Antagonistic roles of canonical and Alternative-RPA in disease-associated tandem CAG repeat instability.

Expansions of repeat DNA tracts cause >70 diseases, and ongoing expansions in brains exacerbate disease. During expansion mutations, single-stranded DNAs (ssDNAs) form slipped-DNAs. We find the ssDNA-binding complexes canonical replication protein A (RPA1, RPA2, and RPA3) and Alternative-RPA (RPA1, RPA3, and primate-specific RPA4) are upregulated in Huntington disease and spinocerebellar ataxia type 1 (SCA1) patient brains. Protein interactomes of RPA and Alt-RPA reveal unique and shared partners, including modifiers of CAG instability and disease presentation. RPA enhances in vitro melting, FAN1 excision, and repair of slipped-CAGs and protects against CAG expansions in human cells. RPA overexpression in SCA1 mouse brains ablates expansions, coincident with decreased ATXN1 aggregation, reduced brain DNA damage, improved neuron morphology, and rescued motor phenotypes. In contrast, Alt-RPA inhibits melting, FAN1 excision, and repair of slipped-CAGs and promotes CAG expansions. These findings suggest a functional interplay between the two RPAs where Alt-RPA may antagonistically offset RPA's suppression of disease-associated repeat expansions, which may extend to other DNA processes.

27-gene Immuno-Oncology (IO) Score is Associated With Efficacy of Checkpoint Immunotherapy in Advanced NSCLC: A Retrospective BC Cancer Study.

BACKGROUND: Immune checkpoint inhibitors (ICI) are standard of care in advanced non-small cell lung cancer (NSCLC). However, not all patients benefit, even among PD-L1 tumor proportional score (TPS) ≥50%, indicating an unmet need for additional biomarkers such as those assessing the tumor immune microenvironment (TIME). DetermaIO is a 27-gene assay that classifies TIME and has previously demonstrated association with ICI response. METHODS: FFPE samples were selected from BC Cancer and West Clinic Cancer Center patients with performance status (PS) ≤2 who received at least 2 cycles of ICI monotherapy in the first (1L) or second line (2L). IO scores were generated and analyzed for association with PFS and OS. RESULTS: In the entire cohort (N=147), IO score was significantly associated with OS (HR=0.68, 95%CI 0.47-0.99, P = .042) and PFS (HR=0.62, 95%CI 0.43-0.88, P = .0069). In 1L treated patients (PD-L1≥50%, N=78), IO score was significantly associated with PFS (HR=0.55, 95%CI 0.32-0.94, P = .028). In exploratory analyses, IO score was associated with benefit in 1L PS2 patients for OS (HR = 0.26, 95%CI 0.091-0.74, P = .012) and PFS (HR = 0.27, 95%CI 0.098-0.72, P = .0095) which was confirmed in PFS subgroup analysis in the independent West Cancer Center study (N=13 HR=0.14, 95%CI 0.027-0.76, P = .023). CONCLUSION: These data confirm the association of DetermaIO with ICI clinical benefit in NSCLC, and expand on previous studies by demonstrating that first line treated PD-L1≥50% patients can further be stratified by IO score to identify efficacy. Exploratory analysis suggested that the IO score identifies benefit in patients with poor PS.

Recent Trends and Advances in Porous Metal-Organic Framework Nanostructures for the Electrochemical and Optical Sensing of Heavy Metals in Water.

With the expansion and advancement in agricultural and chemical industries, various toxic heavy metals such as lead, cadmium, mercury, zinc, copper, arsenic etc. are continuously released into the environment. Intake of sources contaminated with such toxic metals leads to various health issues. Keeping the serious effects of these toxic metal ions in view, various organic-inorganic nanomaterials based sensors have been exploited for their detection via optical, electrochemical and colorimetric approaches. Since a chemical sensor works on the principle of interaction between the sensing layer and the analytes, a sensor material with large surface area is required to enable the largest possible interaction with the target molecules and hence the sensitivity of the chemical sensor. However, commonly employed materials such as metal oxides and conducting polymers tend to feature relatively low surface areas, and hence resulting in low sensitivity of the sensor. Metal-Organic Frameworks (MOFs) nanostructures are another category of organic-inorganic materials endowed with large surface area, ultra-high and tunable porosity, post-synthesis modification features, readily available active sites, catalytic activity, and chemical/thermal stability. These properties provide high sensitivity to the MOF based sensors due to the adsorption of large number of target analytes. The current review article focuses on MOFs based optical and electrochemical sensors for the detection of heavy metals.

