OD Values Research Articles

Evaluating the probiotic potential of Yeasts isolated from Ethiopian traditionally fermented foods and dairy products

Fermented foods were most prepared products for their benefits of health such as medicinal, therapeutic and probiotic effects. These effects were due to potential probiotic microorganisms such as lactic acid bacteria and yeasts. The purpose of this study was to evaluate the probiotic characteristics of yeasts isolated from Ethiopian traditionally fermented foods and dairy products. The isolates identified and subjected to analyses to determine their probiotic properties. From the selected isolates ten were found to be resistant and showed good growth at pH 2, 2.5and 3for 24h and were able to grow at both 25, 37 and 45°C but best growth was found at body temperature 370C. The three isolates namely Y66, Y82 andY36 showed maximum growth at pH 2.5 with corresponding mean OD value of 1.45, 1.40 and 1.09. From the finding insolate Y77 and Y82 performed well at pH 2.5 and 3 compared to other isolates. The three isolates GB48, Y77 and Y82 shown highest growth rate with mean OD630nm value of 2.25, 1.85 and 1.78 respectively at 370C. Isolates were also investigated for bile salt tolerance, cholesterol assimilation, co aggregation and antioxidant abilities. IsolateY76 and Y82 were found to be with highest bile salt tolerant and cholesterol assimilation of 46.56, 56.20% and 96.11, 92.11% of assimilation respectively. The two isolates Y77 and Y82 also showed good survival rate of 44.59 and 36.56% in gastrointestinal fluid and antioxidant scavenging ability of 44.59 and 36.00% respectively. Among these isolates Y82, Y76 and Y77 have been found to bear with promising desirable probiotic potentials properties.

Analysis of Patients Mislabeled with Heparin Induced Thrombocytopenia

Introduction: Heparin-induced thrombocytopenia (HIT) is a rare diagnosis, however commonly tested. We hypothesize that the 4T score, a diagnostic tool to diagnose HIT, is underutilized by clinicians. This may subsequently lead to unnecessary testing for HIT, administration of costly alternative anticoagulants, and mislabeling of heparin as allergy. We present preliminary data of our quality improvement study. Our aim was to analyze patients previously labeled with heparin allergy and analyze the 4T's scores, optical density values (OD), and serotonin release assay (SRA) results.Methods: We conducted a retrospective study of patient at two tertiary care hospitals who were evaluated for HIT from October 2015- present. We were notified by pharmacy when a HIT panel was sent and alternative anticoagulation was started. Patients with unknown heparin administration from outside institutions were excluded from the study. We calculated the 4T's score and further categorized low (0-3), intermediate score (4-5), and high (6-8) score. We considered the patients to have “true HIT” if they had 4T's score > 5 or optical density values >0.4 and positive SRA.Results: We analyzed 50 patients who were evaluated for HIT and placed on alternative anticoagulation. We observed that 64% (32/50) of the patients were listed as having heparin allergy secondary to HIT, while 36% (18/50) were not listed to have HIT as allergy on electronic health record. 19 of the 50 (38%) had scored had low 4T's score, of that two tested for HIT with a score of 0 (4%, n=50), 22 (44%) had intermediate, and 9 (18%) had high. When we utilized the 4T's score as pre-test clinical score, 0/13 patients with low score (6 not tested for SRA) and 0 /18 with intermediate score (4 were not tested) had positive SRA. 4 of the 9 (67%) high score were positive (3 were not tested). When we evaluated SRA results for patients with OD value >0.4, we observed 3 with OD value >2 and 1 with OD value of 0.798 had positive SRA. Interestingly, 1 with OD value of 2.136 had negative SRA. 8 patients with OD values > 1 had negative SRA.Conclusion: Preliminary results show that patients are frequently tested for HIT, misdiagnosed with HIT, and mislabeled with HIT in electronic health record. When 4T's is used as clinical score, patients with low probability should not be further tested for HIT. Our study has different results for compared to study of Lo GK et al who utilized 4T's score as pre-test clinical score. 1According to the study patients with low score had <5% of probability of HIT and intermediate score had 8%-29% probability of HIT. However, based on our preliminary data, none of the patients with low and intermediate score had true HIT when SRA was tested. With the intent to practice cost-effective medicine and prevent unwanted bleeding events, we plan to implement 4T score as pre-test diagnostic tool in the EMR for physicians prior to ordering alternative anticoagulation for patients with low 4T's score. We also aim to decrease hospital cost by not only preventing unnecessary HIT panel tests, but also reduce cost spent on costly alternative anticoagulation. Whether SRA is a valuable test for patients with OD values between 1-2 remains uncertain. Limitations include small sample size and incomplete chart data given retrospective study.Reference: Lo GK, Juhl D, Warkentin TE, Sigouin CS, Eichler P, Greinacher A. Evaluation of pretest clinical score (4 T's) for the diagnosis of heparin-induced thrombocytopenia in two clinical settings. J Thromb Haemost 2006; 4: 759-65. DisclosuresNo relevant conflicts of interest to declare.

