Potential Screening Method Research Articles

Single molecule tracking based drug screening

The single-molecule tracking of transmembrane receptors in living cells has provided significant insights into signaling mechanisms, such as mobility and clustering upon their activation/inactivation, making it a potential screening method for drug discovery. Here we show that single-molecule tracking-based screening can be used to explore compounds both detectable and undetectable by conventional methods for disease-related receptors. Using an automated system for a fast large-scale single-molecule analysis, we screen for epidermal growth factor receptor (EGFR) from 1134 of FDA approved drugs. The 18 hit compounds include all EGFR-targeted tyrosine kinase inhibitors (TKIs) in the library that suppress any phosphorylation-dependent mobility shift of EGFR, proving the concept of this approach. The remaining hit compounds are not reported as EGFR-targeted drugs and do not inhibit EGF-induced EGFR phosphorylation. These non-TKI compounds affect the mobility and/or clustering of EGFR without EGF and induce EGFR internalization, to impede EGFR-dependent cell growth. Thus, single-molecule tracking provides an alternative modality for discovering therapeutics on various receptor functions with previously untargeted mechanisms.

Abstract A045: Pan-viral serological characterization in African Americans and European Americans with Hepatocellular Carcinoma

Abstract Hepatocellular carcinoma (HCC) is often diagnosed at late-stages, particularly among African Americans (AA). Poor screening and diagnosis are key factors that contribute to worse prognoses and disproportionately high mortality rates. We propose investigating viral exposure history as a potential useful screening method. Additionally, by accounting for neighborhood deprivation index (NDI), a metric for differences in socio-economic statuses, we can potentially mitigate this disparity. With NDI and viral exposure history, our study aims to assess HCC prognoses in racially disparate populations. We used a technique called VirScan, which conjugates Phage-Immunoprecipitation (PhIP) and DNA-sequencing, to profile the antibody repertoire against a viral epitope library. Analyzing serum and plasma samples from population controls (PC), individuals at high-risk for developing HCC (HR), and HCC cases, we performed VirScan on 524 AA (272 PC, 205 HR, and 47 HCC) and 342 European Americans (EA) (141 PC, 130 HR, and 71 HCC) participants from the NCI-UMD cohort between 2009-2020 from the greater Baltimore area (NCT00913757; clinicaltrials.gov). We used XGBoost, a machine- learning model, to classify viral features that can discriminate HCC cases from PC group. XGBoost predicted 31 important viral features which distinguish HCC from PC. Of the 31 viral features, 6 viruses were different between AA and EA (FDR < 0.05). To determine the extent to which socio-economic factors contribute to the cancer disparities, we gathered variables from the 1999 census tract as contributors to deprivation. We used Principal Component Analysis (PCA) to determine loadings of these contributor variables; composite of the final variables is derived as NDI. Comparing across groups, we find that average NDI is highest in HR group (x = 0.33), followed by HCC group (x = 0.21), then PC group (x = 0.19). We find that NDI for AA is significantly higher in HCC group (p = 1.3e-09), HR group (p < 2.2e-16), and PC group (p = 5.0e-10) when compared to that of EA. Thus, we demonstrate that both the viral exposure history and NDI of an individual might be useful in prognoses of HCC. This approach could be used to examine the disparities we see in both incidence and mortality of HCC between AA and EA. Citation Format: Theressa Ewa, Whitney Leet, Limin Wang, Marshonna Forgues, Xin Wei Wang. Pan-viral serological characterization in African Americans and European Americans with Hepatocellular Carcinoma [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr A045.

Predictive value of soluble suppression of tumorigenicity 2 in atrial fibrillation: a systematic review and meta-analysis.

