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Microfluidic concentration gradient generators are useful in drug testing. drug screening, and other cellular applications to avoid manual errors, save time, and labor. However, expensive fabrication techniques make such devices prohibitively costly. Here, in the present work, we developed a microfluidic concentration gradient generator (μCGG) using a recently proposed non-conventional photolithography-less method. In this method, ceramic suspension fluid was shaped into a square mesh by controlling Saffman Taylor instability in a Multiport Lifted Hele-Shaw Cell (MLHSC). Using the shaped ceramic structure as template, the microfluidic concentration gradient generator (μCGG) was prepared by soft lithography. The concentration gradient was characterized and effect of the flow rates were studied usingCOMSOL simulations. The simulation result was further validated by creating fluorescein dye (Fluorescein isothiocanate, FITC) gradient in the fabricated μCGG. To demonstrate the use of this device for drug testing, we created various concentrations of an anticancer drug - curcumin - using the device and determined its inhibitory concentration on cervical cancer cell-line HeLa. We found that the IC50 of curcumin for HeLa to be 28.6 ± 6.1 μM which matched well with the conventional muti-well drug testing method (34.9 ± 1.7 μM). This method of μCGG fabrication has multiple advantages over conventional photolithography such as: i) the channel layout and inlet-outlet arrangements can be changed by simply wiping the ceramic fluid before it solidifies, (ii) it is cost effective, (iii) large area patterning is easily achievable, and (iv) the method is scalable. This technique can be utilised to achieve broad range of concentration gradient to be used for various biological and non-biological applications.Table of Content

SOX2 is an oncogenic transcription factor overexpressed in nearly half of the basal-like triple-negative breast cancers associated with very poor outcomes. Targeting and inhibiting SOX2 is clinically relevant as high SOX2 mRNA levels are positively correlated with decreased overall survival and progression-free survival in patients affected with breast cancer. Given its key role as a master regulator of cell proliferation, SOX2 represents an important scaffold for the engineering of dominant-negative synthetic DNA-binding domains (DBDs) that act by blocking or interfering with the oncogenic activity of the endogenous transcription factor in cancer cells. We have synthesized an interference peptide (iPep) encompassing a truncated 24 amino acid long C-terminus of SOX2 containing a potential SOX-specific nuclear localization sequence, and the determinants of the binding of SOX2 to the DNA and to its transcription factor binding partners. We found that the resulting peptide (SOX2-iPep) possessed intrinsic cell penetration and promising nuclear localization into breast cancer cells, and decreased cellular proliferation of SOX2 overexpressing cell lines. The novel SOX2-iPep was found to exhibit a random coil conformation predominantly in solution. Molecular dynamics simulations were used to characterize the interactions of both the SOX2 transcription factor and the SOX2-iPep with FGF4-enhancer DNA in the presence of the POU domain of the partner transcription factor OCT4. Predictions of the free energy of binding revealed that the iPep largely retained the binding affinity for DNA of parental SOX2. This work will enable the future engineering of novel dominant interference peptides to transport different therapeutic cargo molecules such as anti-cancer drugs into cells.

BackgroundHormone-dependent cancers (HDC) are among the leading causes of death worldwide among both men and women. Some of the established risk factors of HDC include unhealthy lifestyles, environmental factors, and genetic influences. Numerous studies have been conducted to understand gene–cancer associations. Transcriptome-wide association studies (TWAS) integrate data from genome-wide association studies (GWAS) and gene expression (expression quantitative trait loci – eQTL) to yield meaningful information on biological pathways associated with complex traits/diseases. Recently, TWAS have enabled the identification of novel associations between HDC risk and protein-coding genes.MethodsIn the present study, we performed a TWAS analysis using the summary data-based Mendelian randomization (SMR)–heterogeneity in dependent instruments (HEIDI) method to identify microRNAs (miRNAs), a group of non-coding RNAs (ncRNAs) associated with HDC risk. We obtained eQTL and GWAS summary statistics from the ncRNA-eQTL database and the National Human Genome Research Institute–European Bioinformatics Institute (NHGRI-EBI) GWAS Catalog.ResultsWe identified 13 TWAS-significant miRNAs at cis regions (±1 Mb) associated with HDC risk (two, five, one, two, and three miRNAs for prostate, breast, ovarian, colorectal, and endometrial cancers, respectively). Among them, eight novel miRNAs were recognized in HDC risk. Eight protein-coding genes targeted by TWAS-identified miRNAs (SIRT1, SOX4, RUNX2, FOXA1, ABL2, SUB1, HNRNPH1, and WAC) are associated with HDC functions and signaling pathways.ConclusionOverall, identifying risk-associated miRNAs across a group of related cancers may help to understand cancer biology and provide novel insights into cancer genetic mechanisms. This customized approach can be applied to identify significant miRNAs in any trait/disease of interest.

Fluorescent voltage indicators are an attractive alternative for studying the electrical activity of excitable cells; however, the development of indicators that are both highly sensitive and low in toxicity over long-term experiments remains a challenge. Previously, we reported a fluorene-based voltage-sensitive fluorophore that exhibits much lower phototoxicity than previous voltage indicators in cardiomyocyte monolayers, but suffers from low sensitivity to membrane potential changes. Here, we report that the addition of a single vinyl spacer in the fluorene molecular wire scaffold improves the voltage sensitivity 1.5- to 3.5-fold over fluorene-based voltage probes. Furthermore, we demonstrate the improved ability of the new vinyl-fluorene VoltageFluors (v-fVFs) to monitor action potential kinetics in both mammalian neurons and human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Addition of the vinyl spacer between the aniline donor and fluorene monomer results in indicators that are significantly less phototoxic in cardiomyocyte monolayers. These results demonstrate how structural modification to the voltage sensing domain have a large effect on improving the overall properties of molecular wire-based voltage indicators.

