Mean Optical Density Research Articles

High-Density Integration of Ultrabright OLEDs on a Miniaturized Needle-Shaped CMOS Backplane.

Direct deposition of organic light-emitting diodes (OLEDs) on silicon-based complementary metal-oxide-semiconductor (CMOS) chips has enabled self-emissive microdisplays with high resolution and fill-factor. Emerging applications of OLEDs in augmented and virtual reality (AR/VR) displays and in biomedical applications, e.g., as brain implants for cell-specific light delivery in optogenetics, require light intensities orders of magnitude above those found in traditional displays. Further requirements often include a microscopic device footprint, a specific shape and ultrastable passivation, e.g., to ensure biocompatibility and minimal invasiveness of OLED-based implants. In this work, up to 1024 ultrabright, microscopic OLEDs are deposited directly on needle-shaped CMOS chips. Transmission electron microscopy and energy-dispersive X-ray spectroscopy are performed on the foundry-provided aluminum contact pads of the CMOS chips to guide a systematic optimization of the contacts. Plasma treatment and implementation of silver interlayers lead to ohmic contact conditions and thus facilitate direct vacuum deposition of orange- and blue-emitting OLED stacks leading to micrometer-sized pixels on the chips. The electronics in each needle allow each pixel to switch individually. The OLED pixels generate a mean optical power density of 0.25mWmm-2, corresponding to >40000cdm-2, well above the requirement for daylight AR applications and optogenetic single-unit activation in the brain.

Methodological Principles of Nasal Food Challenge.

Thanks to their valuable assessment possibilities (subjective complaints and changes in nasal patency during the examination), nasal provocation tests may serve as an alternative tool for oral food challenges in the future. However, this test requires successive attempts to regulate its methodology in order to develop a standardized lyophilisate form and determine the threshold dose for a positive result. The study objective was to present the methodological foundation for nasal food allergen provocation tests induced by freeze-dried powdered chicken egg whites. A control group of 25 individuals with no history of allergy to chicken eggs or any other allergy was included in the study. Optical rhinometry and visual analog scales were used to assess the response of nasal mucosa to local allergen challenges. Minor variations in nasal flows, as measured by optical rhinometry, were observed in the provocation tests. The mean optical density measurements (as measured regardless of the allergen dose used) varied from positive to negative values and vice versa, e.g., amounting to 0.018 OD (standard deviation 0.095) at 15 min and -0.011 OD (standard deviation 0.090) at 30 min. No significant differences were observed concerning the perceived nasal discomfort using the visual analog scale. Due to the absence of nasal mucosal reactivity, nasal challenge is an excellent methodological tool for implementing food allergen tests.

Expression of early growth responsive gene-1 in the lateral geniculate body of kittens with amblyopia caused by monocular form deprivation.

The expression of early growth responsive gene-1 (Egr-1) in the lateral geniculate body in the normal kittens and those affected with amblyopia caused by monocular visual deprivation was compared to explore the potential significance of Egr-1 in the pathogenesis of amblyopia. A total of 30 healthy kittens were equally and randomly divided into the control (n = 15) and the deprivation group (n = 15). The kittens were raised in natural light and the right eyes of the deprived kittens were covered with a black opaque covering. Pattern visual evoked potential (PVEP) was measured before and 1, 3, and 5 weeks after covering. Five kittens from each group were randomly selected and euthanized with 2% sodium pentobarbital (100 mg/kg) during the 1st, 3rd and 5th week after covering. The expression of Egr-1 in the lateral geniculate body in the two groups was compared by performing immunohistochemistry and in situ hybridization. After three weeks of covering, PVEP detection indicated that the P100 wave latency in the deprivation group was significantly higher than that in the control group (P < 0.05), whereas the amplitude decreased markedly (P < 0.05). The number of the positive cells (P < 0.05) and mean optical density (P < 0.05) of Egr-1 protein expression in the lateral geniculate body of the deprivation group were found to be substantially lower in comparison to the normal group, as well as the number (P < 0.05) and mean optical density of Egr-1 mRNA-positive cells (P < 0.05). However, with increase of age, positive expression of Egr-1 in the control group showed an upward trend (P < 0.05), but this trend was not noted in the deprivation group (P > 0.05). Monocular form deprivation can lead to substantially decreased expressions of Egr-1 protein and mRNA in the lateral geniculate body, which in turn can affect the normal expression of neuronal functions in the lateral geniculate body, thereby promoting the occurrence and development of amblyopia.

