bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of three different methods for the detection of human papillomavirus infected in cervical lesions
Authors: Ahn W. , Shin J. , Park D. , Lee S. , Wen L. , Kim K. , Kim Y.
Published: August 2020

e22166

Background: Several techniques are in use for HPV DNA detection: Hybrid Capture IIR(HCIIR) assay, PCR-based DNA microarray and Luminex-based assay. This study was aimed to examine the performances of three validated HPV assays. We compared the concordance between the three methods for detecting HPV in cervical specimen. Methods: Cervical samples of 189 Korean women (21∼77 ages, average 41.2 age) were tested for HPV detection in this study. We examined the performances of three validated HPV assays according to each method's instructions : 1) Hybrid Capture IIR (HCIIR) assay (HCIIR; Digene Corporation, USA) was used to test for detection of 18 types of HPV (high-risk types: HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68 and low-risk types types: HPV 6, 11, 42, 43, 44). 2) HPV DNA Chip, PCR-based DNA microarray system (MyGene Co., Seoul, South Korea) was used to test for genotyping of 24 types of HPV: 15 types of high-risk types (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68) and 9 types of low-risk types (HPV6, 11, 34, 40, 42, 43, 44, 54, 70). 3) Luminex-based HPV test (YBT, Seoul, Korea) was performed for genotyping 26 types of HPV including 15 high-risk HPV types (HPV-16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68) and 11 low-risk types (HPV-6, 11, 34, 40, 42, 43, 44, 54, 55, 67, 70). Results: We compared HPV detection results obtained by the HC II assay with those from the Luminex assay. 51 cases were discordant: 29 samples were HC II negative/Luminex positive and 22 samples were HCII positive/Luminex negative. We also compared HPV detection results obtained by the DNA chip assay with those from the Luminex assay. 37 cases were discordant: 5 samples were DNA chip negative/Luminex positive and 32 samples were DNA chip positive/Luminex negative. Conclusions: The distribution of the HPV detection results obtained for 91 samples using three different methods suggested that the results of DNA chip assay are more often discordant (17.9% instead of 5.1% for Luminex assay and 5.1% for the HC II assay).

No significant financial relationships to disclose.
Effect of CCL2 on survival of breast cancer MDA-MB-231 cells via inhibition of apoptosis
Authors: Alva A. S. , Varsos Z. , Roca H. , Pienta K.
Published: August 2020

e22176

Background: The cytokine CCL2 (Chemokine (C-C) motif ligand 2 or monocyte chemo attractant protein-1 MCP-1) has been shown to play an important role in breast cancer progression by promoting angiogenesis and migration. CCL2 is over-expressed by breast cancer cells as well as by macrophages and mesenchymal stem cells in the tumor micro-environment. We studied the effects of CCL2 and IL-6 on survival of breast cancer MDA-MB-231 cells. Methods: Cytokine expression including of CCL2 and IL- 6 were analyzed in supernatants from MDA-MB-231 cells using cytokine antibody array (Human Antibody Array 3; Ray Biotech, Inc). Serum starved MDA-MB-231 cells were treated with CCL2 (75 ng/mL) and IL-6 (100 ng/mL; both from Apollo Cytokine Research). Cell viability in response to treatment was analyzed at serial time points by cell proliferation assay WST-1 (Roche Applied Science). Protein expression of genes of interest was studied by Western blot. Results: In contrast to primary breast tumor cells, MDA-MB-231 cells express minimal CCL2 at baseline. MDA-MB-231 cells also exhibit low expression of IL-6. Stimulation of MDA-MB-231 cells with either exogenous CCL2 or exogenous IL-6 did not increase levels of the other cytokine. In MDA-MB-231 cells, CCL2, alone and in combination with IL-6, promotes survival in serum starved conditions. Treatment of MDA-MB-231 cells with CCL2 leads to survivin up-regulation beginning at 48 hours as determined by immunoblotting analysis. CCL2 in combination with IL-6 inhibits the cleavage of PARP, lamin and caspase3. Conclusions: There is minimal endogenous expression of CCL2 in MDA-MB-231 cells. CCL2 from the tumor micro-environment promotes survival of MDA-MB-231 cells associated with up-regulation of survivin. IL-6 may play a syngeristic role with CCL2 in promoting survival of MDA-MB-231 cells. We plan to explore the role of survivin up-regulation in protecting cells from cell death. Our findings raise the possibility of utilizing neutralizing antibodies against CCL2 in therapy of breast cancer.