Background: Secondaries of brain (Brain metastases, BM) are most common central nervous system (CNS) tumors and it may be initial manifestation of systemic malignancies. Histomorphology of brain metastatic tumors along with immunohistochemistry (IHC) will helpful in the detection of unknown primary site (UPS) of brain metastases. Aim: To evaluate unknown primary site of brain metastases with help of immunohistochemistry. Material and Method: This study was conducted at the department of onco-pathology of our institute. Five years data from 2014 to 2018 were obtained from online Laboratory Information System (LIS), department of onco-pathology of our institute. IHC was performed on basis of histomorphology. Results: Total 128 brain metastatic tumors were retrieved. Of which the present study have applied IHC on 107 cases and in remaining 21 cases, systemic malignancies were already diagnosed. Out of 107 cases, primary site was detected in 92.53% cases (n=99) and in 7.47% cases (n = 8), not able to detect primary site even after IHC. Lung was the most common primary site detected (n = 73) followed Gastrointestinal tract (GIT, n = 9). Conclusion: Lung followed by breast are the most common primary malignancies for secondaries of brain (BM). Systemic approach using immunohistochemistry is really helpful in detection of unknown primary site of secondaries of brain (BM).

The DNA repair capacity of human cells declines with age, in a process that is not clearly understood. Mutation of the nuclear envelope protein barrier-to-autointegration factor 1 (Banf1) has previously been shown to cause a human progeroid disorder, Néstor–Guillermo progeria syndrome (NGPS). The underlying links between Banf1, DNA repair and the ageing process are unknown. Here, we report that Banf1 controls the DNA damage response to oxidative stress via regulation of poly [ADP-ribose] polymerase 1 (PARP1). Specifically, oxidative lesions promote direct binding of Banf1 to PARP1, a critical NAD+-dependent DNA repair protein, leading to inhibition of PARP1 auto-ADP-ribosylation and defective repair of oxidative lesions, in cells with increased Banf1. Consistent with this, cells from patients with NGPS have defective PARP1 activity and impaired repair of oxidative lesions. These data support a model whereby Banf1 is crucial to reset oxidative-stress-induced PARP1 activity. Together, these data offer insight into Banf1-regulated, PARP1-directed repair of oxidative lesions., Mutation of the nuclear envelope protein, barrier-to-autointegration factor 1 (Banf1), has previously been associated with the development of ageing associated diseases in a human progeria syndrome. Here, the authors reveal the functional link between Banf1-regulated, PARP1-directed repair of oxidative lesions.

Brucellosis is an important zoonosis worldwide. In livestock, it frequently causes chronic disease with reproductive failures that contribute to production losses, and in humans, it causes an often-chronic febrile illness that is frequently underdiagnosed in many low- and middle-income countries, including India. India has one of the largest ruminant populations in the world, and brucellosis is endemic in the country in both humans and animals. In November 2017, the International Livestock Research Institute invited experts from government, national research institutes, universities, and different international organizations to a one-day meeting to set priorities towards a "One Health" control strategy for brucellosis in India. Using a risk prioritization exercise followed by discussions, the meeting agreed on the following priorities: collaboration (transboundary and transdisciplinary); collection of more epidemiological evidence in humans, cattle, and in small ruminants (which have been neglected in past research); Economic impact studies, including cost effectiveness of control programmes; livestock vaccination, including national facilities for securing vaccines for the cattle population; management of infected animals (with the ban on bovine slaughter, alternatives such as sanctuaries must be explored); laboratory capacities and diagnostics (quality must be assured and better rapid tests developed); and increased awareness, making farmers, health workers, and the general public more aware of risks of brucellosis and zoonoses in general. Overall, the meeting participants agreed that brucellosis control will be challenging in India, but with collaboration to address the priority areas listed here, it could be possible.

Energy detection based non-cooperative and cooperative spectrum sensing methods are considered in this paper for a full-duplex (FD) cognitive radio (CR) network. The CRs are assumed to be mobile and the wireless channels between CR(s) and primary transmitter (PT) are considered as time-selective. Such time-selective channels are characterized using Nakagami-m fading. The effect of mobility is captured using Jake’s model. The expressions for the false alarm and detection probabilities are derived under the influence of Nakagami-m distributed residual self-interference (RSI). Moreover, with double threshold at CR, we analyze the receiver operating characteristics (ROC), area under ROC curve (AUC) and the total error rate performances. All the analytical results are validated through Monte Carlo simulations.