Comparison of human immunodeficiency virus and syphilis results in pooled sera versus individual samples of blood donors attending suez canal university hospital

Egypt is one of the countries where sexually transmitted diseases like human immunodeficiency virus (HIV) and syphilis are least prevalent. HIV and syphilis count less than one percent of total Egyptian population. An ELISA protocol for pooling serum samples is simple and may provide a way to reduce the cost and time needed for analysis. This study aimed to investigate the applicability and reliability of testing pooled sera of blood donors for HIV and syphilis compared to testing their individual sera and to assess the cost-effectiveness of this procedure. The study included 75 sera from randomly selected blood donors attending Suez Canal University hospital. Sera were screened by two ELISA kits, HIV Ag-Ab ELISA kit, and syphilis total antibody ELISA kit. Screening protocols were done by two sequential steps. At first, samples in pools of five were screened for both HIV and syphilis then, samples in positive pools were individually retested. There was no significant difference between the mean optical density for samples tested HIV and syphilis positive either individually or in pooled sera. There was no difference between the number of individual sera, tested positive for both HIV and syphilis and their pooled sera results (100 % positivity). There was significant decrease of the mean cost in one pool of 5 samples (16.5 L. E) in comparison to 5 individual samples (82.5 L. E) by HIV ELISA. Also, there was significant decrease of the mean cost in one pool of 5 samples (16 L.E) in comparison to 5 individual samples (80 L.E) by syphilis ELISA. In conclusion, the studied pooling protocol appeared reliable and can save up to 80 % of the cost for testing either HIV or syphilis by regular procedures.

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.

Thyroxine restores hippocampal neurogenesis and synaptogenesis in a male rat model of carbimazole-induced hypothyroidism: a histological study

BackgroundAdult-onset hypothyroidism has a deleterious effect on hippocampal cognitive and memory functions. This study was performed to evaluate the possible therapeutic effect of thyroxine on hippocampus degeneration in an adult male rat model of carbimazole-induced hypothyroidism and the potentiality of spontaneous recovery. Thirty-two adult male albino rats were divided equally into four groups, as follows: I (control group), II (hypothyroidism group) received carbimazole (20 mg/kg) orally once daily for 4 weeks; III (recovery group) rats were managed as in group II, then left untreated for an additional 4 weeks to assess spontaneous recovery; and IV (thyroxine-treated group): hypothyroidism was induced as in group II, then rats received levothyroxine (20 µg/kg/day) orally for 4 weeks. Rats and their corresponding controls were sacrificed after 4 weeks in group II and after 8 weeks in groups III and IV. The levels of T3, T4, and TSH were measured. Hematoxylin and Eosin staining of thyroid and hippocampal sections was performed. Additionally, toluidine blue staining and immunohistochemical staining for PCNA, GFAP, and synaptophysin were applied to hippocampus sections. Both morphometric measurements and statistical analysis were performed.ResultsComparison of thyroxine-treated group with hypothyroidism and recovery groups revealed a significant reduction in TSH level and an increase in T3 and T4 levels, as well as improved histological architecture in both the thyroid and hippocampal sections. Hippocampal sections revealed a significant decrease in the mean area percent of GFAP, a significant increase in the mean number of PCNA-positive cells in the subgranular zone (SGZ); a niche for the adult neural stem cells (NSCs) in the hippocampus; and a significant increase in the mean area percent as well as the mean optical density of synaptophysin.ConclusionHippocampal degeneration is induced by hypothyroidism and can be restored by thyroxine replacement therapy, probably through neuronal cell preservation, synaptogenesis, and stimulation of neurogenesis in SGZ. On the other hand, spontaneous recovery from this degeneration was inadequate.

Expression and significance of cyclinD1, cyclin-dependent kinase4 and cyclin-dependent kinase inhibitor P27 in patients with non-neoplastic epithelial disorders of the vulva.