No significant financial relationships to disclose.
Evaluation of the association of CXCR4 expression with HER2 status or mTOR expression levels
Authors: Smith M. H. , Li B. D. , Sun G. , Chu Q. D.
Published: August 2020

e22192

Introduction: CXCR4 is a chemokine receptor that is linked to breast cancer metastasis. The putative mechanism involves activation of the HER-2/PI3K/PTEN/Akt/mammalian target of rapamycin (mTOR) pathway, resulting in enhanced synthesis of downstream proteins such as CXCR4 and eukaryotic initiation factor 4E (eIF4E). Overexpression of eIF4E has been shown to be an independent predictor of breast cancer outcome. Because HER-2/mTOR/CXCR4 pathway has mainly been observed in vitro, it deserves scrutiny to determine whether it is also seen in human breast cancers. We hypothesize that HER-2/mTOR axis is not significantly associated with CXCR4 regulation in human breast cancers. Method: 107 stages 1 - 3 breast cancers from a prospective breast cancer database were analyzed. Samples were quantified for mTOR, CXCR4, and eIF4E levels by Western Blot. mTOR, and eIF4E levels were quantified as fold-over benign breast tissues and CXCR4 levels were quantified as fold-over Hela cells. CXCR4 overexpression was defined as low (< 6.0 fold) or high (≥ 6.0 fold), and eIF4E expression level was separated into tertiles. Primary end points were cancer recurrence and death. Statistical analyses performed were independent student t-test, Spearman correlation, Kaplan-Meier survival curves, log-rank test, and Cox proportional hazard model. Results: All 59 HER-2 (-) and 48 HER-2 (+) breast cancer specimens overexpressed CXCR4 (mean=6.18 ±4.4), mTOR (mean=1.24 ±1.58), and eIF4E (mean=9.53 ±6.42). Unlike in vitro data, there was no difference in mean CXCR4 expression level between HER-2 (-) and HER-2 (+) tumors (mean=6.55, 5.72, respectively) (p=0.33). While mTOR correlated significantly with eIF4E level (p=0.0006, r = 0.34), it did not correlate with CXCR4 level (p=0.713, r=0.036). Only nodal status (p=0.04) and eIF4E level (p=0.03) were independent predictors of breast cancer death. Conclusions: We demonstrate that CXCR4 regulation in human breast cancers is more complex than previously thought. There was no correlation between mTOR and CXCR4 levels, nor was there a difference in mean CXCR4 overexpression levels between HER-2 (+) and HER-2 (-) tumors. Pathways regulating CXCR4 expression warrant further investigation.

No significant financial relationships to disclose.
Establishment and molecular characteristics of PC-9 derived erlotinib resistant cell lines
Authors: Jida M. , Yamamoto H. , Kubo T. , Hayashi T. , Otani H. , Sano Y. , Toyooka S.
Published: August 2020

