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Background: Gold nanoparticles (GNPs) are among the most promising radiosensitive materials in radiotherapy. Studying the effective sensitizing factors such as nanoparticle size, concentration, surface features, radiation energy and cell type can help to optimize the effect and possible clinical application of GNPs in radiation therapy. In this study, the radiation sensitive polymer gel was used to investigate the dosimetric effect of GNP size in megavoltage (MV) photon beam radiotherapy. Material and Methods: GNPs with the size of 30nm, 50nm and 100nm in diameter were used. Transmission electron microscope (TEM) and dynamic light scattering (DLS) were applied to analyze the size of nanoparticles. The MAGICA polymer gel was synthesized and impregnated with different sizes of GNPs. The samples were irradiated with 6MV photon beam and 24 hours after irradiation, they were read using a Magnetic Resonance Imaging (MRI) scanner. Macroscopic Dose Enhancement Factor (DEF) was measured to compare the effect of GNP size. The MAGICA response of the 6MV x-ray beam was verified comparing Percentage Depth Dose (PDD) curve extracted from polymer gel dosimetry and Treatment Planning System (TPS). Results: MAGICA polymer gel dose response curve was linear in the range of 0 to 10 Gy. DEFs by adding 30nm, 50nm and 100nm GNPs were 1.1, 1.17 and 1.12, respectively. PDD curves of polymer gel dosimeter and treatment planning system were in good agreement. Conclusion: The results indicated a substantial increase in DEF uses a MV photon beam in combination with GNPs of different sizes and it was inconsistent with previous radiobiological studies. The maximum DEF was achieved for 50nm GNPs in comparison with 30nm and 100nm leading to the assumption of self-absorption effect by larger diameters. According to the outcomes of this work, MAGICA polymer gel can be recommended as a reliable dosimeter to investigate the dosimetric effect of GNP size and also a useful method to validate the current radiobiological and simulation studies.

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Background: The use of medicinal plants due to their lower side effects and the various efficient components has been increased in recent years. The aim of this study was to evaluate the antinociceptive effect of hydroalcoholic extract of green tea in male mice using the hot plate and formalin tests. Materials and Methods: In this experimental study, 56 NMRI male mice (weight, 28±3 g) were assigned to the control and experimental groups. The experimental group received the green tea extract (75, 150 and 300 mg/kg) and 30 min after the injection, the hot plate and formalin tests were used to assess the acute and chronic pain. Results: Results showed that hydroalcoholic extract of green tea (75mg/kg) could induce antinociception in phase 2, not in phase 1 and interphase of the formalin test. Hydroalcoholic extract of green tea (150 and 300 mg/kg) could also induce antinociception in phase 1, interphase and phase 2 of the formalin test. Moreover, hydroalcoholic extract of green tea caused a delay on the painful behaviors in hot-plate test. Conclusion: Hydroalcoholic extract of green tea has analgesic properties and can be used as a substitute for chemical analgesic drugs. [ABSTRACT FROM AUTHOR]

Background: Diabetes is the sixth main cause of mortality in the world, and its both types are accompanied with eye complications in a way that the risk of blindness in diabetic patients is 25 times more than normal people. Hence, this study was conducted to determine diabetic patients' awareness of the eye complications associated with this disease. Materials and Methods: In a cross-sectional study, 200 diabetic patients referring to the eye clinic of Amir Kabir Hospital of Arak in 2008 were evaluated. Data collection was done through questionnaire and referral interviews. Then some of the factors affecting patients' awareness of eye complications associated with diabetes were analyzed. Results: The level of awareness of eye complications associated with diabetes in 40% of the patients was average. Also, significant relationships were observed between the level of awareness and age, job, education, type of retinopathy, type of treatment, and the history of referring to ophthalmologist (p<0.001), whereas no significant relationships were observed between the level of awareness and gender, familial history of diabetes, and controlling the diet (p>0.05). Conclusion: This study showed that a higher level of awareness in diabetic patients does not necessitate that they observe a proper diet, but it can lead to patients' greater care in preventing the development of diabetes. [ABSTRACT FROM AUTHOR]

