Head and Neck Diseases by Alexandros G.Sfakianakis: 04/12/17

Τετάρτη 12 Απριλίου 2017

Anisotropic tough double network hydrogel from fish collagen and its spontaneous in vivo bonding to bone

Publication date: July 2017
Source:Biomaterials, Volume 132
Author(s): Md. Tariful Islam Mredha, Nobuto Kitamura, Takayuki Nonoyama, Susumu Wada, Keiko Goto, Xi Zhang, Tasuku Nakajima, Takayuki Kurokawa, Yasuaki Takagi, Kazunori Yasuda, Jian Ping Gong
Soft supporting tissues in the human body, such as cartilages and ligaments, are tough materials and firmly fixed to bones. These soft tissues, once injured, cannot regenerate spontaneously invivo. Developing tough and biocompatible hydrogels as artificial soft supporting tissues would substantially improve outcomes after soft tissue injury. Collagen is the main rigid component in soft connective tissues which is organized in various hierarchical arrays. We have successfully developed a novel class of collagen fibril-based tough hydrogels based on the double network (DN) concept using swim bladder collagen (SBC) extracted from Bester sturgeon fish. The DN hydrogels, SBC/PDMAAm, are composed of physically/chemically crosslinked anisotropic SBC fibril as the first network and neutral, biocompatible poly(N,N′-dimethylacrylamide) (PDMAAm) as the second network. The anisotropic structure of SBC fibril network, which is well retained in the DN hydrogels, is formed by free injection method, taking advantage of the excellent fibrillogenesis capacity of SBC. The denaturation temperature of collagen is improved in the DN hydrogels. These DN gels possess anisotropic swelling behavior, exhibit excellent mechanical properties comparable to natural cartilage. The 4 weeks implantation of the gels in the osteochondral defect of rabbit knee also shows excellent biomechanical performance in vivo. Furthermore, the hydroxyapatite (HAp) coated DN gels, HAp/SBC/PDMAAm gels, strongly bond to bone after 4 weeks. This new class of collagen-based hybrid DN gels, as soft and elastic ceramics, having good biomechanical performance and strong bonding ability with bone would expand the choice for designing next-generation orthopedic implants such as artificial cartilage, bone defect repair material in the load-bearing region of the body.

Graphical abstract

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Activatable near infrared dye conjugated hyaluronic acid based nanoparticles as a targeted theranostic agent for enhanced fluorescence/CT/photoacoustic imaging guided photothermal therapy

Publication date: July 2017
Source:Biomaterials, Volume 132
Author(s): Xiaolong Liang, Lei Fang, Xiaoda Li, Xu Zhang, Fan Wang
Targeted theranostic nano-system integrating functions of both diagnosis and therapy shows great potential for improving diagnosis and therapeutic efficacy. Herein, multifunctional nanoparticle based on activatable hyaluronic acid (HA) conjugating two near-infrared (NIR) dyes of Cy5.5 and IR825 was successfully designed and fabricated, and simultaneously used as a carrier for encapsulating perfluorooctylbromide (PFOB). In this system, PFOB showed good capability to absorb the X-rays, Cy5.5 on the outer surface acted as a fluorescent dye activatable by hyaluronidases (Hyals) in the tumor, and IR825 in the core as a photothermal agent. The obtained nanoparticles (NPs) of PFOB@IR825-HA-Cy5.5 can be utilized for triple X-ray computed tomography (CT), fluorescence and photoacoustic imaging. When PFOB@IR825-HA-Cy5.5 NPs were intravenously injected into the mice bearing HT-29 tumor, efficient tumor accumulation was clearly observed, as revealed by the triple modal imaging. An in vivo tumor treatment experiment was conducted by combination of PFOB@IR825-HA-Cy5.5 and near-infrared laser irradiation, achieving effective tumor ablation in mice. Therefore, PFOB@IR825-HA-Cy5.5 NPs is a safe, efficient, imageable photothermal nanoprobe, showing great potential for cancer theranostics.

Graphical abstract

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Peptide valency plays an important role in the activity of a synthetic fibrin-crosslinking polymer

Publication date: July 2017
Source:Biomaterials, Volume 132
Author(s): Robert J. Lamm, Esther B. Lim, Katie M. Weigandt, Lilo D. Pozzo, Nathan J. White, Suzie H. Pun
Therapeutic polymers have the potential to improve the standard of care for hemorrhage, or uncontrolled bleeding, as synthetic hemostats. PolySTAT, a fibrin-crosslinking peptide-polymer conjugate, has the capacity to rescue fibrin clot formation and improve survival in a model of acute traumatic bleeding. PolySTAT consists of a synthetic polymer backbone to which targeting fibrin-binding peptides are linked. For translation of PolySTAT, the optimal valency of peptides must be determined. Grafting of fibrin-binding peptides to the poly(hydroxyethyl methacrylate)-based backbone was controlled to produce peptide valencies ranging from 0 to 10 peptides per polymer. PolySTATs with valencies of ≈4 or greater resulted in increased clot firmness, kinetics, and decreased breakdown as measured by thromboelastometry. A valency of ≈4 increased clot firmness 57% and decreased clot breakdown 69% compared to phosphate-buffered saline. This trend was characterized by neutron scattering, which probed the structure of clots formed in the presence of PolySTAT. Finally, PolySTAT with valencies of 4 (100% survival; p = 0.013) and 8 (80% survival; p = 0.063) improved survival compared to an albumin control in a femoral artery injury model (20% survival). This work demonstrates tunability of hemostatic polymers and the ability of in vitro assays to predict in vivo efficacy.

Graphical abstract

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Determination of forces on a split palatal screw after rapid maxillary expansion

Abstract

Objectives

Aim of this study was to develop a finite element model of the forces that patients with rapid maxillary expansion bear and to validate it by a mechanical test.

Methods

Computer-aided design models of the metallic screw and polymeric splint were modelled and discretized. Two forces were generated and considered independently: F1 at the temporary molar (2.5 N) and F2 at the permanent molar (2.5 N). The results of the finite element analysis were used to define the strain values which the anterior and posterior arms of the rapid maxillary expansion appliance bore as a linear function of F1–F2 by calculating the strain–force coefficient δ _ij . Two strain gauge rosettes were attached to an appliance which was placed in an XY motorized stage to reproduce the same forces used in the finite element analysis. Once the system was validated, the matrix was inverted to determine forces F1 and F2 that a group of 40 patients underwent (median age 8.33 years, standard deviation 1.86 years) for 75 days, using their strain values. The parents of the patients activated a quarter turn (0.20 mm) twice a day until 50% transversal overcorrection was achieved.

Results

Finite element analysis showed that the effects of the forces on stress at the location of the arms were notably different. There was a satisfactory correlation between finite element analysis predictions and in vitro values. Dissipation of F1 and F2 in patients was predicted to be 62.5 and 80%, respectively, after 75 days of retention.

Conclusion

These results back the finite element analysis model for force prediction.

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International Journal of Otolaryngology and Head & Neck Surgery Vol.6,No.2 (March 2017)

Reinke's Oedema Presenting as Stridor: Implications for Otolaryngologists in Difficult Airway
Reinke's Oedema, Stridor, Difficult Airway and Micro Laryngeal Surgery
Paper Information Full Paper: PDF (Size:340KB)
DOI: 10.4236/ijohns.2017.62003

A Pedunculated Fibrolipomatous Polyp of Tonsil: A Rare Case Report
Palatine Tonsil, Lipoma, Fibrolipoma
Paper Information Full Paper: PDF (Size:765KB)
DOI: 10.4236/ijohns.2017.62004

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International Journal of Otolaryngology and Head & Neck Surgery Vol.6,No.2 (March 2017)

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Prophylactic atropine administration attenuates the negative haemodynamic effects of induction of anaesthesia with propofol and high-dose remifentanil: A randomised controlled trial.

BACKGROUND: Induction of anaesthesia with propofol and remifentanil often induces unwanted bradycardia and hypotension, raising concerns regarding tissue oxygenation. The electrophysiological cardiac effects of remifentanil can be reversed by atropine. OBJECTIVE: To investigate if prophylactic administration of atropine can attenuate the negative haemodynamic effects of propofol and a high dose of remifentanil during induction of anaesthesia. DESIGN: A double-blind, randomised controlled trial. SETTING: Single-centre, University Medical Center Groningen, The Netherlands. PATIENTS: Sixty euvolaemic patients scheduled for surgery under general anaesthesia. INTERVENTIONS: Anaesthesia was induced and maintained with a target-controlled infusion of propofol with a target effect-site concentration (Ce) of 2.5 [mu]g ml-1, remifentanil (target-controlled infusion), (Ce 8 ng ml-1) and cis-atracurium. Methylatropine (500 [mu]g) or 0.9% saline was administered at immediately before induction of anaesthesia. MAIN OUTCOME MEASURES: The changes ([DELTA]) in mean arterial pressure (MAP), heart rate (HR), cardiac index (CI), rate pressure product, cerebral tissue oxygenation and peripheral tissue oxygenation between induction of anaesthesia (T0) and 10 min later (T10). RESULTS: Atropine significantly attenuated the changes in the outcome measures between T0 and T10. Median (inter-quartile range) changes were MAP, [DELTA] = -24 (-40 to -21) vs. [DELTA] = -37 mmHg (-41 to -31) (P = 0.02); HR, [DELTA] = 0 +/- 13 vs. -19 +/- 11 bpm (P

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Effects of different fresh gas flows with or without a heat and moisture exchanger on inhaled gas humidity in adults undergoing general anaesthesia: A systematic review and meta-analysis of randomised controlled trials.

BACKGROUND: The minimum inhaled gas absolute humidity level is 20 mgH2O l-1 for short-duration use in general anaesthesia and 30 mgH2O l-1 for long-duration use in intensive care to avoid respiratory tract dehydration. OBJECTIVE: The aim is to compare the effects of different fresh gas flows (FGFs) through a circle rebreathing system with or without a heat and moisture exchanger (HME) on inhaled gas absolute humidity in adults undergoing general anaesthesia. DESIGN: Systematic review and meta-analyses of randomised controlled trials. We defined FGF (l min-1) as minimal (0.25 to 0.5), low (0.6 to 1.0) or high (>=2). We extracted the inhaled gas absolute humidity data at 60 and 120 min after connection of the patient to the breathing circuit. The effect size is expressed as the mean differences and corresponding 95% confidence intervals (CI). DATA SOURCES: PubMed, EMBASE, SciELO, LILACS and CENTRAL until January 2016. RESULTS: We included 10 studies. The inhaled gas absolute humidity was higher with minimal flow compared with low flow at 120 min [mean differences 2.51 (95%CI: 0.32 to 4.70); P = 0.02] but not at 60 min [mean differences 2.95 (95%CI: -0.95 to 6.84); P = 0.14], and higher with low flow compared with high flow at 120 min [mean differences 7.19 (95%CI: 4.53 to 9.86); P

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Prophylactic atropine administration attenuates the negative haemodynamic effects of induction of anaesthesia with propofol and high-dose remifentanil: A randomised controlled trial.