e22193

Background: EGFR mutation is closely associated with tumor responsiveness to EGFR tyrosine kinase inhibitors (TKIs), gefitinib or erlotinib. However, many non-small cell lung cancer (NSCLC) cases with EGFR mutation that had responded to EGFR-TKIs finally acquire resistance to EGFR-TKIs. It has been reported that T790M mutation and MET amplification caused resistance to EGFR-TKIs . Recent study suggested that overexpression of HGF was also related to resistance to EGFR-TKIs. However, the whole mechanism of acquired resistance to TKIs is still not fully known. Methods: We established 5 clones of elrotinib resistant PC-9 cell line (harboring EGFR exon 19 deletion) (designated as PC-9ER1–5) by exposing PC-9 to low-dose erlotinib. The protein expression of EGFR-related molecules was examined by Western blotting. Array comparative genomic hybridization (aCGH) or real time quantitative PCR (qPCR) assay was performed to examine copy numbers of EGFR-related genes. Results: In MTS assay, the IC50 value for the parental PC-9 cells was 0.02 μmol/L. By contrast, those for PC-9ER1–5 were more than 33μmol/L. All 5 resistant cell lines retained exon19 deletions and did not obtain T790M mutation. The qPCR assay and aCGH showed no MET amplification in PC-9 or PC-9ER1–5. HGF protein was not overexpressed in PC-9ER1–5 by ELISA. The protein expressions of several molecules in the EGFR-Akt signaling pathway were examined by western blotting after erlotinib treatment. Although phospho-EGFR was suppressed in both PC-9 and PC-9ER1–5 with 2-μmol/L erlotinib, phospho-Akt was not suppressed in PC-9ER1–5. PTEN expression was not down-regulated in PC-9ER1–5. The combination of erlotinib and PHA-665752, MET tyrosine kinase inhibitor, did not suppress phospho-Akt or cell proliferation in PC-9ER1–5. Conclusions: As common molecular features of our EGFR-TKI resistant cell lines, phospho-Akt was not suppressed with exposure to erlotinib. These cell lines did not show previously reported features of resistant cell lines including T790M mutation, MET amplification or HGF overexpression. Our results indicated that other mechanisms leading to Akt activation caused resistance to EGFR-TKIs.

No significant financial relationships to disclose.
Proteome analysis of malignant pleural effusion
Authors: Lee K. , Bang J. , Kim K.
Published: August 2020

e22194

Background: Because pleural effusion contains proteins of potential diagnostic value, a comprehensive proteomic analysis of the pleural effusion is worthy to discover a new biomarker. Malignant pleural effusion and tuberculous pleural effusion are sometimes diagnositc challenge due to their similarity like lymphocyte-dominant effusion. Herein, we tried to identify differentially expressed proteins in both effusion using proteomic anlaysis. Methods: Twenty microliters of pleural effusions(PEs) from 3 patients with non-small cell lung cancer(NSCLC) and 3 patients with tuberculous pleurisy(TBC) were used for proteome analysis. After depletion of high abundant proteins including albumin, IgG with MARS Hu-6(Agilent), proteins were separated by SDS-PAGE and subject to in-gel tryptic digestion. The resulting peptides were analyzed by liquid chromatography/tandem mass spectrometry (LC/MS/MS). The MS/MS spectra were analyzed by Spectrum Mill against normal and reversed human protein databases. Results: In the total of 6 samples, 90 proteins were identified with more than 2 peptides and less than 1% of false positive rate. Among the identified pleural proteins, 57 proteins were detected both in PEs of NSCLS and TBC, 19 and 14 proteins were identified only in the PE of NSCLC and TBC, respectively. We analysed molecular functions, molecular composition and molecular processes of identified proteins with FindGo software. Among the identified proteins, we found the biomarker candidates that significantly have different expression levels in malignant effusion; apolipoprotein B precursor, vitronectin, complement factor B, histidine-rich glycoprotein precursor, coagulation factor II precursor variant. Conclusions: We found that several pleural effusion proteins may serve as potential biomarker candidates for differential diagnosis between maligant and tuberculous pleural effusion. We'll confirm these proteins through the proteomic method(MRM) and immunological method(Western bolt).

No significant financial relationships to disclose.
Candidate treatments of brain metastases suggested fromin silico gene expression analysis of archival primary breast tumor tissue and brain metastases (BM)
Authors: Iyengar T. , Tran N. L. , Armstrong B. A. , Berens M. E.
Published: August 2020

