Mean Optical Density Research Articles

Comparison of 1,3-β-d-glucan with galactomannan in serum and bronchoalveolar fluid for the detection of Aspergillus species in immunosuppressed mechanical ventilated critically ill patients

PurposeInvasive pulmonary aspergillosis (IPA) is an important cause of morbidity/mortality in immunocompromised critically ill patients. New diagnostic strategies for early detection of IPA include the noninvasive biomarkers 1,3-β-d-glucan (BDG), serum, and bronchoalveolar (BAL) fluid galactomannan (GM). The aim of this study was to compare these markers for early detection of IPA in immunosuppressed critically ill patients. MethodsBetween December 2014 and December 2015, 49 immunosuppressed patients with respiratory failure were treated at our intensive care unit (ICU). We compared the BDG Fungitell assay with GM Platelia assay in serum and BAL for early detection of IPA. All tests were performed initially after admission at the ICU. ResultsIn our study with 49 patients, 13 (26%) had probable IPA. These patients had a higher Acute Physiology And Chronic Health Evaluation II score (28 vs 23, P<.001), Sequential Organ Failure Assessment score (16 vs 14, P<.001), more neutropenia (77% vs 30%, P<.001), worse Horowitz Index (99 vs 73 P<.020), a longer ICU stay (26 vs 17 days, P<.044), and a higher mortality rate (77% vs 58%, P<.001) as compared with patients without probable IPA.The used biomarker BDG presented in patients with probable IPA showed significantly higher levels as compared with patients without probable IPA (375 [103-1000 pg/mL; P<.001] vs 64 [30-105 pg/mL; P < .001]).Comparison of BDG with GM showed that positive serum GM could be detected in only 4 (30%), whereas positive BAL GM could be detected in 12 (92%; mean optical density index, 3.7) of 13 probable IPA cases.These results can be expressed as an overall sensitivity of 88% and a specificity of 82% for probable IPA using the BDG Fungitell assay, a sensitivity of 35% and a specificity of 70% using the serum GM Platelia assay, and a sensitivity of 70% and a specificity of 94% using the BAL GM Platelia assay. The negative predictive values of the used tests were 94% for the BDG Fungitell assay, 94% for the serum GM Platelia assay, and 90% for the BAL GM Platelia assay. Conclusion1,3-β-d-Glucan may be a useful marker for patients under surveillance at risk for IPA. In critically ill patients with immunosuppression, early diagnosis of IPA may be improved by BDG as compared with serum GM. However, diagnostic performance and accuracy increase when BDG is run in parallel with GM from BAL; moreover, the association of the 2 parameters has also the advantage of detecting early and reliable IPA.

English

Cyanobacteria are a cheap source of nitrogen and quite suitable for farmers of developing countries. Although, they live in a diverse range of environments, different environmental variables influence their nitrogen fixing ability. Thus, this study evaluated the effect of pond depth and lining plastic colors on nitrogen fixing capacity of Anabaena species strain E3. Factorial combinations of four pond lining plastic colors and two depths were laid out in a complete randomized design with three replications. The ANOVA results revealed that the 20 cm depth pond had a higher mean growth rate (0.063 OD day -1 ), dissolved oxygen (17.63 mg L -1 ), dry biomass (0.58 g L -1 ) and total nitrogen (27.77 mg L -1 ) than the 40 cm depth. The highest mean growth rate (0.089 OD day -1 ), dissolved oxygen (19.07 mg L -1 ), dry biomass (0.66 g L -1 ) and total nitrogen (41.60 mg L -1 ) were registered in the treatment with a transparent lining plastic color. Moreover, it was noted that, the test strain under the treatment combinations of 20 cm depth and transparent plastic lining expressed the highest mean optical density (1.227), heterocyst frequency (2.92%) and pH (10.28). Therefore, for mass-production of E3-strain-based biofertilizer, the strain should be grown in ponds of 20 cm depth with transparent plastic. Keywords: Cyanobacteria, dry biomass, growth rate, shallow pond, total nitrogen

The expression and significance of DDR2 and MMP-13in human middle ear cholesteatoma

