Morphologically Changed Research Articles

Leveraging deep learning for detecting red blood cell morphological changes in blood films from children with severe malaria anaemia.

In sub-Saharan Africa, acute-onset severe malaria anaemia (SMA) is a critical challenge, particularly affecting children under five. The acute drop in haematocrit in SMA is thought to be driven by an increased phagocytotic pathological process in the spleen, leading to the presence of distinct red blood cells (RBCs) with altered morphological characteristics. We hypothesized that these RBCs could be detected systematically and at scale in peripheral blood films (PBFs) by harnessing the capabilities of deep learning models. Assessment of PBFs by a microscopist does not scale for this task and is subject to variability. Here we introduce a deep learning model, leveraging a weakly supervised Multiple Instance Learning framework, to Identify SMA (MILISMA) through the presence of morphologically changed RBCs. MILISMA achieved a classification accuracy of 83% (receiver operating characteristic area under the curve [AUC] of 87%; precision-recall AUC of 76%). More importantly, MILISMA's capabilities extend to identifying statistically significant morphological distinctions (p < 0.01) in RBCs descriptors. Our findings are enriched by visual analyses, which underscore the unique morphological features of SMA-affected RBCs when compared to non-SMA cells. This model aided detection and characterization of RBC alterations could enhance the understanding of SMA's pathology and refine SMA diagnostic and prognostic evaluation processes at scale.

The combination of TRAIL and the Smac mimetic LCL-161 induces an irreversible phenotypic change of MCF-7 breast cancer cells

IntroductionBreast cancer is the most common malignancy affecting women. Although the prognosis generally is good, a substantial number of patients still suffer from relapse, emphasizing the need for novel treatments. Smac mimetics were developed to facilitate cell death by blocking inhibitor of apoptosis proteins (IAPs). It has been suggested that TNF-related apoptosis inducing ligand (TRAIL) can be used together with Smac mimetics to induce cancer cell death. MethodsCell viability was studied with Trypan blue staining and Annexin V assay, siRNA was used to downregulate specific proteins, protein levels were estimated with Western blot, and mRNA levels were analyzed with qPCR, microarray and RNA-seq. For global expression, groups were compared with principal component analysis and the limma package in R. Gene enrichment was analyzed with Fisher's test. For other experiments, significance of difference was tested by one-way ANOVA, followed by Tukey's HSD test. ResultsThe combination of Smac mimetic LCL-161 and TRAIL induces an irreversible change in phenotype, but not cell death, of luminal MCF-7 breast cancer cells. The cells become small and circular and dissociate from each other and the effect could not be reversed by returning the cells to regular growth medium. The morphology change could be prevented by caspase inhibition using z-VAD-FMK and downregulation of caspase-8. Caspase-7 is also indicated to be of importance since downregulation of this caspase resulted in fewer morphologically changed cells. Enrichment analyses of changes in global gene expression demonstrated that genes associated with estrogen receptor (ER) signaling are downregulated, whereas nuclear factor kappa B- (NF-κB) and interferon- (IFN) driven genes are upregulated in altered cells. However, inhibition of these pathways did not influence the change in morphology. Induction of IFN-induced genes were potentiated but NF-ĸB-driven genes were slightly suppressed by caspase inhibition. ConclusionsThe results demonstrate that LCL-161 and TRAIL can irreversibly alter the MCF-7 breast cancer cell phenotype. However, the changes in morphology and global gene expression are mediated via separate pathways.

Prevalence and Natural Course of Transverse Ligament of the Atlas Calcification in Asymptomatic Healthy Individuals.