Effects of different fresh gas flows with or without a heat and moisture exchanger on inhaled gas humidity in adults undergoing general anaesthesia: A systematic review and meta-analysis of randomised controlled trials.

Differential Regulation of chTLR3 by Infectious Bursal Disease Viruses with Different Virulence In Vitro and In Vivo

Viral Immunology , Vol. 0, No. 0.

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The epigenetic mechanism for discordance of autoimmunity in monozygotic twins

Publication date: Available online 12 April 2017
Source:Journal of Autoimmunity
Author(s): Zhongyuan Xiang, Yuanqing Yang, Christopher Chang, Qianjin Lu
Monozygotic twins share an identical DNA sequence but are not truly "identical". In fact, when it comes to health and disease, they may often display some level of phenotypic discordance. The cause of this discordance is often unknown. Epigenetic modifications such as DNA methylation, histone modification, and microRNAs-mediated regulation regulate gene expression and are sensitive to external stimuli. These modifications may be seen to bridge the gap between genetics and the environment. Over the years, the importance of epigenetics as a primary mechanism for the role that the environment plays in defining phenotype has been increasingly appreciated. Mechanisms of epigenetics include DNA methylation, histone modifications and microRNAs. Discordance rates in monozygotic twins vary depending on the specific condition, from 11% in SLE to 64% in psoriasis and 77% in PBC. Other autoimmune diseases in which discordance is found among monozygotic twins has also been studied include type 1 diabetes, multiple sclerosis, rheumatoid arthritis, dermatomyositis and systemic sclerosis. In some cases, the differences in various epigenetic modifications is slight, even though the concordance rate is low, suggesting that epigenetics is not the only factor that needs to be considered. Nonetheless, the study of phenotypic discordance in monozygotic twins may shed light on the pathogenesis of autoimmune diseases and contribute to the development of new methodologies for the diagnosis and treatment of these diseases.

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Antimicrobial and anti-inflammatory activities, wound-healing effectiveness and chemical characterization of the latex of Jatropha neopauciflora Pax

Publication date: 23 May 2017
Source:Journal of Ethnopharmacology, Volume 204
Author(s): A.B. Hernandez-Hernandez, F.J. Alarcon-Aguilar, J.C. Almanza-Perez, O. Nieto-Yañez, J.M. Olivares-Sanchez, A. Duran-Diaz, M.A. Rodriguez-Monroy, M.M. Canales-Martinez
Ethnopharmacological relevanceJatropha neopauciflora Pax is an endemic species to Mexico, and its latex is used in traditional medicine to treat mouth infections when there are loose teeth and to heal wounds. In this research, we evaluated the antimicrobial activity, wound healing efficacy and chemical characterization of J. neopauciflora latex in a murine model.Materials and methodsThe antibacterial activity was determined using Gram positive and negative strains, the antifungal activity was determined using yeast and filamentous fungi, and the wound healing efficacy of the latex was determined using the tensiometric method. The anti-inflammatory activity was evaluated using the plantar oedema model in rats, administering the latex orally and topically. Cytotoxic activity was determined in vitro in two different cell lines. Antioxidant capacity, total phenolics, total flavonoids, reducing carbohydrates and latex proteins were quantified. The latex analysis was performed by High Performance Liquid Chromatography (HPLC). Finally, molecular exclusion chromatography was performed.ResultsThe latex demonstrated antibacterial activity. The most sensitive strains were Gram positive bacteria, particularly S. aureus (MIC=2mg/mL), and the latex had bacteriostatic activity. The latex did not show antifungal activity. The latex demonstrated a wound-healing efficacy, even the positive control (Recoveron). The orally administered latex demonstrated the best anti-inflammatory activity and was not toxic to either of the 2 cell lines. The latex had a high antioxidant capacity (SA50=5.4µg/mL), directly related to the total phenolic (6.9mg GAE/mL) and flavonoid (12.53µg QE/mL) concentration. The carbohydrate concentration was 18.52µg/mL, and fructose was the most abundantly expressed carbohydrate in the latex (14.63µg/mL, 79.03%). Additionally, the latex contained proteins (7.62µg/mL) in its chemical constitution. As secondary metabolites, the HPLC analysis indicated the presence of phenols and flavonoids.ConclusionsThe J. neopauciflora latex promotes the wound healing process by avoiding microorganism infections, inhibiting inflammation and acting as an antioxidant.

Graphical abstract

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Paeoniflorin and liquiritin, two major constituents in Chinese herbal formulas used to treat hyperprolactinemia-associated disorders, inhibits prolactin secretion in prolactinoma cells by different mechanisms

Publication date: 23 May 2017
Source:Journal of Ethnopharmacology, Volume 204
Author(s): Yuanyi Wei, Lei La, Lili Wang, Robert Batey, Chunxia Wang, Yuhao Li
Ethnopharmacological relevancePaeoniflorin and liquiritin are major constituents in some Chinese herbal formulas, such as Yiru Tiaojing (YRTJ) Granule (a hospitalized preparation) and Peony-Glycyrrhiza Decoction, used for hyperprolactinemia-associated disorders.Aim of the studyTo investigate the effect of paeoniflorin and liquiritin on prolactin secretion.Materials and methodsThe effect of YRTJ Granule on metoclopramide-induced hyperprolactinemia was tested in rats. Paeoniflorin and liquiritin in the YRTJ Granule extract were identified and quantified by HPLC. The effects of paeoniflorin and liquiritin on prolactin secretion were examined in prolactinoma cells that were identified morphologically and by Western blot. The concentration of prolactin was determined by ELISA. The gene expression was analyzed by Western blot.ResultsYRTJ Granule ameliorated metoclopramide-induced hyperprolactinemia in rats. The contents of paeoniflorin and liquiritin in YRTJ Granule were 7.43 and 2.05mg/g extract, respectively. Paeoniflorin, liquiritin and bromocriptine (a dopamine D2 receptor (D2R) agonist) decreased prolactin concentration in MMQ cells expressing D2R. However, the effect of liquiritin and bromocriptine was abolished in GH3 cells lacking D2R expression. Interestingly, paeoniflorin still decreased prolactin concentration in GH3 cells in the same manner. Furthermore, paeoniflorin suppressed prolactin protein expression, and was without effect on D2R protein expression in both MMQ and GH3 cells.ConclusionsThe present results suggest that paeoniflorin and liquiritin play a role in YRTJ Granule-elicited improvement of hyperprolactinemia. While the effect of liquiritin is D2R-dependent, paeoniflorin D2R-independently inhibits prolactin secretion in prolactinoma cells that may especially benefit the hyperprolactinemic patients who are refractory to dopaminergic therapies.

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Effects of Gnaphalium affine D. Don on hyperuricemia and acute gouty arthritis

Publication date: 5 May 2017
Source:Journal of Ethnopharmacology, Volume 203
Author(s): Hong-jian Zhang, Ling-nan Li, Jie Zhou, Qin-qin Yang, Pei-gang Liu, Pan Xu, Wei-qing Liang, Lin Cheng, Ying-qi Zhang, Jin-bao Pu, Yi-juan Hu
Ethnopharmacological relevanceThe Gnaphalium affine D. Don is used in China as a folk medicine to treat gout, anti-inflammatory, antitussive and expectorant activities. The aim of this study was to evaluate the potential of the extract of G. affine to treat hyperuricemia and acute gouty arthritis in animal model.Materials and methodsG. affine extract was evaluated in an experimental model with potassium oxonate (PO) induced hyperuricemia in mice which was used to evaluate anti-hyperuricemia activity and xanthine oxidase (XO) inhibition. Therapies for acute gouty arthritis was also investigated on monosodium urate (MSU) crystal induced paw edema model.ResultsG. affine extract showed expressive results on active in reducing serum uric acid (Sur) through effect renal mGLUT9 and mURAT1 mainly and inhibit XO activity in vivo. The extract of G. affine also showed significant anti-inflammatory activity and reduced the paw swelling on MSU crystal-induced paw edema model. Meanwhile, eight major compounds were identified by HPLC-ESI-QTOF-MS/MS.ConclusionsThe extract of G. affine showed significant effect on evaluated models and therefore may be active agents for the treatment of hyperuricemia and acute gouty arthritis.

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An overview on Leucosidea sericea Eckl. & Zeyh.: A multi-purpose tree with potential as a phytomedicine

Publication date: 5 May 2017
Source:Journal of Ethnopharmacology, Volume 203
Author(s): Tshepiso C. Mafole, Adeyemi O. Aremu, Thandekile Mthethwa, Mack Moyo
Ethnopharmacological relevanceLeucosidea sericea (the sole species in this genus) is a tree species found in southern Africa and possesses several therapeutical effects against infectious diseases in humans and livestock. This review aims to document and summarize the botany, phytochemical and biological properties of Leucosidea sericea.Materials and methodsUsing the term 'Leucosidea sericea', we systematically searched literature including library catalogues, academic dissertations and databases such as PubMed, SciFinder, Web of Science, Google Scholar and Wanfang. Taxonomy of the species was validated using 'The Plant List' (www.theplantlist.org).ResultsLeucosidea sericea remains a widely used species among the different ethnic groups in southern Africa. The species is a rich source of approximately 50 essential oils and different classes of phytochemicals (phenolics, phloroglucinols, cholestane triterpenoids, alkaloids and saponins) which may account for their diverse biological properties. Documented biological activities which were mainly observed under in vitro systems included antimicrobial, anti-parasitic, antioxidant, acetylcholinesterase (AChE) inhibition and anti-inflammatory properties. Preliminary safety tests on Leucosidea sericea extracts suggest moderate cytotoxic effects based on a few cell lines that were investigated.ConclusionsLeucosidea sericea possesses diverse medicinal potential with the antimicrobial, anti-parasitic, antioxidant and anti-inflammatory activities being the most prominent. The relative abundance and tendency of Leucosidea sericea to evade in nature suggest an abundant reservoir of raw materials for potential commercialization (upon validation of its pharmacological potential). However, more stringent investigations on the extracts (and isolated bioactive compounds) focusing on the mode of actions, which will inevitably unravel their pharmacokinetics, pharmacodynamics and clinical relevance remain pertinent.

Graphical abstract

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The senile functional evolution of the larynx after supracricoid reconstructive surgery.

The senile functional evolution of the larynx after supracricoid reconstructive surgery.

Eur Arch Otorhinolaryngol. 2016 Dec;273(12):4359-4368

Authors: Serra A, Maiolino L, Di Mauro P, Licciardello L, Cocuzza S

Abstract
Supracricoid partial laryngectomy is a relevant organ-preserving surgical technique used for the treatment of early or locally advanced laryngeal tumors with an oncologic outcome comparable with that of total laryngectomy. This reconstructive surgery is certainly technically feasible also in the elderly, but the problem is postoperatively as the patient may lack the will and strength to be successfully rehabilitated. The goal was to evaluate functional outcomes, in a maximum follow-up period of ten years from the end of the postoperative follow-up, in a cohort of elderly patients to look at the senile evolution of the laryngeal post surgical function comparing the amount of resection and grade of possible impairment of swallowing and phonatory parameters. A group of 33 patients were selected for a retrospective study aimed at evaluating the long-term evolution of laryngeal functionality after surgery, with the baseline at 5 years from the end of follow-up. All the patients had three-monthly visits for a period up to ten years, during which the objective and subjective swallowing and phonatory parameters were recorded, analyzed and, at the end of the study, compared with baseline. The results suggest that long-term post surgical functional problems may develop also in the neolarynx where the effects of the reconstructive surgery, on speech and swallowing, are largely stabilized. The presence of arytenoid resection had significantly impacted on the occurrence of aspiration even if the overall number of dysphagic patients was not statistically significant compared to baseline.

