Mean Optical Density Values Research Articles

Biodegradation potentials of soil-borne fungal cultures grown on haloxyfop R methyl ester herbicide medium

The heterotrophic and haloxyfop-R methyl ester utilizing fungal counts associated with two top soil samples was determined. Five (5) fungal isolates; Aspergillus niger, Candida sp., Trichoderma sp., Fusarium sp., and Alternaria sp. Were cultured from the soil samples using serial dilution as well as pour plate techniques. These isolates were sub-cultured and screened for their ability to use haloxyfop-R methyl ester as sole source of metabolic energy as well as using the turbidimeteric method. Two of the screened fungal isolates which exhibited maximal optical density (OD) difference values; A. niger and Fusarium sp. Were selected and utilized for the subsequent degradation/shake flask experiment. Growth profile parameters which included; pH, OD, dissolved CO2 and weighed dry fungal biomass were determined during the growth profiling test at a 96 hour interval for 16 days using appropriate methods and equipment. Mean soil pH values were 5.08 ± 0.02 and 4.62 ± 0.02 for samples A and B. The mean total heterotrophic fungal count was 8.7×10³ cfu/g ± 1.5 for A and 1.6 × 10⁴ cfu/g ±1.0 for B. The mean pH value observed for A. niger in the course of the growth profile test ranged from 6.0 ± 8.7 to 7.3 ± 8.0 and 7.3 ± 5.8. The mean OD values observed for A. niger and Fusarium sp. In the course of the shake flask experiment varied from 0.66 ± 7.1 to 1.96 ± 4.5 and 1.62 ± 5.4 to1.99 ± 3.6 respectively. With reference to the detection of mean dissolved CO2 values, utilized as an indirect indicator of herbicide mineralization, the axenic cultured A. niger was the most effective amongst the growth profile fungal cultures.

Preliminary field evaluation of indirect ELISA test using the recombinant antigen rLicNTPDase-2 for serodiagnosis of canine leishmaniasis in Colombia

Leishmaniasis is a significant public health concern, with dogs as the primary reservoir in urban scenarios and facilitating transmission. Diagnosing infected dogs is a crucial step for public health interventions, and the development of new diagnostic platforms can significantly enhance efforts in various regions worldwide. Given the limited availability of diagnostic methods in Colombia, this study evaluates the effectiveness of an Indirect Enzyme-Linked Immunosorbent Assay (ELISA) based on the recombinant protein rLicNTPDase-2 to detect Leishmania in infected dogs. Serum samples were collected from dogs in both endemic and non-endemic areas and classified as natural standards based on prior parasitological diagnoses. The results revealed 24 true positives (TP) and 9 true negatives (TN). Subsequently, the test was then validated with samples from symptomatic and asymptomatic animals, alongside the standards, yielding a specificity of 96 %, a sensitivity of 81 %, efficiency of 90.6 %, a positive predictive value of 92.8 %, and a negative predictive value of 89.6 %. The positive likelihood ratio (RV+) was 20, while the negative likelihood ratio (RV-) was 0.19, indicating high relevance and a robust clinical utility. The area under the curve (AUC) was 1.00, suggesting that the test has excellent discriminatory ability, significantly deviating from the reference diagonal. This is further supported by the significant difference(p < 0.0001) between TN and TP results determined by Fisher's exact test. Involving 163 animals showed 47 % positive and 46 % negative results with a significant difference (p < 0.05) in the mean optical density (OD) values between positive and negative samples. These findings indicate that the ELISA test effectively differentiates between positive and negative samples based on OD values. This study suggests that ELISA based on the recombinant antigen rLicNTPDase-2 could serve as a viable alternative for the serodiagnosis of leishmaniasis in canines in Colombia.

Photoactivated disinfection procedure for denture stomatitis in diabetic rats.

To study the efficacy of PADTM Plus-based photoactivated disinfection (PAD) for treating denture stomatitis (DS) in diabetic rats by establishing a diabetic rat DS model. The diabetic rat DS model was developed by randomly selecting 2-month-old male Sprague-Dawley rats and dividing them into four groups. The palate and denture surfaces of rats in the PAD groups were incubated with 1 mg/mL toluidine blue O for 1 min each, followed by a 1-min exposure to 750-mW light-emitting diode light. The PAD-1 group received one radiation treatment, and the PAD-2 group received three radiation treatments over 5 days with a 1-day interval. The nystatin (NYS) group received treatment for 5 days with a suspension of NYS of 100,000 IU. The infection group did not receive any treatment. In each group, assessments included an inflammation score of the palate, tests for fungal load, histological evaluation, and immunohistochemical detection of interleukin-17 (IL-17) and tumor necrosis factor (TNF-α) conducted 1 and 7 days following the conclusion of treatment. One day after treatment, the fungal load on the palate and dentures, as well as the mean optical density values of IL-17 and TNF-α, were found to be greater in the infection group than in the other three treatment groups (P < 0.05). On the 7th day after treatment, these values were significantly higher in the infection group than in the PAD-2 and NYS groups (P < 0.05). Importantly, there were no differences between the infection and PAD-1 groups nor between the PAD-2 and NYS groups (P > 0.05). PAD effectively reduced the fungal load and the expressions of IL-17 and TNF-α in the palate and denture of diabetic DS rats. The efficacy of multiple-light treatments was superior to that of single-light treatments and similar to that of NYS.