Association of a novel 27-gene immuno-oncology assay with efficacy of immune checkpoint inhibitors in advanced non-small cell lung cancer.

BACKGROUND: Immune checkpoint inhibitor (ICI) therapies represent a major advance in treating a variety of advanced-stage malignancies. Nevertheless, only a subset of patients benefit, even when selected based on approved biomarkers such as PD-L1 and tumor mutational burden. New biomarkers are needed to maximize the therapeutic ratio of these therapies. METHODS: In this retrospective cohort, we assessed a 27-gene RT-qPCR immuno-oncology (IO) gene expression assay of the tumor immune microenvironment and determined its association with the efficacy of ICI therapy in 67 advanced-stage NSCLC patients. The 27-gene IO test score (IO score), programmed cell death ligand 1 immunohistochemistry tumor proportion score (PD-L1 TPS), and tumor mutational burden (TMB) were analyzed as continuous variables for response and as binary variables for one-year progression free survival. The threshold for the IO score was prospectively set based upon a previously described training cohort. Prognostic implications of the IO score were evaluated in a separate cohort of 104 advanced-stage NSCLC patients from The Cancer Genome Atlas (TCGA) who received non-ICI therapy. RESULTS: The IO score was significantly different between responders or non-responders (p = 0.007) and associated with progression-free survival (p = 0.001). Bivariate analysis established that the IO score was independent of PD-L1 TPS and TMB in identifying patients benefiting from ICI therapy. In a separate cohort of late-stage NSCLC patients from TCGA, the IO score was not prognostic of outcome from non-ICI-treated patients. CONCLUSIONS: This study is the first application of this 27-gene IO RT-qPCR assay in a clinical cohort with outcome data. IO scores were significantly associated with response to ICI therapy and prolonged progression-free survival. Together, these data suggest the IO score should be further studied to define its role in informing clinical decision-making for ICI treatment in NSCLC.

Renal Cell Carcinoma Presenting With Combined Cervical Lymphadenopathy and Cardiac Metastasis Without Inferior Vena Cava Involvement.

Renal cell cancer is the third most common urological malignancy following prostate and bladder malignancies. Cardiac metastases to the right side of the heart without inferior vena cava (IVC) involvement are exceedingly rare, with only a handful of cases described in the literature. Metastasis to the head and neck region is also rare, occurring in an estimated 1% of cases. Here we present a case of a patient with recurrent syncopal events secondary to renal cell carcinoma without IVC involvement, with metastases both to the right ventricle and cervical lymph nodes. To our knowledge, this is the first case that presents with both of these rare findings together and that highlights cancer screening in patients with high risk factors and new exam findings in patients with syncopal events having negative initial workup.

FAN1, a DNA Repair Nuclease, as a Modifier of Repeat Expansion Disorders.

FAN1 encodes a DNA repair nuclease. Genetic deficiencies, copy number variants, and single nucleotide variants of FAN1 have been linked to karyomegalic interstitial nephritis, 15q13.3 microdeletion/microduplication syndrome (autism, schizophrenia, and epilepsy), cancer, and most recently repeat expansion diseases. For seven CAG repeat expansion diseases (Huntington's disease (HD) and certain spinocerebellar ataxias), modification of age of onset is linked to variants of specific DNA repair proteins. FAN1 variants are the strongest modifiers. Non-coding disease-delaying FAN1 variants and coding disease-hastening variants (p.R507H and p.R377W) are known, where the former may lead to increased FAN1 levels and the latter have unknown effects upon FAN1 functions. Current thoughts are that ongoing repeat expansions in disease-vulnerable tissues, as individuals age, promote disease onset. Fan1 is required to suppress against high levels of ongoing somatic CAG and CGG repeat expansions in tissues of HD and FMR1 transgenic mice respectively, in addition to participating in DNA interstrand crosslink repair. FAN1 is also a modifier of autism, schizophrenia, and epilepsy. Coupled with the association of these diseases with repeat expansions, this suggests a common mechanism, by which FAN1 modifies repeat diseases. Yet how any of the FAN1 variants modify disease is unknown. Here, we review FAN1 variants, associated clinical effects, protein structure, and the enzyme's attributed functional roles. We highlight how variants may alter its activities in DNA damage response and/or repeat instability. A thorough awareness of the FAN1 gene and FAN1 protein functions will reveal if and how it may be targeted for clinical benefit.