In view of easy availability and increasing trend of consumption of fast foods and sugar sweetened beverages (fruit juices and drinks, carbonated drinks, energy drinks) in Indian children, and their association with increasing obesity and related non-communicable diseases, there is a need to develop guidelines related to consumption of foods and drinks that have the potential to increase this problem in children and adolescents. To review the evidence and formulate consensus statements related to terminology, magnitude of problem and possible ill effects of junk foods, fast foods, sugar-sweetened beverages and carbonated drinks; and to formulate recommendations for limiting consumption of these foods and beverages in Indian children and adolescents. Process: A National Consultative group constituted by the Nutrition Chapter of the Indian Academy of Pediatrics (IAP), consisting of various stakeholders in private and public sector, reviewed the literature and existing guidelines and policy regulations. Detailed review of literature was circulated to the members, and the Group met on 11th March 2019 at New Delhi for a day-long deliberation on framing the guidelines. The consensus statements and recommendations formulated by the Group were circulated to the participants and a consensus document was finalized. The Group suggests a new acronym ‘JUNCS’ foods, to cover a wide variety of concepts related to unhealthy foods (Junk foods, Ultra-processed foods, Nutritionally inappropriate foods, Caffeinated/colored/carbonated foods/beverages, and Sugar-sweetened beverages). The Group concludes that consumption of these foods and beverages is associated with higher free sugar and energy intake; and is associated with higher body mass index (and possibly with adverse cardiometabolic consequences) in children and adolescents. Intake of caffeinated drinks may be associated with cardiac and sleep disturbances. The Group recommends avoiding consumption of the JUNCS by all children and adolescents as far as possible and limit their consumption to not more than one serving per week. The Group recommends intake of regional and seasonal whole fruits over fruit juices in children and adolescents, and advises no fruit juices/drinks to infants and young children (age 5–18 y, their intake should be limited to 125 mL/day and 250 mL/day, respectively. The Group recommends that caffeinated energy drinks should not be consumed by children and adolescents. The Group supports recommendations of ban on sale of JUNCS foods in school canteens and in near vicinity, and suggests efforts to ensure availability and affordability of healthy snacks and foods. The Group supports traffic light coding of food available in school canteens and recommends legal ban of screen/print/digital advertisements of all the JUNCS foods for channels/magazines/websites/social media catering to children and adolescents. The Group further suggests communication, marketing and policy/taxation strategies to promote consumption of healthy foods, and limit availability and consumption of the JUNCS foods.

Dengue virus (DENV) is the most prevalent mosquito-borne flavivirus that infects humans. At present, there are no specific antiviral drugs to treat DENV infection and vaccine development has met with challenges. DENV encodes two glycosaminoglycan (GAG) binding proteins; Envelope (E) and non-structural protein 1 (NS1). While previous work has validated the use of GAG analogues as inhibitors of E mediated virus-cell attachment, their potential for antiviral intervention in NS1 protein toxicity has not yet been explored. Here, we investigate the potential of the heparan sulfate mimetic PG545 as a dual purpose compound to target both DENV virion infectivity and NS1 function. In comparison to a non-sulfated analogue, we show that PG545 potently inhibits DENV infectivity with no cytotoxic effect. Against NS1, PG545 completely blocks the induction of cellular activation and abolishes NS1-mediated disruption of endothelial monolayer integrity. Furthermore, PG545 treatment moderately improves survival from lethal DENV challenge in a murine model. At peak disease, PG545-treated mice have lower viremia, circulating NS1 and serum TNF-α. Consistent with anti-NS1 activity, PG545 treatment also reduces systemic vascular leakage caused by DENV infection in vivo. Taken together, these findings demonstrate that the dual targeting of DENV virions and NS1 using GAG analogues offers a new avenue for DENV drug development.

© 2019 Elsevier B.V. Inhaled nanoparticles (NPs) are experienced by the first biological barrier inside the alveolus known as lung surfactant (LS), a surface tension reducing agent, consisting of phospholipids and proteins in the form of the monolayer at the air-water interface. The monolayer surface tension is continuously regulated by the alveolus compression and expansion and protects the alveoli from collapsing. Inhaled NPs can reach deep into the lungs and interfere with the biophysical properties of the lung components. The interaction mechanisms of bare gold nanoparticles (AuNPs) with the LS monolayer and the consequences of the interactions on lung function are not well understood. Coarse-grained molecular dynamics simulations were carried out to elucidate the interactions of AuNPs with simplified LS monolayers at the nanoscale. It was observed that the interactions of AuNPs and LS components deform the monolayer structure, change the biophysical properties of LS and create pores in the monolayer, which all interfere with the normal lungs function. The results also indicate that AuNP concentrations >0.1 mol% (of AuNPs/lipids) hinder the lowering of the LS surface tension, a prerequisite of the normal breathing process. Overall, these findings could help to identify the possible consequences of airborne NPs inhalation and their contribution to the potential development of various lung diseases.

System xc− (Sxc−), a cystine‐glutamate antiporter, is established as an interesting target for the treatment of several pathologies including epileptic seizures, glioma, neurodegenerative diseases, and multiple sclerosis. Erastin, sorafenib, and sulfasalazine (SSZ) are a few of the established inhibitors of Sxc−. However, its pharmacological inhibition with novel and potent agents is still very much required due to potential issues, for example, potency, bioavailability, and blood–brain barrier (BBB) permeability, with the current lead molecules such as SSZ. Therefore, in this study, we report the synthesis and structure–activity relationships (SAR) of SSZ derivatives along with molecular docking and dynamics simulations using the developed homology model of xCT chain of Sxc− antiporter. The generated homology model attempted to address the limitations of previously reported comparative protein models, thereby increasing the confidence in the computational modeling studies. The main objective of the present study was to derive a suitable lead structure from SSZ eliminating its potential issues for the treatment of glioblastoma multiforme (GBM), a deadly and malignant grade IV astrocytoma. The designed compounds with favorable Sxc− inhibitory activity following in vitro Sxc− inhibition studies, showed moderately potent cytotoxicity in patient‐derived human glioblastoma cells, thereby generating potential interest in these compounds. The xCT‐ligand model can be further optimized in search of potent lead molecules for novel drug discovery and development studies.