A retrospective cross-sectional study. The aim of this study was to assess the prevalence and natural course of calcification of the transverse ligament of the atlas (TLA) in healthy subjects and to investigate the relationship between the calcified lesion and clinical factors. The prevalence and etiology of TLA calcification in healthy individuals remain uncertain. We enrolled 1136 participants undergoing computed tomography (CT) scans in their routine medical checkups. The calcified lesion of the TLA was classified into two types: type 1, thin calcifications (<2 mm) and type 2, curvilinear lesions (>2 mm). We evaluated the natural course of the lesion and compared each clinical variable among subjects with and without TLA calcifications after adjusting for age and sex using propensity score matching techniques. TLA calcification was present in 40 participants (3.5%; 31 males and 9 females; mean age, 65.3 years), and the frequency increased with age (P = 0.001). Both types 1 and 2 calcifications were observed in 26 (65%) and 14 cases (35%), with mean ages of 63.0 and 69.4 years (P = 0.033), respectively. Of these, 37 (93%) underwent another medical checkup afterward, and the morphological changes were further assessed. Five cases (14%) showed enlargement [one case (4%) in type 1 vs. four cases (33%) in type 2], whereas six cases (16%) had disappearance or decrease [six cases (24%) in type 1 vs. 0 cases in type 2, which was statistically significant (P = 0.019)]. A one-to-one propensity score matching analysis revealed no significant differences in clinical variables among subjects with and without TLA calcifications. The prevalence of the TLA calcification was not rare in normal subjects. Our results suggest that the calcified lesion could be morphologically changed on a yearly basis and may be associated with the normal aging process. 3.

Evaluation of valve function in antireflux biliary metal stents

BackgroundTo overcome duodenobiliary reflux induced by biliary stents, antireflux valve (ARV) biliary stents have been developed and showed improvement in stent patency. However, negative study results have also been reported because stent patency may be decreased by the malfunction of ARV itself. Given such mixed results, the true efficacy of ARV remains unknown and the mechanism of its dysfunction needs to be clearly elucidated. The aim of this study was to investigate the exact mechanism of ARV dysfunction using in vitro phantom models.MethodsTwo experimental models were designed to evaluate two important environmental factors suspected to cause ARV malfunction, i.e. bile flow and pH. Three types of ARV metal stents from different companies were used for the experiments: a funnel type ARV, a windsock type ARV, and a wine glass-shaped ARV. Ten stents of each type were tested (five stents in the bile flow phantom model, and another five stents in the duodenal pH environmental model). To determine ARV malfunction, ARV-induced flow resistance was measured using a custom-made testing device. All stents from the two models were removed every 2 weeks for 12 weeks after stent insertion and were evaluated on morphological and functional changes of the ARV.ResultsOnly ARV of wine glass-shaped ARV was morphologically changed due to silicone bond detachment in the bile flow model. All types of ARV were morphologically changed in the pH model. The morphological changes of ARV influenced the flow resistance. The antegrade pressure gradients were increased over time in the pH model (p < 0.05).ConclusionsMorphological change of the ARVs may induce dysfunction of ARV metal stents, which is mainly due to duodenal pH environment. In the future, development of new ARV that is not affected by duodenal environmental factors can be expected to improve stent patency.

Relationship of arctic fox (Alopex lagopus L.) sperm morphology with age of males, sperm concentration, ejaculate volume and acrosin activity

Abstract The aim of the study was to determine the relationship of sperm morphology with age of males, ejaculate concentration and volume, as well as with acrosin activity determined in sperm acrosome extracts. The study used manually collected ejaculates from 9 male arctic foxes, including 6 young males aged one year and 3 older males (between 3 and 5 years of age). All of the 39 ejaculates used in the study were classified as normal based on motility exceeding 70%. The ejaculates collected from the foxes were evaluated for volume, sperm concentration and frequency of morphological changes including primary and secondary defects. The spermiograms of the male arctic foxes were classified according to a six-grade subjective scale. In addition, acrosin activity was determined in the sperm acrosome extracts. The data were analysed using the criteria of male age, sperm concentration, ejaculate volume, and acrosin activity. The morphology of arctic fox spermatozoa was dependent on the age of the male. A greater number of morphologically altered spermatozoa tended to occur in the ejaculates of young foxes, which were in their first breeding season. In addition, statistical analysis revealed positive relationships between the frequency of morphological changes in sperm and their ejaculate concentration. In contrast, there were no significant correlations between the percentages of morphologically changed spermatozoa and the ejaculate volume and the content of acrosin, which is an indicator of acrosomal integrity. Semen quality is dependent on the number of sperm in ejaculate with morphological defects which prevent oocyte fertilization. Therefore, morphological assessment of semen, which covers both the number and type of morphological changes, is highly useful when selecting appropriate males for reproduction.