Non-neoplastic epithelial disorders of the vulva (NNEDV) are prevalent and refractory gynecological diseases. However, the underlying pathogenesis of these diseases remain unclear. The present study aimed to investigate the expression and significance of cyclin D1, cyclin-dependent kinase 4 (CDK4) and cyclin-dependent kinase inhibitor P27 (P27) in patients with NNEDV and provide a reference for clinical diagnosis and treatment. Normal vulvar skin samples from patients with perineum repair (control group, n=20) and skin samples from the vulvar lesions of patients with NNEDV (NNEDV group, n=36) were collected. Expression levels of cyclin D1, CDK4 and P27 were assessed in the samples using immunohistochemistry. The expression of each protein was evaluated based on the mean optical density (MOD). The MODs of cyclin D1 and CDK4 were significantly higher in samples of the three pathological types of NNEDV, namely squamous hyperplasia (SH), lichen sclerosus (LS) and mixed SH and LS lesions, compared with those of the control group. The MOD of P27 was lower in samples of the three pathological types of NNEDV than in the control group, although the difference was not statistically significant. No significant differences in the MOD of cyclin D1, CDK4 and P27 were detected among the three pathological types of NNEDV. The ratios of the MOD of cyclin D1 and CDK4 in the prickle cell layer to those in the basal cell layer were significantly higher in the NNEDV group than in the control group. However, the ratio of the MOD of P27 in the prickle cell layer to that in the basal cell layer exhibited no significant difference between the NNEDV and control groups. NNEDV has the potential for malignant transformation. The occurrence and development of NNEDV may be associated with the acceleration of cell proliferation, in which cyclin D1, CDK4 and P27 contribute to regulation of the cell cycle. Therefore, cyclin D1, CDK4 and P27 may be potential targets in the development of new clinical therapeutic drugs for patients with NNEDV.

The cytotoxic effect of cysteamine and its combinations with various endodontic intracanal medications on fibroblast cells: in vitro study

BackgroundThis study was established to assess cysteamine’s cytotoxic effect alone and in combination with various intracanal medications on fibroblast cells, because the biocompatibility of intracanal medication is considered one of the main factors that affect the selection of specific medication for usage near vital periodontal tissues.MethodsAll tested medications were prepared in a solution form. Cysteamine preparation was prepared at 200 mg/ml concentration in distilled water. The chlorhexidine–cysteamine combination was prepared by dissolving 10 mg/ml of cysteamine in chlorhexidine. Calcium hydroxide–cysteamine combination was prepared by dissolving 10 mg/mL of cysteamine in a saturated solution of calcium hydroxide (CaOH). Triple antibiotic paste (TAP)–cysteamine combination was prepared by dissolving 10 mg/mL of cysteamine in triple antibiotic paste (TAP). BHK cells were seeded in well-microtiter plates. The testing materials were filtrated using a 0.22 μm syringe filter. BHK-21 cells precultured well plates were treated with descending 12-fold serially diluted medications at 37 °C for 24 h. Residual living cells were treated with 25 μl of MTT dye. MTT was discarded, and then, dimethyl sulfoxide was added as 50 μl/well. The absorbance was conducted at 570 nm. The mean optical density and 50% cell growth inhibition (IC50) were calculated. Cell viability data showed parametric distribution, so they were analyzed using one-way ANOVA followed by Tukey’s post hoc test for intergroup comparisons and repeated measures ANOVA followed by Bonferroni’s post hoc test for intragroup comparisons. The significance level was set at p ≤ 0.05.ResultsViability % and IC50 results showed that triple antibiotic paste (TAP)–cysteamine combination had the lowest cytotoxicity level compared to other intracanal combinations followed by cysteamine and the highest cytotoxicity was with chlorhexidine–cysteamine combination.ConclusionsTriple antibiotic paste (TAP)–cysteamine combination was the safest drug compared to other drug combinations with cysteamine, so it needs more research to detect its acceptance with stem cells and its effect on defense mechanisms during healing.

Role of the Medial Canthus Fibrous Band in Forming Moderate and Severe Epicanthal Folds in Asians and Its Clinical Application.