Yttrium monosilicate (YMS) is a candidate environmental barrier coating material. It is a line compound, and its synthesis requires strict stoichiometric control of the starting materials. In this work, YMS was synthesised by the sol-gel method. Yttrium oxide and tetraethyl orthosilicate (TEOS) were used as precursors. The ratio of water to ethanol was varied to study the role of water in the formation of phase-pure YMS. The formation of secondary phases during synthesis was attributed to the complex interaction of TEOS with water in the presence of ethanol. It was seen that water and ethanol play a significant role in aqueous sol-gel processes involving silica than merely being present as solvents. The results are helpful in identifying the suitable sol-gel parameters for the synthesis of phase-pure yttrium monosilicate.

Amorphous alumina-yttria powders with 20, 25, 30 and 37.5 mol% yttria were synthesised by a co-precipitation method using the respective nitrates as precursors for identifying a composition with the best amorphous phase stability. Thermal analysis showed that calcination to yield amorphous oxides completed at around 800 °C. Crystallisation behaviour investigated by x-ray diffraction (XRD) showed that Al2O3–25 mol% Y2O3 (25Y) had the highest crystallisation temperature of 900 °C. Crystallisation involved the appearance and disappearance of metastable hexagonal yttrium aluminate (H-YAlO3) below 1000 °C. The phase evolution sequences for all the compositions were studied in detail. All the phases were nanocrystalline, establishing the efficacy of amorphous phase crystallisation as a viable route for the synthesis of nanocrystalline ceramics. Solution combustion synthesis (SCS) was selected to synthesis 25Y with enhanced amorphous phase stability using different proportions of citric acid and ethylene glycol. Thermal analysis revealed that the powder was free of organic residue after heat treating at 650 °C. The powder produced with 10 vol% of fuel (citric acid + ethylene glycol) yielded the highest crystallisation temperature of 925 °C and the appearance and disappearance of H-YAlO3 between 950 and 1100 °C. Fourier transform infrared (FT-IR) spectroscopy revealed that the co-precipitation synthesis might have residual NH4+ and NO3- radicals whereas SCS did not contain any residual carbonate above 700 °C.

Objective: This article describes the preparation of a 170Tm source by chemical deposition technique, its encapsulation in a titanium holder, and preliminary quality evaluation for potenti...

Nanotechnology has quickly emerged as a promising research field with potential effects in disease treatments. For example, gold nanoparticles (AuNPs) have been extensively used in diagnostics and therapeutics. When administrated into human tissues, AuNPs first encounter extracellular matrix (ECM) molecules. Amongst all the ECM components, collagen is the main tension-resisting constituent, whose biofunctional and mechanical properties are strongly dependent on its hierarchical structure. Therefore, an in-depth understanding of the structural response of collagen to the presence of gold nanosurfaces (AuNS) and AuNPs is crucial in terms of clinical applications of AuNPs. However, detailed understanding of the molecular-level and atomic-level interaction between AuNS/AuNPs and collagen in the ECM is elusive. In this study, comprehensive molecular dynamics (MD) simulations have been performed to investigate the molecular behaviour of a collagen molecule segment (CMS) in the presence of AuNS/AuNPs in explicit water, aiming to explore the interaction of AuNS/AuNPs with collagen triple helices at the molecular and atomic levels. The results show that the CMS forms a rapid association with AuNS/AuNPs and undergoes a severe unfolding upon adsorption on AuNS/AuNPs, indicating an unfolding propensity of gold surfaces. We conclude that collagen triple helices unfold readily on AuNS and bare AuNPs, due to the interaction of gold surfaces with the protein backbone. The revealed clear unfolding nature and the unravelled atomic-level unfolding mechanism of collagen triple helices onto AuNPs contribute to the development of AuNPs for biomedical and therapeutic applications, and the design of gold-binding proteins.

A continuous, sealed endothelial membrane is essential for the blood-brain barrier (BBB) to protect neurons from toxins present in systemic circulation. Endothelial cells are critical sensors of the capillary environment, where factors like fluid shear stress (FSS) and systemic signaling molecules activate intracellular pathways that either promote or disrupt the BBB. The brain vasculature exhibits complex heterogeneity across the bed, which is challenging to recapitulate in BBB microfluidic models with fixed dimensions and rectangular cross-section microchannels. Here, a Cayley-tree pattern, fabricated using lithography-less, fluid shaping technique in a modified Hele-Shaw cell is used to emulate the brain vasculature in a microfluidic chip. This geometry generates an inherent distribution of heterogeneous FSS, due to smooth variations in branch height and width. hCMEC/D3 endothelial cells cultured in the Cayley-tree designed chip generate a 3D monolayer of brain endothelium with branching hierarchy, enabling the study of the effect of heterogeneous FSS on the brain endothelium. The model is employed to study neuroinflammatory conditions by stimulating the brain endothelium with tumor necrosis factor-α under heterogeneous FSS conditions. The model has immense potential for studies involving drug transport across the BBB, which can be misrepresented in fixed dimension models.