Sensitive impedimetric detection of troponin I with metal-organic framework composite electrode.

Metal-organic frameworks (MOFs) are promising materials for biosensing applications due to their large surface to volume ratio, easy assembly as thin films, and better biocompatibility than other nanomaterials. Their application in electrochemical biosensing devices can be realized by integrating them with other conducting materials, like polyaniline (PANI). In the present research, a composite of a copper-MOF (i.e., Cu3(BTC)2) with PANI has been explored to develop an impedimetric sensor for cardiac marker troponin I (cTnI). The solvothermally synthesized Cu3(BTC)2/PANI composite has been coated as a thin layer on the screen-printed carbon electrodes (SPE). This electroconductive thin film was conjugated with anti-cTnI antibodies. The above formed immunosensor has allowed the impedimetric detection of cTnI antigen over a clinically important concentration range of 1-400 ng mL-1. The whole process of antigen analysis could be completed within 5 min. The detection method was specific to cTnI even in the co-presence of other possibly interfering proteins.

Advancements in 2D Materials Based Biosensors for Oxidative Stress Biomarkers.

The imbalance in generation of reactive oxygen species and its depletion causes oxidative stress. Because of its importance, there is a need to explore the role of oxidative stress biomarkers. The limitations of the conventional methods cause the researchers look for other alternatives. Biosensors are highly promising candidates for the detection of trace quantities for various analytes with high specificity, selectivity, and quick response time. Nanomaterial based matrices are the most popular choice while fabricating a biosensor. Two-dimensional (2D) materials such as graphene, transitional metal dichalcogenides, and various metal oxides have been used for the biosensing of different oxidative stress analytes. High electron mobility, good optical properties, tunable properties, high yields, easy synthesis, and availability make these materials the first choice. In this review, we have comprehensively discussed various biomarkers associated with oxidative stress. The review will provide information related to different kinds and various synthesis procedures employed for 2D nanomaterials. The major focus of the review is to elaborate the role of 2D nanomaterial based structures for the optical and electrochemical methods for the detection of oxidative stress. This review is an effort to help the researchers better understand various 2D based transducers available for the detection of oxidative stress biomarkers.

White graphene quantum dots as electrochemical sensing platform for ferritin.

The use of hexagonal boron nitride quantum dots (hBN QDs) as an electrochemical sensor for ferritin is reported for the first time. These QDs were synthesized using a simple liquid exfoliation method. The synthesized material was characterized using analytical techniques such as UV-visible, Fourier transform infrared (FTIR) and Raman spectroscopy, X-ray diffraction (XRD), and high-resolution transmission electron microscopy (HR-TEM) to study different aspects of the QDs. These QDs were explored for their plausible application as a platform for the electrochemical detection of ferritin. For this, electrochemical impedance spectroscopy was used as a sensing technique and disposable hBN QD functionalized screen printed electrodes were used as a sensing platform. The developed immunosensor had a dynamic linear range from 10-2000 ng mL-1 of ferritin concentration with a limit of detection of 1.306 ng mL-1. The immunosensor was highly selective, did not deviate in the presence of interfering agents and was also highly reproducible.

Advancement in biosensors for inflammatory biomarkers of SARS-CoV-2 during 2019-2020.