PMID: 27363403 [PubMed - indexed for MEDLINE]

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"Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi"[jour]; +16 new citations

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Food labeling issues in patients with severe food allergies: solving a hamlet-like doubt.

Purpose of review: We review the laws on labeling in the international community, the difficulties they pose to the food manufacturers to prepare the food labels and the methodologies to determine the concentration of potential allergens in foods. Recent findings: European Food Safety Authority and International Life Sciences Institute Europe are evaluating strategies to identify the threshold level of allergen that can trigger a reaction in individuals. The most used techniques to detect the presence of protein in food are Enzyme-linked immunosorbent assay, polymerase chain reaction and real time polymerase chain reaction. Researchers are now trying to apply proteomics to estimate the amount of protein within the food. In order to protect the health of consumers, the Codex Alimentarius Commission updates constantly the list of allergens. In response to these regulations, some industries have also added some precautionary allergen labeling (PAL). It was generally agreed that PAL statements needed to be visible, simple, and safe. It was suggested that PAL be standardized, an action that would occur if the 'Voluntary Incidental Trace Allergen Labelling' process was made mandatory. Summary: So far, no laboratory technique is able to reassure the consumers about the composition of foods found on the packaging. International authorities produced increasingly stringent laws, but more is still to do. Copyright (C) 2017 Wolters Kluwer Health, Inc. All rights reserved.

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Drug use among transgender people in Ontario, Canada: Disparities and associations with social exclusion

Publication date: September 2017
Source:Addictive Behaviors, Volume 72
Author(s): Ayden I. Scheim, Greta R. Bauer, Mostafa Shokoohi
IntroductionWe identified the prevalence and correlates of past-year illicit drug use among transgender people in Ontario, Canada, and disparities with the age-standardized non-transgender population.MethodsData on transgender persons aged 16+ (n=406) were obtained from Trans PULSE, a respondent-driven sampling (RDS) survey (2009–2010). Overall and sex-specific estimates of past-year drug use (cocaine and amphetamines, based on data availability) in the reference population were obtained from Ontario residents aged 16+ (n=39, 980) in the Canadian Community Health Survey (2009–2010), and standardized to the overall and gender-specific transgender age distributions. For regression analyses with Trans PULSE data, past-year drug use included drug types associated with high risk of physical, psychological, and social harm to the user, and RDS-II weights were applied to frequencies and prevalence ratios (PR) derived from blockwise logistic regression models.ResultsAn estimated 12.3% (95% CI: 7.7, 17.0) of transgender Ontarians had used at least one of the specified drugs in the past year, with no significant difference by gender identity. Transgender Ontarians were more likely to use both cocaine (standardized prevalence difference; SPD=6.8%; 95% CI=1.6, 10.9) and amphetamines (SPD=SPD=1.3%, 95% CI=0.2, 3.1) as compared to the age-standardized non-transgender population. History of transphobic assault, homelessness or underhousing, and sex work were associated with greater drug use among transgender persons.ConclusionsThe prevalence of cocaine and amphetamine use among transgender people in Ontario, Canada was higher than in the age-standardized reference population. Social exclusion predicted within-group variation in drug use among transgender persons.

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Variation of Housekeeping Genes in Clinical Isolates and Vaccine Strains of Bordetella pertussis.

Variation of Housekeeping Genes in Clinical Isolates and Vaccine Strains of Bordetella pertussis.

Clin Lab. 2017 Apr 01;63(4):809-815

Authors: Fathi M, Haghighi F, Shahcheraghi F, Abbasi E, Eshraghi SS, Ghourchian S, Zeraati H, Yaseri M, Douraghi M, Shokri F

Abstract
BACKGROUND: Bordetella pertussis causes serious contagious infections, primarily in childhood. A whole-cell vaccine, diphtheria-tetanus-whole cell pertussis (DTwP), has been used to protect against pertussis in children in Iran, but the pertussis cases have been increasing during recent years. We determined the allelic variation level of housekeeping genes in isolates recovered from pertussis patients and vaccine strains used in national vaccination program.
METHODS: Five clinical isolates, 2 vaccine strains and a Tohama I strain were studied through multilocus sequence typing (MLST) of housekeeping genes. The relatedness between STs, the founder, single- and double-locus variants (SLVs, DLVs) was determined using eBURST algorithm. The concordance between the type assignments by MLST and PFGE was determined.
RESULTS: In the 5 clinical isolates, 2 STs were identified, ST2 and ST79. The vaccine strains displayed two distinct allelic profiles assigned to ST1 and ST2. ST2 was predicted as founder and the remaining STs were SLVs of ST2. MLST and PFGE type assignments were 86.6% concordant.
CONCLUSIONS: The clinical isolates of B. pertussis were different from vaccine strains used in the national vaccination program. This study confirms the low level of variation in housekeeping genes of B. pertussis. MLST of virulent antigenic genes needs to be applied as a complementary method for the characterization of new ST-harboring isolates that may predominate periodically. The combination of these data allows rapid and efficient surveillance of currently circulating isolates. These data might elucidate the future trends and considerations for vaccine formulation and design.

PMID: 28397465 [PubMed - in process]

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Enforced miR-144-3p Expression as a Non-Invasive Biomarker for the Acute Myeloid Leukemia Patients Mainly by Targeting NRF2.

Enforced miR-144-3p Expression as a Non-Invasive Biomarker for the Acute Myeloid Leukemia Patients Mainly by Targeting NRF2.

Clin Lab. 2017 Apr 01;63(4):679-687

Authors: Sun X, Liu D, Xue Y, Hu X

Abstract
BACKGROUND: MicroRNAs (miRNAs) are endogenous non-coding RNAs of 19-25 nucleotides in length. Abnormal miRNA expression has been identified in various types of cancer. However, the aberrant expression of miR-1244 has not been reported in acute myeloid leukemia.
METHODS: First, the expression of miR-144-3p was explored in the bone marrow and peripheral blood of AML patients and healthy control. Then, we also evaluated the level of miR-144-3p in HL-60 cells. The possible target gene of miR-144-3p was predicted using TargetScan. Dual luciferase reporter assay was applied to validate the target gene of miR-144-3p. Cell viability and apoptosis on miR-144-3p was explored. Western blot analysis was applied to identify the downstream signaling of miR-144-3p.
RESULTS: Our data showed that miR-144-3p was markedly increased in both the peripheral blood and bone marrow of AML patients compared with healthy controls. Moreover, we also found an increased expression of miR144-3p in HL-60 cells. In comparison, NRF2 protein expression was significantly decreased in HL-60 cells. Dual luciferase reporter assays demonstrated that miR-144-3p significantly suppressed the relative luciferase reporter activity of a pmirGLO-NRF2-3'UTR. In accordance with the downstream effects of NRF2 overexpression, inhibition of miR-144-3p reduced cell viability and prompted apoptosis. More importantly, we found that the inhibition of miR-144-3p in HL-60 cells could not enhance Caspase-3 activation when NRF2 protein expression was silenced.
CONCLUSIONS: These findings suggest a potential oncogenic function of miR-144-3p in HL-60 cells, which is mainly achieved by targeting NRF2.

PMID: 28397476 [PubMed - in process]

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Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry (MALDI- TOF MS) for Early Identification of Septic Patients.

Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry (MALDI- TOF MS) for Early Identification of Septic Patients.

Clin Lab. 2017 Apr 01;63(4):839-844

Authors: Sekercioglu AO, Cekin Y, Ogunc D, Ongut G, Baysan BO, Colak D, Gunseren F, Donmez L

Abstract
BACKGROUND: Matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a powerful technique for the rapid identification of bacteria from growing colonies in routine cultures. In this study, we evaluated the feasibility of a 5-hour incubation on solid medium after sub-cultivation of positive blood culture broth without any preparation steps in order to speed up the identification of bacteria.
METHODS: In addition to standard laboratory protocols, a Columbia agar plate with 5% sheep blood was inoculated with 1 drop from the blood culture broth. After a 5-hour incubation period, a colony from the culture plate was submitted to MALDI-TOF MS.
RESULTS: A total of 1351 positive blood cultures (1299 monomicrobial and 51 polymicrobial) were analyzed. When compared to routine identification procedure results for positive blood cultures, 79.3% of isolates were correctly identified to the species level. When manufacturer-recommended score values were taken into account, MALDITOF MS correctly identified 98.4% of the isolates to the species level with a score of > 2.0, 89.1% with a score between 1.7 and 2.0, and 75.4% with a score of < 1.7.
CONCLUSIONS: ln our evaluation, a large majority of the S. aureus (91.5%) and Enterobacteriaceae (87.6%) were correctly identified at the species level. A 5-hour incubation period was found to be associated with moderate identification results for CoNS, Enterococcus spp., and nonfermentative gram negative bacilli, with failure being mostly observed with Streptococcus spp., Candida spp., and other gram positive bacteria. We believe that the performance of MALDI-TOF MS identification after short-term culture is directly related to the sufficient growth of microorganisms at 5 hours.

PMID: 28397470 [PubMed - in process]

http://ift.tt/2oBCnBJ

Association of Serum Interleukin-28B with Clinical Features, Laboratory Values and Radiographic Score in Rheumatoid Arthritis Patients.

Association of Serum Interleukin-28B with Clinical Features, Laboratory Values and Radiographic Score in Rheumatoid Arthritis Patients.

Clin Lab. 2017 Apr 01;63(4):757-764

Authors: Lv C, Chang QJ, Zhao F, Bi LQ, Li P

Abstract
BACKGROUND: Interleukin-28B (IL-28B) is a member of the interferon lambda family (also known as type III interferons), it has already been studied in many diseases, but it has almost never been studied in rheumatoid arthritis (RA). With this background, we determined the serum levels of IL-28B in RA patients and investigated its clinical significances.
METHODS: IL-28B levels were measured by enzyme-linked immunosorbent assay (ELISA) in 80 RA patients and 80 healthy controls. Radiographs were scored for total Sharp score (TSS).
RESULTS: Serum IL-28B levels were significantly lower in the RA than in the control (p = 0.02). Anticyclic citrullinated peptide (anti-CCP) antibodies and rheumatoid factor (RF) were negatively correlated with serum IL-28B levels. There was no association between serum IL-28B levels and TSS at baseline, but there was a significant difference in change of IL-28B during 6 months follow up between progressors and non-progressors. The disease activity of RA patients decreased, but no difference was observed in serum IL-28B levels between before and after disease-modifying anti-rheumatic drugs (DMARDs) therapy.
CONCLUSIONS: These findings indicated a role of IL-28B in RA and it may contribute to avoiding osteoclasia in RA patients.

