Average OD Research Articles

Effect of electroacupuncture of "Jiaji" (EX-B2) on NOD-like receptor protein 3 mediated pyroptosis in spinal cord tissue of rats with acute spinal cord injury.

To observe the effect of electroacupuncture (EA) stimulation of "Jiaji"(EX-B2) on motor function, histomorphology, and expression of NOD-like receptor protein 3 (NLRP3) and N-terminal domain of gasdermin D (GSDMD-N) in the spinal cord tissue of rats with spinal cord injury (SCI), so as to explore its mechanism underlying improvement of SCI. Forty eight female SD rats were randomly divided into sham surgery (sham), SCI model (model), EA, and NLRP3 agonist (monosodium urate, MSU) combined with Jiaji EA (MSU+EA) groups, with 12 rats in each group which were further divided into 3 d and 7 d subgroups, with 6 rats at each time point. Two EA groups received EA stimulation of EX-B2 with a frequency of 100 Hz, electrical current of 1-2 mA for 30 min, once a day for 3 or 7 days. After 5 min, 6 h, and 24 h of modeling, rats of the MSU+EA group received intraperitoneal injection of MSU (200 μg/kg, 200 μg/mL) . The motor function was evaluated using Basso-Beattie-Bresnahan (BBB) scale, the morphological structure of rat spinal cord tissue was observed by H.E. staining. The expression of pyroptosis related factors NLRP3, cleaved Caspase-1 and GSDMD-N of the spinal cord was observed by using immunohistochemistry and Western blot separately, the expression and localization of Iba-1 and GSDMD-N in the spinal cord tissue were observed using immunofluorescence double staining method. Compared with the sham group, the BBB scores after modeling and on day 3 and 7 were decreased (P<0.05), while the average OD values (immunoactivity) and expression levels of NLRP3, cleaved Caspase-1 and GSDMD-N proteins, and the immunofluorescence intensity of Iba-1/GSDMD-N (co-expression) of the spinal cord tissues on day 3 and 7 were significantly increased in the model group (P<0.05). In comparison with the model group, the BBB scores on day 3 and 7 were obviously increased (P<0.05), while the immunoactivity and expression levels of NLRP3, cleaved Caspase-1 and GSDMD proteins, and the immunofluorescence intensity of Iba-1/GSDMD-N on day 3 and 7 significantly down-regulated in the EA group (P<0.05) but not in the MSU+EA group (P>0.05), suggesting an elimination of the effects of EA after administration of NLRP3 agonist (MSU). H.E. staining showed obvious bleeding area in the spinal cord tissue, loose tissue and inflammatory cell infiltration on day 3 after modeling, and basic absorption of the bleeding, loose tissue, obvious vacuolar changes of the white matter area, loss and contraction of neurons with infiltration of a large number of inflammatory cells, which was milder in the EA group but not in the MSU+EA group. EA of EX-B2 can improve the motor function of SCI rats, which may be related to its functions in inhibiting pyroptosis of microglia mediated by NLRP3/Caspase-1 signaling pathway.

Biofilm Formation From Listeria monocytogenes Isolated From Pangasius Fish-processing Plants

Biofilm formation of Listeria monocytogenes in food processing environments cause potential source of cross-contamination to foodstuffs; hence, the control of biofilm is currently addressed to find effective solutions for preventing biofilm formation or eliminating the established one. Forty-five strains of Listeria monocytogenes isolated from Pangasius fish-processing plants were studied for their capability to form a biofilm on 96-well microtiter plate by using the conventional crystal violet staining. Additionally, the inhibitory effect of biofilm formation by food additives including monascus pigment and ε-polylysine was examined. The average OD value showing biofilm mass of all 45 strains L. monocytogenes increased with an increasing temperature and time (p < 0.05). Monascus pigment and ε-polylysine significantly decreased biofilm formation by 80 ± 5.5% and 20 ± 5.9%, respectively, at the tested concentration (p < 0.05) Further, the effects of lysozyme (0.1 mg/mL) alone or in combination with slightly acidic hypochlorous water (SAHW) with 40 mg/L available chlorine or sodium hypochlorite (NaOCl) with 100 mg/L available chlorine against 7-d established biofilm of L. monocytogenes were investigated. The results indicated that slightly acidic hypochlorous water alone exhibited significant antibacterial activity (p < 0.05), decreasing the viable count by 5.2 ± 0.5 log CFU/mL. It seems that sequential treatment of lysozyme and SAHW showed an additional efficacy against biofilm of L. monocytogenes on polystyrene plate surface, reducing 70% of biomass of biofilm and 7.6 ± 0.3 log of biofilm viable cells (p < 0.05). Additionally, SAHW exhibited greater bactericidal activity against viable biofilm cells than NaOCl did. This result reveals that SAHW is a promising disinfectant agent against L. monocytogenes and the potential alternative to NaOCl in practice.