Atrial fibrosis is the main pathological basis for the pathogenesis and progression of atrial fibrillation (AF). Soluble suppression of tumorigenicity 2 (sST2) is involved in fibrosis. Recent studies have explored its predictive value in AF outcomes. We performed this study to assess whether sST2 is an independent biomarker of AF outcomes and explore the potential mechanism. PubMed, Web of Science, EMBASE, and Cochrane Library databases were searched systematically from inception through July 1, 2023, to identify relevant studies. Outcomes of interest included occurrence, recurrence, and major adverse cardiac events (MACEs) of AF. This meta-analysis was reported following the criteria outlined in PRISMA 2020, and the protocol was registered in PROSPERO (number: CRD42023459789). All statistical analyses were performed using the STATA version 16. Twenty four studies with 14,755 patients were included in the meta-analysis. The meta-analyses found that sST2 was significantly associated with the risk of occurrence [HR:1.04, 95% CI: 1.02-1.07, P < 0.01; I2 = 67.8%], recurrence [HR:1.09, 95% CI: 1.02-1.16, P < 0.01; I2 = 89.5%], and MACEs (HR:1.60, 95% CI: 1.13-2.27, P < 0.01; I2 = 82.0%) of AF. Furthermore, patients with AF showed higher sST2 than controls without AF (SMD: 0.41, 95% CI: 0.27-0.54, P < 0.01; I2 = 0%), and AF patients with recurrence after catheter ablation (CA) showed significantly higher sST2 than those without recurrence (SMD: 0.81, 95% CI: 0.33-1.28, P < 0.01; I2 = 83.9%). Sensitivity analyses showed that the outcomes were stable. Higher sST2 was association with an increased risk of occurrence, recurrence, and MACEs of AF. Assessing sST2 can be used as a potential screening method to predict AF outcomes. PROSPERO (CRD42023459789).

Trajectories of health conditions and their associations with the risk of cognitive impairment among older adults: insights from a national prospective cohort study

BackgroundThe associations between trajectories of different health conditions and cognitive impairment among older adults were unknown. Our cohort study aimed to investigate the impact of various trajectories, including sleep disturbances, depressive symptoms, functional limitations, and multimorbidity, on the subsequent risk of cognitive impairment.MethodsWe conducted a prospective cohort study by using eight waves of national data from the Health and Retirement Study (HRS 2002–2018), involving 4319 adults aged 60 years or older in the USA. Sleep disturbances and depressive symptoms were measured using the Jenkins Sleep Scale and the Centers for Epidemiologic Research Depression (CES-D) scale, respectively. Functional limitations were assessed using activities of daily living (ADLs) and instrumental activities of daily living (IADLs), respectively. Multimorbidity status was assessed by self-reporting physician-diagnosed diseases. We identified 8-year trajectories at four examinations from 2002 to 2010 using latent class trajectory modeling. We screened participants for cognitive impairment using the 27-point HRS cognitive scale from 2010 to 2018 across four subsequent waves. We calculated hazard ratios (HR) using Cox proportional hazard models.ResultsDuring 25,914 person-years, 1230 participants developed cognitive impairment. In the fully adjusted model 3, the trajectories of sleep disturbances and ADLs limitations were not associated with the risk of cognitive impairment. Compared to the low trajectory, we found that the increasing trajectory of depressive symptoms (HR = 1.39; 95% CI = 1.17–1.65), the increasing trajectory of IADLs limitations (HR = 1.88; 95% CI = 1.43–2.46), and the high trajectory of multimorbidity status (HR = 1.48; 95% CI = 1.16–1.88) all posed an elevated risk of cognitive impairment. The increasing trajectory of IADLs limitations was associated with a higher risk of cognitive impairment among older adults living in urban areas (HR = 2.30; 95% CI = 1.65–3.21) and those who smoked (HR = 2.77; 95% CI = 1.91–4.02) (all P for interaction < 0.05).ConclusionsThe results suggest that tracking trajectories of depressive symptoms, instrumental functioning limitations, and multimorbidity status may be a potential and feasible screening method for identifying older adults at risk of cognitive impairment.

Rapid non-invasive prenatal screening test for trisomy 21 based on digital droplet PCR

Non-invasive prenatal tests for the detection of fetal aneuploidies are predominantly based on the analysis of cell-free DNA (cfDNA) from the plasma of pregnant women by next-generation sequencing. The development of alternative tests for routine genetic laboratories is therefore desirable. Multiplex digital droplet PCR was used to detect 16 amplicons from chromosome 21 and 16 amplicons from chromosome 18 as the reference. Two fluorescently labeled lock nucleic acid probes were used for the detection of reaction products. The required accuracy was achieved by examining 12 chips from each patient using Stilla technology. The plasma cfDNA of 26 pregnant women with euploid pregnancies and 16 plasma samples from pregnancies with trisomy 21 were analyzed to determine the cutoff value for sample classification. The test was validated in a blind study on 30 plasma samples from pregnant patients with a risk for trisomy 21 ranging from 1:4 to 1:801. The results were in complete agreement with the results of the invasive diagnostic procedure (sensitivity, specificity, PPV, and NPV of 100%). Low cost, and speed of analysis make it a potential screening method for implementation into the clinical workflow to support the combined biochemical and ultrasound results indicating a high risk for trisomy 21.