PMID: 28397468 [PubMed - in process]

http://ift.tt/2oBlRll

Efficiency of Leukocyte Differential Using Flow Cytometry with CytoDiff in Different Workflows.

Efficiency of Leukocyte Differential Using Flow Cytometry with CytoDiff in Different Workflows.

Clin Lab. 2017 Apr 01;63(4):659-668

Authors: Qu C, Wang J, Pu C, Xing Y, Shang K, Dong N, Li X

Abstract
BACKGROUND: To set up the review criteria for flow cytometry with CytoDiff (FCC) and evaluate the efficiency of different workflows using combinations of FCC with a hematology analyzer and microscopy.
METHODS: Leucocyte differentials of the samples from 995 clinical specimens and 278 specimens from healthy donors were analyzed using a hematology analyzer, FCC, and microscopy.
RESULTS: The correlations between the hematology analyzer, FCC, and microscopy were good for neutrophils, lymphocytes, and monocytes, but not in the case of basophils (r = 0.464, 0.358, 0.33) and eosinophils (r = 0.69, 0.67, 0.621). As a reference method of WBC differential using microscopy, the threshold of blasts for FCC was defined by a ROC curve at 1% (specificity 97.9%, sensitivity 97.5%, AUC 0.989). The optimal cutoff of immature granulocytes for FCC was 1% (specificity 85.8%, sensitivity 76.5%, AUC 0.866). The optimal cutoff of lymphocytes, neutrophils, and monocytes were 50%, 85% and 12%, respectively. According to the review criteria, the workflow of CBC after FCC and then microscopy had the highest sensitivity (97.07%), lower false negative rate (2.93%), and higher accuracy (80.3%) compared with others.
CONCLUSIONS: Our study integrated FCC into a WBC differential workflow in a routine laboratory and, for the first time, demonstrates the efficiency of different workflows. It can be used for reference in the selection of different hematology workflows.

PMID: 28397483 [PubMed - in process]

http://ift.tt/2o8OpyJ

Genetic Polymorphism of Epidermal Growth Factor rs4444903 Influences Susceptibility to HCV-Related Liver Cirrhosis and Hepatocellular Carcinoma in a Chinese Han Population.

Genetic Polymorphism of Epidermal Growth Factor rs4444903 Influences Susceptibility to HCV-Related Liver Cirrhosis and Hepatocellular Carcinoma in a Chinese Han Population.

Clin Lab. 2017 Apr 01;63(4):845-850

Authors: Zhang S, Qiao K, Trieu C, Huo Z, Dai Q, Du Y, Lu W, Hou W

Abstract
BACKGROUND: Genetic polymorphism in the epidermal growth factor (EGF, rs4444903) gene has been demonstrated to be associated with the clinical deterioration in hepatitis C virus (HCV)-related liver cirrhosis (LC) and the development of hepatocellular carcinoma (HCC). Whether this single nucleotide polymorphism (SNP) influences susceptibility to HCV-related LC and HCC in the Chinese Han population is largely unknown.
METHODS: In this case-control study, a total of 187 Chinese Han patients with chronic HCV infection were enrolled, including 62 HCV-related LC patients, 46 HCV-related HCC patients, and 79 chronic hepatitis C (CHC) patients without LC and HCC, and the genetic polymorphism was genotyped via a matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) assay. The logistical regression analysis was employed to determine the correlation between the genetic polymorphism and risk of HCV-related LC and HCC.
RESULTS: The distribution of EGF rs4444903 genotypes and alleles significantly differed between LC patients and CHC subjects (p = 0.045, p = 0.043, respectively). Under the recessive model, the GG genotype was significantly associated with a two-fold risk of HCV-related LC compared to the AA+AG genotype after an adjustment for age, gender, body mass index (BMI), duration of HCV infection, and HCV RNA level (OR = 2.188; 95% CI = 1.072 - 4.465; p = 0.031). Significant association was observed as well between the GG genotype and increased HCV-related HCC risk (OR = 3.104; 95% CI = 1.319 - 7.307; p = 0.010).
CONCLUSIONS: The EGF rs4444903 GG genotype is associated with higher susceptibility to HCV-related LC and HCC in the Chinese Han population. Screening of host genetic polymorphisms might be helpful in designing effective and efficient LC and HCC surveillance programs for chronic HCV-infected patients.

PMID: 28397482 [PubMed - in process]

http://ift.tt/2o8yLDG

Diagnosis of Hepatitis C Virus Infection Using a Novel Mouse Monoclonal Antibody to Detect Serum HCV E1/E2 Antigens.

Diagnosis of Hepatitis C Virus Infection Using a Novel Mouse Monoclonal Antibody to Detect Serum HCV E1/E2 Antigens.

Clin Lab. 2017 Apr 01;63(4):669-678

Authors: Touni I, Tabll AA, Sobieh SS, Hewedy MA, Abd YS, Viazov S

Abstract
BACKGROUND: Hepatitis C virus (HCV) is a major cause of liver disease worldwide and in Egypt. The aim of this study was to detect HCV E1/E2 antigens using a novel mouse monoclonal antibody (mAb) designated (7G9) as a diagnostic and alternative approach for HCV detection.
METHODS: The detection of HCV-E1/E2 antigens in 138 patients positive for HCV infection tested by RT-PCR and 25 healthy individuals negative for HCV as control group was done by an optimized in-house ELISA and DotELISA (based on the molecular mimicry of E2 to immunoglobulins).
RESULTS: The mAb (7G9) was found to be IgM (heavy-chain)/kappa (light-chain) and characterization by western blot revealed two bands at 63 kDa for E2 and 31 kDa for E1. ELISA peptide mapping showed high reactivity with peptide derived from HCV E1 (a.a. 315 - 323) and low reactivity to peptides derived from HCV E2 (a.a. 517 - 531) and HCV E2 (a.a. 412 - 419). The mAb (7G9) showed no reactivity with HBV Ag, S. typhi or B. Abortus Ag proving high specificity. AUC for HCV-E1/E2 detection was 0.96 for all HCV patients with sensitivity 87% (119/137), specificity 88% (22/25) and efficiency 87%. The HCV-E1/E2 antigens detection by Dot-ELISA showed 76.8% sensitivity, 88% specificity and the efficiency of the assay was 78.5%. Furthermore, no correlation was found between serum HCV viral load and HCV E1/E2 antigens detection.
CONCLUSIONS: The ELISA and Dot-ELISA based on the monoclonal antibody (7G9) are reliable, rapid, easy and economic diagnostic assays for HCV infection.

PMID: 28397481 [PubMed - in process]

http://ift.tt/2o8OvWV

Genetic Polymorphism of MTHFR C677T Influences Susceptibility to HBV-Related Hepatocellular Carcinoma in a Chinese Population: a Case-Control Study.

Genetic Polymorphism of MTHFR C677T Influences Susceptibility to HBV-Related Hepatocellular Carcinoma in a Chinese Population: a Case-Control Study.

Clin Lab. 2017 Apr 01;63(4):787-795

Authors: Qiao K, Zhang S, Trieu C, Dai Q, Huo Z, Du Y, Lu W, Hou W

Abstract
BACKGROUND: Methylene tetrahydrofolate reductase (MTHFR) is the key enzyme of folic acid metabolism and the C677T mutation is associated with decreased enzyme activity. Several studies have shown its regulatory role in carcinogenesis and tumor growth. HBV (hepatitis B virus)-related HCC (hepatocellular carcinoma) is one of the most common liver cancers worldwide. Therefore, the present case-control study aimed to investigate the role of genetic polymorphism of MTHFR C677T in the development and progression of HBV-related HCC in a Chinese population.
METHODS: Subjects enrolled included 204 HBV-related HCC patients and 211 HBV infected patients without HCC. MTHFR C677T polymorphism was genotyped via a DNA microarray-based assay. The relationship between the MTHFR C677T polymorphism and HBV-related HCC was analyzed.
RESULTS: The genotype frequencies of MTHFR C677T were statistically different between the HCC and control groups (p = 0.025). The TT genotype was associated with elevated risk of HBV-related HCC in a Chinese population under different genetic models after an adjustment for age, gender, HBV infection duration, and HCC family history (T vs. C, OR = 1.462, 95% CI: 1.090 - 1.962, p = 0.011; TT vs. CC, OR = 2.151, 95% CI: 1.143 - 4.049, p = 0.018; TT vs. CC+CT, OR = 1.918, 95% CI: 1.215 - 3.026, p = 0.005). When stratified with the known duration of HBV infection, subjects with HBV infection duration of more than 20 years and carrying the homozygous TT genotype had a higher susceptibility to HCC than those with the C allele (CC/CT) (OR = 2.568, 95% CI: 1.244 - 5.303; p = 0.011). There was no significant association between MTHFR C677T genotypes and HCC stages based on BCLC staging system.
CONCLUSIONS: MTHFR C677T polymorphism with TT genotype could be a factor that increases the risk of HBVrelated HCC in a Chinese population, especially those with HBV infection duration of more than 20 years.

PMID: 28397480 [PubMed - in process]

http://ift.tt/2o8MGt2

Estimation of Diagnostic Markers in Rheumatoid Arthritis and Ankylosing Spondylitis.

Estimation of Diagnostic Markers in Rheumatoid Arthritis and Ankylosing Spondylitis.

Clin Lab. 2017 Apr 01;63(4):797-800

Authors: Dashti N, Javadi Z, Safari F, Farahvash S, Zarebavani M

Abstract
BACKGROUND: Laboratory examination is a great value to confirm a diagnosis and estimate disease activity of Rheumatoid Arthritis (RA) and Ankylosing Spondylitis (AS). Since little research related to the diagnosis of RA and AS has been done in Iran, the aim of the present study was to evaluate the diagnosis markers in patients with RA and AS.
METHODS: This study was conducted among 104 patients with RA and 42 patients with AS in Iran during 2016. Inclusion criteria were according to the American College of Rheumatology (ACR) and European League against Rheumatism (EULAR) criteria. Five mL blood samples were collected from all patients. Laboratory studies consisted of ESR and anti-CCP tests and also determination of the presence of RF and CRP in these patients. Finally, the statistical analysis test was conducted using SPSS version 16.0.
RESULTS: The study patients with RA included 23 males (22.1%) and 81 females (77.9%) with the average age of 50.25 ± 14.34. In RA patients, 82.7% were RF positive and 17.3% were negative. Also, 49% were CRP positive and 51% were negative. The mean ESR in RA patients was 27.76 ± 20.17 mm/hour. The mean levels of anti-CCP were 109.15 ± 90.55 IU/mL. The mean ages of 42 patients with AS, including 30 male (71.4%) and 12 females (28.6%), were 38.69 ± 11.82 years. Among them, 24 patients (57.1%) were positive for CRP and 18 patients (42.9%) were negative. Only 2 patients (4.8%) were RF positive. 24 patients (57.1%) were positive for HLA-B27 and 18 patients (42.9%) were negative. The mean levels of ESR in AS patients were 30.30 ± 26.98 mm/hour.
CONCLUSIONS: Early diagnosis of the Rheumatoid arthritis and Ankylosing Spondylitis can help to prevent complications and its progression and help patients to recover more quickly.