Bacillus subtilis alleviates excessive apoptosis of intestinal epithelial cells in intrauterine growth restriction suckling piglets via the members of Bcl-2 and caspase families.

The intestine is a barrier resisting various stress responses. Intrauterine growth restriction (IUGR) can cause damage to the intestinal barrier via destroying the balance of intestinal epithelial cells' proliferation and apoptosis. Bacillus subtilis has been reported to regulate intestinal epithelial cells' proliferation and apoptosis. Thus, the purpose of this study was to determine if B. subtilis could regulate intestinal epithelial cells' proliferation and apoptosis in intrauterine growth restriction suckling piglets. Compared with the normal birth weight group, the IUGR group showed greater mean optical density values of Ki-67-positive cells in the ileal crypt (P < 0.05). IUGR resulted in higher ability of proliferation and apoptosis of intestinal epithelial cells, by upregulation of the messenger RNA (mRNA) or proteins expression of leucine rich repeat containing G protein coupled receptor 5, Caspase-3, Caspase-7, β-catenin, cyclinD1, B-cell lymphoma-2 associated agonist of cell death, and BCL2 associated X (P < 0.05), and downregulation of the mRNA or protein expression of B-cell lymphoma-2 and B-cell lymphoma-2-like 1 (P < 0.05). However, B. subtilis supplementation decreased the mRNA or proteins expression of leucine rich repeat containing G protein coupled receptor 5, SPARC related modular calcium binding 2, tumor necrosis factor receptor superfamily member 19, cyclinD1, Caspase-7, β-catenin, B-cell lymphoma-2 associated agonist of cell death, and Caspase-3 (P < 0.05), and increased the mRNA expression of B-cell lymphoma-2 (P < 0.05). IUGR led to excessive apoptosis of intestinal epithelial cells, which induced compensatory proliferation. However, B. subtilis treatment prevented intestinal epithelial cells of IUGR suckling piglets from excessive apoptosis. © 2024 Society of Chemical Industry.

Improving Access to Anti-HDV Testing: Development and Validation of an Affordable In-House ELISA Assay.

In certain low-income nations, the hepatitis Delta virus and hepatitis B virus (HBV) pose a serious medical burden, where the prevalence of hepatitis B surface antigen (HBsAg) is greater than 8%. Especially in rural places, irregular diagnostic exams are the main restriction and reason for underestimation. Utilizing serum samples from a Pakistani isolate, an internal ELISA for the quick identification of anti-HDV was created, and the effectiveness of the test was compared to a commercial diagnostic kit. HDV-positive serum samples were collected, and a highly antigenic domain of HDAg antigen was derived from them. This antigenic HDAg was expressed in a bacterial expression system, purified by Ni-chromatography, and confirmed by SDS-PAGE and Western blot analysis. The purified antigen was utilized to develop an in-house ELISA assay for anti-HDV antibody detection of the patient's serum samples at very low cost. Purified antigens and positive and negative controls can detect anti-HDV (antibodies) in ELISA plates. The in-house developed kit's efficiency was compared with that of a commercial kit (Witech Inc., USA) by the mean optical density values of both kits. No significant difference was observed (a P value of 0.576) by applying statistical analysis. The newly developed in-house ELISA is equally efficient compared to commercial kits, and these may be useful in regular diagnostic laboratories, especially for analyzing local isolates.

Evaluation of Cytotoxic Effects of Purified Mercury in Human Gingival Fibroblasts-In vitro Study.