Ferroptosis related gene signature in T cell-mediated rejection after kidney transplantation

BackgroundT cell-mediated rejection is an important factor affecting early transplant kidney survival. Ferroptosis has been shown to play a pathogenic role in a variety of diseases, which was not reported in TCMR. Here we developed a model for assessing activation of ferroptosis-related genes in TCMR to find a better screening method and explore the contribution of ferroptosis in TCMR.MethodsWe performed unsupervised consensus clustering according to expression of ferroptosis-related genes based on RNA-seq data from kidney transplant biopsies, and developed an assessment model characterized by ferroptosis gene expression through PCA, which was evaluated in multiple external datasets as well as blood and urine samples. Pathway enrichment and immune cell infiltration analysis were used to explore the possible targets and pathways involved in ferroptosis and TCMR.ResultsA ferroptosis gene expression scoring model was established. The diagnostic specificity and sensitivity of TCMR in renal biopsy samples were both over 80%, AUC = 0.843, and AUC was around 0.8 in multi-dataset validation, and was also close to 0.7 in blood and urine samples, while in predicting of graft survival at 3 years, scoring model had a good prognostic effect as well. Pathway enrichment and PPI network speculated that TLR4, CD44, IFNG, etc. may be the key genes of ferroptosis in TCMR.ConclusionsFerroptosis scoring model could better diagnose TCMR and predict graft loss, and could be used as a potential screening method in blood and urine samples. We speculate that ferroptosis plays an important role in TCMR.

Bile acids as drivers and biomarkers of hepatocellular carcinoma.

The prevalence of hepatocellular carcinoma (HCC) is rapidly increasing, driven not least in part by the escalating prevalence of non-alcoholic fatty liver disease. Bile acid (BA) profiles are altered in patients with HCC and there is a developing body of evidence from in vitro human cellular models as well as rodent data suggesting that BA are able to modulate fundamental processes that impact on cellular phenotype predisposing to the development of HCC including senescence, proliferation and epithelial-mesenchymal transition. Changes in BA profiles associated with HCC have the potential to be exploited clinically. Whilst excellent diagnostic and imaging tools are available, their use to screen populations with advanced liver disease at risk of HCC is limited by high cost and low availability. The mainstay for HCC screening among subjects with cirrhosis remains frequent interval ultrasound scanning. Importantly, currently available serum biomarkers add little to diagnostic accuracy. Here, we review the current literature on the use of BA measurements as predictors of HCC incidence in addition to their use as a potential screening method for the early detection of HCC. Whilst these approaches do show early promise, there are limitations including the relatively small cohort sizes, the lack of a standardized approach to BA measurement, and the use of inappropriate control comparator samples.

The sonographic quantitative assessment of the deltoid muscle to detect type 2 diabetes mellitus: a potential noninvasive and sensitive screening method?