Dehydropeptidase 1 Promoted Survival and Cell Cycle Progression through Regulation of CREB and P53 Pathways in B-Cell ALL

Background: Outcome of adults with B-cell acute lymphoblastic leukemia (ALL) remains poor and relapse is the major cause of treatment-failure. Identifying new biomarkers in B-cell ALL and studying their clinical significance and biological function will be helpful for risk-stratification, treatment decision and targeted therapy. Dehydropeptidase 1 (DPEP1) is a zinc-dependent metalloproteinase that is expressed aberrantly in several cancers. The role of DPEP1 in cancer remains controversial. In our previously conducted microarray analysis, DPEP1 was identified as one of most differentially expressed genes which were up-regulated in B-cell ALL more than 20-fold compared with normal bone marrow. However, the expression and biological function of DPEP1 in B-cell ALL remains unknown.Aims: To investigate the expression levels of DPEP1 in adults with B-cell ALL and to explore the biological function of DPEP1 in the cell line derived from B-cell ALL.Methods: A real-time quantitative RT-PCR was used to examine DPEP1 expression in 11 malignant hematological disorders cell lines and bone marrow samples from 146 adult acute leukemia and 26 health donors. We constructed 2 B-cell lines: DPEP1 over-expressing (OE) and DPEP1 knocked-down (KD) BV173 cell line to study the biological role of DPEP1 in B-cell ALL. The cell proliferation and colony formation were detected by CCK-8 and colony formation assay. The cell apoptosis and cell cycle were analysed by flow cytometry. The cell migration ability was assayed by the transwell assay. Western blot was used to determine the protein level of signaling pathway.Results: These results showed that the relative levels of DPEP1 gene expression in marrow from the 126 newly diagnosed B-cell ALL(median 136%; range 0-1066%) were significantly higher than those of marrow from the 26 healthy donors(8.4%; 0-20.8%; p <0.01), whereas no significant difference was detected in newly diagnosed AML patients compared with health donors (Figure A). DPEP1 gene expression was higher in BV-173 cells derived from B-cell ALL, but nearly undetectable in cell lines derived from AML(KG-1, NB4, HL60, MEG-01), CML(K562), MM(MOLP2, SKO-007), lymphoma(U937, Raji, Ramos) and the normal B cell line IM-9. Proliferation was significantly decreased in DPEP1-KD cells compared with cells transfected with control lentivirus (OD value: 1.90±0.05 vs 1.68±0.06, p < 0.05). In contrast, DPEP1 over-expression markedly increased proliferation of transfected cells (OD value: 1.32±0.06 vs 1.72±0.03, p < 0.05) (Figure B). Silencing of DPEP1 expression significantly decreased numbers of colony-forming units compared with controls (65.7±10.4 vs 109.7±8.3, p <0.01), whereas DPEP1 over-expression had a converse effect (116.7±15.5 vs 106.7±6.6, p <0.05). Data from flow cytometry analyses showed DPEP1 expression promoted cell-cycle progression: DPEP1-OE BV173 had a higher percentage cells in S phases (43.19±0.35% vs 38.58±0.29%, p <0.01), whereas DPEP1-KD BV173 cells significantly prone to accumulate in G0/G1 phases compared with control cells (57.75±1.66% vs 50.78±0.39%, p <0.01) (Figure C). Migration and invasion activity were significantly enhanced in DPEP1-OE BV173 cells but attenuated in DPEP-KD BV173 cells as detected by transwell assay. However, DPEP1-KD significantly increased the apoptosis in BV173 cells (11.68±0.18% vs 1.72±0.12%, p <0.001) (Figure C). Then we sought to identify the pathway and molecular mechanisms that mediated the effect of DPEP1.The p-CREB were significantly up-regulated in DPEP1-OE cells but down-regulated in DPEP1-KD cells. And expression of p53 was increased in DPEP1-KD cells (Figure D).Conclusion: These findings suggest that abnormal expression of DPEP1 in leukemia may be involved in the pathomechanism of B-cell ALL. DPEP1 improves the proliferation, migration, invasion and promote the survival in B-cell ALL cells, which might be mediated via CREB and p53 pathways. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Biodegradation of polypropylene films by Bacillus paralicheniformis and Lysinibacillus fusiformis isolated from municipality solid waste contaminated soil

The fossil fuel or petroleum derived plastics are applied in our routine life because of their easy availability. Distribution and contamination of the plastics in the landfills are the major reasons for these biodegradation study. This current study reveals the biodegradation of polypropylene films and the growth of Bacillus paralicheniformis and Lysinibacillus fusiformis isolated from plastic contaminated soil collected from municipality solid waste management site. The degradation rate of PP films was confirmed by the results of biodegradation analysis. The growth of Bacillus paralicheniformis and Lysinibacillus fusiformis had shown OD values at 600nm after the degradation period of 4 weeks increasing from 0.131 to 0.334 and 0.148 to 0.213 respectively. The viable cell count increased from 8×104cells/ml to 12×104cells/ml and 10.1×104 cells/ml to 15.2×104 cells/ml respectively. The physical and chemical changes of PP films were confirmed by FT-IR and XRD analysis. These analysis confirmed that the bacterial strains have the ability to change the chemical and physical nature of PP films and can utilize the PP films as sole carbon source.