Objective:To ascertain the expression level of discoidin domain receptors 2(DDR2) and matrix metalloproteinase-13(MMP-13) in human middle ear cholesteatoma tissues and further investigate their roles in bone destruction and correlation.Method:Using immunohistochemical S-P method to detect the expression level of DDR2 ,MMP-13 in 30 specimens with middle ear cholesteatoma epithelial tissue and 10 specimens with normal ear epithelial tissues．At the same time,the computer image analysis system was used to detect the expression of the two indexes by the quantitative analysis,analyzing their expression in middle ear cholesteatoma epithelial tissue and the correlation between the extent of bone destruction．Result:The expression of DDR2 and MMP-13 were confirmed in human middle ear cholesteatoma epithelial tissues and normal ear epithelial tissues． The mean optical density of DDR2 and MMP13 in human middle ear cholesteatoma epithelial tissues which were tested by the computer image quantitative analysis system were higher than those in normal ear epithelial tissues(P<0．05).The expression of DDR2 and MMP-13 in middle ear cholesteatoma epithelial tissues were positively correlated(r=0.738,P<0.01).In addition,the two indexes were associated and relative to the extent of bone destruction,the wider the extent of bone destruction was,the higher the expression of both indexes(P<0．05)．Conclusion:DDR2 and MMP-13 may play important roles in bone destruction of human middle ear cholesteatoma．.

OPTICAL DENSITY OF CORTICAL BONE MATRIX IS DIMINISHED IN EXPERIMENTALLY INDUCED OSTEOPOROSIS

Osteoporosis is characterized by low bone mineral density (BMD) and abnormalities in bone structural and material properties, with unexplained low trauma fractures. The aim of the present study was to quantify the optical density of cortical bone matrix in rats with experimentally induced osteoporosis by ovariectomy. The experimental group was divided in two equal subgroups, the first sacrificed in the third month after ovariectomy and second sacrificed in the fifth month after ovariectomy. After decalcification, on routinely stained histopathologic sections optical density (OD), standard deviation of OD, mode OD, minimal and maximal OD of cortical bone matrix were estimated. Mean optical density and mode optical density of cortical bone were statistically higher in the control than in the experimental group (p<0.05). Maximal optical density of cortical bone was significantly lower in rats three months after ovariectomy than in other groups. Obtained results indicate that in experimentally induced osteoporosis the optical density of cortical bone matrix is diminished, similarly to low bone mineral density. Acta Medica Medianae 2016;55(2):35-39.

Preeclampsia is associated with low placental transthyretin levels

ObjectiveTo investigate the relationship between placental transthyretin (TTR) level and preeclampsia. Materials and MethodsPlacental tissues from uncomplicated and preeclamptic pregnancies were analyzed using immunohistochemistry and image analysis. We measured the mean optical density (OD) of immunohistochemical staining of TTR across multiple sections using Image Pro Plus 6.0. To avoid bias, we used placental tissue array, which contained preeclamptic placentas (n=8) and the control placentas (n=6) on the same slide. ResultsThe mean TTR OD of the syncytiotrophoblast layer of placentas (95% confidence interval) from the first trimester was higher than those from the second/third trimester, and term placentas [0.149 (0.014–0.285) for the 1st trimester, 0.037 (0.000–0.073) for the 2nd/3rd trimester, and 0.011 (0.035–0.056) for term; p<0.01]. Although the OD of the second/third trimester placentas appeared greater than that of term placentas, this was not statistically significant. The mean TTR OD of the syncytiotrophoblast layer of the severe preeclampsia group was lower than that of controls [0.010 (0.005–0.016) vs. 0.027 (0.013–0.041), p<0.05]. ConclusionThe immunohistochemical expression of TTR in the syncytiotrophoblast layer of the placenta decreased significantly after 12 weeks of gestation, paralleling the changing demands of thyroid hormone uptake into the placenta. The reduced TTR expression in the syncytiotrophoblast layer of the preeclamptic placenta might impair thyroid hormone uptake and contribute to the pathophysiology of the disease.

IgE anti-Haemophilus influenzae type b (Hib) antibodies detected in serum of Hib-vaccinated asthmatic and non-asthmatic pediatric patients.

Haemophilus influenzae type b (Hib) bacterium causes severe illness in infants and children, but has largely been eliminated by introducing a universal Hib conjugate vaccine. While effects of certain vaccinations on atopic disease have been studied, little is known about the relationship between Hib vaccination and diseases of altered immunoglobulin E (IgE) regulation (asthma or atopy). As such, it is necessary to provide more evidence concerning Hib vaccination as a possible risk factor for atopic disease. Total serum IgE and IgE-and IgG-anti-Hib antibody responses were studied in Hib vaccinated asthmatic (N.=14) and non-asthmatic children (N=26) (VaccZyme™ Human Anti Hib Enzyme Immunoassay Kit). Data are reported as mean optical density (OD) values. We found that: 1) total serum IgE levels were higher in asthmatic compared with non-asthmatic subjects (389±125 vs. 125±129, P<0.001); 2) IgE and IgG anti-Hib antibody responses were similar in both asthmatic and non-asthmatic subjects (0.722±0.279 and 0.681±0.280, respectively; P=0.65; 0.450±0.505 and 0.573±0.779, respectively; P=0.580). The universal Hib vaccine antigen did not result in either increased IgE, or IgG anti-Hib antibody responses in asthmatic or non-asthmatics subjects. Thus, in this cohort, no association between Hib vaccination and asthma status was identified.