BackgroundIn our previous published study, we demonstrated that a qualitatively assessed elevation in deltoid muscle echogenicity on ultrasound was both sensitive for and a strong predictor of a type 2 diabetes (T2DM) diagnosis. This study aims to evaluate if a sonographic quantitative assessment of the deltoid muscle can be used to detect T2DM.MethodsDeltoid muscle ultrasound images from 124 patients were stored: 31 obese T2DM, 31 non-obese T2DM, 31 obese non-T2DM and 31 non-obese non-T2DM. Images were independently reviewed by 3 musculoskeletal radiologists, blinded to the patient’s category. Each measured the grayscale pixel intensity of the deltoid muscle and humeral cortex to calculate a muscle/bone ratio for each patient. Following a 3-week delay, the 3 radiologists independently repeated measurements on a randomly selected 40 subjects. Ratios, age, gender, race, body mass index, insulin usage and hemoglobin A1c were analyzed. The difference among the 4 groups was compared using analysis of variance or chi-square tests. Both univariate and multivariate linear mixed models were performed. Multivariate mixed-effects regression models were used, adjusting for demographic and clinical variables. Post hoc comparisons were done with Bonferroni adjustments to identify any differences between groups. The sample size achieved 90% power. Sensitivity and specificity were calculated based on set threshold ratios. Both intra- and inter-radiologist variability or agreement were assessed.ResultsA statistically significant difference in muscle/bone ratios between the groups was identified with the average ratios as follows: obese T2DM, 0.54 (P < 0.001); non-obese T2DM, 0.48 (P < 0.001); obese non-T2DM, 0.42 (P = 0.03); and non-obese non-T2DM, 0.35. There was excellent inter-observer agreement (intraclass correlation coefficient 0.87) and excellent intra-observer agreements (intraclass correlation coefficient 0.92, 0.95 and 0.94). Using threshold ratios, the sensitivity for detecting T2DM was 80% (95% CI 67% to 88%) with a specificity of 63% (95% CI 50% to 75%).ConclusionsThe sonographic quantitative assessment of the deltoid muscle by ultrasound is sensitive and accurate for the detection of T2DM. Following further studies, this process could translate into a dedicated, simple and noninvasive screening method to detect T2DM with the prospects of identifying even a fraction of the undiagnosed persons worldwide. This could prove especially beneficial in screening of underserved and underrepresented communities.

Insertional Profiling of Pyrroloquinoline Quinone Glucose Dehydrogenase for Two-Component Biosensor Engineering

Colorectal cancer (CRC) is the third most common cancer worldwide and the second leading cause of cancer death. Currently, it is both expensive and invasive to screen for CRC. Developing a potential screening method that is more affordable and less invasive than endoscopy and biopsy would be beneficial to vulnerable communities. This method should also be able to diagnose and monitor the development and progression of GI cancers in real-time. A sensor that is capable of sending out an electrical signal in the presence of a cancer biomarker would alleviate the physical and economic costs of endoscopy while offering a significant amount of information about the gastrointestinal environment. Pyrroloquinoline quinone (PQQ) glucose dehydrogenase (PQQ-GDH), a redox enzyme, is capable of using glucose as a substrate for electron transfer. Engineering PQQ-GDH by insertion of a ligand-binding domain that is specific to a cancer biomarker would create an allosteric enzymatic switch that only produces current in the presence of the biomarker. To begin building this platform, I am creating a comprehensive insertion library to identify functionally important residues and domains of the PQQ-GDH. From there, I will profile which locations are capable of small primary, secondary, and/or tertiary structure disruptions. After identifying these regions, I will then create two-component protein switches capable of detecting cancer biomarkers. Creating an oxidoreductase switch that controls current transfer in the presence of gastrointestinal cancer biomarkers will advance the development of a novel, portable bio-recognition device capable of real-time, continuous sensing.

Serum pepsinogen: A potential non-invasive screening method for moderate and severe atrophic gastritis among an asian population

BackgroundSerum pepsinogen has been approved and used widely as an effective biomarker in diagnosis of atrophic gastritis and gastric cancer; however, its validity and appropriate cut-off values vary among different populations. This study aimed to initially assess the diagnostic value of the serum pepsinogen in diagnosis of moderate and severe atrophic gastritis for Vietnamese population. Materials and methodsA cross-sectional study enrolled 273 participants from June 2008 to November 2019. All participants underwent a gastroscopy procedure and three tests including serum PG test, pathology test, and Hp-Igg Elisa test. The Kimura-Takemoto classification and OLGA system were used to classify the mild versus moderate-severe atrophic gastritis. Receiver Operating Characteristic curve was used to assess the value of PGI, PGII and PGR. ResultsBased on Kimura-Takemoto classification, the AUC of PGI and PGR was 0.635 (p = 0.008, 95% CI 0.554–0.716) and 0.766 (p < 0.001, 95% CI 0.676–0.857) respectively. The best cut-off values were PGI ≤69.0 and PGR ≤4.6 (sensitivity: 73%, specificity: 83.9%, positive predictive value: 41.5%, negative predictive value: 95.2%, accuracy: 82.4%). According to the OLGA system, the AUC of PGI and PGR was 0.612 (p = 0.004, 95% CI 0.540–0.684) and 0.689 (p < 0.001, 95% CI 0.621–0.758) respectively. The best cut-off values were PGI ≤63.5 and PGR ≤5.2 (sensitivity: 49.4%, specificity: 82.1%, positive predictive value: 52.1%, negative predictive value: 80.5%, accuracy: 72.9%). ConclusionsThe serum pepsinogen II and pepsinogen I/II ratio had reliable diagnostic value for screening of moderate and severe atrophic gastritis among Vietnamese population. Further research was recommended to focus on larger scale to improve the diagnostic yield and to continue finding the cut-off values for diagnosis of gastric cancer among Vietnamese population.