Phosphorylation of the neuronal microtubule-associated Tau protein plays a critical role in the aggregation process leading to the formation of insoluble intraneuronal fibrils within Alzheimer’s disease (AD) brains. In recent years, other posttranslational modifications (PTMs) have been highlighted in the regulation of Tau (dys)functions. Among these PTMs, the O-β-linked N-acetylglucosaminylation (O-GlcNAcylation) modulates Tau phosphorylation and aggregation. We here focus on the role of the PHF-1 phospho-epitope of Tau C-terminal domain that is hyperphosphorylated in AD (at pS396/pS404) and encompasses S400 as the major O-GlcNAc site of Tau while two additional O-GlcNAc sites were found in the extreme C-terminus at S412 and S413. Using high resolution NMR spectroscopy, we showed that the O-GlcNAc glycosylation reduces phosphorylation of PHF-1 epitope by GSK3β alone or after priming by CDK2/cyclin A. Furthermore, investigations of the impact of PTMs on local conformation performed in small peptides highlight the role of S404 phosphorylation in inducing helical propensity in the region downstream pS404 that is exacerbated by other phosphorylations of PHF-1 epitope at S396 and S400, or O-GlcNAcylation of S400. Finally, the role of phosphorylation and O-GlcNAcylation of PHF-1 epitope was probed in in-vitro fibrillization assays in which O-GlcNAcylation slows down the rate of fibrillar assembly while GSK3β phosphorylation stimulates aggregation counteracting the effect of glycosylation.

The purpose of this study was to evaluate the effect of race, insurance status, and socioeconomic status on successful or unsuccessful healing of osteochondritis dissecans (OCD) lesions in the pediatric knee. The authors retrospectively reviewed patients younger than 18 years who were treated for a knee OCD lesion between 2006 and 2017. Patients were required to have at least 6 months of clinical and radiographic follow-up to be included, unless complete healing was achieved sooner. The primary outcome of interest was healing of the OCD lesion based on radiographic and clinical examination. A total of 204 OCD lesions in 196 patients with a mean follow-up of 15.8±6.4 months were included. The mean age at initial presentation was 12.4±2.8 years. At most recent follow-up, 28 (13.7%) lesions did not show radiographic or clinical evidence of healing. Nonhealing lesions were found in 25.0% of Black children compared with 9.4% of White children ( P =.02). After controlling for age, sex, sports participation, lesion size and stability, skeletal maturity, and operative vs nonoperative treatment in a multivariate model, Black children had 6.7 times higher odds of unsuccessful healing compared with their White counterparts (95% CI, 1.1–41.7; P =.04). In this study, Black children with OCD of the knee were significantly less likely to heal than were White patients, even when controlling for numerous other factors in a multivariate model. Although the exact etiology of this finding is unclear, future work should focus on the social, economic, and cultural factors that may lead to disparate outcomes. [ Orthopedics . 2021;44(3):e378–e384.]

Platinum-based chemotherapy remains the cornerstone of treatment for most non-small cell lung cancer (NSCLC) cases either as maintenance therapy or in combination with immunotherapy. However, resistance remains a primary issue. Our findings point to the possibility of exploiting levels of cell division cycle associated protein-3 (CDCA3) to improve response of NSCLC tumours to therapy. We demonstrate that in patients and in vitro analyses, CDCA3 levels correlate with measures of genome instability and platinum sensitivity, whereby CDCA3high tumours are sensitive to cisplatin and carboplatin. In NSCLC, CDCA3 protein levels are regulated by the ubiquitin ligase APC/C and cofactor Cdh1. Here, we identified that the degradation of CDCA3 is modulated by activity of casein kinase 2 (CK2) which promotes an interaction between CDCA3 and Cdh1. Supporting this, pharmacological inhibition of CK2 with CX-4945 disrupts CDCA3 degradation, elevating CDCA3 levels and increasing sensitivity to platinum agents. We propose that combining CK2 inhibitors with platinum-based chemotherapy could enhance platinum efficacy in CDCA3low NSCLC tumours and benefit patients., Kildey et al find that high levels of mitotic regulator CDCA3 correlates with sensitivity to platinum agents in non-small cell lung cancer patients and cell lines. They show that interfering with CDCA3 degradation through CK2 inhibition enhances CDCA3 levels and increases sensitivity to platinum agents suggesting a therapeutic route.

Genomic stability is critical for normal cellular function and its deregulation is a universal hallmark of cancer. Here we outline a previously undescribed role of COMMD4 in maintaining genomic stability, by regulation of chromatin remodelling at sites of DNA double-strand breaks. At break-sites, COMMD4 binds to and protects histone H2B from monoubiquitination by RNF20/RNF40. DNA damage-induced phosphorylation of the H2A-H2B heterodimer disrupts the dimer allowing COMMD4 to preferentially bind H2A. Displacement of COMMD4 from H2B allows RNF20/40 to monoubiquitinate H2B and for remodelling of the break-site. Consistent with this critical function, COMMD4-deficient cells show excessive elongation of remodelled chromatin and failure of both non-homologous-end-joining and homologous recombination. We present peptide-mapping and mutagenesis data for the potential molecular mechanisms governing COMMD4-mediated chromatin regulation at DNA double-strand breaks., Amila Suraweera et al. use a range of biochemical and in vitro cellular assays to examine the role of the COMMD4 in DNA repair. Their results suggest that COMMD4 interacts with the histone H2A-H2B during repair of double-stranded DNA breaks, thereby maintaining genomic stability by regulating chromatin structure.