Detection of Salmonella infection in slaughter cattle using meat juice and serum

Monitoring for Salmonella in slaughter cattle is important to enable targeted control measures to be applied on problem farms and at the abattoir. The aim of this study was to determine whether meat juice could be used as much as serum to identify slaughter cattle with a high prevalence of infection. Samples of meat juice &amp; serum were taken from 100 slaughter cattle and comparisons were made between the results of individual Enzyme-linked immunosorbent assay (ELISA) tests on serum and meat juice. The ELISA tests showed a statistically significant Serum mean optical density (O.D.) with meat juice mean optical density (O.D.) from seven animals. All but one of the seven positive individual sample serum O.D. and sample/positive control (S/P) ratio results correlated significantly with the results of the meat juice O.D. ELISA. The results show a generally good correlation between serological results of individual serum and meat juice samples in slaughter cattle. Thus, ELISA could be used to flag up potentially hazardous Salmonella contaminated meat from slaughter cattle or herds which are more likely to be in need of improved Salmonella control. This can then be confirmed by bacteriological sampling at the abattoir or farm level and a control plan imposed.

Effect of the Fructus Ligustri Lucidi extract and its monomers quercetin and oleanolic acid on the adhesion and migration of melanocytes and intracellular actin.

The present study aimed to investigate the effects of the Fructus Ligustri Lucidi (FLL) extract and its monomers quercetin and oleanolic acid on the adhesion and migration of human epidermal melanocytes (MCs) and intracellular actin. The human epidermal MCs were cultured and identified. The cells were treated with different concentrations of FLL extract, quercetin and oleanolic acid. The adhesion and migration abilities of the cells were determined by the fibronectin-coated culture experiment and Transwell assay, respectively. The structure and distribution of intracellular actin were observed by confocal laser microscopy, with semi-quantitative analysis. Results showed that compared with the control group, 0.0375-0.3 mg/ml of the FLL extract and 40 µM quercetin significantly improved the adhesion rate of MCs (P<0.05). The numbers of MCs permeating the microporous membrane in the 0.15 mg/ml FLL extract and 12 µM oleanolic acid groups were 43.7 and 30.3, respectively, significantly higher compared to the control group (P<0.01). In the control group, the intracellular actin was less, and the stress fiber structure was not clear. In the 0.15 mg/ml FLL extract, 12 µM oleanolic acid and 40 µM quercetin groups, there were numerous bunched stress fibers, indicating the aggregation of filamentous fibrous actin. The mean optical densities of actin expression in the 0.15 mg/ml FLL extract, 12 µM oleanolic acid and 40 µM quercetin groups were significantly higher compared to the control group (P<0.05). The FLL extract has a significant stimulatory effect on the adhesion and migration of human epidermal MCs. The mechanism may be associated with the promotion of intracellular actin cytoskeleton aggregation.

Influence of serum antibody on occurrence of Streptococcus zooepidemicus in the nasopharynx of weanling foals

Abstract Streptococcus zooepidemicus (Sz) is a tonsillar commensal of horses that opportunistically invades the respiratory tract of weanling foals causing purulent rhinitis and tracheobronchitis. Ten weanling foals in an isolated herd with a history of Sz-associated respiratory disease in recent years were immunized with recombinant proteins SzPNC78, endopeptidase, and hyaluronidase prior to weaning to determine the effect of serum antibodies on populations of nasopharyngeal Sz and mucopurulent nasal discharges. Levels of serum antibodies to all three proteins were significantly ( P ≤ .01) increased compared to levels in 10 nonimmunized control foals at weaning (day 0). Numbers of Sz in nasopharyngeal washes of control foals were initially higher on day 35 than in washes of immunized foals. However, this difference was not maintained, and counts in washes of both groups were similar over the following 7 weeks. By day 104 postweaning, Sz was no longer detected in washes of six foals in each group. Streptococcus zooepidemicus (ST-45) expressing a specific protective protein (SzP) phenotype dominated from days 35 to 104 in both groups. A second clone, ST-10, was detected intermittently in five immunized and two control foals. Mucopurulent nasal discharges of seven immunized and eight control foals on days 78 and 85 were associated with counts of Sz that ranged as high as 10 6 colony-forming units/mL and with significantly increased levels of SzP-specific IgG in washes. Increase in specific IgG but not IgA was temporary suggesting transudation through inflamed mucosa. This increase was generally greater in washes of immunized (mean enzyme-linked immunosorbent assay optical density [ELISA OD] = 1.73) than control (mean ELISA OD = 1.08) foals suggesting a benefit of systemic immunization may be therapeutic by enhancing clearance of Sz following inflammation provoked by local invasion.