Context.--The immunohistochemical identification of thyroid transcription factor-1 (TTF-1) is regarded as the presence of a nuclear pattern of staining and is used to identify tumors of thyroid or pulmonary origin. Although there have been reports of cytoplasmic expression of TTF-1, the significance of this pattern has not been studied in detail. Objectives.--To determine the incidence at which cytoplasmic immunostaining for TTF-1 occurs and to analyze the diagnostic value of this pattern of immunoreactivity. Design.--Histologic sections of 361 consecutive cases of neoplasms stained for TTF-1 were reviewed, and those showing cytoplasmic staining in the tumor cells or in non-neoplastic cells were selected. Clinical correlation was obtained on the latter cases regarding the origin of the tumor. Setting.--An immunohistochemistry laboratory in a tertiary-care institution. Results.--The 361 tumors were obtained from 29 organ sites and corresponded to primary and metastatic neoplasms. Twenty-three (6.3%) tumors showed cytoplasmic staining for TTF-1. In 13 of these, the primary site of origin was established with certainty: 7 were lung carcinomas (3 primary lung adenocarcinomas, 1 primary large cell carcinoma, 1 metastatic small cell carcinoma to the liver, 1 metastatic adenocarcinoma to a neck lymph node, and 1 metastatic adenocarcinoma to thigh soft tissue), 3 colonic adenocarcinomas (2 metastases to vertebrae and 1 to lung), 1 metastatic breast ductal adenocarcinoma to femur, 1 metastatic laryngeal squamous cell carcinoma to liver, and 1 meningioma involving the orbit bone. There were 3 lung carcinomas with concomitant nuclear immunostaining. Nonneoplastic liver tissue was the most frequent non-tumoral tissue in which the cytoplasm stained with TTF-1 antibody. Conclusion.--Occasional cytoplasmic staining for TTF-1 in tumors is seen, but it is a nonspecific finding; when present, it should be disregarded for diagnostic purposes. [ABSTRACT FROM AUTHOR]

Introduction: The finding of a mixture of both 46,XX and 46,XY cells in amniotic fluid culture has been frequently described. In the great majority of cases, the finding is followed by the birth of a normal male infant, leading to a consensus that the finding is the result of contamination with maternal cells in a normal male fetus. There are, however, several other possible explanations including the presence of cells from an undiagnosed twin pregnancy, cross contamination in the laboratory, the presence of cells from a "vanished" male twin, and true fetal chimerism. An accurate obstetric history and thorough sonography can be of great value in correct assessment of such situations. Materials and Methods: A 29 Years-old woman was referred for genetic counseling. Her gestational age was 16 weeks and maternal biochemical serum screening test indicated high risk for Down Syndrome. She underwent amniocentesis for chromosome study using standard high resolution GTG banding technique. Results: Karyotype result was 46,XY and sonography revealed a female fetus with normal internal genital organs. A normal baby girl was borned with 46,XX karyotype. Upon reviewing of amnio GTG slides both 46,XY and 46,XX cell lines were detected. The patient admitted that she had experienced a miscarriage and heavy bleeding early in pregnancy. The sonography at the time of amniocentesis had shown a shrinking cyst next to the fetus. These findings strengthened the vanished or resorbed twin as the reason for such a finding. Conclusion: The fetus was most likely a non-identical twin. Most of grown cells were from the vanished male fetus which led to misdiagnosis. Thorough genetic counseling with the view of obstetric history is of great value in such situations. [ABSTRACT FROM AUTHOR]

Background: Depression is one of the most important psychological problems in society that reduces their health. This study aimed to compare the effectiveness of cognitive-behavioral therapy with behavioral activation therapy on learned helplessness and cognitive emotion regulation in individuals with depressive symptoms. Materials and Methods: The present study was a quasi-experimental pretest and posttest with two experimental groups and a control group. The statistical population of the study included all people with depression in Babol who referred to psychotherapy clinics in 2021. Forty-five subjects who scored above-average on the Beck Depression Inventory were randomly selected and randomly assigned to three groups of 15 responding to the Garnfsky Emotion Cognitive Regulation and Queens and Nelson Learned Helplessness Questionnaire. Analysis of covariance and Benferoni test using SPSS-23 software were used to analyze the results. Results: The results showed that there was no significant difference between the effect of two methods of behavioral methodology and behavioral activation on learned helplessness (P