e22203

Background: Patients with metastatic breast cancer are living longer due to advances in therapy. Interestingly, there are times when systemic disease is well controlled but progression continues within the central nervous system (CNS). We report our retrospective analysis of gene expression data from the GEO database. Methods: We interrogated the NCBI/GEO database using the following search terms: breast cancer, gene expression, and brain metastases. We identified a set of tissue samples and compared gene expression arrays from 4 separate BM specimens to 7 primary breast cancer specimens; all specimens were Her-2 neu +/ER - tumors except one (information unavailable). Using a t-test, a two-fold or greater change in gene expression was considered significant if the p-value was <0.05. Results: Analysis of the gene expression profiles revealed several gene candidates over expressed in the BM specimens as compared to primary tumor specimens. Of interest are the motility-associated genes (ICAM1, cortactin, DOCK11, and myosin; p<0.03), pro-survival gene (AKT3; p<0.03), the FGF-growth factor signaling pathway (FGF1; p<0.02), and genes positively regulating the epithelial-to-mesenchymal transformation (EMT) process [TGFB1, SNAI2 (SLUG), NOTCH2; p<0.03]. Notably, the TGFB/Slug pathway is influenced by Sonic hedgehog (SHH). SHH inhibitors are currently being investigated in phase I clinical trials. Conclusions: To the best of our knowledge, this is the first analysis to suggest a role for TGFB and SHH signaling as it relates to the metastatic potential of breast cancer. Our findings argue for gene expression analysis on a larger number of BM specimens for discovery and validation of gene candidates important for this malignant progression. As there are drugs already available to target these pathways, the goal would be to accrue more patients with primary or secondary CNS malignancies to assess response.

No significant financial relationships to disclose.
Clinicopathologic predictors of the 21-gene assay in a Hispanic cohort
Authors: Khan A. , Rodriguez C. , Tovar Y. E. , Rajabi B. , Mulla Z. D. , Hakim M. N.
Published: August 2020

e22209

Background: The 21-gene recurrence score (RS) assay became commercially available (Oncotype DX) in January 2005. It is used to predict the likelihood of distant recurrence in women with estrogen-receptor (ER)-positive, lymph node-negative invasive breast cancer treated with adjuvant tamoxifen. We aimed to identify clinicopathologic predictors of an elevated recurrence score in a group of women with invasive breast cancer living in a predominantly Hispanic population on the Texas-Mexico border. Methods: A retrospective cohort study of 60 consecutive patients with primary ER-positive, lymph node-negative invasive breast cancer who had an Oncotype DX assay performed by Genomic Health, Inc. on paraffin-embedded tumor between March 2006 and November 2008. ER, PR and HER-2 was performed by immunohistochemistry. Patients were classified as low RS (<18), intermediate RS (18–30), or high RS (≥31). Ordinal logistic regression was used to calculate odds ratios (OR) for an increasing RS. Patient age and tumor pathological features were documented in each group. There was no selection bias in ordering the assay. Results: 95% of the cohort was Hispanic. Average age was 55.5 years. 29 patients (48%) had a low RS, 15 (25%) had an intermediate RS, while 16 (27%) had a high RS. HER-2 was amplified in 8 patients (13%). HER-2 positivity was strongly associated with a high RS (p<0.0001). Tumor grade and Ki-67 proliferative index were correlated with one another in a positive fashion (weighted kappa=0.46, p<0.05) and in univariate analyses both factors were significant predictors of a higher RS: tumor grade 3 vs. 1 OR=26.6 (p<0.0001) and Ki-67 proliferative index high vs. low OR=6.59 (p=0.0033). After adjustment, neither OR was significant. A negative progesterone receptor (PR) status was a strong risk factor for a higher RS (adjusted OR=21.64, p=0.0002). Univariate and multivariate ORs for higher RS for invasive ductal vs. invasive lobular carcinoma were, respectively, 4.3 (p=0.051) and 2.23 (p=0.4). Conclusions: Initially, a tumor grade of 3 and a high Ki-67 proliferative index were strong risk factors for a higher RS. However, after controlling for multiple factors, neither tumor grade nor Ki-67 proliferative index had an impact on RS in this predominantly Hispanic cohort.