Since the introduction of amalgam for tooth fillings, there have been concerns that mercury toxicity could pose unacceptable health risks. Rasa shastra is an ancient medical discipline that focuses on the utilization of metals and minerals for the treatment of diseases. Nevertheless, these minerals cannot be directly administered to the human body in their natural state due to their potential adverse effects. Hence, for medicinal purposes, these metals and minerals need to undergo purification (Shodhana) to eliminate impurities and modify their physical, chemical, and biological characteristics. Human gingival fibroblasts (HGF) were exposed to commercially available mercury (CA-Hg) and ayurvedically purified mercury (AP-Hg) at concentrations of 6.25 μM, 12.5 μM, 25 μM and 50 μM. The unexposed HGF cultured in basal media was considered a control. All the samples were cultured for 24 hours and 48 hours, and the cytotoxicity was analyzed by MTT assay. Cell viability between the control and experimental groups varied at 24 hours, however, the results were not statistically significant (p>0.05). At 48 hours, cell viability was higher in the AP-Hg group as compared to the CA-Hg group at the concentration of 6.25 μM, and the difference was statistically significant (p<0.05). The cell proliferation assay results demonstrated a statistically significant difference in the mean optical density values (p<0.05) between CA-Hg and AP-Hg at 12.50 μM, 25 μM, and 50, μM concentrations observed at 24 hours. At 48 hours, a statistically significant difference in the mean OD values (p<0.05) between CA-Hg and AP-Hg at all four concentrations was observed. Conclusion: AP-Hg at a concentration of 6.25 μM demonstrated higher cell viability at 48 hours. Further, the cell proliferation rate was also higher for AP-Hg at all concentrations at 24 and 48 hours. These results indicated a less cytotoxic effect of AP-Hg than CA-Hg in HGF and hence could be employed for dental amalgam preparations.

Comparative evaluation of the antibacterial efficacy of two experimental calcium silicate-based intracanal medicaments: An in-vitro study.

Success of endodontic treatment relies on minimizing microbial load by chemo-mechanical preparation and intra-canal medication(ICM). Calcium hydroxide based ICMs have known disadvantages. Calcium silicate-based cements(CSC) exhibit antibacterial activity, thus promoting researchers to experiment with their formulations to use them as ICMs. Evaluation and comparison of the antimicrobial efficacy of two experimental CSC (MTA & Biodentine + 2%chlorhexidine) and Bio-C Temp against E.faecalis. Test materials were divided into four groups namely Group1-Bio-C Temp, Group2-UltraCAL XS, Group3-Biodentine+2%CHX and Group4-MTA+2%CHX. Direct contact test was done by placing a standardized suspension of E.faecalis on test materials and bacterial growth was assessed spectrophotometrically using ELISA at one, three and seven days. Data was analysed using one-way ANOVA, Tukey's multiple post hoc test and paired-t test. Results: Intragroup comparison revealed decreased mean optical density(OD) in groups 1, 2, and 4; no significant difference in group 3. Intergroup comparison showed statistical differences in mean OD values between groups (3 and 4); groups (1 and 2) at days one(p-0.018) and three(p-0.035), but no difference individually. Group 4 showed the highest antimicrobial efficacy on day seven. MTA+2%CHX & Biodentine+2%CHX showed better antimicrobial efficacy and hence could be used as potential ICMs.

Associations of Gastrointestinal Tract Tumor Necrosis Factor Receptor-Associated Factor 6 Expression with Clinical Features and Prognosis of Eosinophilic Gastroenteritis.

Few studies have been conducted to explore the expression of tumor necrosis factor receptor-associated factor 6 in eosinophilic gastroenteritis patients. Therefore, the expression profile of tumor necrosis factor receptor-associated factor 6 in the gastrointestinal tract of eosinophilic gastroenteritis patients and its associations with clinical features were explored in this study. Thirty-four eosinophilic gastroenteritis patients who presented in Ruijin Hospital from December 2012 to May 2019 and had accepted gastrointestinal endoscopic examinations were recruited. Medical records and endoscopic biopsies were collected, and the prognosis was followed up by telephone. Healthy persons were selected as the control group. Hematoxylin and eosin and immunohistochemical staining were performed in both eosinophilic gastroenteritis patients and healthy persons. The final results were analyzed by skilled pathologists, and mean optical density values of tumor necrosis factor receptor-associated factor 6 were calculated by Image J software. Final results were analyzed by Statistical Package for the Social Sciences software 22.0. Thirty-four patients (mean age: 25.56 ± 21.14 years, 61.76% males) were recruited for this study. There was no significant difference in tumor necrosis factor receptor-associated factor 6 mean optical density values of gastric tissues in eosinophilic gastroenteritis patients and healthy people (0.22 ± 0.16 vs. 0.14 ± 0.05, P > .05). Eosinophilic gastroenteritis patients had a significantly lower level of intestinal tumor necrosis factor receptor-associated factor 6 mean optical density values than that of healthy people (0.16 ± 0.05 vs. 0.23 ± 0.06, P < .05). Intestinal tumor necrosis factor receptor-associated factor 6 mean optical density values negatively linearly correlated with serum interleukin-10 level (r = -0.618, P = .043 < .05). There were no differences between eosinophilic gastroenteritis patients with or without relapse regarding the expression level of intestinal tumor necrosis factor receptor-associated factor 6 (P = .227 > .05). Patients with eosinophilic gastroenteritis might have a deficiency of intestinal tumor necrosis factor receptor-associated factor 6 compared to healthy controls.