Potential of Lignocellulosic Waste for Laccase Production by Trametes versicolor under Submerged Fermentation.

&lt;b&gt;Background and Objective:&lt;/b&gt; Laccase is one of the ligninolytic enzymes classified as a multicopper oxidoreductase group that has the ability in oxidizing phenolic compounds and has widespread use in both food and non-food industries. This enzyme is extracellularly secreted by white-rot fungi, &lt;i&gt;Trametes versicolor&lt;/i&gt; (L.) Lloyd in the media containing lignocellulose, for example, kapok banana peels and sawdust. The objective of this study was to evaluate lignocellulosic substrate that able to produce the highest activity of the laccase from the &lt;i&gt;T. versicolor &lt;/i&gt;(L.) Lloyd. &lt;b&gt;Materials and Methods:&lt;/b&gt; Three substrate variations used in this work included the cultivation media with the addition of either kapok banana peels or sawdust and without using both materials. The inducer (CuSO&lt;sub&gt;4&lt;/sub&gt;) was added to each substrate variation and the laccase activity was subsequently measured. &lt;b&gt;Results:&lt;/b&gt; The qualitative test result for laccase detection showed that &lt;i&gt;T. versicolor &lt;/i&gt;(L.) Lloyd&lt;i&gt; &lt;/i&gt;was able to produce this enzyme indicated with a reddish-brown surrounding fungal colony. The fungi cultivated in media with the content of sawdust and 1 mM CuSO&lt;sub&gt;4&lt;/sub&gt; yielded the highest laccase activity, reaching 573.6 U L&lt;sup&gt;&lt;/sup&gt;&lt;sup&gt;1&lt;/sup&gt; with an OD value of 0.5567&lt;i&gt; &lt;/i&gt;and a pH of 5.3 after 7 days of incubation. Meanwhile, the addition of kepok banana peels and 1 mM CuSO&lt;sub&gt;4&lt;/sub&gt;, showed the maximum laccase activity (297.7 U L&lt;sup&gt;&lt;/sup&gt;&lt;sup&gt;1&lt;/sup&gt;) with the OD value of 0.6932 and a pH of 5 after incubation for 6 days. &lt;b&gt;Conclusion:&lt;/b&gt; The white-rot fungi of &lt;i&gt;T. versicolor&lt;/i&gt; (L.) Lloyd&lt;i&gt; &lt;/i&gt;could produce optimal laccase activity by adding sawdust substrate and 1 mM CuSO&lt;sub&gt;4 &lt;/sub&gt;inducer on submerged fermentation.

Expression of angiogenesis-related proteins in bone marrow mesenchymal stem cells induced by osteoprotegerin during osteogenic differentiation in rats

This study aimed to discuss the expression of angiogenesis-related proteins in bone marrow mesenchymal stem cells (BMSCs) induced by osteoprotegerin (OGP) during osteogenic differentiation in rats, and to analyze the effect of fracture healing inflammatory factor TNF-ɑ on the osteogenic differentiation of BMSCs of rats. BMSCs isolated and cultured from the third generation rats were taken as the research object. According to the addition amount of OGP, BMSCs were divided into control group, OGP (10−7 mol/L) group, OGP (10−8 mol/L) group, and OGP (10−9 mol/L) group. The cell growth and morphological characteristics of each group were observed by inverted phase contrast microscope, the cell proliferation rate was measured by MTT method, angiogenesis-related markers (platelet growth factor (VEGF), cingulate protein 5 (Fbln5), and angiogenin-like protein 4 (Angptl4)) were quantitatively detected by Western blot, and the effect of TNF-ɑ on osteogenic differentiation was detected by CCK. Compared with the control group, MTT results showed that the value-added rate of cells in the OGP (10−8 mol/L) group reached the maximum at 9 days (P < 0.05). The ALP activity in osteoblasts in the OGP (10−8 mol/L) group reached the maximum at 9 days (P < 0.01). The OGP (10−8 mol/L) group had the highest expression of vascular regeneration proteins (VEGF, Fbln5, and Angptl4) (P < 0.05). CCK analysis showed that the TNF-ɑ (1.0 ng/mL) group showed a significant increase in absorbance compared with the control group on 6 days (P < 0.05), and the OD value of the TNF-ɑ (10 ng/mL) group decreased at all time points (P < 0.05). Overall, 10−8 mol/L OGP can induce the proliferation and osteogenic differentiation of MSCs, and promote the expression of angiogenesis-related proteins (VEGF, Fbln5, and Angptl4) during osteogenic differentiation. Besides, 1.0 ng/mL of TNF-ɑ can also promote osteogenesis differentiation of BMSCs in the short term.