No significant financial relationships to disclose.
Identification of potential diagnostic markers in bronchial fluid of patients with non small cell lung cancer (NSCLC)
Authors: Lopez-Vivanco G. , Muñoz A. , Carrera S. , Zalacain R. , Uribarri M. , Algorta J. , Calvo B. , Aresti U. , Buque A. , Mielgo X.
Published: August 2020

e22216

Background: Early diagnosis in lung cancer could improve its treatment and, subsequently, its prognosis and survival. New proteomic techniques can identify potential diagnostic and prognostic markers. The aim of this study was to find protein markers in bronchial fluid which could enable early diagnosis in NSCLC. Methods: We have included 96 patients with NSCLC diagnosed using bronchoscope (64 scamous/29 adenocarcinoma/3 others) and 49 consecutive patients with non pathological bronchoscope. Bronchial fluid was obtained from each patient and potential protein markers were studied. Bronchial fluid was centrifuged and supernatant proteins were analysed using bidimensional electrophoresis with poliacrilamid gel stained with silver nitrate. Gel was scanned and analysed with Progenesis PG6220 program, which measures intensity of each spot. Resultant intensities in each group of patients (NSCLC/non pathological bronchoscope) were compared using T-Student method. We selected as potential markers those spots with a p value of less than 0.05. We calculated “fold change” of each spot as the ratio between mean intensity in NSCLC bronchoscopes samples and non pathological bronchoscopes samples. Results: We analysed 300 spots in each sample and we found 31 potential markers whose fold-change ranges from 1.49 to 7.41; 15 of the markers were expressed in a higher level in NSCLC samples and the other 16 were expressed in a lower level. Conclusions: We have identified 31 differential protein markers in bronchial fluid among our patients. These results could lead in an early diagnostic test which must be validated in future studies.

No significant financial relationships to disclose.
The prognostic importance of triple negative breast cancer: A population based study in India
Authors: Ajaikumar B. S. , Rao R. , Prabhu J. , Kulkarni J. D. , K P. , V Patil G. , Nr N. , Babu C. , Sahoo R.
Published: August 2020

e22219

Background: Triple-negative (ER-negative, PR-negative, HER2/neu negative) breast cancer has distinct clinical and pathologic features, and is a clinical problem because of its typically high grade, relatively poor prognosis, aggressive behavior and lack of targeted therapies leaving chemotherapy as the mainstay of treatment. This study envisaged to analyse the influence of triple negativity status on survival and disease free survival in prospective cohort of breast cancer patients. Methods: Breast tumors of 215 women aged 30–75, diagnosed from 2004 were tested for ER, PR and HER2 positivity by immunohistochemistry and correlated with clinical outcomes such as recurrence, disease free survival and overall survival using Kaplan Meiers Survival analysis and Coxs regression analysis. The study cohort was followed up for 60 months or until death whichever was earlier. Results: Triple negativity significantly influenced disease free survival (46 ± 3, 41, 52) vs. non triple negative cohort (mean ± SE; 95%CI, 37 ± 2; 32, 40) and log rank = 2.1, p = 0.04. However triple negativity did not influence overall survival in months (56 ± 0; 55, 56) vs. non triple negative cohort (43 ± 1; 42, 45), (log rank = 1.78, p = 0.16). However, the mean disease free survival was (45 ± 7; 32, 58) months for patients >40 years age vs (37 ± 4; 33, 39) for patients < 40 years of age (log rank = 2.87, p =0.02). Stage of disease, node status, grade and menopausal status did not influence disease free survival significantly. However, Cox regression analysis did not predict significant effects of triple negativity on overall survival or disease free survival when controlled for confounding factors such as age, node status, stage etc Conclusions: Our observations suggest that triple negativity can significantly affect progression of breast cancer in Indian breast cancer patients and longer follow up is necessary (10 years) to determine its effects on survival.

No significant financial relationships to disclose.
Exploratory study of the subcutaneous fat gene expression profile in patients with metastatic pancreatic carcinoma treated with standard gemcitabine chemotherapy regimen
Authors: Castellano D. E. , Gomez-Martin C. , Cubedo Cervera R. , Garcia Lopez J. , Gomez-Sanz R. , Gravalos C. , Nuñez Sobrino J. , Cortes-Funes H.
Published: August 2020