Purpose/objective(s) Retrospective and prospective studies in patients with oligometastatic non-small-cell lung cancer (NSCLC) suggested clinical benefit after complete metastatic ablation (CMA). So far, no studies have addressed the toxicity and efficacy of CMA to oligometastatic lesions in patients that progressed during immune checkpoint inhibitors (ICIs) therapy. Here, we report results of a single arm- radiotherapy (RT) to oligometastatic lesions from our ongoing phase II trial (NCT02710253). Materials/methods This study enrolled patients that progressed within 6 months of ICI therapy and developed oligo-metastases. Treatment intervention allowed for any dose and fractions of RT per the treating physician to all metastatic sites, with irradiation for up to 6 sites. Primary endpoints of this study are time to new lesion development, and disease control rate (DCR) by RECIST 1.1. Secondary endpoints are toxicity, progression-free survival (PFS), overall survival (OS) and absolute lymphocyte counts (ALC). Results Of the 30 patients initially enrolled in this study, 26 patients (N = 21 NSCLC; N = 5 melanoma) were included in the analysis as they had completed imaging follow-up. A total of 64 lesions were irradiated with a mean of 2.5 lesions per patient. Most frequent location of irradiated was lung (n = 45), followed by adrenal (n = 7), liver (n = 4), bone (n = 4) and others (n = 4); 50% of patients received SBRT. At a median follow-up time of 23.0 months (range, 5.0-44.7 months), 12 patients had no new lesions, while 14 patients developed new lesions in lung (50%), bone (25%), brain (21%). Median time to new lesion development was 14.6 months. DCR was 77% (complete response 19%, partial response 19%, stable disease 38%); median PFS was 10.1 months and median OS was not reached. No toxicities greater than grade 2 was observed. Pre-RT ALC is an independent risk factor for time to new lesion development (P = 0.006, HR = 0.1, 95% CI, 0.02-0.51). Patients without new lesion development had significantly higher baseline ALC (P = 0.011) and more ALC after RT at 1 month (P = 0.025), 3 months (P = 0.006), 6 months (P = 0.002) and 12 months (P = 0.045) compared to patients with new lesions. Other factors including age, gender, SBRT, baseline tumor size, performance status and tumor histology did not affect the time to new lesion development. Conclusion In patients with oligometastatic lesions that progressed on ICI, CMA may be a valid treatment option that generates clinical benefits. By eradicating gross sites of disease by converting area sites into in-situ vaccines, T cells are able to access microscopic metastasis. Larger randomized trials are indicated to further define the role of CMA for ICIs resistance.

PURPOSE/OBJECTIVE(S) Radiation therapy (XRT) induces lymphocyte infiltration of primary tumors that is increased when using NBTXR3 radioenhancer nanoparticle, leading to improved response to combination immunotherapy (IT). However, treatment effects on the metastatic tumor microenvironment are poorly understood. We studied the effects of NBTXR3 injection into primary tumors plus XRT plus dual-agent immunotherapy on secondary tumor-infiltrating T cells in a mouse model of lung cancer. The hypothesis is that intratumoral primary tumor treatment with NBTXR3 nanoparticle enriches conserved T cell receptor repertoires among metastatic tumor-infiltrating lymphocytes. MATERIALS/METHODS Four groups of 4 mice each were inoculated subcutaneously with anti-PD1-resistant 344SQR murine lung cancer cells in each hind leg, 3 days apart, to establish 'primary' (right) and 'secondary' (left, which represented metastasis) tumors. All mice received intraperitoneal anti-PD1 and anti-CTLA-4 (IT) on days 4, 7, 10, and 13, and 12-Gy high-dose XRT (HD) to the primary tumors on days 7, 8 and 9. Primary tumors in groups 2 and 4 also received intratumoral NBTXR3 on day 6. Secondary tumors in groups 3 and 4 were irradiated with 1-Gy low-dose XRT (LD) on days 12 and 13. Treatment groups were designated as 1 = HD+IT, 2 = NBTXR3+HD+IT, 3 = HD+LD+IT, and 4 = NBTXR3+HD+LD+IT. All secondary tumors were harvested on day 19 for tumor-infiltrating T-cell RNA extraction and sequencing on an Illumina MiSeq. T-cell receptor (TCR) beta complementarity-determining region 3 (CDR3b) repertoires were analyzed with MiXCR v3.0.12 software, and CDR3b analyses and Circos plots were generated using software. P < 0.05 from Mann-Whitney U tests were considered statistically significant. RESULTS NBTXR3 significantly increased the proportion of conserved CDR3b clonotypes within the secondary tumors among mice that received only HD XRT to the primary tumors (group 2 vs 1, P = 0.0476). When the secondary tumors were exposed to LD XRT, NBTXR3 still increased the frequency of overlapping clonotypes although not significantly (group 4 vs 3, P = 0.3095). Most interestingly, there were significantly more conserved CDR3b repertoires from the secondary tumors that were shared between groups 2 and 4 that received NBTXR3 compared to those shared between groups 1 and 3 (P < 0.0001). Comparisons of CDR3b normalized Shannon clonality indices and Circos plots did not reveal any difference in repertoire diversity between NBTXR3-treated and non-NBTXR3-treated groups. CONCLUSION NBTXR3 radioenhancer nanoparticle, given with immunoradiation, might reshape the TCR repertoire of metastatic lung cancer tumor-infiltrating T cells by driving the selection of conserved CDR3b clonotypes.