Serine/Arginine Repetitive Matrix 2 Antisense RNA 1 Negatively Regulates miR-370-3p and Promotes Hyperplasia, Migration, and Aggression of the Colon Cancer Cell Line

The purpose of this study is to explore the effect and possible machine-processing of the long non-coding RNA (lncRNA) SRRM2-AS1 in the development and pathogenesis of colorectal cancer. LncRNA plays an important role in tumorigenesis and development. LncRNA can regulate gene transcription and translation, cell proliferation, differentiation and apoptosis by affecting gene expression pathways of various coding proteins. SRRM2-AS1 is a kind of lncRNA. Studies have confirmed that the expression of SRRM2-AS1 is increased in colon adenocarcinoma tissues of colon cancer patients and is closely related to the prognosis of patients. However, the influence and molecular mechanism of SRRM2-AS1 on the malignant biological behavior of colon cancer cells are no yet clear. SRRM2-AS1 may interact with miR-370-3p. Studies have confirmed that overexpression of miR-370-3p can inhibit the proliferation and epithelial-mesenchymal transition of colon cancer cells in vitro. However, it is not yet clear whether SRRM2-AS1 can target miR-370-3p to affect the occurrence and development of tumors. In this study, RT-qPCR was employed to detect levels of SRRM2-AS1 and miRNA-370-3p in carcinoma tissues and corresponding paracarcinoma tissues from 41 patients with colon cancer. SW1116 colon cancer cells were cultured in vitro and separated into 4 groups: (1) si-NC group, (2) si-SRRM2-AS1 group, (3) si-SRRM2-AS1+anti-miRNA-NC group, and (4) si-SRRM2-AS1+anti-miRNA-370-3p group. The CCK-8 assay and colony formation experiment was employed to gauge cell proliferation. The scratch test was used to detect cell migration while the transwell assay was used to detect cell invasion. Finally, Western blot analysis was employed to detect levels of Ki67, E-cadherin, and N-cadherin proteins in colorectal cancer cells. The dual-luciferase reporter gene experiment verified that SRRM2-AS1 regulates miRNA-370-3p. The study found that compared to paracarcinoma tissue, levels of SRRM2-AS1 in colon cancer tissues was increased (P &lt; 0.05). Compared to the si-NC group, the SW1116 cell OD value, number of colonies formed, scratch healing rate, number of invasive cells, and expression levels of Ki67 and N-cadherin protein in the si-SRRM2-AS1 group were all decreased (P &lt; 0.05). However, E-cadherin protein levels were elevated (P &lt; 0.05). SRRM2-AS1 negatively regulates levels of miRNA-370-3p in SW1116 cells. Compared to the si-SRRM2-AS1+anti-miRNA-NC group, SW1116 cell OD value, number of colonies formed, scratch healing rate, number of invasive cells, and Ki67 and N-cadherin protein levels were increased (P &lt; 0.05) in the si-SRRM2-AS1+anti-miRNA-370-3p group. Conversely, E-cadherin protein levels were decreased (P &lt; 0.05). These findings indicate that SRRM2-AS1 is predominately expressed in cancerous colon tissues. Attenuating expression of SRRM2-AS1 may curb the hyperplasia of colon carcinoma cell line SW1116 and promote cell apoptosis by regulating miRNA-370-3p expression.

Effects of different implant surface properties on the biological behavior of Schwann cells.

This study investigated the effects of different implant surface properties on the biological behavior of Schwann cells. Schwann cells (SCs) were cultured on three types of implant surfaces including smooth polished (SMO), sand-blasted, large grit, acid-etched (SLA), and chemically-modified SLA (modSLA). At different time points, the morphology and adhesion of SCs on the implant surfaces were observed by scanning electron microscope. Cell proliferation activity was detected by MTT method. The expression levels of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) were detected by enzyme-linked immunosorbent assay. Changes in the mRNA levels of NGF and BDNF were detected by real-time fluorescent quantitative polymerase chain reaction (PCR). SCs adhered, stretched, and proliferated well on the three types of implant surfaces. On the 3rd, 5th, and 7th days, the OD values of the SMO group were higher than those of the SLA group and the modSLA group, and the difference was statistically significant (P<0.05). On the 3rd day, the expression and mRNA levels of NGF and BDNF in the SLA group and the modSLA group were higher than those in the SMO group (P<0.05); in particular, the levels in the modSLA group were higher than those in the SLA group (P<0.05). Different implant surface properties have different effects on the biological behavior of SCs. Proliferation of SCs is significantly promoted by smooth surface, while secretion and gene expression of neurotrophic factors are significantly promoted by modSLA surface at early stage.

Anti-cancer effects of baicalein on cervical carcinoma cells through down-regulation of the ERK/p38/MAPK pathway.