e22220

Background: Most clinical trials are designed to assess the antitumor effect of the chemoterapeutic intervention. There are few examples where the endpoint is to assess the biology of the host response to the treatment of the tumor. A large number of patients with pancreatic cancer present features of the cachexia syndrome and specially a marked weight loss. It has been postulated that a “cytokine storm” is the cause of the profound effect that this cancer has on distant tissues. This trial analyzed changes in the subcutaneous fat gene expression profile in relation with the clinical benefit variable with standard gemcitabine (G) treatment. Methods: Patients with histology confirmed advanced pancreatic cancer, adequate organ function and written informed consent. Eligible pts were intended for a subcutaneous fat biopsy pretreatment and after 7 weeks of gemcitabine 1000 mg/m2 together with response assessment. Clinical benefit (CB) (pain, analgesic consumption, Karnofsky and weight), QLQ-C30, serum cytokines and tumor markers were evaluated pretreatment, at 4 and 8 weeks. Fat gene expression profile was analyzed using Affimetrix U133Plus2.0 with the corresponding bioinformatic software. Serum cytokines where analyzed with xMAP technology with the Luminex 200 platform. Results: 16 pts [8 m, 8 f, median age 62 yrs (range 47–72)]. Median weight change -0.75 kg (range -4.5 to 2). Nine pts had pre and post treatment biopsies and 7 only pretreatment. Three pts achieved CB at 8 weeks. Objective responses: 0 CR, 0 PR, 31% SD and 68%PD. Toxicity was similar to the one reported in gemcitabine's label. It was possible to extract quality RNA for microarray from subcutaneous fat use from all samples but 1. The limited number of samples precluded to obtain genes clearly involved in cachexia, however the IL-8 expression (p0.03) was significantly correlated with CB response either to gene and serum profile. Conclusions: It is feasible to study prospectively the impact of cancer treatment on different tissue biomarkers and correlated with standard antitumor evaluation system. The reduced number of samples in this exploratory trial precludes producing significant biological conclusions.

No significant financial relationships to disclose.
Effect of erlotinib on platelet activation in patients with non-small cell lung cancer
Authors: Yamaguchi K. , Kanazawa S. , Sasada S. , Hirashima T. , Imamura F. , Nomura S.
Published: August 2020

e22232

Background: Previously we reported that serum levels of soluble P-selectine and thromboxane (TX) B2 elevated in the patients with lung cancer treated with gefitinib and low dose aspirin improved skin eruption. We measured the serum levels of TXB2 in patients with NSCLC. Methods: Serial blood samples were withdrawn from each patient before and after the administration of erlotinib. TXB2 was measured by enzyme-linked immuno solvent assay. The kit was used according to the manufacturer's instructions. Results: The serum levels of TXB2 were elevated after treatment with erlotinib. Two cases developed skin lesion then treated with low dose aspirin. Skin lesions were better with improving itching without disease progression. Conclusions: We think that platelets activity, COX-1, COX-2 and PGI2 / TXA2 ratio are one of factor of anti-cancer and adverse effect for EGFR-TKI. And low dose aspirin may be useful for adverse effect of erlotinib and other EGFR-TKI.

No significant financial relationships to disclose.
Adjuvant radiotherapy and regional lymph node field control in melanoma patients after lymphadenectomy: Results of an intergroup randomized trial (ANZMTG 01.02/TROG 02.01)
Authors: Henderson M. A. , Burmeister B. , Thompson J. F. , Di Iulio J. , Fisher R. , Hong A. , Scolyer R. , Shannon K. , Hoesktra H. , Ainslie J.
Published: August 2020

LBA9084

The full, final text of this abstract will be available in Part II of the 2009 ASCO Annual Meeting Proceedings, distributed onsite at the Meeting on May 30, 2009, and as a supplement to the June 20, 2009, issue of the Journal of Clinical Oncology.

No significant financial relationships to disclose.
Impact of Preconception Treatment Initiation for Hypothyroidism on Neurocognitive Function in Children
Authors: Zhou Qiongjie , Wang Chunfang , Xu Huan , Li Xiaotian
Published: August 2020

Context

Levothyroxine treatment for hypothyroidism in early pregnancy has inconsistent outcomes.

Objective

This study evaluated children’s cognitive function to ascertain the impact of preconception maternal hypothyroidism treatment on fetal neurodevelopment.

Design and Setting

This prospective, single-center cohort study was conducted at a tertiary-care hospital.

Patients

Women were assigned to the before conception (BC) and after conception (AC; 8-14 gestational weeks) groups by time point of hypothyroidism diagnosis and treatment.