Epicanthal folds (EFs) are skin folds located at the medial canthus in Asians. However, the anatomical structure of EFs remains unclear. The authors discovered a fibrous band connected to the medial canthal tendon (MCT) and referred to it as the medial canthal fibrous band (MCFB). This study aimed to verify whether the MCFB is different from the MCT and whether its unique anatomical relationship with the MCT plays an important role in EF formation. Forty patients who underwent epicanthoplasty from February of 2020 to October of 2021 were included. EFs from 11 patients underwent biopsy and were stained with hematoxylin and eosin, Masson trichrome, and Weigert stains to reveal their composition. Expression of collagens I and III and elastin was determined through immunohistochemical staining, and their mean optical density was measured. Preoperative and immediate exposed lacrimal caruncle area (ELCA) was measured after removing the MCFB. The MCFB is a fibrous tissue located in the EF and above the MCT. The orientation and composition of collagen fibers of the MCFB are different from those of the MCT ( P < 0.001). The MCFB also has more elastin fibers than the MCT ( P < 0.05). Immediate ELCA was significantly higher than before ELCA ( P < 0.001) once the MCFB was removed. The MCFB is composed of collagen fibers different from those in the MCT and plays a role in EF formation. Removing the MCFB during epicanthoplasty can result in a more attractive appearance postoperatively. Therapeutic, IV.

Curcumin alleviates spleen immunotoxicity induced by decabrominated diphenyl ethers (BDE-209) by improving immune function and inhibiting inflammation and apoptosis in broilers

This study was conducted to assess the mitigating effects of curcumin (Cur) on immunotoxicity in the spleen of broilers induced by the polybrominated diphenyl ether BDE-209. Eighty one-day-old broilers were allocated to the following four groups: control group, BDE-209 (0.4 g/kg) group, BDE-209 (0.4 g/kg) + Cur (0.3 mg/kg) group, and Cur (0.3 mg/kg) group. Growth performance, immunological function, inflammation, and apoptosis were assessed after 42 days of treatment. The findings demonstrate that firstly, Cur restored spleen damage caused by BDE-209 by increasing body weight, decreasing feed-to-gain ratio, correcting the spleen index, and improving the histopathological structure of the spleen. Secondly, Cur relieved BDE-209-induced immunosuppression by increasing the levels of the immunoglobulins IgG, IgM, and IgA in the serum, as well as the levels of white blood cells and lymphocytes. The levels at which GATA binding protein 3, T-box expressed in T cells, interferon-γ, and interleukin (IL)− 4 are expressed were controlled. The ratio of T helper (Th) type 1 (Th1) to Th2 cells in the spleen of broilers was also controlled. Thirdly, Cur reduced the expression of Toll like receptor (TLR) 2, TLR4, nuclear factor (NF)-κB, IL-8, IL-6, and IL-1β, which alleviated BDE-209-induced inflammation in broilers. Cur reduced BDE-209-induced apoptosis by increasing the expression of the bcl-2 protein, decreasing the expression of cleaved caspase-3 and bax proteins, decreasing the bax/bcl-2 protein ratio, and decreasing the mean optical density of TUNEL. These results suggest that Cur protects broiler spleens from BDE-209-induced immunotoxicity via modulating humoral immunity, the equilibrium between Th1 and Th2 cells, the TLRs/NF-κB inflammatory pathway, and the apoptotic pathway.

Investigating persistent factor VIII-specific IgM in the humoral immune response to factor VIII

Abstract Hemophilia A is an X-linked bleeding disorder resulting in bleeding events due to insufficient levels of factor VIII (FVIII). The most significant complication in the management of hemophilia A is the formation of polyclonal neutralizing IgG (i.e., inhibitors) which impede FVIII procoagulant activity. In a hemophilia A (FVIII KO) murine model injected with multiple doses of FVIII, FVIII-specific IgM were found to persist despite an established IgG response. In attempt to characterize the role of IgM in FVIII immunity, 18 IgMs were purified from immortal FVIII-specific hybridomas and FVIII binding was confirmed via ELISA. IgMs demonstrated weak binding to FVIII in a static ELISA model with mean optical densities (OD) at 405 nm of 0.33 ± 0.43 compared to anti-FVIII IgG with a mean OD of 1.85 ± 0.02. A novel fluid-phase ELISA, developed in our lab, utilizing N-hydroxysuccinimide (NHS)-activated magnetic beads indirectly conjugated to FVIII increased IgM-FVIII binding with mean OD of 1.15 ± 0.60. IgM binding also demonstrated porcine FVIII cross-reactivity. To evaluate the effect of IgM on antibody titers in vivo, FVIII KO mice received 5 weekly injections of either FVIII or 4 separate immune complexes (IC) of IgM-FVIII. Mice injected with IgM-FVIII IC showed slightly higher median IgG ELISA titers than FVIII alone. These findings suggest a potential role for persistent anti-FVIII IgM in propagating FVIII inhibitor development. Further studies determining the effect of IgM-FVIII IC on complement activation, as well as anti-FVIII IgM epitope specificity and clonality utilizing computational methods of RNA-sequencing, may provide additional mechanistic insight into the role of persistent IgM in the humoral immune response to FVIII. Hemostasis and Thrombosis Research Society- Student Research Award (EY), NIH K99HL150595 (GB), and Hemophilia of Georgia Clinical Scientist Development Grant (GB).