The objective of this study was to investigate the effects of baicalein on apoptosis of HeLa human cervical cancer (CC) cells and to elucidate the underlying mechanism. HeLa cells were treated with 20, 50, 100, or 200 μmol/L baicalein for 24, 36, and 48 hours, and CCK-8 assays were used to detect cell viability, and flow cytometry was performed to assess apoptosis rate. Reverse-transcription quantitative PCR was used to measure ERK1/2, p38, and JNK mRNA levels in HeLa cells, and western blotting was performed to measure ERK1/2, p38, and JNK protein levels. The CCK-8 assay showed that the OD value of HeLa cells gradually decreased with increasing baicalein concentrations (P < 0.01) and treatment time (P < 0.01). These results indicated a negative time- and dose-dependent effect of baicalein on HeLa cells. Baicalein treatment of HeLa cells significantly increased apoptosis rate (P < 0.01). In HeLa cells treated with 50 or 200 μmol/L baicalein for 24 h, expression levels of ERK1/2 and p38 mRNA were significantly reduced, whereas that of JNK mRNA was increased (P < 0.01). The levels of phosphorylated ERK1/2 and p38 were significantly reduced, and the level of JNK protein was increased (P < 0.01). Taken together, baicalein appeared to exert anti-cancer effects on HeLa cells through induction of apoptosis and regulation of the ERK/p38/mitogen-activated protein kinase pathway.

Serum Samples from Co-Infected and Domestic Cat Field Isolates Nonspecifically Bind FIV and Other Antigens in Enzyme-Linked Immunosorbent Assays.

We evaluated enzyme-linked immunosorbent assay (ELISA) specificity for measuring seroantibody responses to two types of retroviral infections in domestic cats: feline immunodeficiency virus (FIV) and feline foamy virus (FFV). We compared the seroreactivity of specific pathogen-free (SPF) cat sera, sera from SPF cats inoculated with either FIV or FFV, and field isolates (e.g., shelter or privately owned cats). Sera from SPF cats experimentally infected with the cognate virus had significantly lower background in both FIV and FFV ELISAs compared to sera from negative field isolates. ELISA values for SPF cats exposed to either FIV or FFV tended to have higher OD values on the opposite ELISA antigen plate. FIV nonspecific background absorbance was greater than that of FFV, and 10 of 15 sera samples from FIV seronegative field samples were measured in the indeterminant range. These findings highlight that exposure to off-target pathogens elicit antibodies that may nonspecifically bind to antigens used in binding assays; therefore, validation using sera from SPF animals exposed during controlled infection results in the setting of a cutoff value that may be inappropriately low when applied to field samples. Our work also suggests that infection of domestic cats with pathogens other than FIV results in antibodies that cross-react with the FIV Gag antigen.

Inhibitory effect of lignans from Ailanthus altissima on proliferation of human lung cancer cells and its mechanism.

e15059 Background: Plants are thought to contain compounds that inhibit the proliferation of cancer cells in vitro, and many attempts have been made to isolate anti-cancer drugs from plants. Ailanthus altissima is a family of Simaroubaceae, the bark is gray to grayish-black, because of the smell of the gland patches at the base of the leaves. 3,5,3-trimethoxy-8,1-neoligna-4,2,7,9,9-pentol (TNP) isolated from Ailanthus altissima. Methods: (1) Apoptosis was detected by flow cytometry: the cells were incubated with 10 mmol/L TNP for 24 h, and the cells were collected. The cell concentration was adjusted to 5×105 ̃ 5×106/ L. While propionate iodide was added for DNA fluorescence staining (Propidium Iodide, PI: 50 mg/L，triton-x100 1.0%). (2) The changes of p53, Bax and caspase-3 were detected by Western blot. The cells were cultured in 10 μmol/L cisplatin and TNP medium for 24 hours. Image J image analysis software is used for semi quantitative analysis. The data were expressed by mean standard deviation, and the mean values of each group were compared by analysis of variance and the least significant difference. There was a significant difference in P&lt; 0.05. (3) A549 cells were treated with 10 μmol/L cisplatin and TNP. A549 cells were incubated with cisplatin and TNP, and then treated with solvent control (final concentration 0.1% dimethyl sulfoxide) with or without caspase inhibitor for 48 hours The OD value was determined by MTT method. Cell survival rate (%) = (OD value of control group /OD value of control group) 100%. Results: (1) The reverse effect of caspase inhibitor on the proliferation of A549 cells: the cell survival rates of A549 cells treated with 0.1, 1 and 10 μmol/L cisplatin and TNP were 96.11%, 72.36%, 27.57% and 71.42%, 28.97% and 4.34% respectively. The survival rate of A549 cells incubated with cisplatin and TNP 20 μmol/L increased to 100.00%, 81.92%, 57.54% and 85.69%, 48.57% and 16.95% respectively. Caspase inhibitor reversed the inhibitory effect of TNP on A549 cells. (2) The effect of TNP on apoptosis of A549 cells: after treatment with 10 μmol/L TNP for 24 h, the apoptosis rate of A549 cells was (24.01)%, which was significantly higher than that of the blank control group (6.79)% ( P&lt; 0.05). (3) The results showed that the up regulation of p53, Bax and caspase-3 protein by TNP was 10 μmol/L. Conclusions: TNP can induce apoptosis of human lung cancer cell line A549, and it can up regulate the expression of p53, Bax and Caspase-3, which may be one of the mechanisms of TNP inducing apoptosis of A549 cells. The experiment also proves that caspase inhibitor can reverse the effect of TNP. TNP inhibits the proliferation of A549 cells.