COVID-19 pandemic has affected everyone throughout the world and has resulted in the loss of lives of many souls. Due to the restless efforts of the researchers working hard day and night, some success has been gained for the detection of virus. As on date, the traditional polymerized chain reactions (PCR), lateral flow devices (LFID) and enzyme linked immunosorbent assays (ELISA) are being adapted for the detection of this deadly virus. However, a more exciting avenue is the detection of certain biomarkers associated with this viral infection which can be done by simply re-purposing our existing infrastructure. SARS-CoV-2 viral infection triggers various inflammatory, biochemical and hematological biomarkers. Because of the infection route that the virus follows, it causes significant inflammatory response. As a result, various inflammatory markers have been reported to be closely associated with this infection such as C-reactive proteins, interleukin-6, procalcitonin and ferritin. Sensing of these biomarkers can simultaneously help in understanding the illness level of the affected patient. Also, by monitoring these biomarkers, we can predict the viral infections in those patients who have low SARS-CoV-2 RNA and hence are missed by traditional tests. This can give more targets to the researchers and scientists, working in the area of drug development and provide better prognosis. In this review, we propose to highlight the conventional as well as the non-conventional methods for the detection of these inflammatory biomarkers which can act as a single platform of knowledge for the researchers and scientists working for the treatment of COVID-19.

Chemoresistive Room-Temperature Sensing of Ammonia Using Zeolite Imidazole Framework and Reduced Graphene Oxide (ZIF-67/rGO) Composite.

The present work demonstrates the application of a composite of the zeolite imidazole framework (ZIF-67) and reduced graphene oxide (rGO), synthesized via a simple hydrothermal route for the sensitive sensing of ammonia. The successful synthesis of ZIF-67 and rGO composite was confirmed with structural and spectroscopic characterizations. A porous structure and a high surface area (1080 m2 g-1) of the composite indicate its suitability as a gas sensing material. The composite material was coated as a thin film onto interdigitated gold electrodes. The sensor displays a change in its chemoresistive property (i.e., resistance) in the presence of ammonia (NH3) gas. A sensor response of 1.22 ± 0.02 [standard deviation (sd)] is measured for 20 ppm of NH3, while it shows a value of 4.77 ± 0.15 (sd) for 50 ppm of NH3. The fabricated sensor is reproducible and offers a stable response, while also providing tolerance against humidity and some other volatile compounds. The average response and recovery times of the sensor, for 50 ppm NH3 concentration, are found to be 46.5 ± 2.12 (sd) and 66.5 ± 2.12 (sd) s, respectively. The limit of detection of the sensor was found to be 74 ppb.

Highly Sensitive Optical Detection of Escherichia coli Using Terbium-Based Metal-Organic Framework.

Metal-organic frameworks (MOFs) are envisaged as highly useful for the development of biosensors. Herein, for the first time, we report the optical detection of Escherichia coli using a water-dispersible terbium MOF (Tb-BTC; BTC, 1,3,5-benzenetricarboxylic acid). The successful synthesis of Tb-BTC is verified using spectroscopic and morphological techniques like UV-vis, fluorescence and FTIR spectroscopy, X-ray diffraction analysis, and electron microscopy. Tb-BTC has been bio-interfaced with anti-E. coli antibodies and then investigated as a biosensor for E. coli. The biosensor displays detection ability in an analyte concentration range of 1.3 × 102 to 1.3 × 108 cfu/mL with a detection limit of 3 cfu/mL, having a response time of 5 min and a total analysis time of about 20-25 min. The results are also found to be reproducible and specific in the presence of some other interfering bacterial species. As demonstrated, the present sensor provides highly sensitive and specific detection of E. coli in fruit juice sample. To the best of our knowledge, this is the first report to showcase the potential of the MOF-based fluorescent biosensor for the detection of E. coli.

Association Between Pesticide Use and Incidence of Diffuse Large B-Cell Lymphoma.