Association between glomerular mTORC1 activation and crescents formation in lupus nephritis patients.

This study aims to explore the association between glomerular mammalian target of rapamycin complex 1 (mTORC1) pathway activation and crescents' degree in lupus nephritis (LN) patients. A total of 159 biopsy-proven LN patients were enrolled in this retrospective study. The clinical and pathological data of them were collected at the time of renal biopsy. mTORC1 pathway activation was measured by immunohistochemistry, expressed by the mean optical density (MOD) of p-RPS6 (ser235/236), and multiplexed immunofluorescence. The association of mTORC1 pathway activation with clinico-pathological features especially renal crescentic lesions, and the composite outcomes in LN patients was further analyzed. mTORC1 pathway activation could be detected in the crescentic lesions and was positively correlated with the percentage of crescents (r=0.479, P<0.001) in LN patients. Subgroup analysis showed mTORC1 pathway was more activated in patients with cellular or fibrocellular crescentic lesions (P<0.001), but not fibrous crescentic lesions (P=0.270). The optimal cutoff value of the MOD of p-RPS6 (ser235/236) was 0.0111299 for predicting the presence of cellular-fibrocellular crescents in >7.39% of the glomeruli by the receiver operating characteristic curve. Cox regression survival analysis showed that mTORC1 pathway activation was an independent risk factor for the worse outcome (defined by composite endpoints of death, end-stage renal disease and a decrease of >30% in eGFR from baseline). Activation of mTORC1 pathway was closely associated with the cellular-fibrocellular crescentic lesions and could be a prognostic marker in LN patients.

MP04-15 MULTI-REGION SAMPLING OF LOCALIZED RENAL CELL CARCINOMA TUMORS DEMONSTRATES PD-L1 EXPRESSION HETEROGENEITY AND PROGNOSTIC ABILITY TO IDENTIFY RISK OF METASTATIC PROGRESSION