СТРУКТУРНО-ФУНКЦИОНАЛЬНАЯ ПЕРЕСТРОЙКА НЕЙРОНОВ ГИППОКАМПА ПОСЛЕ ОПЕРАЦИИ ПОД АНЕСТЕЗИЕЙ СЕВОФЛУРАНОМ (ЭКСПЕРИМЕНТАЛЬНОЕ ИССЛЕДОВАНИЕ)

Hippocampus plays an important role in the cognitive mnestic functions. It coordinates emotional process, defines the intensity and specificity of behavioral, neuron and hormone reactions. Structural changes in hippocampus neurons can result in learning and memory disorders. The objective: to investigate morphometric parameters of hippocampus fields in rats after abdomen surgery with exposure to sevoflurane. Subjects and methods. The experimental research was performed using Wistar rats (n = 15). Rats from the experimental group (n = 7) had abdomen surgery with anesthesia with sevoflurane with the consequent continuous exposure to it (6 hours, 2 vol. % of sevoflurane, air flow – 1 l/min.). During 21 days rats from both groups had a number of behavioral tests. In 12 hours after the last behavioral test (on the 22-th day after the start of the experiment) the rats were decapitated with consequent brain extraction. The obtained materials were fixed in 10% neutral formalin on phosphate buffer (pH 7.4) for 24 hours minimum. Preparations of CA1, Ca2, Ca3, and Ca4 fields were studied under light microscope DM-750 (Leica, Germany) using the computer image analysis software of ImageScope M with 400-fold magnification. Results. The tests showed that cytoarchitectonics of hippocampus field was intact, pyramidal neurons had a large rounded nucleus with one, two or more distinct nucleoli. No pericellular and perivascular edema was detected. In the experimental group, all fields of the hippocampus had structural and functional changes. It manifested through both quantitative and qualitative signs of neuronal damage, especially in the Ca1 field. Segments of pyramidal neurons were disorganized, morphologically modified neurons were found: hyperchromatic shriveled neurons with no nucleus or poorly visible nucleus of the irregular shape. The appearance of morphologically changed neurons and disorganization of hippocampal layers led to changes in the width of pyramidal neurons segments. Conclusion. Morphological changes of hippocampal structures of rats after surgery and anesthesia may be the cause of cognitive functions decline during the postoperative period.

Clinical significance of interval changes in breast lesions initially categorized as probably benign on breast ultrasound.

The aims of this study were to determine the malignancy rate of probably benign lesions that show an interval change on follow-up ultrasound and to evaluate the differences seen on imaging between benign and malignant lesions initially categorized as probably benign but with interval change on follow-up breast ultrasound.We retrospectively reviewed 11,323 lesions from ultrasound-guided core-biopsies performed between June 2004 and December 2014 and identified 289 lesions (266 patients) with an interval change from probably benign (Breast Imaging Reporting and Data System [BI-RADS] category 3) in the previous 2 years. Malignancy rates were compared according to the ultrasound findings and the characteristics of the interval changes, including changes in morphology and/or diameter.The malignancy rate for probably benign lesions that showed an interval change on follow-up ultrasound was 6.9% (20/289). The malignancy rate was higher for clustered cysts (33.3%) and irregular or noncircumscribed masses (12.7%) than for circumscribed oval masses (5%) or complicated cysts (5%) seen on initial ultrasound (P = 0.043). Fifty-five percent of the malignancies were found to be ductal carcinoma in situ and there was 1 case of lymph node metastasis among the patients with invasive disease in whom biopsy was delayed by 6 to 15 months. The extent of invasiveness was greater in missed cases. There was a significant difference in the maximal diameter change between the 20 malignant lesions and the 269 benign lesions (4.0 mm vs 2.7 mm, P = 0.002). The cutoff value for maximal diameter change per initial diameter was 39.0% for predicting malignancy (sensitivity 95%, specificity 53.5%). The malignancy rate for morphologically changed lesions was significantly higher than for morphologically stable lesions (13.6% vs 4.9%; P = 0.024)Our 6.9% of probably benign lesions that showed an interval change finally turned out to be malignancy was mostly DCIS. The sonographic features, interval changes in sonographic features, and lesion size might help in the recategorization of these lesions.