Chiral amino acids (AAs) are essential in metabolism and understanding physiological processes, and they could be used as biomarkers for the diagnosis of different diseases. In this study, chiral Cdots@Van were prepared by postmodifying an achiral Cdots core with vancomycin for recognizing and determining the enantiomeric excess (ee) of tyrosine (Tyr) enantiomers. The fluorescence response of Cdots@Van is based on an "on-off" strategy, with different quenching percentages for d- and l-tyrosine. Interestingly, the circular dichroism (CD) spectrum of Cdots@Van responded to only one form of Tyr enantiomer, specifically d-Tyr, and remained nearly unchanged upon the addition of l-Tyr. Quantum mechanical (QM) calculations were in excellent agreement with the experimental results, confirming the stronger binding affinity of Cdots@Van for d-Tyr compared to l-Tyr. We further investigated the chiral recognition ability of the interconnected vancomycin particles, which was synthesized using the EDC/NHS coupling reaction between vancomycin molecules without a Cdots core. Surprisingly, unlike free vancomycin molecules, interconnected vancomycin displayed an enantiomeric recognition ability by CD spectroscopy, similar to what was observed for Cdots@Van. Crucially, this chiral probe has been successfully utilized for cell imaging applications.

Purpose: To evaluate how risk factors impact success rates of initial probing and nasolacrimal duct (NLD) tube intubation in children over 18 months of age with congenital nasolacrimal duct obstruction (CNLDO)., Methods: This cohort study included 98 CNLDO patients aged 18 months to 10 years who underwent NLD probing with stent insertion. We employed the multivariate frailty model as our final model to conceptually elaborate on our correlated eye data, with the primary outcome measure evaluating the success rates of probing and tube intubation. Factors such as age, probing complexity, tube type, prior surgeries, and passive smoking were considered in the evaluation., Results: The study involved 98 patients (54 males, 44 females) with a mean age of 41.46 months and an average follow-up of 98.37 days (95 % CI 87.65-109.1). Out of the 110 eyes that underwent surgery, 13 (11.8 %) experienced failure while 97 (88.2 %) were censored. Kaplan-Meier analysis indicated significant differences in age category and probing (P-value = 0.03 and 0.006 respectively), but not tube type (P-value = 0.8). Multivariable analysis confirmed that older age and complex probing were associated with higher failure rates in CNLDO cases, with each monthly increase correlating to a two percent higher likelihood of intubation failure., Conclusions: Patient age and probing complexity influence CNLDO treatment, impacting surgical techniques and outcomes. Tube type, prior surgery, and passive smoking have no significant impact on treatment success., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

High-sensitivity nanoflow liquid chromatography (nLC) is seldom employed in untargeted metabolomics because current sample preparation techniques are inefficient at preventing nanocapillary column performance degradation. Here, we describe an nLC-based tandem mass spectrometry workflow that enables seamless joint analysis and integration of metabolomics (including lipidomics) and proteomics from the same samples without instrument duplication. This workflow is based on a robust solid-phase micro-extraction step for routine sample cleanup and bioactive molecule enrichment. Our method, termed proteomic and nanoflow metabolomic analysis (PANAMA), improves compound resolution and detection sensitivity without compromising the depth of coverage as compared with existing widely used analytical procedures. Notably, PANAMA can be applied to a broad array of specimens, including biofluids, cell lines, and tissue samples. It generates high-quality, information-rich metabolite-protein datasets while bypassing the need for specialized instrumentation., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Aberrant activation of GLI transcription factors has been implicated in the pathogenesis of different tumor types including pancreatic ductal adenocarcinoma. However, the mechanistic link with established drivers of this disease remains in part elusive. In this study, using a new genetically engineered mouse model overexpressing constitutively active mouse form of GLI2 and a combination of genome-wide assays, we provide evidence of a novel mechanism underlying the interplay between KRAS, a major driver of pancreatic ductal adenocarcinoma development, and GLI2 to control oncogenic gene expression. These mice, also expressing KrasG12D, show significantly reduced median survival rate and accelerated tumorigenesis compared with the KrasG12D only expressing mice. Analysis of the mechanism using RNA sequencing demonstrate higher levels of GLI2 targets, particularly tumor growth-promoting genes, including Ccnd1, N-Myc, and Bcl2, in KrasG12D mutant cells. Furthermore, chromatin immunoprecipitation sequencing studies showed that in these cells KrasG12D increases the levels of trimethylation of lysine 4 of the histone 3 (H3K4me3) at the promoter of GLI2 targets without affecting significantly the levels of other major active chromatin marks. Importantly, Gli2 knockdown reduces H3K4me3 enrichment and gene expression induced by mutant Kras. In summary, we demonstrate that Gli2 plays a significant role in pancreatic carcinogenesis by acting as a downstream effector of KrasG12D to control gene expression., (©2024 The Authors; Published by the American Association for Cancer Research.)