Antibodies to microtubule-associated protein-2 in the cerebrospinal fluid are a useful diagnostic biomarker for neuropsychiatric systemic lupus erythematosus

Objective: Previous reports indicate that serum anti-microtubule-associated protein 2 (MAP-2) antibodies are common in sera from patients with neuropsychiatric systemic lupus erythematosus (NPSLE). Differential diagnosis of NPSLE is occasionally difficult because of differential diagnosis which can mimic NPSLE. Therefore, specific biomarkers for NPSLE are needed. We conducted this study to clarify whether cerebrospinal fluid (CSF) anti-MAP-2 antibodies are a useful diagnostic biomarker for NPSLE.Methods: Enzyme-linked immunosorbent assay was conducted to measure CSF concentrations of anti-MAP-2 and anti-ribosomal P antibodies and of IL-6 in NPSLE patients (n = 24) and non-NPSLE controls (n = 17). The non-NPSLE controls consisted of systemic lupus erythematosus patients with neuropsychiatric symptoms caused by non-NPSLE conditions (n = 10) and patients with other connective tissue diseases (n = 7).Results: Significantly higher anti-MAP-2 antibody titers were found in the CSF of patients with NPSLE versus non-NPSLE controls. The prevalence of anti-MAP-2 antibodies was 33.3% (8/24) in NPSLE patients when a positive cutoff value was 3 standard deviations above the mean optical density of non-NPSLE controls. None of the controls had anti-MAP-2 antibodies in their CSF. Both anti-ribosomal P antibody titers and concentration of IL-6 in the CSF were significantly higher in patients with NPSLE having anti-MAP-2 antibodies than in patients with non-NPSLE controls.Conclusion: Anti-MAP-2 antibodies could be detected in the CSF of 33.3% of patients with NPSLE, and its presence was highly specific for NPSLE. We propose that CSF anti-MAP-2 antibodies are a novel and useful diagnostic biomarker for NPSLE.

MAGI-2 in prostate cancer: an immunohistochemical study

Membrane-associated guanylate kinase, WW and PDZ domain-containing protein 2 (MAGI-2) is a scaffolding protein that links cell adhesion molecules, receptors, and signaling molecules to the cytoskeleton and maintains the architecture of cell junctions. MAGI-2 gene rearrangements have recently been described in prostate cancer. We studied the immunohistochemical expression of MAGI-2 protein in prostate tissue. Seventy-eight radical prostatectomies were used to construct 3 tissue microarrays consisting of 512 cores, including benign tissue, benign prostatic hyperplasia, high-grade prostatic intraepithelial neoplasia (HGPIN), and adenocarcinoma, Gleason patterns 3 to 5. Immunohistochemistry for phosphatase and tensin homologue (PTEN) and double-stain MAGI-2/p63 was performed and analyzed by visual and image analysis, the latter as percent of analyzed area (%AREA), and mean optical density multiplied by %AREA (STAIN). By visual and image analysis, MAGI-2 was significantly higher in adenocarcinoma and HGPIN compared with benign (benign versus HGPIN P < .001; benign versus adenocarcinoma, P < .001). HGPIN and adenocarcinoma did not significantly differ by either modality. Using visual intensity to distinguish benign tissue and adenocarcinoma, a receiver operating curve yielded an area under the curve of 0.902. A STAIN threshold of 1470 yielded a sensitivity of 0.66 and specificity of 0.96. There was a significant correlation between PTEN and MAGI-2 staining for normal and benign prostatic hyperplasia, but this was lost in HGPIN and cancer. We conclude that MAGI-2 immunoreactivity is elevated in prostate cancer and HGPIN compared with normal tissue, and suggest that MAGI-2 may contribute to prostate carcinogenesis. This is the first report of MAGI-2 staining by immunohistochemistry in prostate cancer.