Putative stem cells and epithelial-mesenchymal transition revealed in sections of ovarian tumor in patients with serous ovarian carcinoma using immunohistochemistry for vimentin and pluripotency-related markers

BackgroundThe mechanism of aggressive character of ovarian cancer and unsuccessful treatment of women with this deadly disease has been recently explained by the theory of cancer stem cells (CSCs). It has been reported that ovarian carcinogenesis and progression of disease is associated with epithelial-mesenchymal transition (EMT). EMT, a physiological cell process during embryonic development and later in life during regeneration, could, when induced in pathological condition, generate CSCs-like cells. Until now EMT in the ovarian tissue has been mainly studied in cell cultures in vitro. The aim of this study was to focus on in situ morphological changes in the ovarian surface epithelium of tumor tissue in women with epithelial ovarian cancer after we applied the antibodies for markers of EMT vimentin and pluripotency-related markers NANOG, SOX2 and SSEA-4.MethodsWe analyzed ovarian tissue sections of 20 women with high grade serous ovarian carcinoma. After eosin and hematoxylin staining, used in standard practice, immunohistochemistry was performed for vimentin and markers of pluripotency: NANOG, SSEA-4 and SOX2. We focused on the ovarian surface epithelium in order to observe morphological changes in tumor tissue.ResultsAmong epithelial cells of the ovarian surface epithelium in women with serous ovarian carcinoma we observed a population of small NANOG-positive cells with diameters of up to 5 μm and nuclei, which filled almost the entire cell volumes. These small NANOG-positive cells were in some cases concentrated in the regions with morphologically changed epithelial cells. In these regions, a population of bigger round cells with diameters of 10–15 μm with large nuclei, and positively stained for vimentin, NANOG and other markers of pluripotnecy, were released from the surface epithelium. These cells are proposed as CSCs, and possibly originate from small stem cells among epithelial cells. They formed typical cell clusters, invaded the tissue by changing their round shape into a mesenchymal-like phenotype, and contributed to the manifestation of ovarian cancer.ConclusionsOur findings show morphological changes in the ovarian surface epithelium in tumor slides of high grade serous ovarian carcinoma and provide a new population of putative CSCs.

Morphological changes of olivine grains reacted with amino acid solutions by impact process

Early oceans on Earth might have contained certain amounts of biomolecules such as amino acids, and they were subjected to meteorite impacts, especially during the late heavy bombardment. We performed shock recovery experiments by using a propellant gun in order to simulate shock reactions among olivine as a representative meteorite component, water and biomolecules in oceans in the process of marine meteorite impacts. In the present study, recovered solid samples were analyzed by using X-ray powder diffraction method, scanning electron microscopy, electron probe microanalysis, and transmission electron microscopy with energy-dispersive X-ray spectrometry. The analytical results on shocked products in the recovered sample showed (1) morphological changes of olivine to fiber- and bamboo shoot-like crystals, and to pulverized grains; and features of lumpy surfaces affected by hot water, (2) the formation of carbon-rich substances derived from amino acids, and (3) the incorporation of metals from container into samples. According to the present results, fine-grained olivine in meteorites might have morphologically changed and shock-induced chemical reactions might have been enhanced so that amino acids related to the origin of life may have transformed to carbon-rich substances by impacts.

A long period grating optical fiber sensor with nano-assembled porphyrin layers for detecting ammonia gas

In this study, the ability of a long period grating (LPG) optical fiber sensor coated with a multilayer film of poly(diallyldimethyammonium chloride) (PDDA) and tetrakis(4-sulfophenyl)porphine (TSPP) to detect ammonia gas was demonstrated. The nano-assembled thin film was prepared using a layer-by-layer deposition technique. This combination of an LPG and TSPP could allow highly sensitive optical sensors that specifically bind ammonia to be fabricated. An LPG with a period of 100μm and a PDDA/TSPP coating produced by depositing fifteen layers afforded a limit of detection of 0.67ppm for ammonia gas. TSPP molecules in the film acted as ammonia receptors because the TSPP morphologically changed from J- or H-aggregates to free base monomers when it was deprotonated by being exposed to ammonia. Interestingly, HCl vapor could be used to increase the relative amount of J-aggregation in the TSPP and to restore the sensor response. The reversibility of the morphological change in the TSPP allowed reversible changes to occur in the properties of the coating, including the refractive index, film thickness and density, and electrostatic interactions; these influenced the transmission spectrum of the LPG.