Screening and specificity study of aptamer of common proteins in colorectal cancer

Over the years, the case fatality rate of colorectal cancer is increasing. It is increasingly important to discover new tumor markers for early diagnosis of colorectal cancer. Aptamers obtained by SELEX method can be used as molecular probes to specifically bind tumor markers. Therefore, Med-SELEX was used in this study. HT29 and SW480 cell culture medium were used as positive screen, while NCM460 cell culture medium was used as reverse screen. After 14 rounds of screening, 4 candidate aptamers were selected, which were Apt-C2, Apt-C4, Apt-C15 and Apt-C27. Their affinity to HT29 cell culture medium were 9.059 ± 2.511, 10.36 ± 3.451, 6.646 ± 1.651, 8.127 ± 1.926 nmol/L, respectively. The affinity to SW480 cell culture medium was 5.324 ± 2.341, 7.854 ± 1.611, 8.239 ± 2.666, 4.164 ± 0.9671 nmol/L, respectively. Q-PCR and quantum dot method verified the high specificity of Apt-C4. Q-PCR results showed that the average Ct value of Apt-C4 binding to colorectal cancer serum was 9.921, and the average OD value of quantum dot was 49.498. Serum stability test showed that Apt-C4 did not degrade within 60h. Therefore, Apt-C4 aptamer was confirmed to have potential as a serum probe, providing a new idea for discovering new tumor markers of colorectal cancer.

Correlation analysis of serum CTRP3 and PGC-1α expression with traumatic fracture healing

To investigate the relationship between serum C1q/tumor necrosis factor-related protein-3(CTRP3) and peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α) on predictive value of expression level on fracture healing. From January 2019 to January 2020, 80 patients with traumatic tibial plateau fractures were treated by internal fixation with support plates through the posterior approach of the knee joint. The patients were followed up for 12 months. According to the criteria for delayed fracture healing, the patients were divided into two groups:54 patients in fracture healing group included 24 males and 30 females, aged 29 to 75 years old with an average of (52.36±13.17) years;In the delayed healing group, there were 26 cases, 13 males and 13 females, aged from 29 to 75 with an average od (53.82±13.52) years. The serum levels of CTRP3, PGC-1αand 25 hydroxyvitamin D3[25(OH)D3] in patients with traumatic fracture were detected by enzyme-linked immunosorbent assay(ELISA);Blood phosphorus and calcium levels were measured by automatic biochemical analyzer, and the product of calcium and phosphorus was calculated;Pearson's method was used to analyze the correlation between serum CTRP3, PGC-1αand bone biochemical indexes in patients with delayed union one week after operation;The predictive value of serum levels of CTRP3 and PGC-1αon traumatic fracture healing was analyzed by receiver operating characteristic curve(ROC curve). PGC-1α, calcium phosphorus product and 25(OH)D3 in the fracture healing group were higher than those in the delayed healing group at 1 and 4 weeks after operation(P<0.05). Serum CTRP3 was positively correlated with PGC-1α(r=0.637, P<0.05) and positively correlated with calcium phosphorus product and 25(OH)D3(P<0.05). The areas under the curve(AUC) of serum ctrp3 and PGC-1α levels in predicting traumatic fracture healing were 0.845 and 0.855, respectively. The cutoff values were 188.678 pg/ml and 2.697 ng/ml, respectively. The specificity was 96.2% and 80.8%, and the sensitivity was 53.7% and 77.8%;The predicted AUC was 0.904, the specificity was 88.5%, and the sensitivity was 81.5%. The serum levels of CTRP3 and PGC-1 in patients with delayed union of traumatic fracture at 1 and 4 weeks after operation α The expression level is of certain reference value to predict the fracture healing status of patients.

The Effects of Probiotic-Prebiotic on the Biomass and Protein Content of Spirulina platensis in Different Temperatures and Illuminations

The blue-green algae Spirulina platensis Geitler 1925 has been used by people since ancient times due to its high protein content. This study was conducted to determine the effect of a Probiotic-Prebiotic (P-P) product, which contains Bifidobacterium infantis, Bifidobacterium thermophilum, Leuconostoc mesenteroides as probiotic bacteria, and beta-glucan and oligosaccharide as a prebiotic mixture on S. platensis. In the study, four groups were one of which was a control. In this study, we investigated whether the P-P product contributed to the biomass development and protein production of S. platensis under different temperatures and lighting conditions. In terms of biomass, the P-P product gave different responses at different temperatures and light intensities, but its results were positive in all groups in terms of protein ratio. The most positive effect on biomass and protein content was in the group where 0.1 ml/L P-P product was added, at 40°C and 3200 lux with 68.71% protein content and an average OD of 0.727.