Evaluation of a mimotope of the Rickettsia outer membrane protein A (OmpA) as an antigen in enzyme-linked immunosorbent assay to detect rickettsiosis in capybaras (Hydrochoerus hydrochaeris), horses (Equus caballus), and opossums (Didelphis sp.).

Rickettsia rickettsii is the etiological agent of Rocky Mountain spotted fever, which is an important tick-borne zoonosis and, in Brazil, it causes Brazilian spotted fever, which has high lethality rate. This study aimed to evaluate a synthetic peptide corresponding to a segment of the outer membrane protein A (OmpA) as an antigen in a serological test for the diagnosis of rickettsial infections. The amino acid sequence of the peptide was selected by predicting B cell epitopes using B Cell Epitope Prediction (Immune Epitope Database and Analysis Resource) and Epitopia and OmpA sequences of Rickettsia rickettsii strain 'Brazil' and Rickettsia parkeri strains 'Maculatum 20' and 'Portsmouth'. A peptide with amino acid sequence common to both Rickettsia species was synthesized and arbitrarily named OmpA-pLMC. To evaluate this peptide in enzyme-linked immunosorbent assay (ELISA), serum samples of capybara (Hydrochoerus hydrochaeris), horse (Equus caballus), and opossum (Didelphis albiventris) that had been previously tested by indirect immunofluorescence assay (IFA) for rickettsial infection were separated into IFA-positive and IFA-negative groups and used in the assay. There were no significant differences in ELISA optical density (OD) values between IFA-positive and IFA-negative groups with horse samples. The mean OD values were significantly higher in the IFA-positive capybara serum samples (IFA-pos vs. IFA-neg = 2.389 ± 0.761 vs. 1.760 ± 0.840). However, receiver operating characteristic (ROC) curve analysis did not show significant diagnostic parameters. On the other hand, 12 out of 14 (85.7%) opossum samples of the IFA-positive group showed reactivity in ELISA, and this was significantly higher than of the IFA-negative group (0.7196 ± 0.440 vs. 0.2318 ± 0.098, respectively; 85.7% sensitivity, 100% specificity). Therefore, our results show that OmpA-pLMC has a potential to be used in immunodiagnostic assays to detect spotted fever group rickettsial infections.

Evaluation of antimicrobial efficacy of ethanolic extract of Cuminium Cyminium as intracanal medicament on common bacteria of endodontic infections: An in vitro study.

Antimicrobial intracanal medicaments play a vital role in successful outcome of any endodontic procedure. One such plant extract Cuminium cyminium, as intracanal medicaments needs to be researched. The purpose of this study was in vitro assessment of the antibacterial activity of ethanol extract of C. Cyminium in comparison to Calcium hydroxide (Ca[OH]2) as intracanal medicament against the pathogens of endodontic infection, at an interval 1 h, 24 h, 48 h, and 72 h. The study was conducted in the central research laboratory of our institute. Freshly prepared C. cyminium extract was procured from AYUSH approved laboratory and direct contact test (DCT) was utilized, which is based on turbidometric determination of microbial growth in a 96-well microplate, carrying 6 times for each bacteria. Three groups were assigned for each material in a 96 microwell plate for DCT. Bacterial growth kinetics was monitored at intervals of 1 h, 24 h, 48 h, and 72 h using spectrophotometer at 595 nm. The optical density of T2 (Test group), P2 (Positive control), and N2 (Negative control) was considered. After compiling the data, based on the normality of data, further statistical analysis was conducted using Kolmogorov-Smirnov test, Paired t-test, and pairwise comparisons by Turkey's multiple post hoc procedures. The level of statistical significance was set at P = 0.05. The comparison of mean optical density values of C. cyminium in comparison with Ca(OH)2 against the microorganisms of endodontic origin showed a statically significant decrease in bacterial viability at the end of 24 h, 48 h, and 72 h. Based on the results of the study, it can be concluded that C. cyminium has significant antibacterial action against endodontic origin, at interval of 24 h, 48 h, and 72 h.

A cohort study using IL-6/Stat3 activity and PD-1/PD-L1 expression to predict five-year survival for patients after gastric cancer resection.