Abstract 2368: Differentiated gastric cancer cells have a potential to induce cancer-associated fibroblasts

Abstract Gastric cancer is one of the most common causes of cancer death. Stromal fibroblasts in the tumor microenvironment have recently been implicated in tumor growth, invasion and metastasis. Cancer-associated fibroblasts (CAFs) have been suggested to be responsible for progression of undifferentiated scirrhous gastric cancers with vast fibrous stroma. However, the relationship between differentiated gastric cancer cells and CAFs in tumor progression remains unclear. In this study, we investigated whether differentiated and/or undifferentiated human gastric cancer cells induce CAFs. We used two differentiated human gastric cancer cells (MKN-7, MKN-74) and two undifferentiated human scirrhous gastric cancer cells (KATO-III, NUGC-4). Two types of human normal fibroblasts (WI-38, FEF-3) were used for induction of CAFs. The conditioned media (CM) from all cancer cells were obtained 48 hours after serum starvation. The morphological change and expression of CAF-specific markers (α-smooth muscle actin (α-SMA), fibroblast activation protein-α (FAP)) were analyzed by microscopy and Western blot analysis, respectively, in fibroblasts treated with CM for 120 hours. Moreover, the effect of CM from CAFs in the migration ability of cancer cells was evaluated using in vitro Migration assay. The CM from differentiated MKN-7 and MKN-74 cells induced the morphological change like CAFs in normal human fibroblasts, whereas CM from undifferentiated KATO-III and NUGU-4 cells did not induce. In fact, morphologically changed fibroblasts like CAFs showed increased α-SMA expression, whereas FAP expression was not increased. The CM from morphologically changed fibroblasts, but not normal fibroblasts, significantly enhanced the migration ability of MKN-7 cells. These results suggest that differentiated gastric cancer cells have a potential to induce CAFs in the invasion process of gastric cancers. CAF-targeting therapy may be a promising antitumor strategy in differentiated gastric cancers as well as undifferentiated scirrhous gastric cancers with vast fibrous stroma. Citation Format: Yuncheng Li, Hiroshi Tazawa, Nishizaki Masahiko, Yuuri Hashimoto, Naoto Hori, Ryoichi Katsube, Shinji Kuroda, Kazuhiro Noma, Shunsuke Kagawa, Toshiyoshi Fujiwara. Differentiated gastric cancer cells have a potential to induce cancer-associated fibroblasts. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2368. doi:10.1158/1538-7445.AM2015-2368

Abstract LB-271: Inulin inhibits free radical species in HCT 116 adenoma carcinoma and normal human HK (keratinocyte) cell line

Abstract Background: Bile acids have been implicated in oxidative damage via stimulation of reactive species including superoxide and nitric oxide. Superoxide and nitric oxide production are associated with inflammation. NF-κB up- regulation provides a critical link between inflammation and cancer. It has also been shown that there is a possible correlation between total activity of nitric oxide synthase and p53 mutation frequency in lung adenocarcinoma. Inulin is an oligosaccharide, classified as a prebiotic, and shown to improve gut health when used in combination with probiotics. However, there is paucity of data on how prebiotics alone affect gut health and by which mechanism of action. Objectives: To evaluate the potential of inulin in reduction of reactive species and its mechanism of inhibiting cancer cells. Methods: HCT116 adenocarcinoma cells were treated with inulin at different concentrations and stopped at various time ranges (30 minutes - 24 hours). Modified Greiss reagent was used in the study of reactive nitrogen species (RNS). Nitro blue tetrazolium (NBT) assay (Modified), was used in the investigation of superoxide production (SOP) in HCT116 in the presence of deoxycholic acids (DC); a secondary bile acids. Inulin (5%-40%) significantly inhibited DC (100μM-500 μM), generated RNS in HCT 116 cell lines (p&amp;lt;0.05; n = 6). The impact of inulin on RNS was observed to be dose and time-dependent. DC 100μM-300μM was observed to stimulate SOP in HCT116 (p&amp;lt;0.05; n = 3). Inulin (30%) was observed to inhibit SOP in HCT116 (P&amp;lt;0.05). The impact of inulin on RS was not enhanced in the presence of probiotics. Probiotic (Bifidobacterium longum) inhibited nitric oxide production (p&amp;lt;0.05; n = 3).The proliferation (Cyquant assay) of HCT116 was observed to be significantly influenced in the presence of DC; proliferation is inversely proportional to the concentration of DC; 100μM (p&amp;lt;0.05; n = 3) Proliferation of HCT 116 is enhanced in the presence of low concentration of DC &amp;lt;150 μM, while cell death and morphological change into mesenchymal cells is enhanced with &amp;gt;DC150μM. 30% Inulin enhanced cell death of morphologically changed cells in the presence of DC &amp;gt;150μM, with complete cell death occurring at DC300μM.Inulin inhibits proliferation of HCT 116 cancer cells. Inulin inhibits nitric oxide induced by DC p&amp;lt;0.05(n = 3) in normal human HK (keratinocytes) cells. Conclusions: Peroxynitrite is formed by nitric oxide and superoxide; and considered to be cytotoxic, causing DNA damage and leading to carcinogenesis via inactivation of tumour suppressor oncoprotein p53. Possible mechanism of action by which inulin could exert protective effect on colon cells could be via its antioxidant effect. Further work is underway investigating the effect of inulin and bile-acid on NF-κB, p53 and apoptosis in adenocarcinoma and normal dermal HK epithelial cells. Citation Format: Bene Akromaa Ekine-Afolabi, Sandra Appiah, Azra Pachenari, Lucy Ghali. Inulin inhibits free radical species in HCT 116 adenoma carcinoma and normal human HK (keratinocyte) cell line. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr LB-271. doi:10.1158/1538-7445.AM2015-LB-271

