qinling-Huaihe line in central China except UA stones, which were more frequently observed in patients south of the line (p < 0.001).

scholarly journals Comparison of Simulation-Based versus Cadaveric-Tissue-Based Ocular Trauma Training on Novice Ophthalmologists: Repair of Corneal Laceration Model

2021 ◽  
Vol 13 (01) ◽  
pp. e57-e65
Author(s):  
Boonkit Purt ◽  
Timothy Ducey ◽  
Sean Sykes ◽  
Joseph F. Pasternak ◽  
Denise S. Ryan ◽  
...  
Keyword(s):  
Primary Outcome ◽  
Wound Closure ◽  
Ocular Trauma ◽  
Controlled Trial ◽  
Preliminary Evaluation ◽  
Skills Acquisition ◽  
Visual Axis ◽  
Significant Difference ◽  
Skills Laboratory ◽  
Corneal Laceration

Abstract Purpose The aim of this study was to evaluate whether the simulated tissue models may be used in place of animal-based model for corneal laceration repair for surgical skills acquisition. Design Prospective randomized controlled trial. Participants Seventy-nine military and civilian 2nd- and 3rd-year ophthalmology residents and 16 staff ophthalmologists participating in the Tri-Service Ocular Trauma Skills Laboratory at the Uniformed Services University (Bethesda, MD). Methods Resident ophthalmologists underwent preliminary evaluation of their ability to close a 5-mm linear, full-thickness corneal laceration involving the visual axis. They then were randomized to undergo 90 to 120 minutes of either simulator-based (SIM) or swine cadaveric-tissue-based (CADAVER) corneal laceration repair. The same evaluation was performed post training. On a more limited basis, the study was repeated for attending ophthalmologists to act as a pilot for future analysis and test efficacy for “refresher” training. Main Outcome Measures Successful wound closure with secondary outcomes of suture length, tension, depth, and orientation, as graded by attending ophthalmologists. Results No significant difference in CADAVER versus SIM groups in the primary outcome of watertight wound closure of the corneal laceration. CADAVER group performed better than SIM group for certain metrics (suture depth, p = 0.009; length, p = 0.003; and tension, p = 0.043) that are associated with poor wound closure and increased amount of induced corneal astigmatism. For attending ophthalmologists, six of the eight in each group (SIM and CADAVER) retained or improved their skills. Conclusions For resident ophthalmologists, SIM training is sufficient for achieving the primary outcome of watertight wound closure. However, CADAVER training is superior for wound metrics for the ideal closure. For attending ophthalmologists, SIM training may be useful for retention of skills.

scholarly journals A Tumbling Magnetic Microrobot System for Biomedical Applications

Micromachines ◽  
2020 ◽  
Vol 11 (9) ◽  
pp. 861
Author(s):  
Elizabeth E. Niedert ◽  
Chenghao Bi ◽  
Georges Adam ◽  
Elly Lambert ◽  
Luis Solorio ◽  
...  
Keyword(s):  
Ultrasound Imaging ◽  
Biomedical Applications ◽  
Imaging System ◽  
Ex Vivo ◽  
Negative Control ◽  
Circular Path ◽  
Rotational Axis ◽  
Significant Difference

A microrobot system comprising an untethered tumbling magnetic microrobot, a two-degree-of-freedom rotating permanent magnet, and an ultrasound imaging system has been developed for in vitro and in vivo biomedical applications. The microrobot tumbles end-over-end in a net forward motion due to applied magnetic torque from the rotating magnet. By turning the rotational axis of the magnet, two-dimensional directional control is possible and the microrobot was steered along various trajectories, including a circular path and P-shaped path. The microrobot is capable of moving over the unstructured terrain within a murine colon in in vitro, in situ, and in vivo conditions, as well as a porcine colon in ex vivo conditions. High-frequency ultrasound imaging allows for real-time determination of the microrobot’s position while it is optically occluded by animal tissue. When coated with a fluorescein payload, the microrobot was shown to release the majority of the payload over a 1-h time period in phosphate-buffered saline. Cytotoxicity tests demonstrated that the microrobot’s constituent materials, SU-8 and polydimethylsiloxane (PDMS), did not show a statistically significant difference in toxicity to murine fibroblasts from the negative control, even when the materials were doped with magnetic neodymium microparticles. The microrobot system’s capabilities make it promising for targeted drug delivery and other in vivo biomedical applications.