PMID: 28397479 [PubMed - in process]

http://ift.tt/2o8Tbfx

Point of Care Cardiac Troponin Assay Analytical Performances for their Use in Clinical Routine.

Point of Care Cardiac Troponin Assay Analytical Performances for their Use in Clinical Routine.

Clin Lab. 2017 Apr 01;63(4):851-854

Authors: Dupuy AM, Baillet S, Dumont R, Giraud I, Badiou S, Bargnoux AS, Kuster N, Roubille F, Cristol JP

Abstract
BACKGROUND: We report the analytical and clinical performances of the Alere Triage Cardiac3© Panel on the Triage MeterPro© instrument, comparing concordance with hs-cTnT results from central laboratory above the respective 99th percentiles and determining the clinical sensitivity within the framework of AMI.
METHODS: The concordance was obtained with these two methods among unselected patients admitted to both the emergency and cardiology departments.
RESULTS: The LoD of the assay is 0.010 µg/L. At 99th percentile (0.02 µg/L) the CV was found to be 18%, below the clinically acceptable cutoff of 20%. In the overall population, ROC AUC was not significantly different between the central laboratory assay and POC assay, with 0.952 (95% CI, 0.918 - 0.952) for hs-cTnT concentrations at presentation and 0.953 (95% CI, 0.912 - 0.953) for cTnI. Sensitivity and specificity of hs-cTnT vs. cTnI for AMI (n = 32) were 97% and 78% vs. 91% and 86%, respectively. Our results indicated 90.4% concordance between the two methods using the 99th percentile specific for each assay. The Kappa coefficient was higher than 0.75, and the strength of agreement could be considered to be good.
CONCLUSIONS: The results of cTnI Alere assays provide similar clinical classification of patients, particularly for the AMI group as compared to the central laboratory hs-cTnT assay and could be suitable for clinical in accordance with the recommendations of Global Task Force guidelines.

PMID: 28397478 [PubMed - in process]

http://ift.tt/2o8BUTT

Real-time PCR Detection of the Most Common Bacteria and Viruses Causing Meningitis.

Real-time PCR Detection of the Most Common Bacteria and Viruses Causing Meningitis.

Clin Lab. 2017 Apr 01;63(4):827-832

Authors: Akkaya O, Guvenc HI, Yuksekkaya S, Opus A, Guzelant A, Kaya M, Kurtoglu MG, Kaya N

Abstract
BACKGROUND: Central nervous system (CNS) infections require prompt diagnosis, as the clinical condition progresses rapidly and may lead to severe permanent sequelae or death. The causative agents include viruses, bacteria, fungi, and parasites. In this study, samples with the diagnosis of CNS infection based on cerebrospinal fluid (CSF) sent to us from other hospitals/labs, were studied by multiplex real-time Polymerase Chain Reaction (PCR) method. The purpose of this study is to demonstrate, retrospectively, the most common bacteria and viruses causing meningitis and seasonal distribution of these agents using the multiplex real-time PCR method in CSF samples.
METHODS: This study retrospectively evaluated the results of 470 CSF specimens that had been sent to the Molecular Unit of our hospital with a pre-diagnosis of CNS infection and had been tested with the PCR method between January 2014 and December 2015. Specimens were tested using multiplex real-time PCR assay for Adenovirus (AdV), Cytomegalovirus (CMV), Enteroviruses (EV) (Polioviruses, Coxackieviruses, Echoviruses, and other enteroviruses), Epstein- Barr virus (EBV), Herpes simplex virus 1 and 2, Human Herpes virus 6 and 7, Varicella-zoster virus (VZV), Human Parechoviruses and Parvovirus B19, Hemophilus influenzae, Streptococcus pneumoniae or Neisseria meningitidis. (FTD NEURO9 and FTD Bacterial meningitis, multiplex real-time PCR Kit).
RESULTS: A bacterial or viral agent was identified in 98 (21%) of the 470 CSF samples. Of the patients, 85% were children and 15% were adults. Of the 98 positive samples, 22 (22.5%) patients were 15 years or older, and the remaining 76 (77.5%) were younger than 15 years. While Enterovirus (25%) was the most frequently identified agent, Adenovirus ranked second (22%) and Streptococcus pneumoniae ranked third (15%) in total. Positivity was highest in the 0 - 5-year age range. Bacteria were detected with the PCR method in 22 patients: S. pneumonia in 14, and N. meningitidis in 8. In cultures, S. pneumonia grew only in 7 and N. meningitidis in one. EV and AdV were seen in the summer months. The two coexisted in 3 (3%) patients.
CONCLUSIONS: Early diagnosis and treatment of meningitis are very important for reducing its mortality and morbidity. In patients with suspected meningitis, early detection of the responsible agents may be possible with molecular methods, such as PCR. Significant economic benefits may be obtained by preventing unnecessary antibiotic use and hospitalizations through the early detection of the microbial agents.

PMID: 28397477 [PubMed - in process]

http://ift.tt/2o8Bgpk

Evaluation of MALDI-TOF-MS for the Identification of Yeast Isolates Causing Bloodstream Infection.

Evaluation of MALDI-TOF-MS for the Identification of Yeast Isolates Causing Bloodstream Infection.

Clin Lab. 2017 Apr 01;63(4):699-703

Authors: Turhan O, Ozhak-Baysan B, Hernandez OZ, Er H, Ongut G, Ogunc D, Colak D, Cuenca-Estrella M

Abstract
BACKGROUND: Infections due to Candida species are major causes of morbidity and mortality in humans, causing a diverse spectrum of clinical disease ranging from superficial and mucosal infections to invasive disease. Several authors have demonstrated that mortality is closely linked to both timing of therapy and/or source control. The rapid identification of pathogenic species is helpful to start timely and effective antifungal therapy. The aim of this study was to assess the performance of the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) system for the correct and rapid identification of yeast isolates causing bloodstream infection.
METHODS: Between January 2014 and January 2015, a total of 117 yeast like organisms isolated from blood culture samples of 117 episodes from 102 patients who had blood stream infections were included in the study. The isolates were identified by MALDI-TOF MS. The results were compared with those obtained by the standard mycological methods and/or sequence analysis.
RESULTS: One hundred and seventeen yeast isolates including 115 Candida spp and two non-Candida yeasts were analysed. The Biotyper correctly identified 115 (98.3%) isolates to the genus level and 102 (87.2%) isolates to the species level using the manufacturer's recommended cutoff scores.
CONCLUSIONS: The Bruker Biotyper is a rapid, easy, inexpensive, and highly reliable system for the identification of yeast isolates. Early identification with MALDI-TOF MS would save time for determination of antifungal susceptibility and proper treatment strategy. The expansion of the database of the library by addition of less common species will improve the performance of the system.

PMID: 28397475 [PubMed - in process]

http://ift.tt/2oBwkxc

Evaluation of the Three Customized MSI Panels to Improve the Detection of Microsatellite Instability in Gastric Cancer.

Evaluation of the Three Customized MSI Panels to Improve the Detection of Microsatellite Instability in Gastric Cancer.

Clin Lab. 2017 Apr 01;63(4):705-716

Authors: Park J, Shin S, Yoo HM, Lee SW, Kim JG

Abstract
BACKGROUND: We designed and evaluated the suitability of three customized microsatellite instability (MSI) panels using a combination of mono- and dinucleotide markers to improve the detection of MSI status in 56 matched normal and gastric cancer specimens.
METHODS: An MSI analysis was performed to optimize the panel of microsatellite markers to detect instability using two different microsatellite panels: (1) mononucleotide marker panel consisting of mononucleotide markers BAT25, BAT26, BAT40, BAT-RII, NR21, NR22, NR24, and NR27 and (2) dinucleotide marker panel containing D2S123, D5S346, D17S250, D17S261, D17S520, D18S34, and D18S58. The customized panels consisted of five, seven, or ten markers with two, three, or four mononucleotide markers, respectively, among fifteen MSI markers described above to fulfill the MSI-H and MSI-L definition based on the revised Bethesda Guidelines. The "Proposal5" panel consisted of BAT40, BAT26, D18S34, D2S123, and D17S520. "Proposal-7" consisted of "Proposal-5" with BAT25 and D18S58. "Proposal-10" consisted of "Proposal-7" with NR27, D17S250, and D17S261.
RESULTS: Immunohistochemical staining for MMR protein expressions such as mutL homolog 1 (MLH1) and mutS homolog 2 (MSH2) revealed that among 56 matched specimens, 13 had defective DNA mismatch repair (MMR) proteins and 43 had proficient MMR proteins. Out of thirteen specimens with defective MMR expression, eight specimens (62%, 8/13) were classified as MSI-H with an instability at ≥ 6 markers and five (38%, 5/13) were MSIL with instability at ≤ 5 markers using all fifteen MSI markers. On the other hand, the analytical sensitivity and specificity of all three customized panels to detect MMR-deficient specimens were 92% (12/13) and 100% (43/43), respectively. In comparison, the sensitivity and specificity of the Bethesda and QMR panels were 62% (8/13) and 100% (43/43). All customized panels could represent the detection of MSI-L tumors rather than the Bethesda and the QMR panels.
CONCLUSIONS: The increased sensitivity to detect MSI-unstable tumors with customized panels including BAT40 and D18S34 indicates that precise MSI screening to discriminate MSI-H from MSS and MSI-L may be feasible for gastric cancer.

PMID: 28397474 [PubMed - in process]

http://ift.tt/2oBkvqP

Factors Contributing to False-Negative Enzyme-Linked Immunospot Assay for Interferon-Gamma Results in Active Tuberculosis.

Factors Contributing to False-Negative Enzyme-Linked Immunospot Assay for Interferon-Gamma Results in Active Tuberculosis.

Clin Lab. 2017 Apr 01;63(4):773-779

Authors: Lian G, Du F, Wu H, He M, Liu Z

Abstract
BACKGROUND: The enzyme-linked immunospot assay test (T-SPOT.TB) is effective for detecting TB, but there are still some persons with tuberculosis (TB) who have negative results. The aim of this study was to analyze the factors that contribute to false negative results in active TB patients.
METHODS: We investigated the clinical trial and laboratory data of the active TB patients hospitalized in the First Affiliated Hospital of Xi'an Jiaotong University from May 2013 to November 2014.
RESULTS: In 556 active TB patients, 198 infected sites were pulmonary, 358 infected sites were extra pulmonary, and 13.85% (77/556) had false negative T SPOT.TB results. The univariate and multivariate analysis showed that age and bone/joint TB were the factors contributing to the T-SPOT. TB false negative results and pleural TB were related to true positive results.
CONCLUSIONS: The site of TB may significantly affect the sensitivity of T-SPOT.TB. False negative results of TSPOT.TB are associated with older age and bone/joint TB, so in older adult patients or those with bone/joint TB, the exclusion of TB should be regarded with caution when T-SPOT.TB results are negative.