Lipophilic organic pollutants induce changes in phospholipid and membrane protein composition leading to Vero cell morphological change

Membrane damage related to morphological change in Vero cells is a sensitive index of the composite biotoxicity of trace lipophilic chemicals. However, judging whether the morphological change in Vero cells happens and its ratio are difficult because it is not a quantitative characteristic. To find biomarkers of cell morphological change for quantitatively representing the ratio of morphological changed cell, the mechanism of cell membrane damage driven by typical lipophilic chemicals, such as trichlorophenol (TCP) and perfluorooctanesulphonate (PFOS), was explored. The ratio of morphologically changed cells generally increased with increased TCP or PFOS concentrations, and the level of four major components of phospholipids varied with concentrations of TCP or PFOS, but only the ratio of phosphatidylcholine (PC)/phosphatidylethanolamine (PE) decreased regularly as TCP or PFOS concentrations increased. Analysis of membrane proteins showed that the level of vimentin in normal cell membranes is high, while it decreases or vanishes after TCP exposure. These variations in phospholipid and membrane protein components may result in membrane leakage and variation in rigid structure, which leads to changes in cell morphology. Therefore, the ratio of PC/PE and amount of vimentin may be potential biomarkers for representing the ratio of morphological changed Vero cell introduced by trace lipophilic compounds, thus their composite bio-toxicity.

Propidium iodide staining method for testing the cytotoxicity of 2,4,6-trichlorophenol and perfluorooctane sulfonate at low concentrations with Vero cells

In this article, the feasibility of propidium iodide (PI) staining based on cell membrane damage as a sensitive and quantitative method to test the cytotoxicity of typical lipophilic compounds including 2,4,6-trichlorophenol (TCP) and perfluorooctane sulfonate (PFOS) was examined. The sensitivity of PI staining was compared to that of the methylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) assay. We found a good correlation between PI uptake and increasing concentrations of TCP (10–50 μM) and PFOS (20–50 μM) at lower concentrations than those between growth inhibition ratio determined by the MTT assay and increasing concentrations of TCP (150–400 μM) and PFOS (100–500 μM). These findings indicated that the PI staining was more sensitive than the MTT assay. Furthermore, both the amount of PI uptake and the ratio of morphologically changed cells increased with increasing TCP concentrations, suggesting that PI staining may be a suitable quantitative substitute for the original method based on the observation of morphological changes in Vero cells. Our data also suggest that it is possible that the cell membrane damage that resulted in increased PI uptake may be due to variations in the phospholipid and protein content of the membrane, which are affected by interactions between the lipophilic compounds and components of the cell membrane.