Development of n-HA/CS-GM biomimetic nanocomposite for biomedical applications

Nowadays, effective treatment and management of malignant osteomyelitis remain an alarming clinical challenge causing the creation of antimicrobial biomaterials for orthopedic surgeons. The has revived attention in creating antimicrobial biomaterials for orthopedics. The collaboration of nanotechnology and engineered biomaterials will probably provide perception for developing novel and hybrid composites. Because of improved control of the interaction between nanoparticles and polymers, nanohydroxyapatite (n-HA) incorporated nanocomposite would provide versatility in designing specific properties. As a result, the study describes the ethanolic extraction of Guar gum from native Cassia fistula seeds, as well as the development of (n-HA), Chitosan (CS), and Guar gum (GM) nanocomposite via the Co-precipitation method. The nanocomposites were characterized based on their physicochemical and morphological properties, such as XRD, FT-IR, and SEM. The nanocomposites were tested for antibacterial activity against Staphylococcus aureus(S.aureus) ATCC25923 and anticancer activity against MG 63 (osteosarcoma) cancer cell line MTT assay. The antibacterial result confirms that the n-HA/CS-GM hybrid nanocomposites exhibit excellent antibacterial properties against Staphylococcus aureus and average inhibition zones of the different content samples against S. aureus were 15.75 mm for n-HA/CS and 19.75 mm for n-HA/CS-GM hybrid microspheres, respectively. The cytotoxicity result showed that the average OD of cells treated with 7.8 to 1000 µg/mL concentration of n-HA/CS composite varied from 0.479 to 0.297 parallel to 88.70% to 55% cell viability, and the OD of n-HA/CS-GM composite varied from 0.447 to 0.273 corresponding 82.77% to 50.55% cell viability for 7.8µg/mL concentration up to 1000µg/mL.

Carbonylation induced by antibiotic and pesticide residues on casein increases its IgE binding and allergenicity

This work aimed to evaluate the effect of carbonylation induced by tetracyclines, β-lactams, fluoroquinolones, and pyrethroids in caseins of bovine origin on their immunoreactivity and allergenicity. Using a spectrophotometric method, ELISA, dot-blot, and an IgE-mediated milk allergy mouse model, we confirmed that antibiotics and pesticides at their maximum residue limit, promoted the in vitro carbonylation of caseins (among 5.0 ± 0.01 and 67.5 ± 0.70 nmol of carbonyl/mg of protein); furthermore, carbonylations greater than 19 nmol significantly increase the in vitro IgE immunoreactivity of caseins (average OD among 0.63–1.50) regarding the negative control (average OD: 0.56). On the other hand, sensitized mice exposed to oxidized caseins showed increased clinical scores (2–5), positive skin tests, and footpad swelling (0.28–0.59 mm) compared to the negative control (1–2; negative skin tests; 0.1 mm, respectively), denoting increased allergenicity. These results suggest that casein carbonylation increases their IgE immunoreactivity and allergenicity, a fact that could be explained by the resistance to the digestion promoted by carbonylation and by conformational changes in the random coil casein structure, which can expose cryptic epitopes or neoepitopes.

Antibacterial and Antibiofilm Efficacy of Pineapple Hump (Ananas comosus) on Porphyromonas gingivalis in vitro

Background: Periodontal disease is one of the most prevalent oral health problems in Indonesia that affects supporting tissues of the teeth. Porphyromonas gingivalis plays an important role in the pathogenesis of periodontal disease. Alternative therapy with natural plant extracts, including pineapple (Ananas comosus) hump extract may inhibit the growth of bacteria that cause periodontal disease. Objective: To determine the effect of pineapple hump extract on bacterial growth and adhesion of Porphyromonas gingivalis biofilms. Method: The bacterial inhibition test was performed by the agar well diffusion method, and biofilm density measurements were made using the biofilm assay method. Results: Pineapple hump extract can inhibit bacterial growth optimally at a concentration of 100%, with an average zone of inhibition of 7.3 mm. The extract at a 50% concentration can eradicate the biofilms in a 6 h incubation time with an average OD of 0.124. The Shapiro-Wilk method confirmed a normal distribution of the data. Both one-way ANOVA and post hoc test showed a significant difference between the inhibitory ability of pineapple hump extract and between concentrations to restrict formation of Porphyromonas gingivalis biofilms. Conclusion: Pineapple hump extract was able to inhibit bacterial growth and eradicate the adhesion of Porphyromonas gingivalis biofilms in vitro.

PENGARUH EDUKASI GIZI DENGAN MEDIA PUZZLE TERHADAP PENGETAHUAN DAN SIKAP TENTANG SAYUR DAN BUAH PADA ANAK SEKOLAH DASAR

Background: Consumption of vegetables and fruit among school age children is still low due to lack of knowledge and attitudes about vegetables and fruit. Nutrition education can be provided to increase knowledge and attitudes as early as possible. Nutrition education using puzzle media is expected to increase knowledge and form a positive attitude about vegetable and fruit consumption.Objective: To determine the effect of nutrition education using puzzle media towards knowledge and attitudes about vegetables and fruit in elementary school children.Methods: This was a quasi-experimental study with pretest-posttest control group design. Fourty elementary school children were divided into two groups. The treatment group received nutrition education using puzzle media and a control group received nutrition education without puzzle media. Knowledge and attitudes were assessed before and after the study based on scores. Data were analyzed using Paired T-Test and Independent Sample T-Test.Results: The average score od knowledge in the treatment group increase by 18.25 points and the control group scre increased by 12.25 points. The mean score of attitudes in the treatment group increased by 14.45 points and the control group (p=0.014) and the attitude scores between the treatment group and the control group (p=0.003)Conclusion: Nutrition education using puzzle media can increase knowledge and attitudes about vegetables and fruit in elementary school children.Keywords: Nutrition education; Puzzle media; Vegetable and fruit; Knowledge; Attitude.