Abstract 129: Optogenetic Inhibition of Striatal Neurons Improves the Survival of Implanted Neural Stem Cell and Neurological Outcomes After Ischemic Stroke

Background: Administration of neural stem cell (NSC) into peri-infarct region is a promising treatment to improve the outcome after brain ischemia. However, whether the survival of implanted NSC is influenced by endogenous neuronal activity remains unclear. Using optogenetic technique, we regulated the activity of striatal neuron and investigated whether the excitation or the inhibition of striatal neuronal activity could improve NSC survival and overall neurological outcome. Methods: Adeno-assosiated virus carrying ChR2 or ArchT gene under the control of promoter CaMKII was injected into mice striatum two weeks before the surgery of 60 minutes transient middle cerebral artery occlusion (tMCAO). NSCs cultured from transgenic mice expressing fluorescent protein or isometric phosphate buffer saline was transplanted into peri-infarct region at striatum at 4 days after tMCAO. The striatal neurons were stimulated 15 minutes daily by 473nm pulse or 530nm constant laser via implanted optical fiber from 7 to 13 days after tMCAO. Neurological severity score (NSS) was assessed at 3 and 14 days. After sacrificing the animals at 14 days after tMCAO, brain infarct volume was measured with cresyl violet staining. The fluorescence intensity and the diffuse area of implanted NSC were calculated to evaluate the survival of NSC. Results: Mice received NSC transplantation and optogenetic inhibition showed smaller infarct volume compared to that only received NSC transplantation or PBS (p&lt;0.05). The fluorescence diffuse area of transplanted NSC was larger in mice with optogenetic inhibition than that without optogenetic inhibition (p&lt;0.05). Mice received NSC transplantation and optogenetic excitation had larger infarct volume and higher NSS than mice only received NSC transplantation (p&lt;0.05), but showed no differences compared to mice received PBS (p&gt;0.05). The fluorescence diffuse area and the mean optical density of transplanted NSC was smaller in mice with optogenetic excitation than that without optogenetic excitation (p&lt;0.05). Conclusion: Inhibition of striatal neuronal activity at sub-acute phase after ischemia can improve the treatment of implanted NSC. To the opposite, excitation of striatal neuronal activity can reduce the NSC treatment.

A Survey for Contagious Caprine Pleuropneumonia in Agago and Otuke Districts in Northern Uganda

Background: Contagious Caprine Pleuropneumonia (CCPP) is a devastating disease of goats caused by Mycoplasma capricolum subsp. Capripneumoniae (Mccp). The disease was first confirmed in Uganda in 1995 in Karamoja region. Contagious Caprine Pleuropneumonia negatively impacts on goats’ productivity but its extent and magnitude among the local communities in Uganda remain unknown. A cross sectional study was conducted in the districts of Agago and Otuke neighboring Karamoja in Northern Uganda during the months of July and August 2011 to explore for the status of the disease. Methods: Five hundred and four serum samples from goats were obtained from randomly selected unvaccinated herds and 100 goats from vaccinated herds. Serum samples were examined for antibodies against Mycoplasma capricolum subsp. Capripneumoniae (Mccp) by ELISA method. A total of 162 semi-structured questionnaires were administered to selected farmers to obtain information on their understanding of the disease and the risk factors they associated with CCPP. Eight focus group discussions were also conducted with selected farmer groups to obtain detailed qualitative information on CCPP. Results: Among the unvaccinated goats, seroprevalence of CCPP was 32 (17.7%) and 52 (23.3%) for Agago and Otuke districts respectively. Levels of antibodies against Mccp were higher among vaccinated goats than unvaccinated ones (mean optical densities (ODs) of 0.905 and 0.776, p = 0.08). Majority of the farmers 121 (74.7%) had knowledge on CCPP and recognized that CCPP was among the major challenges to goat production in Uganda. Conclusions: This study demonstrated that CCPP was prevalent in Agago and Otuke districts, which are outside but close to Karamoja region where the disease was previously confirmed. There is a need for wider and detailed studies to investigate further CCPP in other districts of Uganda for effective preventive and control of CCPP in Uganda and the neighboring countries.

Histologic Evidence of New Collagen Formulation Using Platelet Rich Plasma in Skin Rejuvenation: A Prospective Controlled Clinical Study.

BackgroundPlatelet-rich plasma (PRP) is an autologous concentration of human platelets contained in a small volume of plasma and has recently been shown to accelerate rejuvenate aging skin by various growth factors and cell adhesion molecules.ObjectiveThis study was conducted to evaluate the efficacy and safety of intradermal injection of PRP in the human facial rejuvenation.MethodsThis study was a prospective, single-center, single-dose, open-label, non-randomized controlled clinical study. PRP injected to the upper site of this right infra-auricular area and all face. Saline was injected to the left infra-auricular area. Histopathological examinations were performed before PRP treatment, 28 days after the PRP, and saline (control) treatments.ResultsTwenty women ranging in age from 40 to 49 years (mean age, 43.65±2.43 years) were enrolled in the study. The mean optical densities (MODs) of collagen in the pre-treatment, control, and PRP-treated area were measured. They were 539±93.2, 787±134.15, 1,019±178, respectively. In the MOD of PRP, 89.05 percent improvement was found when MOD of PRP was compared with MOD of pre-treatment. The mean MOD of collagen fibers was clearly highest on the PRP side (p<0.001). The PRP-to-saline improvement ratio (89.05% to 46.01%) was 1.93:1. No serious side effects were detected.ConclusionPRP increases dermal collagen levels not only by growth factors, but also by skin needling (the mesotherapy technique 'point by point'). PRP application could be considered as an effective (even a single application) and safety procedure for facial skin rejuvenation.