Food-Grade Activated Charcoal for Contrast-Enhanced Photoacoustic Imaging of Aspiration: A Phantom Study.

Aspiration pneumonia has the highest attributable mortality of all medical complications post-stroke, or in individuals with progressive neurological diseases. For optimum health outcomes for individuals with dysphagia, a non-invasive and convenient method for objectively detecting aspiration is needed. This study introduces a potential new aspiration screening method based on photoacoustic imaging (PAI), a medical imaging technology that measures the optical contrast of tissue rather than mechanical or elastic properties. In this preliminary study, a tissue-mimicking neck phantom was designed to test the performance of PAI for aspiration screening with a charcoal solution as a contrast agent. A 1064nm wavelength light source was illuminated on the anterior of the neck phantom to induce the photoacoustic effect. The resulting photoacoustic signal of the charcoal contrast in the mock trachea was detected by a linear transducer array with a 2.25MHz central ultrasound frequency. The phantom results showed that charcoal solution at 10mg/ml exhibited strong photoacoustic signals when flowing into the phantom trachea. By overlaying the photoacoustic signals of the charcoal contrast on top of the ultrasound image, we were able to simultaneously visualize the movement of food contrast and a cross-section of tissue structures during mock swallowing. Moreover, we confirmed the ability to detect the flow of charcoal contrast at a small bolus volume of ~ 7 μl through the phantom, suggesting high sensitivity to detect small aspiration events. The study suggests that PAI holds promise to be developed as an aspiration detection tool with charcoal powder as a contrast agent.

Capillary microscopy is a potential screening method for connective tissue disease in children with Raynaud\u2019s phenomenon

BackgroundNailfold capillary microscopy (NCM) is a cornerstone in the diagnosis of Systemic Sclerosis (SSc) in adulthood. Although Raynaud’s phenomenon (RP) is very common in childhood, studies on diagnostic methods to differentiate between primary RP (PRP) and secondary RP (SRP) at a young age are scarce. The aim of this study was to determine the value of NCM in differentiating between PRP and SRP in children and adolescents with RP.MethodsIn this nested case–control study, 83 patients diagnosed with RP and having underwent NCM in childhood were retrospectively included. Based on whether they were diagnosed with a connective tissue disease (CTD) during follow-up, patients were classified as PRP or SRP. NCM was performed by a vascular technician. PRP and SRP patients were compared on demographics, NCM and serology. Variables associated with SRP were included in a multivariate logistic regression model. Predictive values were calculated for NCM, ANA positivity and the combination of NCM and ANA positivity.ResultsAt the time of the NCM, the mean age of the RP patients was 15.4 ± 2.3 years. Of these patients, 78.3% were classified as PRP and 21.7% as SRP at mean follow-up of 6.4 ± 3.20 years. CTDs were miscellaneous, with only one patient having developed SSc. Of the NCM parameters, only capillary loss was associated with SRP (p = 0.01). In a multivariate logistic regression model including ANA, capillary loss was not a predictor of SRP. In a model without ANAs, capillary loss was an independent predictor (OR = 3.98, CI 95% 1.22–12.99). Capillary loss had a sensitivity of 44.4% and a specificity of 84.4% for SRP. ANA combined with capillary loss had a sensitivity of 66.7% and a specificity of 85.7%.ConclusionWhereas RP in adulthood is most strongly associated with SSc, children with RP seem to be at risk for developing other CTDs with less apparent NCM abnormalities. Of all NCM findings, only capillary loss was predictive for SRP. NCM did not add to the predictive value of ANA screening. However, with a specificity of 84.4% and being non-invasive, NCM shows potential as a screening method for SRP. More research with a larger study population is required before drawing conclusions.