Endoleak Detection after Endovascular Aortic Aneurysm Repair Using Ultrasound Based on Nanoscale Bubble Contrast Agents and Their Effects on Vascular Smooth Muscle Cell Proliferation and Migration

Abdominal aortic aneurysm (AAA) is the most common vascular disease that causes disability and death. Its morbidity is relatively subtle, and the mortality rate is high. Clinically, endovascular aortic aneurysm repair (EVAR) has gradually become the primary treatment of AAA due to its unique advantages such as low trauma and low incidence of short-term complications. However, the outcome of EVAR is greatly compromised by the possible occurrence of endoleaks. Contrast-enhanced ultrasound (CEUS) is a promising alternative technique to detect endoleaks following EVAR due to lack of exposure to ionizing radiation. Traditional ultrasound contrast agents with an overlarge size (microscale) leading to reluctant accumulation in target organs and instability trigger the requirement of nanoscale contrast agents that enter tumor tissues through the enhanced permeability and retention effect. In this study, we used ultrasound based on nanoscale bubble contrast agents to evaluate endoleak detection after endovascular aortic aneurysm repair and analyzed the effects of nanoscale bubble contrast agents on vascular smooth muscle cell (VSMC) proliferation and migration. Among 52 AAA patients a month following EVAR, there were 16 cases of endoleaks after EVAR detected by nanobubble contrast-enhanced ultrasound, including 6 cases of type I endoleak (1 case of type Ia endoleak and 5 cases of type Ib endoleak), 7 cases of type II endoleak, and 3 cases of type III endoleak; there were 12 cases of endoleak after EVAR detected by computed tomography angiography (CTA), including 6 cases of type I endoleak (1 case of type Ia and 5 cases of type Ib), 5 cases of type II endoleak, and 1 case of type III endoleak. Six months after EVAR, 3 cases of type III endoleak were detected by both nanobubble contrast-enhanced ultrasound and CTA. Vascular smooth muscle cells (VSMCs) used for in vitro experiments were subjected to ultrasound irradiation and platelet-derived growth factor (PDGF) treatment with or without the addition of nanobubble contrast agents. After high-intensity and long-term irradiation (0.75 W/cm2 and 1 W/cm2 irradiation for 120 s and 150 s, respectively) by ultrasound with or without the addition of nanobubble contrast agents, PDGF-induced VSMC migration was inhibited ( P &lt; 0.01 ). Low-intensity and short-term ultrasound irradiation did not differ PDGF-induced VSMC migration ( P &gt; 0.05 ), but 0.5 W/cm2 and 90 s ultrasound irradiation could significantly inhibit PDGF-induced VSMC migration without the addition of nanobubble contrast agents ( P &lt; 0.05 ). When VSMCs were irradiated at 1, 0.75, 0.5, and 0.35 W/cm2 for 30 s to 150 s, ultrasound irradiation with or without the addition of nanobubble contrast agents remarkably reduced PDGF-induced VSMC proliferation, as evidenced by reduced OD values ( P &lt; 0.05 ). In conclusion, ultrasound based on nanoscale bubble contrast agents is an effective alternative detection method for the occurrence of AAA patients who are not suitable for CTA.

A revisited two-step microtiter plate assay: Optimization of in vitro multiplicity of infection (MOI) for Coliphage and Vibriophage

A 2-step microtiter plate assay was developed to simultaneously check wide values of MOIs of bacteriophages, ranging between MOI-0.0001 and MOI-10000 in the first step and optimize the most suitable MOI (lowest quantity of phage) for inhibiting the growth of the target bacteria in the second step. The results of the first step revealed that the effective MOI of coliphage-ɸ5 for controlling the growth of antimicrobial resistant (AMR) E. coli was between 4.36 and 43.6 for E.coli-EC-3; between 38.2 and 382 for E.coli-EC-7 and between 81.5 and 815 for E.coli-EC-11. The optimum MOI of coliphage-ɸ5 determined in the second step was 17.44, 191 and 326 for controlling the growth of E.coli-EC-3; E.coli-EC-7 and E.coli-EC-11, respectively. The effective MOI of vibriophage-ɸLV6 for controlling luminescent Vibrio harveyi in the first step was found to be between 18.3 and 183 and the optimum MOI as determined in the second step was 79. The sequential 2-step microtiter plate method yielded faster optimization of MOI and was economical compared to the conventional flask method. The measurement of OD values at 550 nm and 600 nm showed similar trend and replicate data from 5-wells and 3-wells yielded identical pattern indicating that the measuring absorbance data in 3-replicate wells at either OD550 or OD600 is sufficient to generate quantifiable phage lysis data. The 2-step microtiter plate assay finds application in phage therapy in human health care, agriculture and animal agriculture for determining the optimum MOIs for selected bacteriophages.