PMID: 28397473 [PubMed - in process]

http://ift.tt/2oBhPJO

Comparison of Novel Blood Culture System DL-Bt112TM with BacT/Alert 3DTM by Using Clinical Samples.

Comparison of Novel Blood Culture System DL-Bt112TM with BacT/Alert 3DTM by Using Clinical Samples.

Clin Lab. 2017 Apr 01;63(4):781-786

Authors: Altindis M, Demiray T, Koroglu M, Kilic U, Alagoz AN, Ogutlu A, Karabay O

Abstract
BACKGROUND: Blood cultures are the main diagnostic laboratory tool to detect bloodstream infections. Many clinical microbiology laboratories utilize automated blood culture systems to isolate infectious agents from blood samples. The diagnostic performance and time-to-detection values of the novel automated blood culture system, DLBt112TM (DL), was compared with BacT/Alert 3DTM (B3D) in this prospective comparative study with clinical samples.
METHODS: A total of 356 blood culture sets (178 sets for each system) were evaluated over a 6-month period in a university hospital. Two sets of blood culture samples (one for DL and one for B3D) were drawn from intensive care unit patients who were suspected to have bloodstream infections. BacT/ALERT FA FAN® Aerobic/Anaerobic blood culture bottles for B3D and FAN adult anaerobic/aerobic blood culture bottles for DL were used. The Vitek® 2 automated system was used for identification of the isolated bacteria.
RESULTS: We evaluated 178 sets from 105 patients consisting of 712 blood culture bottles in total. In total, 294 negative bottles and 47 positive bottles were detected by both systems. Recovery rate of the B3D (96.7%) was significantly higher than that of DL (79.0) (p < 0.05). We determined significant differences between DL and B3D in terms of time-to-detection values for gram negatives (p = 0.006) and contaminants (p = 0.048). Overall, B3D had shorter time-to-detection mean values.
CONCLUSIONS: The recovery rate of DL was unfavorably low and time-to-detection values for DL were significantly higher than that of B3D. This might result from the ingredients of the culture bottles since the detection technologies of the systems were similar.

PMID: 28397472 [PubMed - in process]

http://ift.tt/2oBnnUm

Interactions of Methylenetetrahydrofolate Reductase Gene Polymorphisms, Folate, and Homocysteine on Blood Pressure in a Chinese Hypertensive Population.

Interactions of Methylenetetrahydrofolate Reductase Gene Polymorphisms, Folate, and Homocysteine on Blood Pressure in a Chinese Hypertensive Population.

Clin Lab. 2017 Apr 01;63(4):817-825

Authors: Li WX, Liao P, Hu CY, Cheng F, Zhang T, Sun YY, Tang L, Wang MM, Liu KS, Liu D, Liu F

Abstract
BACKGROUND: High blood pressure is related to cardiovascular diseases. We aimed to explore the interactions of methylenetetrahydrofolate reductase (MTHTR) gene C677T and A1298C mutations and folate/homocysteine (Hcy) status on blood pressure in a Chinese hypertensive population.
METHODS: The clinical data in the present study derived from a previous trial (NCT00520247). Genotypes in Hcy pathway enzymes were detected by PCR-RFLP methods. Supine blood pressure was measured with a mercury sphygmomanometer. Serum Hcy was measured by high-performance liquid chromatography, and serum folate was measured by chemiluminescent immunoassay.
RESULTS: This study showed that hyperhomocysteinemia independently elevated diastolic blood pressure (DBP) (β (SE): 2.02 (0.85), p = 0.018). Furthermore, individuals with high Hcy and MTHFR1298AC + CC genotypes showed higher DBP than the normal Hcy and 1298AA carriers (β (SE): 1.81 (0.54), p = 0.001). This correlation was verified by the trend test (p = 0.003). However, polymorphisms of MTHFR C677T, MTR A2756G or MTRR A66G do not affect baseline blood pressure level.
CONCLUSIONS: The present study demonstrated that the MTHFR A1298C mutation accompanied by hyperhomocysteinemia jointly elevated DBP. Further studies are necessary to confirm the role of these genotypes and Hcy on blood pressure in a larger population.

PMID: 28397471 [PubMed - in process]

http://ift.tt/2oBcJx2

Intracellular Storage of Duffy Antigen-Binding Chemokines by Duffy-Positive Red Blood Cells.

Intracellular Storage of Duffy Antigen-Binding Chemokines by Duffy-Positive Red Blood Cells.

Clin Lab. 2017 Apr 01;63(4):717-723

Authors: Yamamoto A, Saito N, Ogasawara S, Shiratori T, Kondo J, Itoga M, Kayaba H

Abstract
BACKGROUND: Duffy antigen/chemokine receptor (DARC) is a non-signaling receptor for multiple chemokines. The role of DARC on red blood cells (RBCs) has remained elusive. The purpose of this study was to analyze selective storage of DARC-binding chemokines in RBCs.
METHODS: Peripheral blood from healthy volunteers with DARC-positive blood type was collected in EDTA tubes. The concentration of DARC binding chemokines (i.e., MCP-1, RANTES, eotaxin-1, TARC, and IL-8), DARC nonbinding chemokines (i.e., MIP-1α, IP-10), and several cytokines in the supernatant of purified RBCs before and after hemolysis was measured using Bio-Plex and ELISA assays. Storage of chemokines in RBCs and the expression of DARC were evaluated using flow-cytometry.
RESULTS: The levels of all DARC-binding chemokines except TARC and IL-8 increased significantly after hemolysis. There was no significant increase in any of the DARC non-binding chemokines or in the other cytokines after hemolysis. RANTES, eotaxin-1, and MCP-1 were detectable intracellularly but not on the RBC surface. RANTES was absorbed by RBCs. DARC was expressed intracellularly in RBCs as well as on the surface.
CONCLUSIONS: These data suggested that DARC-positive RBCs store RANTES, MCP-1 and eotaxin-1. DARC on RBC may be internalized from the surface in the process of chemokine absorption.

PMID: 28397469 [PubMed - in process]

http://ift.tt/2oBuoVw

Increase of C3a is Associated with Hemorrhagic Propensity in Patients with Immune Thrombocytopenia.

Increase of C3a is Associated with Hemorrhagic Propensity in Patients with Immune Thrombocytopenia.

Clin Lab. 2017 Apr 01;63(4):765-771

Authors: Ge J, Wang L, Li J, Hu Y, Xu Q, Chen Y, Hou R, Ruan M, Xia R, Zeng Q

Abstract
BACKGROUND: Complement activation is critically involved in multiple autoimmune diseases. Immune thrombocytopenia (ITP) is a hemorrhagic condition with enhanced platelet clearance caused by antiplatelet autoantibodies. However, the roles of complements C3a, C5a, and soluble C5b-9 (sC5b-9) in the hemorrhage of ITP remain unknown.
METHODS: Plasma C3a, C5a, and sC5b-9 levels in ITP patients were measured by enzyme-linked immunosorbent assay (ELISA). Antiplatelet autoantibodies (anti-GPIIb/IIIa and anti-GPIbα) were evaluated by modified monoclonal antibody immobilization of platelet antigen (MAIPA) assay. The severity of bleeding was assessed using the validated bleeding score for each ITP patient at onset.
RESULTS: Levels of C3a, C5a, and sC5b-9 were significantly increased in active ITP patients, compared with those in controls (p < 0.001). However, levels of C3a, C5a, and sC5b-9 were not changed by treatment of HD-DXM. In addition, the C3a levels were correlated with the increase in bleeding scores from the patients with ITP (p < 0.05, r = 0.256). In contrast, neither platelet counts nor antiplatelet autoantibodies (anti-GPIIb/IIIa and anti-GPIbα) showed any correlation with levels of C3a, C5a, and sC5b-9.
CONCLUSIONS: Levels of C3a, C5a, and sC5b-9 are increased in patients with ITP, suggesting a hyperactive complement system. Certain complement components, such as C3a, may contribute to hemorrhage of patients with ITP.

PMID: 28397467 [PubMed - in process]

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Absolute Eosinophil Count was Significantly Associated with Serum (1,3)-β-D-Glucan in Patients with Invasive Fungal Infections.

Absolute Eosinophil Count was Significantly Associated with Serum (1,3)-β-D-Glucan in Patients with Invasive Fungal Infections.

Clin Lab. 2017 Apr 01;63(4):749-756

Authors: Zhu P, Lv J, Kong Y, Xu H, Ming L

Abstract
BACKGROUND: Measurement of serum (1,3)-β-D-glucan has emerged as an important diagnostic strategy for invasive fungal infections (IFI). Previous studies suggested that the eosinophil count would be related with the value of serum (1,3)-β-D-glucan.
METHODS: 572 blood samples obtained from 126 patients at the First Affiliated Hospital of Zhengzhou University were analyzed. The unsuitable samples were excluded according to the patient selection.
RESULTS: In our study, we found that in the 572 cumulative results, the absolute eosinophil count was significantly associated with the value of (1,3)-β-D-glucan (r = 0.57; 95% confidence interval (CI) 0.51 - 0.63) but not with total leukocyte, neutrophils, lymphocytes, monocytes, and basophils. Then, we found that there were 126 patients with invasive fungal infections, 109 patients with candidiasis and 17 patients with aspergillosis. In the 109 patients who had candidiasis, absolute eosinophil count was significantly associated with the value of (1,3)-β-D-glucan (r = 0.59; 95% CI 0.53 - 0.65). Similarly, in the 17 patients who had aspergillosis, the absolute eosinophil count was significantly associated with the value of (1,3)-β-D-glucan (r = 0.65; 95% CI 0.51 - 0.75). Meanwhile, similar findings in different types of Candida species or Aspergillus species in patients were found except in Candida glabrata infection.
CONCLUSIONS: As a routine and broadly used test, serial absolute eosinophil count is related with IFI, and it could be used to evaluate the progress of IFI.

PMID: 28397466 [PubMed - in process]

http://ift.tt/2oBwk0a

Effects of Ulinastatin on In Vitro Storage Lesions of Human Red Blood Cells.

Effects of Ulinastatin on In Vitro Storage Lesions of Human Red Blood Cells.