You have accessJournal of UrologyCME1 Apr 2023MP04-15 MULTI-REGION SAMPLING OF LOCALIZED RENAL CELL CARCINOMA TUMORS DEMONSTRATES PD-L1 EXPRESSION HETEROGENEITY AND PROGNOSTIC ABILITY TO IDENTIFY RISK OF METASTATIC PROGRESSION Daniel Shapiro, Israa Laklouk, Moon Hee Lee, David Jarrard, Tudor Borza, Glenn Allen, Kyle Richards, Wei Huang, and E. Jason Abel Daniel ShapiroDaniel Shapiro More articles by this author , Israa LakloukIsraa Laklouk More articles by this author , Moon Hee LeeMoon Hee Lee More articles by this author , David JarrardDavid Jarrard More articles by this author , Tudor BorzaTudor Borza More articles by this author , Glenn AllenGlenn Allen More articles by this author , Kyle RichardsKyle Richards More articles by this author , Wei HuangWei Huang More articles by this author , and E. Jason AbelE. Jason Abel More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003215.15AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Prior studies have evaluated PD-L1 expression as a prognostic biomarker for renal cell carcinoma (RCC) with mixed results when sampling only one region of large, heterogeneous tumors. The purpose of this study was to investigate PD-L1 expression using multi-region sampling to evaluate intra-tumor heterogeneity and ability to predict metastatic progression following radical nephrectomy for localized disease. METHODS: RCC tumor tissue microarrays (TMAs) were constructed for non-metastatic RCC patients treated with nephrectomy. Two cohorts of patients were selected, those that progressed to metastatic disease and those that did not. The cohorts were matched for age, gender, performance status, pT-stage, grade, tumor diameter and presence of tumor thrombus. From each tumor, a minimum of 10 tissue cores were analyzed. PD-L1 tissue expression was quantified as optical density (OD) per unit area using the Vectra system and InForm software (version 2.4). PD-L1 expression heterogeneity was evaluated with Kernel-density plots and compared between patients who progressed to metastatic RCC vs patients who remained disease free using nested t-tests. RESULTS: TMAs were constructed for 46 patients. There were no differences in matching characteristics between groups. Metastatic progression was identified in 26 patients with median follow-up of 7 years (IQR 5-11). Twenty patients remained disease free with median follow-up of 11 years (IQR 8-15). Median tumor size was 9 cm in both cohorts (p=0.2). Kernel-density plots demonstrated greater variance in PD-L1 expression among recurrent tumors (Figure 1A). Mean and standard deviation of the PD-L1 OD was higher for recurrent patients (mean OD 0.025 vs 0.011, p<0.001) (Figure 1B). When stratified by median PD-L1 OD, the median recurrence free survival for high PD-L1 expressing tumors was 11.7 months and was not reached for low PD-L1 expressing tumors (p<0.001) (Figure 1C). CONCLUSIONS: PD-L1 expression varies substantially throughout individual tumors. Multi-region sampling helps account for PD-L1 tumor heterogeneity, and PD-L1 expression predicted metastatic progression following radical nephrectomy for localized disease. Further studies are required to validate these findings. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e39 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Daniel Shapiro More articles by this author Israa Laklouk More articles by this author Moon Hee Lee More articles by this author David Jarrard More articles by this author Tudor Borza More articles by this author Glenn Allen More articles by this author Kyle Richards More articles by this author Wei Huang More articles by this author E. Jason Abel More articles by this author Expand All Advertisement PDF downloadLoading ...

Integration of Metabolomics and Transcriptomics to Reveal the Metabolic Characteristics of Exercise-Improved Bone Mass

(1) Background: Exercise is effective in promoting and maintaining bone mass. The aim of this study was to detect the exercise-induced metabolic changes in bone tissue of zebrafish. (2) Methods: Thirty-eight zebrafish (Danio rerio, six months old) were analyzed. The exercise group (n = 19) received 8 weeks of counter-current swimming training. The control group (n = 19) was not subjected to exercise. Mineralization was quantified, and alkaline phosphatase (Alp) and anti-tartrate acid phosphatase (Trap) activities were estimated (n = 12). The metabolomics (n = 12) and transcriptomics (n = 14) data of bone tissue were used for the integration analyses. (3) Results: The results showed that the exercise training improved the bone mineralization of zebrafish, e.g., the exercise group (5.74 × 104 ± 7.63 × 103) had a higher mean optical density than the control group (5.26 × 104 ± 8.56 × 103, p = 0.046) for the caudal vertebrae. The amount of mineralized matrix in scales of the exercised zebrafish was also higher (0.156 ± 0.012 vs. 0.102 ± 0.003, p = 0.005). Both histological staining and biochemical analysis revealed increased Alp activity (0.81 ± 0.26 vs. 0.76 ± 0.01, p = 0.002) and decreased Trap activity (1.34 ± 0.01 vs. 1.36 ± 0.01, p = 0.005) in the exercise group. A total of 103 different metabolites (DMs, VIP ≥ 1, fold change (FC) ≥ 1.20 or ≤0.83, p < 0.050) were identified. Alanine, aspartate and glutamate metabolism, β-alanine metabolism, pyrimidine metabolism, and pantothenate and CoA biosynthesis were the significantly enriched metabolic pathways (p < 0.050). A total of 35 genes (q ≤ 0.050 (BH), |Log2FC| ≥ 0.5) were coenriched with the 103 DMs in the four identified pathways. Protein-protein interaction network analysis of the 35 genes showed that entpd3, entpd1, and cmpk2 were the core genes. (4) Conclusions: The results of this study suggest that alanine, aspartate and glutamate metabolism, β-alanine metabolism, pyrimidine metabolism, and pantothenate and CoA biosynthesis contributed to exercise-induced improvements in bone mass.

Network pharmacology, molecular docking, and experimental validation to explore the potential mechanism of Long Mu Qing Xin mixture for the treatment of attention deficit hyperactivity disorder.