A 2 Year of Review of HIT Evaluations at an Urban Medical Center with an Aim Towards Quality Improvement and the Development of Practical Testing Strategies

Introduction:Thrombocytopenia is a common occurrence among inpatients while heparin-induced thrombocytopenia (HIT) is relatively rare and diagnostic strategies can vary. Our goal was to evaluate testing habits over a two year period at an urban tertiary care center.Methods:All cases with HIT panels over a two year period were reviewed to elucidate ordering patterns, test turnaround times, and areas for potential improvement. Sensitivities and specificities were determined for the 4T score, which was retrospectively calculated for all patients, the heparin/PF4 ab immunoassay (H/PF4), the SRA functional assay (SRA) and a combined low risk 4T and H/PF4 OD ratio less than 0.4. Average and median OD ratios were calculated for different levels of risk as determined by 4 T score. Given the lower risk of HIT in patients exposed to low molecular weight heparin (LWMH) (Martel et al. 2005) an exploratory analysis was made by which one point was deducted from the 4T for patients who were only exposed to LMWH during the HIT work-up admission.Results:245 patients received work-ups for HIT that included either a H/PF4 and/or an SRA, with 7 patients ultimately being determined to have HIT (yield of 2.86%). Workup issues included 28 incorrect tests being sent and a lack of 4T score documentation in the charts reviewed (15% with documentation, 38/245). Given that hematology consults were obtained in 60% (147/245) of the cases this was not a problem specific to primary inpatient teams.The sensitivities (SN) and specificities (SP) were as follows: for a low risk 4T score (SN 100%, SP 61.76%), for a low risk 4T after subtracting one point for sole exposure to LMWH (SN 100%, SP 63.03%), for the H/PF4 using Weak Positive as negative (SN 100%, SP 74.84%) for an OD<0.4 as negative (SN 100%, SP 79.05%), for the combined low risk 4T and OD<0.4 (SN 100%, SP 84.62%), for the SRA using Indeterminate as negative (SN 83.33%, SP 100%). Mean and median OD ratios by 4T risk category can be seen in the accompanying table and by score in the accompanying graph.Our calculated turnaround times for these tests were similar; 156 H/PF4 tests ordered for 146 patients took an average of 3.01 days to result, 229 SRA tests ordered for 212 patients took an average of 3.21 days to result. For the 55 HIT negative patients that ultimately received treatment the average length of therapy for those treated with argatroban was 5.11 days and for fondaparinux 7.11 days.Discussion:In an effort to improve the 4T score subtraction of 1 point for exposure to only LMWH was investigated with minimal improvement in specificity. However, 19 of the 23 patients in this category had a 4T score of four or less and this adjustment could have helped to avoid a sizable amount of subsequent testing (18 SRA's), imaging (5 studies) and alternate therapies (9 days total) if it moved these patients from intermediate to low risk.As is true in many hospitals, H/PF4's and SRA's are both send out tests in our institution and thus often ordered simultaneously. High turnaround times are also present in institutions where these tests are “batched” and only performed on certain days. With the small difference in test turnaround times it is worth questioning whether both tests are necessary to send concurrently in every case.One setting to evaluate simultaneous testing is in patients who are low risk by 4T and thus not switched from heparin to alternative agents. In these cases it is practical to not order any testing given the sensitivity of a low risk 4 T score. But when testing is opted for ordering only the H/PF4 and reserving the SRA for cases with OD ratios greater than 0.4 may be prudent. This is supported by the combined low risk 4T H/PF4 testing characteristics (Kim et al 2011). This strategy is already recommended but in institutions with long turnaround times simultaneous testing is much more common.In our cohort, 70 patients with low risk 4T scores had SRA's sent. Of these 70 patients, none of whom had HIT, 57 (81.4%) had H/PF4 ab OD ratios less than 0.40 and would not have needed additional testing. 57 SRA's could have been avoided and given that heparin can almost always be safely continued in these patients the potential costs of continuing alternate therapy are virtually non-existent.Conclusion:The low-cost, highly sensitive 4T remains a vital screening tool for HIT and while a low risk score should obviate the need for further testing our data supports that in cases where more testing is pursued a H/PF4 alone is the correct option. DisclosuresNo relevant conflicts of interest to declare.