Evaluate the Correlation between the TIMI Frame Count, IMR, and CFR in Coronary Microvascular Disease.

Objective To evaluate the correlation between the TIMI frame count, IMR, and CFR in coronary microvascular disease (slow flow phenomenon). Methods TFC and IMR were recorded in the nitroglycerin and ATP administration states, and the relationship between TFC, IMR, and CFR in specific states was analyzed. Results A total of 41 patients with baseline TFC >25 frames on coronary angiography were enrolled, and nitroglycerin reduced TFC by 50% from baseline in 24 (58.54%) patients; 16 of the remaining 17 patients were able to achieve a 50% reduction in TFC by further intracoronary ATP injection. 10 patients were further tested for IMR, and the results showed significant correlations between baseline TFC and IMR (r = 0.775, P=0.008), TFC and IMR after nitroglycerin (r = 0.875, P=0.001), and the minimal TFC and IMR that could be obtained with nitroglycerin or ATP administration (r = 0.890, P=0.001). There was also a significant correlation between the proportional improvement in TFC and CFR before and after nitroglycerin injection (r = 0.685, P=0.029). In addition, we observed a lower IMR measured after nitroglycerin than after ATP in three patients, suggesting that CMD may be dominated by NO-sensitive vascular such as prearterioles and that an extensive analysis of the target site of CMD may be achieved by stepwise drug administration. Conclusion Induction of TFC in different states by a stepwise drug approach may serve as a potential primary screening method for coronary microcirculatory dysfunction, thereby reducing the need for further IMR or CFR testing.

A color channel based on multiple Random Forest coupled with a computer vision technique for the detection and prediction of Sudan dye‐I adulteration in turmeric powder

AbstractAdulterants can cause different health hazards upon prolonged consumption, but it is difficult to detect with human eyes. Non‐destructive turmeric adulteration detection is a challenging research area. The existing adulteration detection processes are largely instrumental and analytical with high accuracy but include limitations like long testing time, expensiveness, and lack of mobility. This work reports a new computer vision framework, which can simultaneously detect the presence of adulteration and predict the possible percentage of adulteration addressing the stated limitations. The scope has been remained to screening of Sudan dye‐I adulteration in turmeric powder. An in‐house database prepared with images of pure and adulterated turmeric powder samples has been used for experimentation. Random Forest algorithm has been employed for both classification and prediction. The model has been validated with standard internal and external validation methods to assess the stability and generalization potential of the model to avoid over‐ and under‐fitting problems. The results of classification show that the presented framework can provide more than 99% accuracy in detection while high correlation coefficient (R2) value in the tune of .99 for prediction. The novelty of the work is its simple histogram‐based color feature extraction, development of ensemble Random Forest prediction model that resulted in high accuracy and development of a faster, non‐invasive, less‐expensive, and validated screening method for adulterated turmeric powder that can be considered as a potential immediate screening method in the supply chain of powdered spices prior to confirmatory testing methods following two‐tiered food fraud testing approach.

A potential screening method for epigenetic drugs: uncovering stress-induced gene silencing in Chlamydomonas

Histone modifications and DNA methylation together govern promoter availability, thereby influencing gene expression. This study queries the unicellular chlorophyte, Chlamydomonas reinhardtii using a three step “epigenetic assay” design to phenotypically track the variegation of a randomly integrated Paromomycin resistance transgene(s) (PmR ). Based on its position of integration, the PmR gene expression hinged on two epigenetic hallmarks: the spreading of heterochromatin, and the transmissible memory of epigenetic states across generations. Using a spot-dilution analysis, the loss of antibiotic resistance phenotype was scored from 0 to 4, four being maximally silenced. Appropriate construct designs were used to demonstrate that the cis-spread of heterochromatin could be interfered with a stronger euchromatic barrier (TUB2 promoter). When assayed for metal ion stress, a combination of Mn deficiency with excess Cu or Zn stress was shown to induce gene silencing in Chlamydomonas. Cu stress resulted in the accumulation of intracellular ROS, while Zn stress elevated the sensitivity to ROS. As proof of functional conservation, mammalian epigenetic drugs demonstrably interfered with stress-induced gene silencing. Finally, a selected group of transgenic clones responsive to HDACi sodium butyrate, when tested in a gradient plate format showed similarity in phenotype to the plant-derived compound cinnamic acid. This indicated a possible commonality in their mode of action, unlike curcumin which might have a different mechanism. Thus, using binned libraries, based on a common set of responses to known drugs, a cost-effective high-throughput screening strategy for epigenetically active compounds from plants or other sources is described.