Interventions

Levothyroxine treatment was adjusted based on results of a monthly thyroid function test.

Main Outcome Measures

The Gesell Development Diagnosis Scale (GDDS; Chinese version) was used to assess neurocognitive development of children at age 6, 12, and 24 months, with total score as the primary outcome.

Results

Of the 466 participants, 187 and 279 were in the BC and AC groups, respectively. Both groups were comparable at baseline antenatal visit, except for a higher proportion of thyroid peroxidase (TPO) antibody-positive participants in the BC group (P < .001). No significant intergroup differences were evident on the GDDS neurodevelopmental assessment at age 6, 12, and 24 months (P > .05), except for unexpected slightly lower motor ability (P = .009) and total (P = .026) scores at 12 months and adaptability at 24 months (P = .037) in the BC group. Differences for motor ability (P < .001) and total score (P = .026) persisted on subgroup analysis for subclinical hypothyroidism, without significant differences in pregnancy and neonatal outcomes by severity and TPO status (P > .05).

Conclusions

Preconception levothyroxine treatment did not induce significantly better cognitive outcomes in children up to age 2 than treatment initiated at 8 to 14 weeks of gestation.
Influence of Obesity in the miRNome: miR-4454, a Key Regulator of Insulin Response Via Splicing Modulation in Prostate
Authors: Herrero-Aguayo Vicente , Jiménez-Vacas Juan M , Sáez-Martínez Prudencio , Gómez-Gómez Enrique , López-Cánovas Juan L , Garrido-Sánchez Lourdes , Herrera-Martínez Aura D , García-Bermejo Laura , Macías-González Manuel , López-Miranda José , Castaño Justo P , Gahete Manuel D , Luque Raúl M
Published: August 2020

Context

Obesity is a major health problem associated with severe comorbidities, including type 2 diabetes and cancer, wherein microRNAs (miRNAs) might be useful as diagnostic/prognostic tools or therapeutic targets.

Objective

To explore the differential expression pattern of miRNAs in obesity and their putative role in obesity-related comorbidities such as insulin resistance.

Methods

An Affymetrix-miRNA array was performed in plasma samples from normoweight (n = 4/body mass index < 25) and obese subjects (n = 4/body mass index > 30). The main changes were validated in 2 independent cohorts (n = 221/n = 18). Additionally, in silico approaches were performed and in vitro assays applied in tissue samples and prostate (RWPE-1) and liver (HepG2) cell-lines.

Results

A total of 26 microRNAs were altered (P < 0.01) in plasma of obese subjects compared to controls using the Affymetrix-miRNA array. Validation in ampler cohorts revealed that miR-4454 levels were consistently higher in obesity, associated with insulin-resistance (Homeostatic Model Assessment of Insulin Resistance/insulin) and modulated by medical (metformin/statins) and surgical (bariatric surgery) strategies. miR-4454 was highly expressed in prostate and liver tissues and its expression was increased in prostate and liver cells by insulin. In vitro, overexpression of miR-4454 in prostate cells resulted in decreased expression levels of INSR, GLUT4, and phosphorylation of AMPK/AKT/ERK, as well as in altered expression of key spliceosome components (ESRP1/ESRP2/RBM45/RNU2) and insulin-receptor splicing variants.

Conclusions

Obesity was associated to an alteration of the plasmatic miRNA landscape, wherein miR-4454 levels were higher, associated with insulin-resistance and modulated by obesity-controlling interventions. Insulin regulated miR-4454, which, in turn may impair the cellular response to insulin, in a cell type-dependent manner (i.e., prostate gland), by modulating the splicing process.
Development, Implementation, and Evaluation of a Telemedicine Preoperative Evaluation Initiative at a Major Academic Medical Center
Authors: Kamdar Nirav V. , Huverserian Ari , Jalilian Laleh , Thi William , Duval Victor , Beck Lauren , Brooker Lindsay , Grogan Tristan , Lin Anne , Cannesson Maxime
Published: August 2020

BACKGROUND:

With health care practice consolidation, the increasing geographic scope of health care systems, and the advancement of mobile telecommunications, there is increasing interest in telemedicine-based health care consultations. Anesthesiology has had experience with telemedicine consultation for preoperative evaluation since 2004, but the majority of studies have been conducted in rural settings. There is a paucity of literature of use in metropolitan areas. In this article, we describe the implementation of a telemedicine-based anesthesia preoperative evaluation and report the program’s patient satisfaction, clinical case cancellation rate outcomes, and cost savings in a large metropolitan area (Los Angeles, CA).