Influence of parity and live weight on the concentration of Pregnancy-Specific Protein B (PSPB) in Kedah Kelantan (KK) cattle

The pregnancy-specific protein B (PSPB) is the protein of binucleated cells in the ruminant’s placenta. It contains several antigenic variants, which contributed to the ability to detect pregnancy from day 20 of pregnancy until parturition. In this research, the main objective is to determine the concentration of PSPB in serum and urine and their relation to Kedah-Kelantan (KK) cattle’s parity and live weight. Ten cattle (n=10) were selected for this study. The blood serum (2 ml) and urine samples were collected at days 0, 24, 42, and 164 post artificial insemination (AI). All the samples were centrifuged at 3000 rpm for 15 minutes at 4°C. The serum and urine were analyzed with competitive ELISA test kit and read at 450 nm wavelength speed. The standard OD value provided by the kit created the equation to calculate the concentration of PSPB in the serum and blood. Then, the correlation and multiple linear regression of parity and live weight were analyzed with a significant level of P < 0.01. Through the finding, the live weight and parity were positive correlate with PSPB in serum. The relationship between live weight and PSPB concentration in urine showed a negative correlation but there no correlation between parity and PSPB concentration in urine. The regression analysis result was 35% and 36% for PSPB concentration in serum and urine, respectively. This study suggests that cattle parity and live weight did not significantly affect the PSPB concentration.

Establishment of an indirect ELISA-based method involving the use of a multiepitope recombinant S protein to detect antibodies against canine coronavirus.

Here, we report the development of an indirect enzyme-linked immunosorbent assay (ELISA) method that involves using multiepitope recombinant S protein (rSP) as the coating antigen to detect antibodies against canine coronavirus (CCoV). rSP was designed by arranging its four S fragments (91–135 aa, S1 gene; 377–434 aa, S2 gene; 647–671 aa, S3 gene; 951–971 aa, S4 gene; 207–227 aa) and two T-cell epitopes in tandem: T–E1–E2–E3–E4–T. This multiepitope antigen, which has a molecular weight of approximately 25 kDa and contains a His-tag, was recognized by a CCoV-positive serum in a Western blot assay. The optimal concentration of rSP as a coating antigen in the ELISA was 2 μg/mL, and the optimal dilution of enzyme-labeled secondary antibody was 1:10,000. The cutoff OD450 value was established at 0.2395. No reactivity was observed with antisera against canine distemper virus, canine parvovirus, or feline calicivirus, indicating that this assay is highly specific. We also tested 64 clinical serum samples using our newly established method, and the positive rate was found to be 82.8%. In conclusion, our assay was found to be highly sensitive and specific for the detection of antibodies against CCoV, and it can therefore serve as a new, efficient diagnostic method.

Building Block Symmetry Relegation Induces Mesopore and Abundant Open-Metal Sites in Metal–Organic Frameworks for Cancer Therapy

Building Block Symmetry Relegation Induces Mesopore and Abundant Open-Metal Sites in Metal–Organic Frameworks for Cancer Therapy

EVALUATION OF IN VITRO ANTICANCER ACTIVITY OF AERIAL PARTS OF AVICENNIA ALBA PLANT METHANOLIC EXTRACT AGAINST HELA AND MCF-7 CELL LINES

Objective: The current study emphasizes the effects of stem and leaf methanolic extract of mangrove plant Avicennia alba on human tumor cell lines HeLa and MCF-7.&#x0D; Method: The collected mangrove plant sample was subjected to extraction with methanol and screened for anticancer activity according to 3-(4,5-dimethythiazol- 2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay method. The ability of the plant extract to inhibit the proliferation of HeLa and MCF-7 cell lines procured from NCCS Pune was compared parallel to inhibitory effect of standard drug Cisplatin. The spectrophotometric optical densities obtained were plotted on graph and consecutive data were obtained to depict the inhibitory effect and IC50 values. The histopathology studies were also done for a clear insight to understand the depth of inhibition microscopically.&#x0D; Results: The MTT assay performed illustrates the plant samples with different concentrations (10, 20, 25, 30, and 50 μg) showed significant inhibitory effect on comparison with the standard drug Cisplatin. The IC50 values and the inhibitory OD values elucidate effective projection of plant as a potential source of anticancer activity based on comparative study. The histopathology studies revealed an insight to a clearer picture of how the cells growth was controlled. It was found that the methanolic extract of A. alba exhibited more activity against HeLa cell lines than MCF-7 cell lines.&#x0D; Conclusion: The synergistic effect of various phytochemicals present in the methanolic plant extract played a significant role in establishing the plant as a significant anticancer source.