Macular Pigment Optical Density in Chinese Primary Open Angle Glaucoma Using the One-Wavelength Reflectometry Method.

Purpose. To investigate macular pigment optical density (MPOD) and its relationship with retinal thickness in primary open angle glaucoma (POAG) patients using the one-wavelength reflectometry method. Methods. A total of 30 eyes from 30 POAG patients (18 males and 12 females, mean age 47.27 ± 16.93) and 52 eyes from 52 controls (27 males and 25 females, mean age 49.54 ± 19.15) were included in this prospective, observational, case-control study. MPOD was measured in a 7-degree area using one-wavelength reflectometry method. Two parameters, max and mean optical density (OD), were used for analyses. Spectral-domain-optical coherence tomography was used to measure retinal thickness, including central retinal thickness (CRT), the macular ganglion cell complex (GCC), and the circumpapillary retinal nerve fiber layer (RNFL). Results. Both maxOD and meanOD were significantly reduced in POAG patients compared with normal subjects (P < 0.001). GCC, CRT, and RNFL thicknesses were also significantly reduced in POAG patients (P < 0.001). GCC thickness had a positive relationship with MPOD. Conclusions. MPOD within the 7-degree area was significantly lower in Chinese POAG patients than in control subjects, and GCC thickness was significantly and positively associated with MPOD. Whether the observed lower MPOD in POAG contributes to the disease process or is secondary to pathological changes caused by the disease (such as loss of ganglion cells) warrants further and longitudinal study.

Effects of Propofol on Several Membrane Characteristics of Cervical Cancer Cell Lines

Background: Although significant advances have been made toward understanding the molecular mechanisms underlying the effect of propofol on tumor cell metastasis, less is known regarding how cell membrane and cytoskeletal ultrastructure are affected in this process. Here, we investigated the relationship between cell morphology and cell size, which are features mainly defined by the cytoskeleton. Methods: To confirm the effects of propofol on the migratory ability of human cervical carcinoma cells, cell migration and invasion were examined through scratch wound healing and transwell membrane assays. Furthermore, HeLa cells cultivated with different concentrations of propofol were examined by confocal microscopy and atomic force microscopy (AFM), and the mean optical density and migration ability of these cells were also assessed. In addition, cell membrane morphology was inspected using AFM. Results: The results of the wound healing and transwell membrane assays indicated that propofol decreases the migratory ability of cervical carcinoma cells compared to control cells. A comparative analysis of the test results revealed that short-term (3 h) exposure to propofol induced marked changes in cell membrane microstructure and in the cytoskeleton in a dose-dependent manner. These morphological changes in the cell membrane were accompanied by cytoskeleton (F-actin) derangement. The present findings demonstrate a close relationship between changes in cell membrane ultrastructure and cytoskeletal alterations (F-actin) in propofol-treated HeLa cells. AFM scanning analysis showed that cell membrane ultrastructure was significantly changed, including a clear reduction in membrane roughness. Conclusion: The influence of propofol on the HeLa cell cytoskeleton can be directly reflected by changes in cellular morphology, as assessed by AFM. Moreover, the use of AFM is a good method for investigating propofol-mediated changes within cytoskeletal ultrastructure.

Immunohistochemical localization of vasoactive intestinal peptide in bursa of fabricius of Chinese yellow quail

This study aimed to reveal the distribution and morphological characteristics of vasoactive intestinal peptide (VIP) at different ages in the bursa of Fabricius (BF) in Chinese Yellow Quail. The distribution and morphological characteristics of VIP of BF in Chinese yellow quails at different ages (0, 1, 3, 5, 9, 13, 17, 21, 25, 29, 32 and 36 weeks) were observed using the immunohistochemical streptavidin-biotin complex method after obtaining the sections and subsequent immunohistochemical staining. Then semi-quantitative analysis of the mean optical density (MOD), integral optical density (IOD) and mean gray value (MGV) for VIP immunoreactivity was performed through Image-Pro Plus 6.0 software. VIP was observed at all ages in Chinese yellow quail’s BF. It formed a long fusiform, oval or granular type structures with clear outline mainly localized in the submucosa, muscular layer and serosal connective tissue. At 25 weeks of age, the MOD value of VIP-positive cells reached the maximum, and then gradually reduced, while the MGV value was the lowest. We observed the localization and morphological characteristics of VIP in BF of Chinese yellow quail. VIP was mainly localized in the submucosa, muscular layer and serosal connective tissue of BF. Findings of this study may provide further understanding of the function of VIP in BF of Chinese yellow quails.