The expression/activation of IL-6, p-Stat3, PD-1 and PD-L1 in gastric cancer (GC) tissues were examined to evaluate their abilities in predicting the survival prognosis in postoperative patients with GC. The clinicopathological data and paraffin-embedded tissues of 205 patients who underwent gastric cancer resection were collected at the First Affiliated Hospital of Shihezi University School of Medicine, and the patients were followed-up annually after surgery. Immunohistochemistry (IHC) was used to detect the expression of IL-6, p-Stat3, PD-1 and PD-L1 proteins using tissue microarrays derived from these patients. Statistical analyses were performed using non-parametric tests, Spearman's correlation, ROC curves, Kaplan-Meier survival analysis, Cox single-factor and multifactor regression models. In comparison, the analyses were also performed for GC patients from public databases (407 patients from TCGA and 433 patients from GEO, respectively). (1) The expression levels of IL-6, p-Stat3, PD-1 and PD-L1 in GC tissues were significantly higher than adjacent normal tissues (ANT) (81.01% vs. 52.78%, P<0.001; 100% vs. 93.41%, P<0.001; 58.58% vs. 40.12%, P<0.001; 38.20% vs. 26.90%, P = 0.025, respectively). The mean optical density (MOD) values of IL-6, p-Stat3, PD-1 and PD-L1 were significantly higher in GC tissues. (2) The higher the levels of IL-6 (P<0.001), p-Stat3 (P<0.001), and PD-L1 (P = 0.003) were, the worse the survival prognoses were observed, respectively, among GC patients. The expression of PD-1 was not correlated with the prognosis of GC patients (P>0.05). The lower the degree of cell differentiation (P<0.001) was, the worse the survival prognoses were observed among GC patients. (3) Independent risk factors for postoperative prognosis in GC patients included age (≥60 years old), poor cell differentiation, invasion depth (T3/T4), lymph node metastasis (N1-3), distant metastasis (M1), and high levels of IL-6 (2+/3+). (4) A multi-factor combination (cell differentiation+IL-6+p-Stat3+PD-1+PD-L1) appeared to be the best survival predictor for GC patients as indicated by AUC (AUC 0.782, 95% CI = 0.709, 0.856, P<0.001). This combination may be the optimal predictor for postoperative survival of GC patients. (5) The levels of IL-6, p-Stat3, PD-1 and PD-L1 correlated with the infiltration levels of various tumor-infiltrating immune cells. (6) The analyses of ROC curves, calibration, DCA and Kaplan-Meier (KM) survival curves in TCGA dataset confirmed that the nomogram model could accurately predict the prognosis in GC patients. (1) The expressed levels of IL-6, p-Stat3, PD-1 and PD-L1 are higher in GC tissues than in adjacent normal tissues. (2) The high levels of IL-6, p-Stat3 and PD-L1 are correlated with poor survival in GC patients. (3) The high levels of IL-6, p-Stat3, PD-1 and PD-L1 have influences in GC tumor microenvironment. (4) The multi-predictor combination of "IL-6+p-Stat3+PD-1+cell differentiation" serves as an optimal survival predictor for postoperative GC patients and better than the TNM staging system. As these molecules can be examined in preoperative biopsies, these observations may provide a useful guide for clinicians to strategize individualized surgical plans for GC patients before surgery.

Anti-PF4 antibodies associated with disease severity in COVID-19

Severe COVID-19 is characterized by a prothrombotic state associated with thrombocytopenia, with microvascular thrombosis being almost invariably present in the lung and other organs at postmortem examination. We evaluated the presence of antibodies to platelet factor 4 (PF4)-polyanion complexes using a clinically validated immunoassay in 100 hospitalized patients with COVID-19 with moderate or severe disease (World Health Organization score, 4 to 10), 25 patients with acute COVID-19 visiting the emergency department, and 65 convalescent individuals. Anti-PF4 antibodies were detected in 95 of 100 hospitalized patients with COVID-19 (95.0%) irrespective of prior heparin treatment, with a mean optical density value of 0.871 ± 0.405 SD (range, 0.177 to 2.706). In contrast, patients hospitalized for severe acute respiratory disease unrelated to COVID-19 had markedly lower levels of the antibodies. In a high proportion of patients with COVID-19, levels of all three immunoglobulin (Ig) isotypes tested (IgG, IgM, and IgA) were simultaneously elevated. Antibody levels were higher in male than in female patients and higher in African Americans and Hispanics than in White patients. Anti-PF4 antibody levels were correlated with the maximum disease severity score and with significant reductions in circulating platelet counts during hospitalization. In individuals convalescent from COVID-19, the antibody levels returned to near-normal values. Sera from patients with COVID-19 induced higher levels of platelet activation than did sera from healthy blood donors, but the results were not correlated with the levels of anti-PF4 antibodies. These results demonstrate that the vast majority of patients with severe COVID-19 develop anti-PF4 antibodies, which may play a role in the clinical complications of COVID-19.