Role of rapid antibody and ELISA tests in the evaluation of serological response in patients with SARS-CoV-2 PCR positivity.

From 7 to 8 days after the onset of symptoms in COVID-19 infection, the sensitivity of serological tests was found to be higher than that of nucleic acid tests. The aims of this study were to investigate antibody levels in patients with SARS-CoV-2 infection, to examine the relationship between antibody levels and virus load, and to evaluate the performance of 2 different commercial kits. A total of 103 patients with confirmed SARS-CoV-2 infection were included in the study. Antibodies against SARS-CoV-2 in serum samples taken from patients were investigated simultaneously with anti-SARS-CoV-2 IgG and IgA ELISAs (Euroimmun) and COVID-19 (SARS-CoV-2) IgG/IgM (Deep Blue) kits. No positivity was detected with any of the test kits in 18 (17.4%) of the 103 samples. In symptomatic patients, 100% of IgM and IgA tests were found to be positive in the group sampled after 10 days, while 100% of IgG tests were found positive after 20 days. The sensitivity of the Deep Blue COVID-19 IgG antibody kit was calculated as 81.48% and the specificity was 97.96%. While there was no statistically significant difference between the PCR CT and ELISA OD values, a positive correlation was found between the ELISA OD values and the days since the date of symptom initiation. The sensitivity and specificity of the rapid antibody test used in this study were found to be quite high. In conditions where ELISA tests cannot be applied, it is thought that it can give an idea in terms of the presence of antibodies as a simple and fast test. Although ELISA tests are valuable in the diagnosis of COVID-19 during the acute period, they are tests that can be used safely in the diagnosis of previous infections and seroepidemiological studies.

Evolutionary Mechanism of Immunological Cross-Reactivity Between Different GII.17 Variants.

Human norovirus is regarded as the leading cause of epidemic acute gastroenteritis with GII.4 being the predominant genotype during the past decades. In the winter of 2014/2015, the GII.17 Kawasaki 2014 emerged as the predominant genotype, surpassing GII.4 in several East Asian countries. Hence, the influence of host immunity response on the continuous evolution of different GII.17 variants needs to be studied in depth. Here, we relate the inferences of evolutionary mechanisms of different GII.17 variants with the investigation of cross-reactivity and cross-protection of their respective antisera using the expression of norovirus P particles in Escherichia coli. The cross-reactivity assay showed that the antisera of previous strains (GII.17 A and GII.17 B) reacted with recent variants (GII.17 C and GII.17 D) at high OD values from 0.8 to 1.16, while recent variant antisera cross-reacting with previous strains were weak with OD values between 0.26 and 0.56. The cross-protection assay indicated that the antisera of previous strains had no inhibitory effect on recent variants. Finally, mutations at amino acids 353–363, 373–384, 394–404, and 444–454 had the greatest impact on cross-reactivity. These data indicate that the recent pandemic variants GII.17 C and GII.17 D avoided the herd immunity effect of previous GII.17 A and GII.17 B strains through antigenic variation.

Dosage Effectiveness of the Combination Calcium Hydroxide-Propolis as an Inhibitor for Aggregatibacter actinomycetemcomitans Biofilm

Objective: Calcium hydroxide is widely used as pulp-capping material, but it is resistant to Aggregatibacteractinomycetemcomitans bacteria. Because of this deficiency, many researchers are considering usingalternative natural ingredients, such as propolis. Propolis has been widely used in dentistry due to itsproperties and because it has active ingredients that can act as antibacterial agents. It is hoped that whencalcium hydroxide and propolis are combined, the combination can overcome the disadvantages. The aim ofthis study was to determine the dosage effectiveness of the combination calcium hydroxide-propolis as aninhibitor for A. actinomycetemcomitans biofilm. Material and methods: The sample was divided into fivegroups. The positive control group is a combination of calcium hydroxide-sterile aquadest, and the negativecontrol group is A. actinomycetemcomitans bacteria. Group I is a combination of calcium hydroxidepropolis with a ratio of 1:1, group II has a ratio of 1:1.5, and group III has a ratio of 1:2. Each group consistsof five replications. Combination of calcium hydroxide-propolis and A. actinomycetemcomitans bacterialsuspension was incubated for 2x24 hours then washed three times using PBS. Furthermore, 2% of crystalviolet was used for staining and covered with aluminum foil and then incubated for 15 minutes. Isopropanol200µL was added, followed by a measurement of OD with an ELISA reader, and the OD values werecalculated using an inhibition biofilm formula. Results: Group II has the largest mean inhibition with 76%compared to groups one and three. Conclusion: A combination of calcium hydroxide and propolis with aratio of 1: 1.5 is more effective than 1:1 or 1:2 to inhibit A. actinomycetemcomitans biofilm.