Background: Long Mu Qing Xin Mixture (LMQXM) has shown potentially positive effects in alleviating attention deficit hyperactivity disorder (ADHD); however, the action mechanism is still not fully understood. This study aimed to predict the potential mechanism of LMQXM for ADHD using network pharmacology and molecular docking, which were then validated using animal experiments. Methods: Network pharmacology and molecular docking techniques were used to predict the core targets and potential pathways of LMQXMQ for ADHD, and KEGG pathway enrichment analysis revealed the potential significance of dopamine (DA) and cyclic adenosine monophosphate (cAMP) signaling pathways. To verify the hypothesis, we conducted an animal experiment. In the animal experiment, the young spontaneously hypertensive rats (SHRs) were randomly divided into the model group (SHR), the methylphenidate hydrochloride group (MPH, 4.22mg/kg), and 3 LMQXM groups (low-dose (LD) group, 5.28ml/kg; medium-dose (MD) group, 10.56ml/kg; and high-dose (HD) group, 21.12ml/kg), and administered by gavage for 4weeks; the WKY rats were set as the control group. The open field test and Morris water maze test were used to evaluate the behavioral performance of rats, high performance liquid chromatography mass spectrometry (LC-MS) was used to analyze DA levels in the prefrontal cortex (PFC) and striatum of rats, ELISA was used to detect cAMP concentrations in the PFC and striatum, and immunohistochemistry and qPCR were used to analyze positive cell expression and mRNA expression for indicators related to DA and cAMP pathways. Results: The results showed that beta-sitosterol, stigmasterol, rhynchophylline, baicalein, and formononetin might be key components of LMQXM for ADHD and that these components bind well to the core targets, DA receptors (DRD1 and DRD2). Furthermore, LMQXM might act through the DA and cAMP signaling pathways. In the animal experiment, we found that MPH and LMQXM-MD controlled hyperactivity and improved learning and memory in SHRs, while LMQXM-HD only controlled hyperactivity in SHRs; meanwhile, MPH and LMQXM-MD upregulated DA and cAMP levels, mean optical density (MOD) of cAMP, and MOD and mRNA expression of DRD1 and PKA in the prefrontal cortex (PFC) and striatum of SHRs, while LMQXM-LD and LMQXM-HD upregulated DA and cAMP levels in the striatum, MOD of cAMP in the PFC, and mRNA expression of PKA in the PFC. However, we did not find a significant regulatory effect of LMQXM on DRD2. Conclusion: To sum up, this study demonstrated that LMQXM may increase DA levels mainly by activating the cAMP/PKA signaling pathway through DRD1, thereby controlling the behavioral disorders of SHRs, which is most effective at moderate doses, and this may be a key mechanism for LMQXM in the treatment of ADHD.

Cancer Genetic Markers Among School Children in Relation to Urogenital Schistosomiasis

Chronic infection with urogenital schistosomiasis can lead to severe complications such as bladder cancer. Hence, this study determined the presence of Cancer Genetic Markers among School Children in relation to Urogenital Schistosomiasis. All 36 S. haematobium positive cases from an earlier study and randomly selected 156 negative samples were used. Immunogens tested for, included glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and fibroblast growth factor receptor (FGFR3). Mean optical density (OD) was 0.73 and 0.79 for GAPDH and FGFR3 respectively. A total of 21 (10.94%) participants, 16 females and 5 males were positive for GAPDH marker. Fifty (26.04%) participants were positive for FGFR3 via ELISA cutoff. Mean OD for S. haematobium positive individuals tested for GAPDH was 0.938, minimum value was 0.119 while maximum value was 4.507. For FGFR3 mean OD was 0.896, Maximum 2.882 and minimum 0.28, while females showed higher GAPDH OD. More females than males who were positive for S. haematobium were also positive for GAPDH (11.1%) and FGFR3 (25%). FGFR3 and GAPDH marker prevalence for Individuals Positive for S. haematobium is 25% and 16.7% respectively. The age group 11-15 years were the only group positive for GAPDH. There was a slight positive correlation between age and FGFR3 and age and GAPDH. The presence of these markers is an indicator that the children in the selected communities maybe at risk of developing bladder cancer in the future if the disease is not properly managed and controlled.

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.