Clin Lab. 2017 Apr 01;63(4):833-838

Authors: Liu DH, Yao YT, Li LH, Huang CM

Abstract
BACKGROUND: The study aimed to investigate the influence of in vitro storage on erythrocyte complement receptor one (E-CR1), cell shrinkage and eryptosis of human red blood cells (RBCs), and to assess the possible effects of ulinastatin (UTI) on them.
METHODS: After collection, RBCs were treated with saline (control group) and different concentrations of UTI (5,000 U/mL, 10,000 U/mL, and 50,000 U/mL in Group C1, Group C2, and Group C3, respectively). E-CR1, cell size, and phosphatidylserine (PS) exposure and intracellular Ca2+ concentration were analyzed by flow cytometer every 7 days up to Day 35.
RESULTS: E-CR1 level and cell size of all groups decreased during storage. In the control group, E-CR1 began to decrease on Day 28 and cells shrank on Day 21. The E-CR1 level of Group C2 was significantly higher than that of the control group beginning on Day 21. The cells of Group C1 and Group C2 began to shrink remarkably on Day 21, and those of Group C3 on Day 35. PS-exposure levels of 4 groups started to increase on Day 7 (p < 0.05), while from Day 14 to 35 those of Group C3 were significantly lower than the control group (p < 0.05). The intracellular Ca2+ levels of the control group started to increase significantly on Day 7, one week earlier than the experimental groups. From Day 21 to 35, the intracellular Ca2+ levels of Group C2 and C3 were significantly lower than those of the control group (p < 0.05).
CONCLUSIONS: RBCs underwent E-CR1 loss, cell shrinkage, and eryptosis during in vitro storage, which could be attenuated by UTI.

PMID: 28397464 [PubMed - in process]

http://ift.tt/2oBxZD5

Food labeling issues in patients with severe food allergies: solving a hamlet-like doubt.

Role of Postoperative Stimulated Thyroglobulin as Prognostic Factor for Differentiated Thyroid Cancer in Children and Adolescents

Thyroid , Vol. 0, No. 0.

http://ift.tt/2pb1RUw

Clear Cell Change in Thyroid Carcinoma: A Clinicopathologic and Molecular Study with Identification of Variable Genetic Anomalies

Thyroid , Vol. 0, No. 0.

http://ift.tt/2oBhEy8

Task-optimal auditory attention set restored as fast in older as in younger adults after distraction

Publication date: Available online 12 April 2017
Source:Biological Psychology
Author(s): Márta Volosin, Zsófia Anna Gaál, János Horváth
The present study investigated how fast younger and older adults recovered from a distracted attentional state induced by rare, unpredictable sound events. The attentional state was characterized by the auditory N1 event-related potential (ERP), which is enhanced for sound events in the focus of attention. Younger (19-26 years) and older (62-74 years) adults listened to continuous tones containing rare pitch changes (glides) and short gaps. Glides and gaps could be separated in 150ms, 250ms, 650ms or longer and the task was gaps detection while ignoring glides. With longer glide-gap separations similar N1 enhancements were observable in both groups suggesting that the duration of the distracted sensory state was not affected by aging. Older adults responded, however, slower at short glide-gap separations which indicated that distraction at subsequent levels of processing may have nonetheless more impact in older than in younger adults.

http://ift.tt/2nGOZZm

Role of Postoperative Stimulated Thyroglobulin as Prognostic Factor for Differentiated Thyroid Cancer in Children and Adolescents

Thyroid , Vol. 0, No. 0.

http://ift.tt/2pb1RUw

Clear Cell Change in Thyroid Carcinoma: A Clinicopathologic and Molecular Study with Identification of Variable Genetic Anomalies

Thyroid , Vol. 0, No. 0.

http://ift.tt/2oBhEy8

Exposure to Extrinsic Stressors, Social Defeat or Bisphenol A, Eliminates Sex Differences in DNA methyltransferase expression in the Amygdala

Abstract

Chemical and psychological stressors can exert long lasting changes in brain function and behavior. Changes in DNA methylation have been shown to be an important mechanism mediating long lasting changes in neural function and behavior, especially for anxiety-like or stress responses. Here we examined the effects of either a social or chemical stressor on DNA methyltransferase (DNMT) gene expression in the amygdala, an important brain region modulating stress responses and anxiety. In adult California mice (Peromyscus californicus) that were naïve to social defeat, females had higher levels of Dnmt1 expression in punch samples of the central amygdala (CeA) than males. In addition, social defeat stress reduced Dnmt1 and Dnmt3a expression in the CeA of females but not males. A second study using more anatomically specific punch samples replicated these effects for Dnmt1. Perinatal exposure, spanning from periconception through lactation, to bisphenol A or ethinyl estradiol (estrogens in birth control pills) also abolished sex differences in Dnmt1 expression in the CeA but not basolateral amygdala. These findings identify a robust sex difference in Dnmt1 expression in the CeA that is sensitive to both psychological and chemical stressors. Our results suggest that future studies should examine the impact of psychological and chemical stressors on DNA methylation in the CeA and that Dnmt1 may have an underappreciated role in plasticity in behavior.

http://ift.tt/2o7RAYf

Coexpressed modular gene expression reveals inverse correlation between immune responsive transcription and aggressiveness in gastric tumours

Abstract

The existing large number of gene expression profiles of tumour samples offers a great advantage for the integrative functional genomic exploration of molecular dysregulation in cancers. The clusters of genes (modules) derived from a gastric cancer (GC) coexpression network were explored to understand their clinical and functional significance. Among the modules derived from the GC mRNA expression network, six modules were relatively highly expressed in diffuse type gastric tumours. Elevated expression of genes related to extracellular matrix (ECM), angiogenesis, collagen and intracellular cytoskeletal components and immune response were identified in these modules. ECM-related modules exhibited an inverse correlation with modules representing the expression of immune response genes. A reduced expression of immune response genes was identified as the key factor associated with the aggressive features of diffuse gastric tumours, which is indicative of tumour progression involving the escape from immune surveillance in diffuse tumours. A part of the identified aggressive factors was common between intestinal and diffuse type tumours. The coexpressed modules and their expression patterns delineate the fine transition involved in cancer progression in the later stages of tumours. The identified modules could serve as surrogate gene-sets, indicating the molecular staging of GC aggressiveness with underlying biological interaction.

http://ift.tt/2p9owAs

Molecular imprinting sensor based on quantum weak measurement

Publication date: 15 August 2017
Source:Biosensors and Bioelectronics, Volume 94
Author(s): Dongmei Li, Qinghua He, Yonghong He, Meiguo Xin, Yilong Zhang, Zhiyuan Shen
A new type of sensing protocol, based on a high precision metrology of quantum weak measurement, was first proposed for molecularly imprinted polymers (MIP) sensor. The feasibility, sensitivity and selectivity of weak measurement based MIP (WMMIP) sensor were experimentally demonstrated with bovine serum albumin (BSA). Weak measurement system exhibits high sensitivity to the optical phase shift corresponding to the refractive index change, which is induced by the specific capture of target protein molecules with its recognition sites. The recognition process can be finally characterized by the central wavelength shift of output spectra through weak value amplification. In our experiment, we prepared BSA@MIP with modified reversed-phase microemulsion method, and coated it on the internal surface of measuring channels assembled into the Mach-Zehnder (MZ) interferometer based optical weak measurement system. The design of this home-built optical system makes it possible to detect analyte in real time. The dynamic process of the specific adsorption and concentration response to BSA from 5×10⁻⁴ to 5×10⁻¹μg/L was achieved with a limit of detection (LOD) of 8.01×10⁻¹²g/L. This WMMIP shows superiority in accuracy, fast response and low cost. Furthermore, real-time monitoring system can creatively promote the performance of MIP in molecular analysis.

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Dialysis membrane-enforced microelectrode array measurement of diverse gut electrical activity

Publication date: 15 August 2017
Source:Biosensors and Bioelectronics, Volume 94
Author(s): Naoko Iwata, Takumi Fujimura, Chiho Takai, Kei Odani, Shin Kawano, Shinsuke Nakayama
A variety of electrical activities occur depending on the functional state in each section of the gut, but the application of microelectrode array (MEA) is rather limited. We thus developed a dialysis membranes-enforced technique to investigate diverse and complex spatio-temporal electrical activity in the gut. Muscle sheets isolated from the gastrointestinal (GI) tract of mice along with a piece of dialysis membrane were woven over and under the strings to fix them to the anchor rig, and mounted on an 8×8 MEA (inter-electrode distance=150µm). Small molecules (molecular weight <12,000) were exchanged through the membrane, maintaining a physiological environment. Low impedance MEA was used to measure electrical signals in a wide frequency range. We demonstrated the following examples: 1) pacemaker activity-like potentials accompanied by bursting spike-like potentials in the ileum; 2) electrotonic potentials reflecting local neurotransmission in the ileum; 3) myoelectric complex-like potentials consisting of slow and rapid oscillations accompanied by spike potentials in the colon. Despite their limited spatial resolution, these recordings detected transient electric activities that optical probes followed with difficulty. In Addition, propagation of pacemaker-like potential was visualized in the stomach and ileum. These results indicate that the dialysis membrane-enforced technique largely extends the application of MEA, probably due to stabilisation of the access resistance between each sensing electrode and a reference electrode and improvement of electric separation between sensing electrodes. We anticipate that this technique will be utilized to characterise spatio-temporal electrical activities in the gut in health and disease.

http://ift.tt/2o8w4C0

One-pot loading high-content thionine on polydopamine-functionalized mesoporous silica nanosphere for ultrasensitive electrochemical immunoassay

Publication date: 15 September 2017
Source:Biosensors and Bioelectronics, Volume 95
Author(s): Guosong Lai, Min Zheng, Wenjie Hu, Aimin Yu
A thionine (TH)-doped mesoporous silica nanosphere (MSN)/polydopamine (PDA) nanocomposite was synthesized for developing a new signal transduction strategy of electrochemical immunoassay. This nanocomposite was synthesized through one-pot loading of TH and in situ formation of PDA-coating on MSN. After antibody labeling, the obtained nanoprobe was used for the signal tracing of sandwich immunoassay at a magnetic bead-assay platform. Based on the specific capture of the MSN-TH/PDA nanoprobes through sandwich immunoreaction to form a magnetic immunocomplex and the following treatment of the immunocomplex with a NaOH solution, the PDA film coated on MSN was destroyed to release TH tags from the nanoprobe. The released TH was then electrochemically measured at a carbon nanotube (CNT)-modified electrode for the signal transduction of the method. Due to the high loading of TH on the nanoprobe for tag release and effective electrochemical signal enhancement by the electrode modification of CNTs, ultrahigh sensitivity was achieved. Using human IgG as a model analyte, this method showed a wide linear range over four orders of magnitude and a low detection limit of 5.8pg/mL. Additionally, the method has excellent specificity, satisfactory reproducibility and stability as well as acceptable reliability. Due to the simple preparation of the nanoprobe and the low cost, convenient operation of the detection strategy, this nonenzymatic immunoassay method possesses great potentials for practical applications.

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InGaN/GaN nanowires as a new platform for photoelectrochemical sensors – detection of NADH

Publication date: 15 August 2017
Source:Biosensors and Bioelectronics, Volume 94
Author(s): M. Riedel, S. Hölzel, P. Hille, J. Schörmann, M. Eickhoff, F. Lisdat
InGaN/GaN nanowire heterostructures are presented as nanophotonic probes for the light-triggered photoelectrochemical detection of NADH. We demonstrate that photogenerated electron-hole pairs give rise to a stable anodic photocurrent whose potential- and pH-dependences exhibit broad applicability. In addition, the simultaneous measurement of the photoluminescence provides an additional tool for the analysis and evaluation of light-triggered reaction processes at the nanostructured interface. InGaN/GaN nanowire ensembles can be excited over a wide wavelength range, which avoids interferences of the photoelectrochemical response by absorption properties of the compounds to be analyzed by adjusting the excitation wavelength. The photocurrent of the nanostructures shows an NADH-dependent magnitude. The anodic current increases with rising analyte concentration in a range from 5µM to 10mM, at a comparatively low potential of 0mV vs. Ag/AgCl. Here, the InGaN/GaN nanowires reach high sensitivities of up to 91µAmM⁻¹cm⁻² (in the linear range) and provide a good reusability for repetitive NADH detection. These results demonstrate the potential of InGaN/GaN nanowire heterostructures for the defined conversion of this analyte paving the way for the realization of light-switchable sensors for the analyte or biosensors by combination with NADH producing enzymes.