Correlation between neuropeptide distribution, cancellous bone microstructure and joint pain in postmenopausal women with osteoarthritis and osteoporosis

ObjectivesTo explore the relationship between the distribution of neuropeptides, cancellous bone microstructure and joint pain in postmenopausal women with osteoarthritis (OA) and osteoporosis (OP). MethodsCancellous bone of the femoral head was obtained at the time of hip arthroplasty from 20 postmenopausal women, 10 with OA and 10 with OP. Pain intensity was evaluated using the visual analog scale (VAS) before the operation. The microstructural parameters were measured with micro-CT and the neuropeptides of the cancellous bone were stained by an immunohistochemical method. ResultsWe observed that BV/TV, Tb.Th and Th.N values in the OP were significantly decreased compared to those in the OA. Immunohistochemical analysis revealed that the mean optical density (MOD) values for SP, CGRP, and VIP in the OA group were significantly higher than those in the OP, and the MOD value for NPY in the OA was significantly lower than that in the OP. We also observed that the MOD values for SP were positively correlated with AD, BV/TV, Tb.Th, Tb.N and Conn.D and negatively with MD, Tb.Sp and SMI in all patients. The MOD values for CGRP were positively correlated with AD, BV/TV and Tb.Th. MOD values for VIP were positively correlated with BV/TV and Tb.Th and negatively with SMI. The VAS score was correlated positively with the MOD values for SP, CGRP, VIP and negatively with NPY in all patients. ConclusionsNeuropeptides play an important role in the pathogenesis of OA and OP, which may cause pain and influence the bone microstructure.

Antiplatelet Glycoprotein Antibodies Profile in Adult Patients with Primary Immune Thrombocytopenia and the Comparison with Regards to the Treatment Response to Corticosteroids

Background: Platelet destruction in immune thrombocytopenia (ITP) is mediated by autoantibodies against platelet antigens. Anti-GPIIb/IIIa and anti-GPIb/IX antibodies are two major antibodies spesific for platelets and megakaryocytes. The diversity of clinical characteristics and therapeutic responses have been thought to be influenced by heterogeneity of spesific glycoprotein complexes. However, there has been no published data regarding either antiplatelet antibodies profile or response rates to corticosteroid, in adult patients with primary ITP.Objectives: To determine antiplatelet glycoprotein antibodies profile in adult patients with primary ITP and to compare the response rate to corticosteroid referring to the profile.Methods: This study is a cross sectional study in adult patients with primary ITP who visited Hematology Clinics in Dr Cipto Mangunkusumo Hospital, Jakarta, Indonesia, in a period of March to October 2013. The tests to determine the auto antibodies against the platelet GPIIb/IIIa and GPIb/IX surface antigens were performed by a direct MAIPA technique.Results: A total of 40 patients who were diagnosed as primary ITP have been enrolled into the study. The subjects of the study were divided in two groups, 10 subjects with newly diagnosed ITP, and 30 subjects with persistent or chronic ITP. Most of the subjects were female (82.5 %) with the median age was 24,5 (17-55) years old.. Seventy (??) subjects had ITP Bleeding Score (IBLS) 2.. In a persistent/chronic ITP group (n= 30), the proportion of anti-GPIIb/IIIa was 20 / 30 (66,67 %) with the median optical density (OD) was 0,461 (0,093-2,116) and the proportion of anti-GPIb/IX was 25/30 (83.33%) with the median OD was 0,507 (0,190-1,924). Meanwhile, in a newly diagnosed ITP group (n=10) the proportion of anti-GPIIb/IIIa was 7/10 (70%) with the mean OD was 0,802 ± 0,71 and the proportion of anti-GPIb/IX was 8/10 (80%) with the mean OD 0,82 ± 0,57. The response rates to corticosteroid were as follows: 42.5 % of the subjects achieved complete response, 45 % achieved response, and 12.5 % achieved no response. Comparison between the subjects with and without anti-GPIIb/IIIa and anti-GPIb/IX antibodies showed similarity response rates to prednisone.Conclusion: Sincemost of patients with primary ITP had anti GPIIb/IIIa and anti GPIb/IX antibodies, those antibodies may support a diagnosis of primary ITP. Since no differences in terms of the response rates to corticosteroid between subjects with and without anti-GPIIb/IIIa and anti-GPIb/IX antibodies, those antibodies can not be used as a predictor of the treatment DisclosuresSukrisman:Division of Hematology Medical Oncology: Other: Colleague.