Isolation of Hexavalent chromium tolerant fungal species from urban vegetable farm soil and effluent waste in Addis Ababa&amp; Rift valley, Ethiopia

Hexavalent chromium is resistant to degradation and harmful toxic substance to environment and community health. Physicochemical treatment methods are demanding high cost, used large quantities of chemicals & energy, release large amount of secondary toxic degradants. Mycoremediation is an eco-friendly alternative treatment method. The main objective of this research is to isolate and characterize chrome (VI) tolerant fungi from farm soil & industry effluent for mycoremedation purpose. The screening and isolation of yeast was carried out on potato dextrose agar media. PDA and broth assay test for fungi tolerance to hexavalent chromium at different concentration, temperature and pH was evaluated. Fungi species was identified biochemically using Biolog Microstation depending on carbon utilization and chemical sensitivity test. The result revealed that 10 yeast species was identified with full ID from effluent waste and farm soil based on their probability ≥ 75% and similarity index ≥ 0.5 as well as their Cr (VI) tolerance ability up to 2500ppm. These are Yarrowia lipolytica (100%, 0.7), Cryptococcus luteolus(100%, 0.64), Rhodotorula aurantiaca A(100%, 0.62), Ustilago maydis(100%, 0.58) Trichosporon beigelii B (100%, 0.51), Cryptococcus terreus A (100%, 0.62), Zygosaccharomyces bailii (98%, 0.65), Nadsoniafulvenscens (90%, 0.62), Schizoblastosporonstarkeyihenricii (89%, 0.56), Endomycopsis vivi (84%, 0.62), Rhodotorula pustula (Sim, 0.59). Two yeast species Yarrowia lipolytica and Nadsoniafulvenscens show the highest growth mean Optical density (OD) measure 0.74 ± 0.2& 0.60 ± 0.2 respectively at pH 7 & 25°C. The highest tolerance index (mm) was recorded by Schizoblastosporon starkey henricii 0.3067 ± 0.152. Cr (VI)-tolerance ability of these yeast strains used in the development of chromium-bioremediation technologies provide an alternative option for chromium sequestration after HPLC analysis& molecular characterization.

Vascular Endothelial Growth Factor A (VEGF-A) and Vascular Endothelial Growth Factor Receptor 2 (VEGFR-2) as Potential Biomarkers for Oral Squamous Cell Carcinoma: A Sri Lankan Study.

The incidence of oral squamous cell carcinoma (OSCC) is very high in South Asia and Vascular endothelial growth factor (VEGF) is one of the key factors essential for cancer growth. The importance of VEGF-A and VEGF Receptor 2(VEGFR-2) in oral cancer pathophysiology is yet to be decided. Vascular Endothelial Growth Factor A (VEGF-A) is the main factor concerned in angiogenesis in tumors, but its role in Oral Squamous Cell Carcinoma (OSCC) is still debatable. Our study aimed to determine the role of VEGF-A and VEGFR-2 in OSCC. Blood from 30 patients with primary OSCC and 1:1 age-sex-matched controls was subjected to qPCR and ELISA to detect VEGF-A gene expression and serum level. Tumors of the 30 patients were investigated for VEGF Receptor-2 (VEGFR-2) expression and were analyzed using Image J software version 1.52 for DAB percentage (DAB-P) area and optical density (OD). VEGF-A relative gene expression among patients was 2.43-fold higher compared to the healthy control group. Well-differentiated had a 1.98-fold increment, while poorly differentiated had a 3.58-fold increment. Serum VEGF-A was significantly elevated among the patients compared to controls (458.7 vs 253.2, p=0.0225). Poorly differentiated had a higher serum VEGF concentration (1262.0±354.7pg/ml) compared with other two. Mean VEGFR-2 DAB-P level in OSCC was 42.41±5.61(p=0.15). Well-differentiated had a DAB-P of 41.20±5.32 while poorly differentiated had DAB-P 46.21±3.78. The mean OD in OSCC was 0.54±0.16. VEGFR-2 OD in well and poorly differentiated OSCC were 0.48±0.12 and 0.68±0.17, respectively. VEGF-A gene expression, serum levels, and tissue VEGFR-2 levels correlated linearly with the stage and grade of the tumor. This study justifies the value of VEGF-A as a potential biomarker in OSCC in early detection of OSCC. More studies are needed to accept the use of VEGF-A.

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.