scholarly journals Translational repression of SLC26A3 by miR-494 in intestinal epithelial cells

2014 ◽  
Vol 306 (2) ◽  
pp. G123-G131 ◽  
Author(s):  
Arivarasu N. Anbazhagan ◽  
Shubha Priyamvada ◽  
Anoop Kumar ◽  
Daniel B. Maher ◽  
Alip Borthakur ◽  
...  
Keyword(s):  
Protein Expression ◽  
Therapeutic Strategy ◽  
Luciferase Activity ◽  
Negative Control ◽  
Intestinal Epithelial ◽  
Diarrheal Diseases ◽  
Regulate Protein ◽  
Putative Binding Site ◽  
Mammalian Intestine ◽  
Relative Luciferase Activity

SLC26A3 [downregulated in adenoma (DRA)] is a Cl−/HCO3− exchanger involved in electroneutral NaCl absorption in the mammalian intestine. Altered DRA expression levels are associated with infectious and inflammatory diarrheal diseases. Therefore, it is critical to understand the regulation of DRA expression. MicroRNAs (miRNAs) are endogenous, small RNAs that regulate protein expression via blocking the translation and/or promoting mRNA degradation. To investigate potential modulation of DRA expression by miRNA, five different in silico algorithms were used to predict the miRNAs that target DRA. Of these miRNAs, miR-494 was shown to have a highly conserved putative binding site in the DRA 3′-untranslated region (3′-UTR) compared with other DRA-targeting miRNAs in vertebrates. Transfection with pmirGLO dual luciferase vector containing DRA 3′-UTR (pmirGLO-3′-UTR DRA) resulted in a significant decrease in relative luciferase activity compared with empty vector. Cotransfection of the DRA 3′-UTR luciferase vector with a miR-494 mimic further decreased luciferase activity compared with cells transfected with negative control. The transfection of a miR-494 mimic into Caco-2 and T-84 cells significantly increased the expression of miR-494 and concomitantly decreased the DRA protein expression. Mutation of the seed sequences for miR-494 in 3′-UTR of DRA abrogated the effect of miR-494 on 3′-UTR. These data demonstrate a novel regulatory mechanism of DRA expression via miR-494 and indicate that targeting this microRNA may serve to be a potential therapeutic strategy for diarrheal diseases.

scholarly journals Hyperhomocysteinaemia in Behçet's Disease

2010 ◽  
Vol 2010 ◽  
pp. 1-5 ◽  
Author(s):  
Amira Hamzaoui ◽  
Olfa Harzallah ◽  
Rim Klii ◽  
Silvia Mahjoub
Keyword(s):  
Behçet’S Disease ◽  
Behcet’S Disease ◽  
Homocysteine Level ◽  
Behçet's Disease ◽  
Negative Control Group ◽  
Negative Control ◽  
Control Group ◽  
Behcet's Disease ◽  
Homocysteine Concentration ◽  
Significant Difference

Objectives. The aim of this study was to investigate if hyperhomocysteinaemia is a contributive risk factor for the pathogenesis and the activity of Behçet's disease (BD).Design and Methods. Fifty four patients fullfiling the criteria of the International Study Group for BD were enrolled. Fifty healthy volunteers matched for age and sex with the BD group were included as a negative control group. Patients, with any condition that might affect plasma homocysteine concentration, were excluded.Results. Mean serum homocysteine concentration was significantly higher in patients with BD than in the healthy controls (), in patients with active disease (), and in masculine gender (). There was no significant difference between homocysteine level and clinical involvement.Conclusions. We demonstrated that plasma total homocysteine level (tHcy) is increased in BD and correlated with disease activity. No association was found between homocysteine levels and clinical involvement.

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