BACKGROUND/AIM: Exposure to pesticides has been reportedly associated with several types of cancer. MATERIALS AND METHODS: In this study, we used data from The United States Geological Survey (USGS), United States Census, and the Surveillance, Epidemiology, and End Results (SEER) database to analyze the association between the area density of specific agricultural pesticides and the county level annual incidence of diffuse large B-cell lymphoma (DLBCL). RESULTS: Incidence of DLBCL was significantly associated with an area density of 14 of the pesticides reported by USGS. CONCLUSION: This highlights the need for further investigation into the safety of the use of these pesticides. The importance of this study comes not only from the significant association it shows between pesticides and the incidence of cancer, but also from the fact that it included all compounds reported to USGS as being used in agriculture. This helps in prioritizing pesticides for further evaluation.

Microfluidic-Based Electrochemical Immunosensing of Ferritin.

Ferritin is a clinically important biomarker which reflects the state of iron in the body and is directly involved with anemia. Current methods available for ferritin estimation are generally not portable or they do not provide a fast response. To combat these issues, an attempt was made for lab-on-a-chip-based electrochemical detection of ferritin, developed with an integrated electrochemically active screen-printed electrode (SPE), combining nanotechnology, microfluidics, and electrochemistry. The SPE surface was modified with amine-functionalized graphene oxide to facilitate the binding of ferritin antibodies on the electrode surface. The functionalized SPE was embedded in the microfluidic flow cell with a simple magnetic clamping mechanism to allow continuous electrochemical detection of ferritin. Ferritin detection was accomplished via cyclic voltammetry with a dynamic linear range from 7.81 to 500 ng·mL-1 and an LOD of 0.413 ng·mL-1. The sensor performance was verified with spiked human serum samples. Furthermore, the sensor was validated by comparing its response with the response of the conventional ELISA method. The current method of microfluidic flow cell-based electrochemical ferritin detection demonstrated promising sensitivity and selectivity. This confirmed the plausibility of using the reported technique in point-of-care testing applications at a much faster rate than conventional techniques.

A Case Report of Disseminated Blastomycosis With Thyroid Involvement in a Pregnant Patient.

Blastomyces dermatitidis is the causal agent of blastomycosis, an invasive and often serious fungal infection. Blastomycosis typically presents as a pulmonary infection, but common extrapulmonary manifestations of blastomycosis include the skin, bones, and reticuloendothelial systems. Disseminated blastomycosis occurs more prominently in immunocompromised individuals, such as organ transplant recipients, HIV patients, and pregnant women. We report here a rare case of disseminated blastomycosis to the thyroid in a pregnant patient. This case emphasizes the unique challenges of diagnosing and treating disseminated fungal infections in pregnancy.

Usefulness of patient-reported outcomes to assess the effectiveness of topical hormonal therapy for gynecologic symptoms after antihormonal treatment for breast cancer.

The aim of this retrospective study was to assess the efficacy of topical hormonal therapy (THT) to relieve vaginal symptoms resulting from antihormonal therapy in women with hormone receptor-positive breast cancer. A total of 74 breast cancer patients who received THT for vaginal complaints were retrospectively identified and statistically matched with 74 control breast cancer patients with vaginal complaints with no documented use of THT. Symptom scores were recorded from the center's proprietary patient-reported outcomes database, Patient Care Monitor (ConcertoHealthAI, Boston). A baseline score was noted at the initiation of antihormonal therapy and was followed at 6 and 12 months. The median differences between baseline, 6-month, and 12-month scores were analyzed. Repeated measures analysis of variance assessed the impact of topical hormonal replacement. There was no statistically significant difference in score change between the two groups at 6 and 12 months. In the active THT group, there were no statistically significant differences in vaginal complaints or sexual problems over time: {F (2, 146) = 0.99, P = 0.369; and F (2, 146) = 1.56, P = 0.217}, respectively. In this study, the use of topical hormonal replacement was not effective in alleviating vaginal symptoms.

Hyperammonemic encephalopathy associated with 5-fluorouracil in a patient with previous orthotopic liver transplantation.

5-Fluorouracil (5-FU)-based chemotherapy is the most common regimen used to treat metastatic or advanced colon cancer. Neurotoxicity is a rare but serious adverse effect of 5-FU-based therapy. We report an unusual case of 5-FU-induced hyperammonemic encephalopathy in a patient with recurrent colon cancer that metastasized to the liver and lung. Two days after receiving the third dose of FOLFOX, he presented with altered mental status, agitation, abdominal pain, nausea, and vomiting. His total ammonia level was 434 µmol/L, for which he was treated with lactulose retention enema and lactulose via nasogastric tube. Over the next 24 hours, his condition improved significantly.