Estimated specific antibody-based true sero-prevalences of canine filariosis in dogs in Central Europe and the UK

Dirofilariosis is a vector-borne disease mainly caused by Dirofilaria immitis and Dirofilaria repens. In contrast to the known endemicity of dirofilariosis in southern and south-eastern Europe, information on the distribution of D. repens in Central-Europe is fragmentary. We tested 8877 serum samples from dogs from Austria, Denmark, Germany, Italy, Lithuania, Poland, Switzerland and the UK using an ELISA detecting filarial-specific antibodies, hypothesising higher occurrence of D. repens. Based on two overlapping frequency distributions, presumed negative samples had a mean optical density (OD) value of 0.097, representing 97.45% of all samples. Presumed positive samples, representing 2.55% of all sera, had a mean OD value of 0.287. Test prevalence based on the calculated cut-off was 3.51% for all sera (4.36% for Austria, 1.94% for Denmark, 1.39% for Germany, 3.37% for Italy, 6.90% for Lithuania, 6.99% for Poland, 0.77% for Switzerland and 0.0% for the UK, respectively). The bimodal distribution, representing overlapping distributions of OD values from positive and negative dogs, enabled the assignment of a probability of true infection status to each dog. Mean probabilities of true infection status across groups, based on the postal codes of origin, allowed us to estimate and map true prevalences. For all countries, except the UK, the true prevalence was lower than the test prevalence. The large number of serum samples and the use of a non-gold standard analytical method allowed us to create a more realistic picture of the distribution of D. repens in Central Europe and the UK.

Programmable and monitorable intradermal vaccine delivery using ultrasound perforation array

Skin disruptions of micrometer- or submillimeter-diameter generated by microneedle or laser ablation can be used for intradermal vaccination. Here, we report a new skin-disruption method that uses highly-focused ultrasound to controllably perforate murine skin. We showed that programming of ultrasound parameters varied skin perforation area from 0.078 ± 0.045 mm2 to 1.295 ± 0.279 mm2. The skin perforation area increased with increasing ultrasound pressure and exposure time, and decreased with increasing ultrasound frequency and incidence angle. Moreover, successful perforation can be monitored using ultrasound pulse-echo imaging. We found that ultrasound perforation elevated local skin expression of heat shock protein 70 and effectively attracted MHC II-positive immune cells after intradermal delivery of Hepatitis B surface antigen (HBsAg). We demonstrated that the antigen dose delivered by ultrasound perforation can be effectively modulated via programmable perforation arrays (comprised of 1 × 2, 1 × 3, 1 × 5 or 3 × 3 areas of exposure). Using a 1 × 3 perforation array for vaccination, the measured mean optical density (OD) value of serum anti-HBsAg IgG was slightly higher than that of intradermal injection. The addition of an adjuvant of recombinant cholera toxin B further increased OD values of anti-HBsAg IgG. This ultrasound perforation method holds great promise for monitorable and programmable intradermal vaccination.

Study on the Correlation between the Levels of HtrA3 and TGF-β2 in Late Pregnancy and Preeclampsia.

Preeclampsia (PE) is a common and proprietary complication during pregnancy. The correlation was found between the levels of HtrA3 and TGF-β 2 and preeclampsia (PE). This study aimed to detect the HtrA3 and TGF-β2 in different parts of the third trimester (maternal serum, placenta). The 102 pregnant women who were eligible for enrollment in the obstetric examination at Tengzhou Maternity and Child Health Hospital from June 2020 to December 2020 were selected as the research objects. 28 cases diagnosed with PE were set up as the observation group 1, and 24 cases diagnosed with severe PE were set up as the observation group 2. Select 50 normal pregnant women as the control group and research the expressions of HtrA3 and TGF-β2 in maternal blood and placental tissues of patients with PE. ELISA was used to measure the concentration of HtrA3 and TGF-β2 in maternal blood. The distribution of HtrA3 and TGF-β2 in the placenta was observed by immunohistochemical techniques (IHC) and mean optical density value (MOD). S/D was measured by using color Doppler ultrasonic. The concentration of HtrA3 and TGF-β2 in the maternal blood and placenta tissue was higher in severe PE compared with PE and normotensive pregnancy, respectively (P < 0.05). There is a negative correlation between the level of HtrA3 and TGF-β2 and the birthweight of newborns both in maternal plasma and placenta tissue in preeclampsia and positive correlation between HtrA3 and TGF-β2 levels and S/D. HtrA3 and TGF-β2 may correlate with severity of PE and their neonatal adverse outcomes.

Evaluation of the In-Vitro Cytotoxicity of Heat Cure Denture Base Resin Modified with Recycled PMMA-Based Denture Base Resin.