A Novel Multimodal Next Generation Sequencing Assay with Total Nucleic Acid Input to Provide Comprehensive Genomic Profiling for Hematologic Malignancies

Background: A cost effective and comprehensive genomic profiling (CGP) approach for diagnosis, risk stratification and therapy would be useful for the evaluation of oncologic specimens. Available approaches involving additive testing for DNA and RNA abnormalities through traditional methods (e.g. Sanger, FISH, cytogenetics, qRT-PCR) are not comprehensive, require multiple different workflows and are sample consuming, often resulting in incomplete testing. While there are next generation sequencing (NGS) assays designed for detecting DNA and RNA abnormalities, they have separate workflows that require twice the amount of sample and effort. To address this, we developed a novel total nucleic acid (TNA) extraction method and single tube workflow utilizing TNA and a custom multimodal chemistry designed for hematologic malignancies. This consolidated workflow enables an efficient discovery based approach for both DNA/RNA abnormalities including single nucleotide variants (SNVs), InDels, copy number variants (CNVs), large structural changes from DNA and gene fusions and gene expression levels from RNA. This method maximizes data derived from valuable samples while delivering a comprehensive profile of the patient's tumor which can help guide therapeutic and clinical decisions. Methods: Total nucleic acid (TNA) was extracted from bone marrow and peripheral blood of 95 patients (CML, CMML, CLL, AML and myeloid disorders). 297 genes that have DNA mutations specific to hematological cancers were targeted, along with 213 genes that were targeted for clinically significant RNA abnormalities. Enriched genomic and transcriptomic regions of interest from 85 patients were successfully sequenced with unique dual indices on an Illumina NovaSeq 6000. DNA variant detection as well as fusion detection from RNA were compared to traditional orthogonal NGS assays that use DNA input or compared to qRT-PCR and Sanger sequencing assays that use RNA as input. Results: In this study, we developed an efficient and high-quality TNA extraction method that can purify enough total nucleic acid from bone marrow, peripheral blood, cytogenetic pellets, flow suspension, and FFPE samples for the downstream NGS assay. The average OD 260/280 value was 1.9 and the OD 260/230 was 2.18. After sequencing, 256/262 (97.7% accuracy) SNV and Indel variants that were candidate pathogenic mutations were concordant from 38 patients. Meanwhile, 100% (7/7) of all BCR/ABL1 gene fusions which had an international scale (IS) value above 6.4% were concordant. In addition, 69 fusion positive samples containing 20 unique gene fusions which had been previously reported by an independent ArcherDX assay designed specifically for gene fusions were also evaluated with this chemistry. Analysis revealed a 92.5% (64/69) concordance. More importantly, the QIAseq multimodal TNA NGS assay detected both DNA and RNA abnormalities in a single tube. For example, in one myeloid leukemia patient, we not only identified pathogenic variants of ASXL1 and JAK2 which had been previously detected by a DNA NGS assay, but also detected a concurrent BCR-FGFR1 fusion which had been previously reported by a FISH assay. Moreover, we were able to provide more comprehensive genomic profiling by investigating many DNA and RNA abnormalities simultaneously. In our study, for 5 patients that previously been tested for BCR-ABL1 fusion only, we are able to assess BCR-ABL1 fusion status from RNA as well as identify pathogenic DNA variants at the same time, including JAK2 p.V617F, U2AF1 p.S34F, ASXL1 p.E635Rfs*15, BRCA p.S1982Rfs*22, and DNMT3A p.S708Vfs*71, which provides valuable information to assist diagnosis and treatment in a cost effective and efficient way. Conclusions: We developed a single tube TNA based workflow with a custom multimodal chemistry that simultaneously detects many DNA and RNA abnormalities in a cost effective and efficient way while reducing sample requirements. This unique TNA NGS assay provides comprehensive genomic profiling for hematologic malignancies and improves the diagnostic testing options for precise patient care. Disclosures Yu: NeoGenomics: Current Employment. Alarcon:NeoGenomics: Current Employment. Mou:NeoGenomics: Current Employment. Jung:NeoGenomics: Current Employment. Nam:NeoGenomics: Current Employment. Thomas:NeoGenomics: Current Employment. Keeler:NeoGenomics: Current Employment. Shinbrot:NeoGenomics: Current Employment. Magnan:NeoGenomics: Current Employment. Bender:NeoGenomics: Current Employment. Jiang:NeoGenomics: Current Employment. Agersborg:NeoGenomics: Current Employment. Weiss:Bayer: Other: speaker; Genentech: Other: Speaker; Merck: Other: Speaker; NeoGenomics: Current Employment. Ye:NeoGenomics: Current Employment. Funari:NeoGenomics: Current Employment.

Optimization of Culture Conditions for Amoxicillin Degrading Bacteria Screened from Pig Manure.