Detection of glucosamine as a marker for Aspergillus niger: a potential screening method for fungal infections

Several species of fungus from the genus Aspergillus are implicated in pulmonary infections in immunocompromised patients. Broad screening methods for fungal infections are desirable, as cultures require a considerable amount of time to provide results. Herein, we developed degradation and detection methods to produce and detect D-glucosamine (GlcN) from Aspergillus niger, a species of filamentous fungus. Ultimately, these techniques hold the potential to contribute to the diagnosis of pulmonary fungal infections in immunocompromised patients. In the following studies, we produced GlcN from fungal-derived chitin to serve as a marker for Aspergillus niger. To accomplish this, A. niger cells were lysed and subjected to a hydrochloric acid degradation protocol. Products were isolated, reconstituted in aqueous solutions, and analyzed using hydrophilic interaction liquid chromatography (HILIC) in tandem with electrospray ionization time-of-flight mass spectrometry. Our results indicated that GlcN was produced from A. niger. To validate these results, products obtained via fungal degradation were compared to products obtained from the degradation of two chitin polymers. The observed retention times and mass spectral extractions provided a two-step validation confirming that GlcN was produced from fungal-derived chitin. Our studies qualitatively illustrate that GlcN can be produced from A. niger; applying these methods to a more diverse range of fungi offers the potential to render a broad screening method for fungal detection pertinent to diagnosis of fungal infections.Graphical abstract

Halitosis in Oral and Maxillofacial Surgery Patients - a Pilot Study.

Halitosis is a relatively inhomogeneous pathology with an extremely high prevalence in the population. Potential risk factors for bad breath include bacterial decomposition of organic material as well as numerous general and systemic diseases. The aim of the present study was to analyze whether certain subgroups of oral and maxillofacial surgery patients have a higher risk of halitosis. Further the impact of halitosis on the patient's quality of life was ascertained. A total of 127 oral and maxillofacial patients aged between 19 and 86 years were enrolled in this study. On account of their underlining disease, patients were divided into five different investigation groups. The dental examination comprised tongue coating, periodontal screening index (PSI), gingival index (GI), PI (plaque index), DMF-T values as well as non-stimulated saliva flow rates. Halitosis was monitored both organoleptically according to Rosenberg and instrumentally by means of a Halimeter®, which records the volatile sulfur compounds (VSC values in ppm). Patients were further asked to fill out questionnaires regarding their medical history and oral hygiene, oral health (OHIP-14), and quality of life (BDI-II). Halitosis values, which were recorded by a Halimeter® correlated with the objective Rosenberg golden standard method. Furthermore, halitosis values correlated with elevated PSI, GI, and DMF-T values as well as the degree of tongue coating. Patients with oral cancer showed significantly higher VSC values compared to all other groups. No difference in VSC values could be found between all other patient groups. The Halimeter® could be validated as a suitable method for determining halitosis in oral and maxillofacial patients. The significantly increased halitosis values in cancer patients as opposed to all other patient groups suggests the potential of halitosis VSC values as a potential screening method. The development of non-invasive breath tests for diagnosis could be subject of future research.