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Prognostic implication of CD274 (PD-L1) protein expression in tumor-infiltrating immune cells for microsatellite unstable and stable colorectal cancer

Abstract

In this study, we investigated the clinical relevance of CD274 (PD-L1) protein expression by tumor cells and tumor-infiltrating immune cells in colorectal cancer (CRC). To this end, 186 microsatellite instability-high (MSI-H) and 153 microsatellite stable (MSS) CRCs were subjected to immunohistochemistry (IHC) analysis for the expression of CD274 and mismatch repair proteins. CD274 expression was evaluated in tumor cells at the center (TC) and periphery (TP), and immune cells at the center (IC) and periphery (IP) of CRC. IHC slides stained for CD3 and CD8 were scanned using an Aperio ScanScope for precise calculation of tumor-infiltrating T cell density. Additionally, samples were screened for the B-Raf (BRAF)-V600E mutation using a Cobas 4800 System and IHC. In total, CD274^TC, CD274^TP, CD274^IC, and CD274^IP were observed in 43 (23.1%), 47 (25.3%), 107 (57.5%), and 102 (54.8%) of the MSI-H CRCs examined, and in three (2.0%), four (2.6%), 47 (30.7%), and 56 (36.6%) of the 153 MSS CRCs tested. Meanwhile, intratumoral heterogeneity of CD274 expression in tumor cells and immune cells was detected in 24 (12.9%) and 47 (25.3%) MSI-H CRCs, respectively. Notably, in both MSI-H and MSS CRC, CD274^IC and CD274^IP were independently associated with improved prognosis (P < 0.05), while BRAF mutation was associated with CD274^TP, poor differentiation, sporadic type, and hMLH1(−)/hMSH2(+)/hMSH6(+)/PMS2(−) in MSI-H CRC (P < 0.006). In conclusion, CD274 expression in tumor-infiltrating immune cells was an independent factor for improved prognosis in CRC patients. A deeper understanding of CD274 status may yield improved responses to future CRC immunotherapies.

http://ift.tt/2nFxPv1

A dynamic sandwich assay on magnetic beads for selective detection of single-nucleotide mutations at room temperature

Publication date: 15 August 2017
Source:Biosensors and Bioelectronics, Volume 94
Author(s): Junxiu Wang, Guoliang Xiong, Liang Ma, Shihui Wang, Xu Zhou, Lei Wang, Lehui Xiao, Xin Su, Changyuan Yu
Single-nucleotide mutation (SNM) has proven to be associated with a variety of human diseases. Development of reliable methods for the detection of SNM is crucial for molecular diagnosis and personalized medicine. The sandwich assays are widely used tools for detecting nucleic acid biomarkers due to their low cost and rapid signaling. However, the poor hybridization specificity of signal probe at room temperature hampers the discrimination of mutant and wild type. Here, we demonstrate a dynamic sandwich assay on magnetic beads for SNM detection based on the transient binding between signal probe and target. By taking the advantage of mismatch sensitive thermodynamics of transient DNA binding, the dynamic sandwich assay exhibits high discrimination factor for mutant with a broad range of salt concentration at room temperature. The beads used in this assay serve as a tool for separation, and might be helpful to enhance SNM selectivity. Flexible design of signal probe and facile magnetic separation allow multiple-mode downstream analysis including colorimetric detection and isothermal amplification. With this method, BRAF mutations in the genomic DNA extracted from cancer cell lines were tested, allowing sensitive detection of SNM at very low abundances (0.1–0.5% mutant/wild type).

http://ift.tt/2ouoWTN

Immunodetection of salivary biomarkers by an optical microfluidic biosensor with polyethylenimine-modified polythiophene-C70 organic photodetectors

Publication date: 15 August 2017
Source:Biosensors and Bioelectronics, Volume 94
Author(s): Tao Dong, Nuno Miguel Matos Pires
This work reports a novel optical microfluidic biosensor with highly sensitive organic photodetectors (OPDs) for absorbance-based detection of salivary protein biomarkers at the point of care. The compact and miniaturized biosensor has comprised OPDs made of polythiophene-C70 bulk heterojunction for the photoactive layer; whilst a calcium-free cathode interfacial layer, made of linear polyethylenimine, was incorporated to the photodetectors to enhance the low cost. The OPDs realized onto a glass chip were aligned to antibody-functionalized chambers of a poly(methyl methacrylate) microfluidic chip, in where immunogold-silver assays were conducted. The biosensor has detected IL-8, IL-1β and MMP-8 protein in spiked saliva with high detection specificity and short analysis time exhibiting detection limits between 80pgmL⁻¹ and 120pgmL⁻¹. The result for IL-8 was below the clinical established cut-off of 600pgmL⁻¹, which revealed the potential of the biosensor to early detection of oral cancer. The detection limit was also comparable to other previously reported immunosensors performed with bulky instrumentation or using inorganic photodetectors. The optical detection sensitivity of the polythiophene-C70 OPD was enhanced by optimizing the thickness of the photoactive layer and anode interfacial layer prior to the saliva immunoassays. Further, the biosensor was tested with unspiked human saliva samples, and the results of measuring IL-8 and IL-1β were in statistical agreement with those provided by two commercial assays of ELISA. The optical microfluidic biosensor reported hereby offers an attractive and cost-effective tool to diagnostics or screening purposes at the point of care.

http://ift.tt/2o8HWnf

Bipolar electrochemiluminescence on thread: A new class of electroanalytical sensors

Publication date: 15 August 2017
Source:Biosensors and Bioelectronics, Volume 94
Author(s): Rui Liu, Cuiling Liu, Huijie Li, Min Liu, Dan Wang, Chunsun Zhang
This paper introduces a new and simple concept for fabricating low-cost, easy-to-use capillary microchannel (CMC) assisted thread-based microfluidic analytical devices (CMCA-μTADs) for bipolar electrochemiluminescence (BP-ECL) application. The thread with patterns of carbon screen-printed electrodes and bare thread zones (BTZs) is embedded into a CMC. Such CMCA-μTADs can produce a strong and stable BP-ECL signal, and have an extremely low cost ($0.01 per device). Interestingly, the CMCA-μTADs are ultraflexible, and can be bent with a 135° bending angle at the BTZ or with a 150° bending angle at the middle of bipolar electrode (BPE), with no loss of analytical performance. Additionally, the two commonly-used ECL systems of Ru(bpy)3²⁺/TPA and luminol/H2O2 are applied to demonstrate the quantitative ability of the BP-ECL CMCA-μTADs. It has been shown that the proposed devices have successfully fulfilled the detection of TPA and H2O2, with detection limits of 0.00432mM and 0.00603mM, respectively. Based on the luminol/H2O2 ECL system, the CMCA-μTADs are further applied for the glucose measurement, with the detection limit of 0.0205mM. Finally, the applicability and validity of the CMCA-μTADs are demonstrated for the measurements of H2O2 in milk, and glucose in human urine and serum. The results indicate that the proposed devices have the potential to become an important new tool for a wide range of applications.

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Aluminium resistant, plant growth-promoting bacteria induce overexpression of Aluminium stress related genes in Arabidopsis thaliana and increase the ginseng tolerance against Aluminium stress

Publication date: Available online 12 April 2017
Source:Microbiological Research
Author(s): Mohamed El-Agamy Farh, Yeon-Ju Kim, Johan Sukweenadhi, Priyanka Singh, Deok-Chun Yang
Panax ginseng is an important cash crop in the Asian countries due to its pharmaceutical effects, however the plant is exposed to various abiotic stresses, lead to reduction of its quality. One of them is the Aluminum (Al) accumulation. Plant growth promoting bacteria which able to tolerate heavy metals has been considered as a new trend for supporting the growth of many crops in heavy metal occupied areas. In this study, twelve bacteria strains were isolated from rhizosphere of diseased Korean ginseng roots located in Gochang province, Republic of Korea and tested for their ability to grow in Al-embedded broth media. Out of them, four strains (Pseudomonas simiae N3, Pseudomonas fragi N8, Chryseobacterium polytrichastri N10, and Burkholderia ginsengiterrae N11-2) were able to grow. The strains could also show other plant growth promoting activities e.g. auxins and siderophores production and phosphate solubilization. P. simiae N3, C. polytrichastri N10, and B. ginsengiterrae N11-2 strains were able to support the growth of Arabidopsis thaliana stressed by Al while P. simiae N3 could not. Plants inoculated with P. simiae N3, C. polytrichastri N10, and B. ginsengiterrae N11-2 showed higher expression level of Al stress related genes, AtAIP, AtALS3 and AtALMT1, compared to non-bacterized plants. Expression profiles of the genes reveal the induction of external mechanism of Al resistance by P. simiae N3 and B. ginsengiterrae N11-2 and internal mechanism by C. polytrichastri N10. Korean ginseng seedlings treated with these strains showed higher biomass, particularly the foliar part, higher chlorophyll content than non-bacterized Al stressed seedlings. According to the present results, these strains can be used in the future for the cultivation of ginseng in Al persisted locations.

http://ift.tt/2oAZ5Ki

Nutrition modulation of human aging: The calorie restriction paradigm

Publication date: Available online 12 April 2017
Source:Molecular and Cellular Endocrinology
Author(s): Sai Krupa Das, Priya Balasubramanian, Yasoma K. Weerasekara
Globally, the aging population is growing rapidly, creating an urgent need to attenuate age-related health conditions, including metabolic disease and disability. A promising strategy for healthy aging based on consistently positive results from studies with a variety of species, including non-human primates (NHP), is calorie restriction (CR), or the restriction of energy intake while maintaining intake of essential nutrients. The burgeoning evidence for this approach in humans is reviewed and the major study to date to address this question, CALERIE (Comprehensive Assessment of the Long-term Effects of Reducing Intake of Energy), is described. CALERIE findings indicate the feasibility of CR in non-obese humans, confirm observations in NHP, and are consistent with improvements in disease risk reduction and potential anti-aging effects. Finally, the mechanisms of CR in humans are reviewed which sums up the fact that evolutionarily conserved mechanisms mediate the anti-aging effects of CR. Overall, the prospect for further research in both NHP and humans is highly encouraging.

http://ift.tt/2o7Pyau

Αρχειοθήκη ιστολογίου

! # Ola via Alexandros G.Sfakianakis on Inoreader

Η λίστα ιστολογίων μου

Τετάρτη 12 Απριλίου 2017

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