Despite the beneficial effects of antibiotics such as chloramphenicol (CAP), they exert some destructive impacts on human health. We designed an electrochemical sensor based on reduced graphene oxide (rGO)/Au/Co 2 CuS 4 nanohybrid for determination of CAP in food and biological samples. The Co 2 CuS 4 was synthesized from binuclear metal-organic framework (CoCu-BDC) through a two-step process. Nanohybrid was characterized by X-ray photoelectron spectroscopy and transmission electron microscopy. The rGO/Au/Co 2 CuS 4 provides more active sites and good electrical conductivity to reduce charge transfer resistance and improve the electrocatalytic activity for determination of CAP. The prepared sensor has a wide linear range from 7 to 141 nM with a limit of detection of 2.5 nM and a limit of quantification of 21.92 nM. It also provided high selectivity and repeatability with a relative standard deviation of 2.6%. Stability studies showed that the electrode has acceptable performance with efficiency of 95% after 33 days., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Infections may contribute to Alzheimer's disease (AD) risk. Limited evidence suggests Toxocara spp. infection/exposure could influence AD development., Methods: We investigated Toxocara seropositivity and AD in Iranian adults using a matched case-control study. Our sample included 90 AD cases and 91 healthy older adults. Anti-Toxocara immunoglobulin G (IgG) antibodies were assessed via enzyme-linked immunosorbent assay. We computed the odds ratios (ORs) and 95% confidence intervals (CIs) through univariable and multivariable analyses, adjusting for potential confounders., Results: There were 33/90 (36.67% [95% CI 26.75 to 47.48]) anti-Toxocara IgG seropositive individuals identified among the AD cases and 21/91 (23.07% [95% CI 14.89 to 33.09]) among the healthy controls. In univariable analysis, a significant association was identified between anti-Toxocara IgG seropositivity and AD (OR 1.93 [95% CI 1.01 to 3.69], p<0.001). Moreover, the association remained significant (OR 2.18 [95% CI 1.05 to 4.49], p<0.001) in multivariable analysis after adjustment for covariates. There was no association between anti-Toxocara IgG seropositivity and the severity of AD (OR 0.75 [95% CI 0.21 to 2.61], p=0.47)., Conclusions: Our findings indicated that Toxocara exposure/infection could be a potential risk factor for development of AD. To better understand a real causality between Toxocara exposure/infection and AD and related dementias, follow-up designed and adequately powered studies are needed., (© The Author(s) 2024. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.)