Aim:The aim of the present study was to evaluate the in-vitro cytotoxicity of heat-cure denture base resin (PMMA) modified with recycled denture base resin at 10%, 20%, 30%, 40%, and 50% (w/w) concentration.Materials and Methods:A total of 30 disk-shaped specimens were prepared and divided into six groups (n = 5). The Control group (R0) consisted of unmodified processed denture base resin, the experimental group consisted of denture base resin processed with substitution of 10% 20%, 30%, 40%, and 50% (w/w) of recycled denture base resin (R10, R20, R30, R40, and R50). Eluates were prepared using five sterile specimens of each group. The mouse fibroblast cell line (L929) was seeded in a 96-well cell plate system at a concentration of 1 × 104 cells/well in the DMEM medium with 1× antibiotic and antimycotic solution and 10% fetal bovine serum at 37°C with 5% CO2 and incubated in a CO2 incubator for 48 h. MTT assay was applied and the absorbance was measured at 570 nm using a microplate reader to assess the in-vitro cytotoxicity. One-way analysis of variance (ANOVA) along with post hoc Scheffe test was used to statistically compare the mean optical density (OD) values and cell survival/viability % amongst the groups.Results:No statistically significant difference was observed in the mean and standard deviation of the optical density and cell viability % of the test groups that were compared.Conclusion:Modification of denture base resin using recycled PMMA does not have a cytotoxic effect on the mouse fibroblast cell line L929.

Comparison of the IOLMaster 700 and the Pentacam in the Analysis of the Lens Nuclear Density Before the Cataract Surgery.

Purpose: To evaluate the difference of the lens nuclear density measured before and after mydriasis by using the IOLMaster 700 (Carl Zeiss Meditec AG, Jena, Germany) and the Pentacam Scheimpflug imaging (Pentacam HR, Oculus Incorporation, Wetzlar, Germany) and investigate the relationship between the measurement data and the phacoemulsification parameters.Methods: Patients with age-related nuclear cataracts diagnosed on the slit-lamp examination were enrolled in the age range of 53–76 years. No patient had a history of ocular surgery, laser treatment, or general disorders affecting vision. The mean optical density (OD) was measured by the IOLMaster 700 by using the Image-Pro® Plus software before and after mydriasis. The Pentacam nucleus densitometry (PND) was obtained automatically from the Pentacam Scheimpflug imaging and compared with OD. The correlation between OD and effective phacoemulsification time (EPT), PND, and EPT were analyzed, respectively.Results: In this study, 53 eyes of 52 patients were evaluated. Before and after mydriasis, the mean OD values were 64.34 ± 23.31 and 63.81 ± 23.21 pixel units, respectively; the mean PND values were 28.51 ± 11.42 and 25.41 ± 11.31, respectively; and the mean EPT value was 6.24 ± 3.49. The Bland–Altman analysis showed that the lens nuclear densities of the two devices were highly consistent. There was no significant difference in the OD values (t = 0.455, p > 0.05) before and after mydriasis, but the difference has existed in the PND values (t = 2.509, p < 0.05). The OD and PND values were positively correlated with EPT before and after mydriasis (rOD−Before = 0.604, rOD−After = 0.593, rPND−Before = 0.701, and rPND−After = 0.891, p < 0.01).Conclusion: The combination of the IOLMaster 700 and the Image-Pro® Plus software can quantitatively evaluate the degree of the cataract lens opacification. It has good consistency with the Pentacam and is positively correlated with the phacoemulsification parameters. It is expected to become a new method to predict the phacoemulsification parameters before and during cataract surgery.

In vivo study of strontium-doped calcium phosphate cement for biological properties