Colloidal nanoparticles, such as gold nanoparticles (AuNPs), are promising materials for the delivery of hydrophilic drugs via the pulmonary route. The inhaled nanoparticle drug carriers primarily deposit in lung alveoli and interact with the alveolar surface known as lung surfactants. Therefore, it is vital to understand the interactions of nanocarriers with the surfactant layer. To understand the interactions at the molecular level, here we simulated model lung surfactant monolayers with phospholipid (PL)-wrapped AuNPs at the vacuum-water interface using coarse-grained molecular dynamics simulations. The PL-wrapped AuNPs quickly adsorbed into the surfactant layer, altered the structural properties of the monolayer, and at high concentrations initiated the compressed monolayer to collapse/buckle. Among the surfactant monolayer lipid components, cholesterol adsorbed to the AuNPs preferentially over PL species. The position of the adsorbed PL-AuNPs within the monolayer, and subsequent monolayer perturbation, vary depending on the monolayer phase, monolayer composition, and species of PL used as a ligand. Information provided by these molecular dynamic simulations helps to rationalize why some colloidal nanoparticles work better as nanocarriers than others and aid the design of new ones, to avoid biological toxicity and improve efficacy for pulmonary drug delivery.

To assess current practice patterns with respect to protocols used for incidental pancreatic cyst follow-up, management guidelines, and template reporting.The Society of Abdominal Radiology Disease Focused Panel on intraductal pancreatic neoplasms distributed an anonymous 14-question survey to its members in June 2018 that focused on current utilization of incidental pancreatic cyst guidelines, protocols, and template reporting.Among the 1,390 email invitations, 323 responded, and 94.7% (306 of 323) completed all questions. Respondents were mainly radiologists (93.8%, 303 of 323) from academic institutions (74.7%, 227 of 304) in North America (93.7%, 286 of 305). Of respondents, 42.5% (136 of 320) preferred 2017 ACR recommendations, 17.8% (57 of 320) homegrown systems, 15.0% (48 of 320) Fukuoka guidelines, and 7.8% (25 of 320) American Gastroenterological Association guidelines. The majority (68.7%, 222 of 323) agreed or strongly agreed that developing a single international consensus recommendation for management was important, and most radiologists preferred to include them in reports (231 of 322, 71.7%); yet only half included recommendations in75% of reports (161 of 321). MR cholangiopancreatography was the modality of choice for follow-up of2.5 cm cysts. Intravenous contrast was routinely used by 69.7% (212 of 304). Standardized reporting templates were rarely used in practice (12.8% 39 of 306).Nearly 7 of 10 radiologists desire a unified international consensus recommendation for management of incidental cystic pancreatic lesions; ACR 2017 recommendations are most commonly used, followed by homegrown systems and Fukuoka guidelines. The majority of radiologists routinely use MR cholangiopancreatography with intravenous contrast for follow-up of incidental cystic lesions, but template reporting is rarely used.

In this research, we report the structure, microstructure and conduction mechanism study of nickel and cobalt co-substituted Mn3O4 with systematic variation in cobalt concentration. Co-substituted Mn3O4 was prepared by coprecipitation technique in an aqueous solution of metal nitrates and ammonia. X-ray diffraction study of nickel and cobalt co-substituted Mn3O4 established the pure and single-phase formations crystallized in tetragonal crystal symmetry. The overall crystallite size estimated by the Scherrer equation was ~ 30 nm to 50 nm after calcination whereas it was ~ 52 nm to 177 nm after sintering. The microstructure analysis was performed from a scanning electron microscope, and the average grain size was estimated between 2.6 µm to 7.8 µm. The DC conductivity of the substitute Mn3O4 was measured between 300 °C and 720 °C using the two-probe technique. The temperature-dependent resistivity measured for all co-substituted Mn3O4 samples showed decreasing resistivity trend confirming the negative temperature coefficient of resistance behavior. The increase in activation energy as a function of Co concentration was attributed to Verwey and de Bohr hopping mechanism.

Graphical abstract, Highlights • High-density lipoprotein subpopulations have unique surface profiles and dynamics. • Relative hydrophobic surface area decreases with increasing lipoprotein size. • Core lipid exposure at the lipoprotein surface decreases with increasing size. • Cholesterol molecules localise near apolipoprotein A-I central helices. • Lipid and protein interactions stabilise multifoil models of apolipoprotein A-I., Human serum high-density lipoproteins (HDLs) are a population of small, dense protein-lipid aggregates that are crucial for intravascular lipid trafficking and are protective against cardiovascular disease. The spheroidal HDL subfraction can be separated by size and density into five major subpopulations with distinct molecular compositions and unique biological functionalities: HDL3c, HDL3b, HDL3a, HDL2a and HDL2b. Representative molecular models of these five subpopulations were developed and characterised for the first time in the presence of multiple copies of its primary protein component apolipoprotein A-I (apoA-I) using coarse-grained molecular dynamics simulations. Each HDL model exhibited size, morphological and compositional profiles consistent with experimental observables. With increasing particle size the separation of core and surface molecules became progressively more defined, resulting in enhanced core lipid mixing, reduced core lipid exposure at the surface, and the formation of an interstitial region between core and surface molecules in HDL2b. Cholesterol molecules tended to localise around the central helix-5 of apoA-I, whilst triglyceride molecules predominantly interacted with aromatic, hydrophobic residues located within the terminal helix-10 across all subpopulation models. The three intermediate HDL models exhibited similar surface profiles despite having distinct molecular compositions. ApoA-I in trefoil, quatrefoil and pentafoil arrangements across the surface of HDL particles exhibited significant warping and twisting, but largely retained intermolecular contacts between adjacent apoA-I chains. Representative HDL subpopulations differed in particle size, morphology, intermolecular interaction profiles and lipid and protein dynamics. These findings reveal how different HDL subpopulations might exhibit distinct functional associations depending on particle size, form and composition.