Urban areas are often hotspots for the dissemination of non-native (invasive) plant species, some of which release (potentially) allergenic pollen. Given the high population density in cities, a considerable number of people can be regularly and potentially intensively exposed to the pollen from these plants. This study delves into the Tree-of-Heaven (Ailanthus altissima, [Mill.] Swingle), native to East Asia, which is known for its high invasiveness in temperate regions worldwide, particularly favoring urban colonization. This study explores the botanical and aerobiological dimensions of this species using the central European metropolitan region of Berlin, Germany, as a case study, and provides a comprehensive global overview of allergological insights. The number of Ailanthus trees decreased markedly from the center to the periphery of Berlin City, following a temperature gradient. The same spatial trend was mirrored by airborne Ailanthus pollen concentrations measured with volumetric spore traps (Hirst-type) at five sites using seven traps. Ailanthus pollen was most abundant around midday and in the afternoon, with concentrations tenfold higher at street level than at roof level. The Ailanthus flowering period in June and July coincided well with the pollen season. To the best of our knowledge this is the first study to investigate Ailanthus altissima pollen production. On average, 5539 pollen grains were found per anther. A literature review on the allergy relevance of Ailanthus altissima pollen indicates the high allergenic potential of pollen from this species. Considering the anticipated expansion of suitable habitats for Ailanthus owing to global warming and the allergological significance of its pollen, it is recommended to include Ailanthus pollen in routine pollen monitoring, particularly in areas colonized by this species. This comprehensive study provides new insights into a pollen taxon whose significance as an emerging aeroallergen should be factored into plant selection and greenspace management in all temperate regions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Knee pain is a common musculoskeletal problem. Lower extremity movement impairments could alter stresses in different planes and contribute to knee pain. Classifying these impairments may be helpful in the diagnosis and treatment of knee problems. Movement system impairment (MSI)-based classification is a system to evaluate movement impairments. Trials that involve this classification are limited. Therefore, it will be of interest to examine the effect of movement system impairment-based classification treatment compared to routine physiotherapy in individuals with tibiofemoral rotation syndrome., Methods: Twenty-two individuals with knee pain aged 18-40 years (2 males, 20 females) diagnosed with tibiofemoral rotation (TFR) syndrome were included. After initial evaluation, individuals were randomly assigned into two treatment groups (MSI-based treatment and routine physiotherapy). Both treatment groups contained 8 treatment sessions over 4 weeks. Alignment and movement impairments data form, a numeric rating scale (NRS), and the Kujala Disability Questionnaire were assessed at baseline and after a four-week intervention. Independent samples t-test and Mann-Whitney U test were used for quantitative variables, and Fisher's exact test was employed for qualitative variables to compare the groups. One-way Analysis of variance (ANOVA) and paired samples t-test were utilized to within-group changes of quantitative variables, and qualitative variables were analyzed with the McNemar test., Results: The results showed that pain intensity and disability significantly decreased within and between groups after intervention (P > 0.05). There were also statistically significant differences between treatment groups for 3 out of 6 alignment and movement impairments (PS-FAdd/IR, Step down-Add/Valgus, and STS-Add/Valgus) (P > 0.05). Within-group differences for alignment and movement impairments were significant only for the MSI-based treatment group (P > 0.05)., Conclusions: The findings suggest that a specific MSI-based treatment, considering a homogenous group of individuals with knee pain, may contribute to pain, disability, and alignment and movement impairments improvement. Therefore, it is important to notice the classification-based treatment for individuals with knee pain., Trial Registration Number (trn) and Date of Registration: The trial was registered at the ( https://www.irct.ir ), (IRCT20210505051181N3) on 17/7/2021., (© 2024. The Author(s).)

Purpose: Our objective was to compare the serum Adropin levels between patients with wet-type Age-Related Macular Degeneration (AMD) and otherwise healthy individuals., Method: The study included 45 patients with wet-type AMD and 45 individuals without age-related macular degeneration. Patients with co-morbidities such as diabetes, hypertension, autoimmune diseases, and a previous history of visual impairment; were excluded. FBS, Hemoglobin A1C (HbA1C), lipid profile, and serum Adropin level were checked., Results: The mean serum Adropin level of patients with wet-type AMD was significantly lower than the control group (P-value < 0.001). Also, the mean High-sensitivity C-reactive protein ( hsCRP) level and High Density Lipoprotein (HDL) were significantly higher in wet-type AMD patients (P-value = 0.031 and < 0.001 respectively)., Conclusions: In our study, wet-type AMD was associated with a lower level of serum Adropin. Because of Adropin involvement in glucose metabolism and age-related changes, it may have a role in the pathogenesis of AMD, but it requires more investigations at the molecular level to elucidate its function., (© 2024. The Author(s).)