Design and development of an optical reflective notch filter using the ion assisted deposition technique with stepwise modulated thickness for avionics applications.

This paper presents research work about the design and fabrication of a 44-layer optical reflective notch filter. The performance of the fabricated notch filter was studied at normal (0°) and oblique (45°) incidence angle. In addition, the paper also discusses a three-layer broadband antireflective coating on both sides of the multilayer stack to suppress the ripples in the passband region. The thickness-modulated reflective stack of the filter was designed by using the materials Al2O3 (1.63) and SiO2 (1.46). Optimization of the multilayer stack was carried out by using the damped least-squares algorithm. The theoretical and experimental results from the ion-assisted e-beam deposited samples for single notch reflective filters are presented. Good agreement in the design and experimental results was observed when the deposition process was controlled by time of evaporation. Further, the filter was characterized for the optical properties by using a UV-VIS-NIR spectrophotometer, surface morphology and protective properties using field emission scanning electron microscopy, a coherence correlation interferometer, and water contact angle.

Unusual recurrent metastasizing benign breast papilloma: a case report.

BACKGROUND: Papillary breast lesions may be benign, atypical, and malignant lesions. Pathological and clinical differentiation of breast papillomas can be a challenge. Unlike malignant lesions, benign breast papillomas are not classically associated with lymph node and distant metastasis. We report a unique case of a recurrent, benign breast papilloma presenting as an aggressive malignant tumor. CASE PRESENTATION: Our patient was a 56-year-old postmenopausal African American woman who was followed in the breast clinic with a long history of multiple breast papillomas. She underwent multiple resections over the course of 7-9 years. After being lost to follow-up for 2 years, she once again presented with a slowly enlarging left breast mass. Subsequent imaging revealed a predominantly cystic mass in the left breast, as well as a suspicious hypermetabolic internal mammary node and a hypermetabolic nodule in the pretracheal space. Biopsy of the internal mammary node demonstrated papillary neoplasm with benign morphology and immunostains positive for estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2/Neu. Due to the clinical picture concerning for malignancy, the patient was then started on endocrine therapy with palbociclib and letrozole before surgery. She then underwent simple mastectomy and sentinel lymph node dissection with negative nodes and pathology once again revealing benign papillary neoplasm. She underwent adjuvant chest wall radiation for 6 weeks and received letrozole following completion of her radiation therapy. She was without evidence of disease 30 months after surgery. CONCLUSIONS: We present an unusual case of multiple recurrent peripheral papillomas with entirely benign histologic features exhibiting malignant behavior over a protracted period of many years, with an invasion of pectoralis musculature and possibly internal mammary and mediastinal nodes. Her treatment course included multiple surgeries (ultimately mastectomy), radiation therapy, and endocrine therapy.

Lysine-Functionalized Tungsten Disulfide Quantum Dots as Artificial Enzyme Mimics for Oxidative Stress Biomarker Sensing.

The color generating from the biochemical reaction between 3,3',5,5'-tetramethylbenzidine and Lysine@WS2 QDs was used a signal for the detection of hydrogen peroxide. The QDs were prepared using a combination of techniques, that is, probe sonication and hydrothermal treatment. Analysis via UV-vis spectroscopy, Fourier transform infrared and Raman spectroscopy, X-ray diffraction, energy-dispersive spectroscopy, and transmission electron microscopy yielded detailed information on the nature and characteristics of these quantum dots. Furthermore, as-synthesized quantum dots were studied for their capability to mimic peroxidase enzyme using 3,3',5,5'-tetramethylbenzidine as a substrate. Consequently, a colorimetric sensor utilizing Lysine@WS2 QDs could detect hydrogen peroxide in a range of 0.1-60 µM with a response time of 5 min. The same material was used for H2O2 detection using impedance spectroscopy, which yielded a dynamic range of 0.1-350 µM with a response time of 30-40 s.