Glucocorticoid-Induced Proliferation and Lack of Differentation in Untreated Pediatric Acute Myeloid Leukemic Blasts

BackgroundAlthough most pediatric acute myeloid leukemia (AML) protocols no longer prescribe glucocorticoids (GCs) as part of antileukemic treatment, several low-income countries may still rely on them. Furthermore these agents are globally being used in the prevention or treatment of chemotherapy-induced nausea, or in the treatment of acute respiratory distress syndrome in children with leukemia. Therefore, to study potential effects of GCs on AML cells remains important.The aim of this study was to evaluate the in vitro responsiveness of pediatric AML cells to GCs in relation to cell biological features.MethodsIn vitro drug resistance or potential proliferation upon GCs (prednisone) was determined using a methyl-thiazol-tetrazolium salt (MTT)-assay. Samples with more than 70% blasts at day 4 in the control wells and a control optical density (OD) >0.050 units were used to calculate the LC50-value, i.e. the PRD concentration lethal to 50% of the AML blasts. Samples were classified as highly sensitive (LC50 PRD ≤0.1 µg/ml), intermediately sensitive (LC50 PRD >0.1-150 µg/ml) or resistant (LC50 PRD ≥150 µg/ml). PRD-induced leukemic cell proliferation was defined as a leukemic cell survival (LCS=(OD drug-exposed well)/(mean OD control wells)) of >105% plus 2 times the standard deviation of the control cell survival in the PRD containing wells.Potential glucocorticoid (GC)-induced differentiation was evaluated using basic morphology. To validate the overall effects of GCs, a subset of randomly selected samples was tested for both prednisone and dexamethasone (DXM).Samples of patients treated according to (at that time) GC-containing protocols, provided by the BFM-Germany and the DCOG, were used to evaluate associations between in vitro effects and clinical outcome, both at an univariate and multivariate level.Within these protocols GCs were only administered during consolidation.ResultsApproximately 70% (n=117) of the 167 available pediatric AML samples could be evaluated for GC resistance and GC-induced proliferation. None of the samples were highly sensitive and only 15% (n=17) were intermediately sensitive to the lytic action of GCs. FAB type M1 samples were significantly more often intermediately sensitive than other FAB types (43% of the M1 samples versus 0-13% in FAB type M0, and M2-M6 samples) (P=0.006). FAB type M7 samples were more intermediately sensitive as well (67%, n=2), but sample numbers were small.GC-induced differentiation was not observed after evaluating 10 unselected samples.Contrary, 27% (n=32) of the samples showed proliferation upon GC-exposure. GC-induced proliferation occurred independently from spontaneous proliferation.Samples with FAB type M5 or activating FLT3 mutations were significantly more prone to this phenomenon: 63% (n=12) of the M5 samples compared to 0-33% of the other samples (P<0.001), and 60% (n=6) of the FLT3itd-positive samples versus 23% (n=22) of the negative samples (P=0.011) showed GC-induced proliferation. There were no significant differences in GC-induced proliferation between samples with favorable or other cytogenetics.The results for the subset of samples tested for both PRD and DXM were similar with regard to GC-resistance, GC-induced proliferation, and GC-differentiation.Probabilities of relapse free survival (pRFS) were lower for patients with GC-induced proliferation at a borderline significant level (P=0.045, univariately). However, in a multivariable COX model, including white blood cell count, cytogenetics, and FLT3 status an independent association between inferior pRFS and GC-induced resistance or GC-induced proliferation could not be confirmed.ConclusionAssuming that the proliferation induced in vitro is paralleled in vivo, it is tempting to speculate that GCs given during consolidation treatment may have an adverse effect on any present minimal residual leukemic cells, and that such exposure may increase the risk of relapse in a significant subset of patients.Although we did not study this in a prospective or in vivo design, given the lack of GC-induced differentiation effects, and the potential of GC-induced proliferation in this study - with at best unknown clinical consequences - the use of GCs in pediatric AML patients should be discouraged in general, and especially in subgroups with FAB type M5 or activating FLT3 mutations. DisclosuresKaspers:Galen Pharmaceuticals: Consultancy; Boehringer Ingelheim: Consultancy; Celgene: Consultancy.