(1) Objective: The objective of this study was to screen amoxicillin (AMX)-degrading bacterial strains in pig manure and optimize the fermentation conditions for these strains to achieve high fermentation rate, which can provide an effective way for the practical application of bacterial strains as antibiotic-degrading bacterial in treating livestock waste for antibiotic residues. (2) Methods: Antibiotic susceptibility tests and high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) were employed to screen AMX-degrading bacterial strains in pig manure. The culture conditions were optimized for AMX-degrading bacterial strains using Plackeet–Burman design (PBD), the steepest ascent design, and the response surface methods, coupled with the Box–Behnken design (BBD). The effects of culture time, temperature, rotator (mixing) speed, inoculum level, and initial pH value on the growth of AMX-degrading strains were investigated. Experimental data obtained from BBD were utilized to generate a second-order polynomial regression model for evaluating the effects of the tested variables on the optical density at 600 nm (OD600) of culture solutions as the growth indicator for the screened AMX-degrading strains. (3) Results: The initial pH, culture time, and the inoculum level had significant effects on the OD600 value (growth) of the screened AMX-degrading strains. The initial pH value was found to be the most critical factor influencing the growth of bacteria. The optimized culture condition for the bacterial growth determined by the response surface methodology was: the initial pH of 6.9, culture time of 52 h, and inoculum level of 2%. The average OD value of 12 different fermentation conditions in the initial fermentation tests in this study was 1.72 and the optimization resulted in an OD value of 3.00. The verification experiment resulted in an OD value of 2.94, which confirmed the adequacy of the optimization model for the determining the optimal culture condition. (4) Conclusions: The growth of the screened strain of AMX-degrading bacteria could be optimized by changing the fermentation conditions. The optimization could be achieved by using the Box–Behnken response surface method and Plackett–Burman experimental design.

Automatic localization of optic disk based on texture orientation voting

Automatic localization of optic disk is a challenging problem due to the interference of multiple factors. In this paper, we aim at the study of the optic disk localization method and propose a simple and efficient method based on texture orientation voting. This method uses multi-directional Gabor filters to detect the texture orientations of fundus images. Before orientation voting, texture orientation extraction and orientation confidence computation are conducted. Only the texture orientations with high confidence could participate in orientation voting. If the texture orientation of a voter pixel points at a receiver pixel, the receiver will get a vote from the voter, which is weighted according to the positional relationship of the voter and the receiver. The pixel with the maximum vote over the image will be taken as the optic disk location. In a variety of difficult environments, an average OD detection accuracy of 96.8% is obtained and the experimental results verify the efficiency and robustness of our proposed method.

The Effects of Intense Pulsed Light on Tear Osmolarity in Dry Eye Disease

Purpose: In this prospective study we show the influence of Intense Pulsed Light Therapy (IPL) on tear osmolarity, an increasingly important metric of dry eye disease. Previous studies have measured the effectiveness IPL has had on other metrics including tear break up time (TBUT), lipid layer grade (LLG), tear evaporation rate (TER), tear meniscus height (TMH), and subjective responses from patients.Methods: Single center prospective study included 16 patients and 32 eyes. Patient ages ranged from 18 to 90 years old with 75% of participants being female. All patients had an at least one eye with a tear osmolarity of 308 mOsm/L or greater, or had an inter-eye difference in tear osmolarity of 11 mOsm/L or greater. Tear osmolarity was measured bilaterally before a single IPL treatment followed by one drop of topical NSAID. Bilateral tear osmolarity was then measured again one month later.Results: Average tear osmolarity pre-treatment OD was 309 mOsm/L and OS was 306.75 mOsm/L. Average tear osmolarity post-treatment was OD 296.75 mOsm/L and OS 296.06 mOsm/L. Paired t tests were performed showing the change in tear osmolarity to be statistically significant in each eye, OD p=0.0029 and OS p=0.0011. Average inter-eye difference in osmolarity went from 11.81 mOsm/L to 6.81 mOsm/L, statistically significant with p=0.0148.Conclusions: A statistically significant decrease in tear osmolarity from abnormal to normal range as well as a reduction in inter-eye difference demonstrates an improvement in tear film quality after one treatment with IPL, thus suggesting IPL's effectiveness at treating dry eye disease.

Seroepidemiological study of Herpes simplex virus 2 in human immuno deficiency virus-positive individuals attending a tertiary care hospital of South India

Background: Herpes Simplex Virus-2 (HSV-2) infection is widely studied and associated disease in human immuno deficiency virus-positive (HIV-positive) individuals. The diagnosis of HSV is difficult as the presentations are atypical. Studies have confirmed that HSV-2 infection increases the risk of HIV acquisition by 2-3 folds. The aim of the study was to estimate the seroprevalence of HSV-2 and its association with age, sex and behavioral characters among HIV-infected individuals attending a tertiary care hospital.Methods: Serum samples from 150 HIV positive patients were collected and screened for HSV-2 Ig G antibodies by indirect ELISA. The cut off index (COI) of each serum sample was determined by dividing the OD which was obtained for that serum sample, by the average OD of the cutoff serum. A COI below or equal to 1 was considered as negative and that above 1.1 was considered as positive.Results: The overall seroprevalence of HSV-2 in the study was 44% (95% CI: 36-52%) with more in men than women. Out of 92 men, 42 (45.65%) were HSV positive and 24 (41.38%) of 58 female were positive. Statistical significance was observed in participants who had past history of sexually transmitted disease and who had multiple sexual partners (≥11) making as risk factors for HSV-2 seropositivity. Genital herpes was reported in 41 of 150 participants and seropositivity was 51.22% indicating the lack of awareness and most of the infections are asymptomatic.Conclusions: Our study demonstrated a clear increase in seroprevalence of HSV-2 in HIV-infected individuals. Findings of our study have relevant public implications and strongly suggest the implementation of interventions in sexually transmitted infections to reduce the prevalence of HSV-2 and associated HIV transmission. As most of the infections of HSV are asymptomatic a continuous program for the screening of HSV in all cases of STD’s must be made to reduce the chances of acquisition of HIV and others.