Meniscal Morphologic Changes on Magnetic Resonance Imaging Are Associated With Symptomatic Discoid Lateral Meniscal Tear in Children

To determine whether meniscal deformation and displacement on magnetic resonance imaging (MRI) are associated with tears by use of arthroscopic findings as a standard of reference in children with discoid lateral meniscus (DLM). We reviewed MRI scans and intraoperative videos of 69 consecutive patients (79 knees) treated arthroscopically for suspicious DLM tears. The mean age at surgery was 10.9 years (range, 4.3 to 17.6 years). Signal changes and morphologic changes (deformation or displacement) of DLM on magnetic resonance (MR) images were graded by 2 independent observers using our modifications of previously described classification schemes, and then the grades were determined by consensus of the observers. Meniscal tears were assessed by an observer, blinded to the MRI studies, based on arthroscopic findings. Signal changes and morphologic changes of DLM on MR images were correlated with tears. Tears were found more frequently in menisci showing morphologic changes on MR images (P = .001). Of the 25 menisci with a grade 3 signal change (linear or band-like signal intensity extending to the superior or inferior meniscal surface), 24 had tears, and a horizontal cleavage was the most commonly associated tear type. Of the 50 menisci with a grade 1 (dot-like intrameniscal signal change), grade 2 (linear or band-like intrameniscal signal change), or diffuse signal change, 34 were morphologically changed on MR images, and 29 of these (85%) were torn, whereas 9 of the 16 menisci (56%) not morphologically changed were torn (P = .036). Preoperative MRI evaluations based on signal intensities do not accurately predict the presence of a DLM tear in children, except when a DLM shows a grade 3 signal change. Meniscal deformation or displacement observed on preoperative MR images suggests a higher risk of meniscal tears, even in menisci with signal changes other than grade 3 changes. Level IV, therapeutic case series.

LONG-TERM EVOLUTION OF MAGNETIC AND DYNAMICAL PROPERTIES IN A SUNSPOT LIGHT BRIDGE

Sunspot light bridges (LBs)—long bright lanes appearing in umbra—sometimes show dynamical behaviors such as plasma ejections, brightenings, and fast gas flows in the photosphere and lower chromosphere, but we have not understood what causes these dynamics. The Hinode Solar Optical Telescope successfully captured the entire period of the evolution of an LB formed at the southeast of the well-developed sunspot in NOAA Active Region 10953, allowing us to track how magnetic and dynamical properties change with time for 3.5 days. The LB produced chromospheric upward ejections intermittently and recurrently on 2007 April 30, and fewer upward ejections were observed on May 1. We found that G-band intensity features morphologically changed from cellar or patchy on April 30 to filamentary on May 1, although there were small changes in the magnetic flux density and inclination. This suggests that the chromospheric activity is related to the change of morphology in the photosphere. Fast gas flows and a pair of strong enhanced vertical electrical currents were also observed in the photosphere after the filamentary structures were dominant. The end of a large Hα filament (or prominence) was extended very close to the LB on May 1, suggesting that the filamentary structures formed along the LB may be magnetically connected to the large Hα filament and the gas flows may originate far from the LB region, although other mechanisms cannot be ruled out.

Sensitivity of Different Cytotoxic Responses of Vero Cells Exposed to Organic Chemical Pollutants and their Reliability in the Bio-toxicity Test of Trace Chemical Pollutants

To find a sensitive cytotoxic response to reflect the bio-toxicity of trace organic pollutants, the sensitivity and reliability of morphological change and proliferation inhibition of Vero cells exposed to 2, 4, 6-trichlorophenol (TCP) and the leachate from products related to drinking water (PRDW) were compared, and the mechanism of the morphological change in Vero cells exposed to chemical pollutants was studied. Vero cells were treated by different concentration of TCP and the leachate from PRDW. Methylthiazol-2-yl-2, 5-diphenyl tetrazolium bromide (MTT) assay was carried out for proliferation inhibition. Bioluminescence method was carried out as another method to test the toxicity of TCP. Flow Cytometry assay was used to test cell Apoptosis and damage of cell-membrane. 0.25 mg/L TCP had an effect on cell morphology, and the proportion of morphologically changed cells increased with increasing TCP concentration. At low TCP concentrations, inhibition of cell proliferation did not seem to correlate to TCP concentration, and was negative when TCP concentration was <1.0 mg/L. After exposure to leachate from PRDW extracted at different temperatures, the percentage of morphologically changed cells increased with extracting temperature, but the inhibition of cell proliferation failed to reflect the correlation between extracting temperature and proliferation inhibition of Vero cells. Although the Sensitivity of bioluminescence method seems to be similar to morphological change in Vero cells, the bacterial in this method is not homologous enough with human body cells to reflect the toxicity to human body. These imply cell morphological change is a more sensitive and reliable method to reflect bio-toxicity of organic pollutants than proliferation inhibition. Flow cytometry analysis and cell rejuvenation experiments indicated cell membrane damage, which results in cell morphological change, was an early and sensitive cytotoxic response comparing with necrosis. These results indicated that the cell membrane toxicity represented by morphological changes is a more sensitive and reliable method to indicate the composite bio-toxicity of trace chemicals than proliferation inhibition, inhibition on bioluminescence and necrosis. Nevertheless, the quantification of morphological change should be studied further.