OBJECTIVE To evaluate the biocompatibility and osteogenic effect of new calcium phosphate cement (CPC) in vivo and to provide experimental basis for its further clinical application. METHODS Thirty New Zealand white rabbits were randomly divided into four groups: CPC group, CPC+Bio-Oss group, Bio-Oss group and blank control group. Bone defect models of 6 mm in diameter and 7 mm in depth were made on the lateral condyle of bilateral hind legs of the rabbits. CPC, Bio-Oss and CPC+Bio-Oss mixture were implanted into the bone defect according to the group, and the mass ratio of CPC and Bio-Oss was 4 ∶ 1. The experimental animals were sacrificed the 4th, 12th and 24th week after operation. The tissue around the bone defect was taken for histological evaluation by H&E staining. Bone ingrowth fraction (BIF) was calculated. The expression of BMP-2 and COL-Ⅰ was detected by immunohis- tochemical staining by calculating the mean optical density (MOD) of the positive area the 4th week after operation, and the bone healing of each group was evaluated at different time points. The measurement data were analyzed by one-way ANOVA and LSD test was used for multiple comparison of the differences between the means by SPSS 19.0. P < 0.05 was considered to be statistically significant. RESULTS The results of H&E staining showed that the BIF values of CPC group, CPC + Bio-Oss group and Bio-Oss group were significantly higher than those of blank control group at the same time point (P < 0.01). The BIF values of CPC group were lower than those of Bio-Oss group and CPC + Bio-Oss group (P < 0.01). There was no significant difference between CPC + Bio-Oss group and Bio-Oss group. Immunohistochemical staining showed that the MOD values of BMP-2 and COL-Ⅰ in CPC group were higher than those in blank control group, but lower than those in Bio-Oss group and CPC+Bio-Oss group (P < 0.01). There was no significant difference between BMP-2 and COL-Ⅰ in CPC+Bio-Oss group and Bio-Oss group. CONCLUSION The new calcium phosphate cement has good biocompatibility and can promote early osteogenesis with stable and long-term effect.

Chronic Wasting Disease Diagnostic Discrepancies: The Importance of Testing Both Medial Retropharyngeal Lymph Nodes.

Chronic wasting disease (CWD) of white-tailed deer (Odocoileus virginianus) is a fatal neurologic disease that is spreading across North America. A common surveillance protocol for CWD currently involves screening with an enzyme-linked immunosorbent assay (ELISA) followed by confirmatory testing with immunohistochemistry (IHC). Medial retropharyngeal lymph nodes (MRPLN) are the tissue of choice to diagnose CWD in free-ranging white-tailed deer. We examined left and right MRPLN from 101 ELISA-positive deer harvested from 2015 to 2019 to determine the prevalence of cases in which prion protein was not detected by IHC as well as differences in IHC labeling between contralateral lymph nodes. Prion protein was not detected using IHC in either MRPLN in 5.9% (6/101) of cases. There was a significant but weak positive relationship between the number of IHC-positive follicles and ELISA optical density values (R2=0.08, P=0.039). Mean optical density values in IHC-positive MRPLN were higher than in IHC-negative MRPLN; however, this was not statistically significant (P=0.260). Failure to confirm ELISA diagnoses with IHC may have been because the methods tested different areas of MRPLN, or that there were differences in test sensitivity or antibody affinity. An additional 5.9% (6/101) of cases had one IHC-positive MRPLN, whereas the contralateral MRPLN was IHC negative. Therefore, testing a single MRPLN for CWD may lead to false-negative results, regardless of methodology, which highlights the importance of collecting and testing both MRPLN.

The effect of sevoflurane on the spatial recall ability and expression of apolipoprotein E and β amyloid in the hippocampus in rats

This study aimed to observe the recent spatial recall ability and the changes of expression of hippocampal apolipoprotein E (ApoE) and amyloid β protein (Aβ) in adult rats after inhaling sevoflurane anesthetic drugs, and to analyze the mechanism of action. For this purpose, a total of 54 adult SD clean-grade rats were selected in this study and were randomly divided into the sevoflurane anesthesia group, carrier gas group and control group, 18 rats in each group. The rats in the carrier gas group were inhaled with 1 L/min of oxygen O2+1 L/min air mixed carrier gas for 2 h, and the rats in the sevoflurane anesthesia group were given 3.2%sevoflurane for 2 hours based on the carrier gas group, the control rats were naturally reared. Before the model was copied, the Morris water maze experiment was performed before the material was taken. Some rat brain tissues were extracted on the first day (T1), the third day (T3), and the seventh day (T7) after model replication. The immunohistochemistry was used to measure the mean optical density (MOD) value of APOE and Aβ in hippocampal CA1, CA3 and DG regions. The indicators above at different time points of each group were compared and analyzed. Results showed that the number of crossing the original platform at each time point, the residence time of the original platform quadrant, the number of entering the original platform quadrant, and the percentage of the original platform quadrant residence time in the sevoflurane anesthesia group and the carrier gas group were compared, and there were no significant differences between two groups (P&gt;0.05). Compared with the carrier gas group, the MOD values of APOE in the hippocampus at T1 and T3 time points in the sevoflurane anesthesia group were decreased (P&lt;0.05), the MOD values of Aβ in the hippocampus at the T7 time point were increased (P&lt;0.05). It concluded that Inhalation of 3.2%sevoflurane has no obvious damage to the recent spatial recall ability of adult rats. Within 7 days after inhalation of 3.2% sevoflurane, it can inhibit hippocampus Aβ deposition through down-regulating APOE expression level. The critical time point for hippocampal Aβ increasing was 7 days after anesthesia.