Visual disturbances in familial forms of multiple sclerosis

Background: According to various studies, with the familial form of the disease opening symptoms are visual, oculomotor and sensory disorders, concordance with respect to the type of the disease.Objective: To explore the visual disturbances in the case of a familial form of MS.Materials and methods: Sister (38 years) with a history of ON, the first manifestation of 22 years of age — ataxia and nystagmus. Brother (27 years) the first manifestation of optic neuritis OD.Methods: RC-5000 «Tomey», OCT on Cirrus HD-OCT; HRT on Humphrey HFA II-750i on program 10-2; Microperimetry on MP-1, MRI of the brain and orbits (Siemens Magnetom Symphony 1.5 Tl).The results: The sister was significant damage to the central nervous system with the significant visual and neurological deficit.Diagnosis: relapsing–remitting MS, cerebrospinal form, optic nerve atrophy. Visus OU 0.01 n|c. OCT — Average RNFL Thickness OD 66 μm, OS 65 μm.Average GCL + IPL thickness OD 53 μm, OS 55 μm. Brother-Diagnosis: MS cerebrospinal form, optic neuritis. Visus 0,1 with corr. According Microperimetry mean Sensitivity OD 5,3 Db, mean Defect — 10,1 Db. OCT Average RNFL Thickness 91 μm, Average GCL + IPL Thickness 83 μm. MRI revealed multiple foci of demyelination and one active focus 8 mm in diameter in the projection of the right optic nerve.Conclusion: At the first manifestation of optic neuritis and family history of MS need to consult a neurologist, brain MRI and orbits, genetic examination. Background: According to various studies, with the familial form of the disease opening symptoms are visual, oculomotor and sensory disorders, concordance with respect to the type of the disease. Objective: To explore the visual disturbances in the case of a familial form of MS. Materials and methods: Sister (38 years) with a history of ON, the first manifestation of 22 years of age — ataxia and nystagmus. Brother (27 years) the first manifestation of optic neuritis OD. Methods: RC-5000 «Tomey», OCT on Cirrus HD-OCT; HRT on Humphrey HFA II-750i on program 10-2; Microperimetry on MP-1, MRI of the brain and orbits (Siemens Magnetom Symphony 1.5 Tl). The results: The sister was significant damage to the central nervous system with the significant visual and neurological deficit. Diagnosis: relapsing–remitting MS, cerebrospinal form, optic nerve atrophy. Visus OU 0.01 n|c. OCT — Average RNFL Thickness OD 66 μm, OS 65 μm. Average GCL + IPL thickness OD 53 μm, OS 55 μm. Brother-Diagnosis: MS cerebrospinal form, optic neuritis. Visus 0,1 with corr. According Microperimetry mean Sensitivity OD 5,3 Db, mean Defect — 10,1 Db. OCT Average RNFL Thickness 91 μm, Average GCL + IPL Thickness 83 μm. MRI revealed multiple foci of demyelination and one active focus 8 mm in diameter in the projection of the right optic nerve. Conclusion: At the first manifestation of optic neuritis and family history of MS need to consult a neurologist, brain MRI and orbits, genetic examination.

Constitutive expression of the sRNA GadY decreases acetate production and improves E. coli growth.

BackgroundEscherichia coli responds to acid stress by applying various physiological, metabolic, and proton-consuming mechanisms depending on the growth media composition, cell density, growth phase, pH, and aerobic or anaerobic growth conditions. It was reported that at mild acidic conditions (pH 5.8), the Hfq-associated sRNA GadY is activated. It was also reported that the two decarboxylase systems—the lysine decarboxylase system (LDS) and the glutamate decarboxylase system (GDS)—are activated to maintain intracellular balance of protons. The purpose of this study was to establish the role of GadY in high density growth of E. coli and to evaluate the possibility of using this small RNA to create an acid-resistant strain suitable for industrial applications.ResultsParental E. coli K-12 and constitutively expressing GadY strains were grown to high cell densities in a bioreactor at pH 7.0 and pH 6.0. At pH 7.0, both strains grew to similar cell densities of 43 OD, but the constitutively expressing GadY strain produced around 6 g/L acetate compared with 10 g/L by the parental strain. At pH 6.0, the parental strain grew to an OD of 20 and produced 10 g/L of acetate while the GadY strain grew to an average OD of 31 and produced 4 g/L acetate. After analyzing 17 genes associated with acid stress, it was found that at pH 7.0 LDS was expressed in the early exponential phase and GDS was expressed in the late exponential phase in both strains. However, at pH.6.0, GDS was expressed in the late exponential phase only in the parental strain and not in the constitutively expressing GadY strain, while there was no difference in the LDS expression pattern; it was expressed in the early exponential phase in both strains. This indicates that GadY affects GDS expression at low pH since the GDS was not detected in the GadY strain at pH 6.0.ConclusionsThe constitutive expression of GadY improves E. coli growth at pH 6.0 by deactivating the expression of the GDS in the late exponential growth phase. The expression of GadY also decreases acetate production regardless of pH, which decreases the inhibitory effect of this acid on bacterial growth.