Bisphenol A (BPA) is one of key raw materials used in the production of epoxy resins and plastics, which has toxicological effects on humans by disrupting cell functions through a variety of cell signaling pathways. Therefore, it is of great significance to develop a simple, rapid, and accurate BPA detection method in real water samples. In this study, a ratiometric fluorescence method based on yellow-emitting surface-functionalized polymer dots (PFBT@L Pdots) and blue-emitting carbon dots (Cdots) was described for the detection of BPA. Pdots as the detecting part were synthesized by using highly fluorescent hydrophobic Poly[(9,9-dioctylfluorenyl-2,7-diyl)-alt-co-(1,4-benzo-(2,1',3)-thiadiazole)] (PFBT) polymer and (R)-5,11,17,23-Tetra-tert-butyl-25,27-bis[(diphenylphosphinoyl)methoxy]-26-(3-oxabutyloxy)-28-[(1-phenylethyl)- carbamoylmethoxy]calix [4]arene (L) functionalizing ligand, and Cdots as internal reference were prepared by hydrothermal treatment of citric acid and urea. In the presence of BPA, chemical binding of the phosphorus atoms of nearby PFBT@L Pdots with BPA hydroxyl functional groups led to the aggregation of the PFBT@L Pdots aggregation and quenching their yellow emission, but the blue emission of Cdots, on the other hand, remained stable. The proposed PFBT@L Pdots probe was successfully applied for the detection of BPA in real water samples, and the results were in good agreement with those obtained by HPLC-FLD. To the best of our knowledge, this is the first report that the calixarene has been utilized to modify Pdots., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Synthesis and catalytic application of polymeric-based nanocomposites are important subjects among researchers due to their high lipophilicity as well as high chemical and mechanical stability. In the present work, a novel nanocomposite material involving ionic liquid and high-density polyethylene supported tungstate (PE/IL-WO 4 = ) is synthesized, characterized and its catalytic application is investigated. The coacervation method was used to incorporate 1-methyl-3-octylimidazolium bromide ([MOIm] [Br]) ionic liquid in high-density polyethylene, resulting in a PE/IL composite. Subsequently, tungstate was anchored on PE/IL to give PE/IL-WO 4 = catalyst. The PXRD, FT-IR, EDX, TGA, and SEM analyses were used to characterize the PE/IL-WO 4 = composite. This material demonstrated high catalytic efficiency in the synthesis of bioactive tetrahydrobenzo[a]xanthen-11-ones under green conditions. The recoverability and leching tests were performed to investigate the stability and durability of the designed PE/IL-WO 4 = catalyst under applied conditions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Mousavi, Elhamifar, Kargar and Elhamifar.)

Background: We hypothesize that the poor survival outcomes of end-stage kidney disease (ESKD) patients undergoing hemodialysis are associated with a low filtering efficiency and selectivity. The current gold standard criteria using single or several markers show an inability to predict or disclose the treatment effect and disease progression accurately. Methods: We performed an integrated mass spectrometry-based metabolomic and proteomic workflow capable of detecting and quantifying circulating small molecules and proteins in the serum of ESKD patients. Markers linked to cardiovascular disease (CVD) were validated on human induced pluripotent stem cell (iPSC)-derived cardiomyocytes. Results: We identified dozens of elevated molecules in the serum of patients compared with healthy controls. Surprisingly, many metabolites, including lipids, remained at an elevated blood concentration despite dialysis. These molecules and their associated physical interaction networks are correlated with clinical complications in chronic kidney disease. This study confirmed two uremic toxins associated with CVD, a major risk for patients with ESKD. Conclusion: The retained molecules and metabolite-protein interaction network address a knowledge gap of candidate uremic toxins associated with clinical complications in patients undergoing dialysis, providing mechanistic insights and potential drug discovery strategies for ESKD., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Lin, Mousavi, Blum, Heckendorf, Moore, Lampl, McComb, Kotelnikov, Yin, Rabhi, Layne, Kozakov, Chitalia and Emili.)