Knowledge, attitudes, and practices of cervical Cancer screening among HIV-positive and HIV-negative women participating in human papillomavirus screening in rural Zimbabwe

BackgroundWomen in low- and middle-income countries are at the highest risk of cervical cancer yet have limited access to and participation in cervical cancer screening programs. Integrating self-collected, community-based screening offers a potential primary screening method in areas of limited resources. In this paper, we present a study evaluating knowledge, attitudes, and practices of cervical cancer and Human Papilloma Virus (HPV) in rural Zimbabwe.MethodsWe performed a community-based cross-sectional knowledge, attitudes and practices of HPV and cervical cancer study in rural Zimbabwe from January 2017–May 2017. Women were selected for the study via random number generation from complete lists of inhabitants in the study area if they satisfied the inclusion criteria (≥30-years-old, ≤65-years-old, not pregnant, intact uterus). If selected, they participated in a 19-question structured knowledge, attitudes and practices survey. The questionnaire included questions on demographics, education, knowledge of HPV, cervical cancer, and risk factors. Chi-squared tests were evaluated comparing knowledge, attitudes and practices relating to HPV and cervical cancer screening with actual infection with HPV. Women were also offered a voluntary HIV and self-collected HPV screening.ResultsSix hundred seventy-nine women were included in the knowledge, attitudes and practices survey. Most women (81%) had heard of cervical cancer while the majority had not heard of HPV (12%). The number of women that had been screened previously for cervical cancer was low (5%). There were no significant differences between and within groups regarding knowledge of cervical cancer and actual overall infection with HR-HPV, HPV 16, and HPV 18/45 test results.ConclusionsMost women in rural Zimbabwe have heard of cervical cancer, but the number that had been screened was low. Extending existing outreach services to include cervical cancer screening, potentially including HPV screening, should include cervical cancer/HPV education and screening triage. This approach would serve to bridge the gap between knowledge and screening availability to address some of the barriers to cervical cancer care still affecting women in many regions of the world.

MLH1 and MSH2 mismatch repair protein profile using immunohistochemistry in Nepalese colorectal cancer patients

BACKGROUND Hereditary nonpolyposis colorectal cancer, or Lynch syndrome, caused by germline mutations or genetic defects in mismatch repair (MMR) genes (MLH1, MSH2, PMS2, MSH6, and epithelial cellular adhesion molecule), is an autosomal dominant condition accounting for 2–5% of all colorectal carcinomas (CRCs). Reports on MMR loss in many populations are available; however, there are no reports on the frequency of MMR protein expression in Nepalese cohorts. Therefore, this study was aimed to assess the expression profiles of MLH1 and MSH2 protein by immunohistochemistry (IHC) in Nepalese CRC patients.&#x0D; METHODS This retrospective study used archived formalin-fixed paraffin-embedded tissue blocks from 43 Nepalese CRC patients. IHC staining was performed using MLH1 and MSH2 antibodies. IHC scoring analysis was assessed using semiquantitative scoring.&#x0D; RESULTS Of the 43 CRC patients, 8 (18.6%) showed loss of staining for MLH1 antibody, 5 (11.6%) showed loss of staining for MSH2 antibody, and 4 (9.3%) showed loss of staining for both MLH1 and MSH2 antibodies.&#x0D; CONCLUSIONS IHC is a potential screening method of determining the MMR expression profile of Nepalese CRC patients. IHC can be performed in local clinical laboratories to find MMR protein defects in selected cases prior to expensive molecular tests.

The Application of an Iterative Structure to the Delay-and-Sum and the Delay-Multiply-and-Sum Beamformers in Breast Microwave Imaging.

Breast microwave imaging (BMI) is a potential breast cancer screening method. This manuscript presents a novel iterative delay-and-sum (DAS) based reconstruction algorithm for BMI. This iterative-DAS (itDAS) algorithm uses a forward radar model to iteratively update an image estimate. A variation of the itDAS reconstruction algorithm that uses the delay-multiply-and-sum (DMAS) beamformer was also implemented (the itDMAS algorithm). Both algorithms were used to reconstruct images from experimental scans of an array of 3D-printed MRI-based breast phantoms performed with a clinical BMI system. The signal-to-clutter ratio (SCR) and signal-to-mean ratio (SMR) were used to compare the performance of the itDAS and itDMAS methods to the DAS and DMAS beamformers. While no significant difference between the itDAS and itDMAS methods was observed in most images, the itDAS algorithm produced reconstructions that had significantly higher SMR than the non-iterative methods, increasing contrast by as much as 19 dB over DAS and 13 dB over DMAS. The itDAS algorithm also increased the SCR of reconstructions by up to 5 dB over DAS and 4 dB over DMAS, indicating that both high-intensity and background clutter are reduced in images reconstructed by the itDAS algorithm.