In vitro and in vivo biocompatibility evaluation of dental zirconia ceramic

Recently, zirconia ceramics have been extensively used as the artificial tooth in modern dentistry on account of their excellent mechanical properties and good esthetics. The purpose of this study was to evaluate biocompatibility of zirconia ceramics using agar diffusion test, acute systemic toxicity, intracutaneous test and local lymph node assay(LLNA). For in vitro test, the agar diffusion assay was used sintered zirconia specimens (In-ceram YZ-55, VITA Co., Bad sackingen, Germany) were performed according to ISO 7405. For in vivo test, three different tests were carried out and samples were prepared according to ISO 10993-12 for all of the test. Acute systemic toxicity test and Intracutaneous test, were performed according to ISO 10993-11 and ISO 10993-10. The murine local lymph node assay test was performed according to OECD guide line 442 B. The overall criterion for the cytotoxicity of zirconia sample was "0". For the acute systemic toxicity test, no symptoms were observed during the visual observation of the animal. For the intracutaneous irritation test, there was no redness and swellings on the reaction sites. The result of LLNA test, the samples extractions in polar solvents showed average OD value of 1.078 and those in non-polar solvents showed average OD value of 1.059. The partially stabilized zirconia materials were proved to be biocompatible. The results of both in vitro and in vivo test concluded that the zirconia materials were biocompatible.

Seroprevalence of enteropathogenic Yersinia spp. in pig batches at slaughter

Enteropathogenic Yersinia spp. are one of the main causes of foodborne bacterial infections in Europe. Slaughter pigs are the main reservoir and carcasses are contaminated during a sub-optimal hygienically slaughtering-process. Serology is potentially an easy option to test for the Yersinia-status of the pig (batches) before slaughter. A study of the variation in activity values (OD%) of Yersinia spp. in pigs and pig batches when applying a serological test were therefore conducted.In this study, pieces of the diaphragm of 7047 pigs, originating from 100 farms, were collected and meat juice was gathered, where after an enzyme-linked immunosorbent assay (ELISA) Pigtype Yopscreen (Labor Diagnostik Leipzig, Qiagen, Leipzig, Germany) was performed. The results were defined positive if the activity values exceeded the proposed cut-off value of 30 OD%. Results at pig level displayed a bimodal-shaped distribution with modes at 0–10% (n=879) and 50–60% (n=667). The average OD% was 51% and 66% of the animals tested positive. The within-batch seroprevalence ranged from 0 to 100% and also showed a bimodal distribution with modes at 0% (n=7) and 85–90% (n=16). On 7 farms, no single seropositive animal was present and in 22 farms, the mean OD% was below 30%.Based on the results obtained at slaughter, 66% of the pigs had contact with enteropathogenic Yersinia spp. at farm level. The latter occurred in at least 93% of the farms indicating that most farms are harboring enteropathogenic Yersinia spp.

Metabolism of Reactive Oxygen Species Involved in Increasing Root Vigour of Cotton Seedlings by Soaking Seeds with Mepiquat Chloride

The plant growth regulator, mepiquat chloride(1,1-dimethylpiperidinium chloride, DPC) has been used worldwide to suppress excessive growth in cotton plants. It also increases the root vigour of cotton plants. To reveal the possible role of reactive oxygen species(ROS) involved, we conducted a experiment to investigate the effect of soaking seed with 200 mg L–1 mepiquat chloride(DPC) on root vigour of cotton(Gossypium hirsutum L.) seedlings by using a cotton cv. Guoxin 3. The results showed that soaking seed with DPC could significantly increase the root vigour of cotton seedlings with two expanded cotyledons. Root viability estimated by 2,3,5-triphenyl tetrazolium chloride(TTC) reduction and root respiratory rate were increased by 167% and 90% in DPC treatment. The average OD of in situ TTC staining in DPC treated root tips was 0.3-fold higher than that of control. At 300 μm from the root apex away, DPC treatment resulted in a 36% increase of net K+ influx. The result of acridine orange staining suggested that seedlings treated with DPC had less apoptotic cells in root elongation zone compared with control. In addition, DPC treatment significantly altered the activities of antioxidant enzyme and ROS accumulation in roots of cotton seedlings. Catalase(CAT), ascorbate peroxidase(APX) and glutathione reductase(GR) were enhanced, but superoxide dismutase(SOD) was reduced, which therefore resulted in 56% less H2O2 accumulation and lighter diaminobenzidine staining(H2O2 histochemical detection in situ) in roots compared with control. Another type of ROS, O2-, its production rate also decreased by 65% in roots treated with DPC. Taken together, we speculated that the increased root vigour of cotton seedlings by soaking seed with DPC may be partially explained by improvement of ROS metabolism.