Sensitivity of morphological change of Vero cells exposed to lipophilic compounds and its mechanism

To find a sensitive cytotoxic response to reflect the toxicity of trace organic pollutants, the sensitivity and reliability of morphological change and proliferation inhibition of Vero cells exposed to lipophilic compounds and the leachate from products related to drinking water (PRDW) were compared, and the mechanism of the morphological change in Vero cells was studied. Results showed the proportion of morphologically changed cells increased with increasing 2,4,6-trichlorophenol (TCP)/perfluorooctane sulfonate (PFOS) concentration. However, at low TCP concentrations, inhibition of cell proliferation did not correlate to TCP concentration. After exposure to the leachate from PRDW extracted at different temperatures, the percentage of morphologically changed cells increased with extracting temperature, but the inhibition of cell proliferation failed to reflect the correlation to extracting temperature. These imply cell morphological change is a more sensitive and reliable method to reflect toxicity of trace organic pollutants than proliferation inhibition. Flow cytometry analysis indicated cell membrane damage was an early and sensitive cytotoxic response comparing with necrosis, resulting in cell morphological change, which may be due to the interference of lipophilic compounds. Lipophilic compound accumulated in cell membrane to interfere the assembly process of membrane protein and phospholipid.

Increase in intracellular Ca2+ concentration is not the only cause of lidocaine-induced cell damage in the cultured neurons of Lymnaea stagnalis

To determine whether the increase in intracellular Ca2+ concentration induced by lidocaine produces neurotoxicity, we compared morphological changes and Ca2+ concentrations, using fura-2 imaging, in the cultured neurons of Lymnaea stagnalis. We used BAPTA-AM, a Ca2+ chelator, to prevent the increase in the intracellular Ca2+ concentration, and Calcimycin A23187, a Ca2+ ionophore, to identify the relationship between increased intracellular Ca(2+) concentrations and neuronal damage without lidocaine. Morphological changes were confirmed using trypan blue to stain the cells. Increasing the dose of lidocaine increased the intracellular Ca2+ concentration; however, there was no morphological damage to the cells in lidocaine at 3 x 10(-3) M. Lidocaine at 3 x 10(-2) M increased the intracellular Ca2+ concentration in both saline (from 238 +/- 63 to 1038 +/- 156 nM) and Ca2+-free medium (from 211 +/- 97 to 1046 +/- 169 nM) and produced morphological damage and shrinkage, with the formation of a rugged surface. With the addition of BAPTA-AM, lidocaine at 3 x 10(-2) M moderately increased the intracellular Ca2+ concentration (from 150 +/- 97 to 428 +/- 246 nM) and produced morphological damage. These morphologically changed cells were stained dark blue with trypan blue dye. The Ca2+ ionophore increased the intracellular Ca2+ concentration (from 277 +/- 191 to 1323 +/- 67 nM) and decreased it to 186 +/- 109 nM at 60 min. Morphological damage was not observed during the 60 min, but became apparent a few hours later. These results indicated that the increase in intracellular Ca2+ concentration is not the only cause of lidocaine-induced cell damage.

Acute Effects of Lingual Nerve Damage on Orofacial Nociceptive-Induced Reflexes

PC12 cells were found to attach on PLGA nerve conduits coated with FNIII9-10 peptide best among all the groups. PC12 cells showed better cellular attachment on nerve conduit in proportion to FNIII9-10 peptide concentration. 3) The SEM view of cell-nerve conduit complexes showed that some cells in all the groups seemed to be morphologically changed from small and round shape to flattened and polygonal shape with elongated cytoplasmic extension after 48 hour incubation. This tendency to change morphologically seemed to be more prominent in Group I and Group III than control and Group II. Conclusion: Further studies will be needed to prove that higher cellular attachment in vitro will lead to better nerve regeneration in vivo. The correlation between cellular morphological changes and their attachments on the nerve conduit also needs further study.