In this work we present two new $\sim10^9$ particle self-consistent simulations of the merger of a Sagittarius-like dwarf galaxy with a Milky Way-like disc galaxy. One model is a violent merger creating a thick disc, and a Gaia-Enceladus/Sausage like remnant. The other is a highly stable disc which we use to illustrate how the improved phase space resolution allows us to better examine the formation and evolution of structures that have been observed in small, local volumes in the Milky Way, such as the $z-v_z$ phase spiral and clustering in the $v_{\mathrm{R}}-v_{\phi}$ plane when compared to previous works. The local $z-v_z$ phase spirals are clearly linked to the global asymmetry across the disc: we find both 2-armed and 1-armed phase spirals, which are related to breathing and bending behaviors respectively. Hercules-like moving groups are common, clustered in $v_{\mathrm{R}}-v_{\phi}$ in local data samples in the simulation. These groups migrate outwards from the inner galaxy, matching observed metallicity trends even in the absence of a galactic bar. We currently release the best fitting `present day' merger snapshots along with the unperturbed galaxies for comparison., Comment: 16 pages, 14 figures, accepted by MNRAS

Innovation in minimally invasive surgical technologies can have a substantial increase in the number of surgeries performed and the patient outcome. Based on the survey done among 100 laparoscopic surgeons throughout India, the need for highly articulated surgical instruments for laparoscopy was put forth irrespective of the experience of the surgeon. The present innovation is directed toward an end-effector mechanism that provides the wristed motion with no singularity in roll, pitch, and yaw. Besides, the end-effector is detachable, making it interchangeable and cost lowering. To obtain the desired yaw and pitch motions, various concepts were analyzed, and the best mechanism in terms of manufacturability, innovation potential, and various design constraints was selected to be tendon-driven rolling joint end-effector mechanism. In this mechanism, each disk is configured to rotate in at least one degree of freedom or DoF for each neighboring disk, which gives either yaw or pitch motion. To control the movements of the disk elements, actuation cables or tendon elements are used to manipulate the end-effector. Each link is in contact with the neighboring disk with a revolute joint. The kinematic model is developed for the novel end-effector using the D-H parameter convention to get the position and orientation of the tip in the fixed reference frame. The end-effector is characterized in terms of instrument workspace and surgical workspace, which is compared with the existing instrument workspace using MATLAB.

Contemporary subspecialization of practice in prostate pathology has seen a transition to complex, nuanced reporting, where a growing number of histopathologic parameters may signal differences in patient management. In this context, the International Society of Urological Pathology (ISUP) and the Genitourinary Pathology Society (GUPS) both published proceedings papers on the grading of prostate cancer in 2019. Overall, the 2 prostate cancer grading manuscripts reached many of the same conclusions and recommendations. Yet, each consensus was conducted somewhat differently, and in a couple of key areas, each reached different conclusions and recommendations. Herein, sourced from the experience and viewpoints of members of both societies, we provide the practicing pathologist a summary of the shared recommendations, and of the discordances. It is anticipated that these 2 documents will inform future iterations of recommendations and guidelines for reporting prostate cancer by organizations such as the College of American Pathologists, the Royal College of Pathologists, and the European Society of Pathology, which will promote best practices for their respective constituents. Our goal is to provide the practicing pathologist a useful catalog of the main points of both, allowing each practitioner to make informed decisions and understand any divergent opinions as may arise between observers for individual cases.

Background: Healthcare workers including nurses, lab technicians and multi-purpose workers are at a higher risk of contracting hepatitis B virus infection than the general population as they are exposed to blood, body fluids and sharps due to their professional activities. The objective of this study was to compare the knowledge about hepatitis B infection, its transmission and prevention among two groups, nurses and lab technicians versus MPWs.Methods: A cross-sectional study was carried out among 257 nurses and lab technicians versus 159 MPWs working at a tertiary care center. After clearance by the institutional ethical committee and informed consent of the participants, they were requested to fill a close ended questionnaire. The data was analyzed statistically using the JASP software.Results: The self-perceived risk of contracting infectious diseases owing to their profession was similar in nurses and lab technicians and MPWs. The awareness about hepatitis B infection, all the correct modes of transmission of the virus and the chances of the infection leading to liver diseases and cancer was higher in nurses and lab technicians than in MPWs. Awareness regarding prevention of infection and the vaccination rates were also higher among nurses and lab technicians. Knowledge about methods of prevention of the infection was found to be significantly more among the nurses and lab technicians.Conclusions: Overall, there is a significant gap in the knowledge about hepatitis B infection, its transmission and prevention in the MPWs. Vaccination coverage is also low in MPWs.