METHODS:

This is a descriptive study of a telemedicine-based preoperative anesthesia evaluation process in an academic medical center within a large metropolitan area. In a 2-year period, we evaluated 419 patients scheduled for surgery by telemedicine and 1785 patients who were evaluated in-person.

RESULTS:

Day-of-surgery case cancellations were 2.95% and 3.23% in the telemedicine and the in-person cohort, respectively. Telemedicine patients avoided a median round trip driving distance of 63 miles (Q1 24; Q3 119) and a median time saved of 137 (Q1 95; Q3 195) and 130 (Q1 91; Q3 237) minutes during morning and afternoon traffic conditions, respectively. Patients experienced time-based savings, particularly from traveling across a metropolitan area, which amounted to $67 of direct and opportunity cost savings. From patient satisfaction surveys, 98% (129 patients out of 131 completed surveys) of patients who were consulted via telemedicine were satisfied with their experience.

CONCLUSIONS:

This study demonstrates the implementation of a telemedicine-based preoperative anesthesia evaluation from an academic medical center in a metropolitan area with high patient satisfaction, cost savings, and without increase in day-of-procedure case cancellations.
Fe65 is the sole member of its family that mediates transcription regulated by the amyloid precursor protein
Authors: Probst Sabine , Krüger Maik , Kägi Larissa , Thöni Sarina , Schuppli Daniel , Nitsch Roger M. , Konietzko Uwe
Published: August 2020

The amyloid precursor protein (APP), a central molecule in Alzheimer's disease (AD), has physiological roles in cell adhesion and signaling, migration, neurite outgrowth and synaptogenesis. Intracellular adapter proteins mediate the function of transmembrane proteins. Fe65 (also known as APBB1) is a major APP-binding protein. Regulated intramembrane proteolysis (RIP) by γ-secretase releases the APP intracellular domain (AICD), together with the interacting proteins, from the membrane. We studied the impact of the Fe65 family (Fe65, and its homologs Fe65L1 and Fe65L2, also known as APBB2 and APBB3, respectively) on the nuclear signaling function of the AICD. All Fe65 family members increased amyloidogenic processing of APP, generating higher levels of β-cleaved APP stubs and AICD. However, Fe65 was the only family member supporting AICD translocation to nuclear spots and its transcriptional activity. Using a recently established transcription assay, we dissected the transcriptional activity of Fe65 and provide strong evidence that Fe65 represents a transcription factor. We show that Fe65 relies on the lysine acetyltransferase Tip60 (also known as KAT5) for nuclear translocation. Furthermore, inhibition of APP cleavage reduces nuclear Tip60 levels, but this does not occur in Fe65-knockout cells. The rate of APP cleavage therefore regulates the nuclear translocation of AICD–Fe65–Tip60 (AFT) complexes, to promote transcription by Fe65.
ACSL3 is a novel GABARAPL2 interactor that links ufmylation and lipid droplet biogenesis
Authors: Eck Franziska , Phuyal Santosh , Smith Matthew D. , Kaulich Manuel , Wilkinson Simon , Farhan Hesso , Behrends Christian
Published: August 2020

While studies of the autophagy-related (ATG) genes in knockout models have led to an explosion of knowledge about the functions of autophagy components, the exact roles of LC3 and GABARAP family proteins (human ATG8 equivalents) are still poorly understood. A major drawback in understanding their roles is that the available interactome data has largely been acquired using overexpression systems. To overcome these limitations, we employed CRISPR/Cas9-based genome-editing to generate a panel of cells in which human ATG8 genes were tagged at their natural chromosomal locations with an N-terminal affinity epitope. This cellular resource was employed to map endogenous GABARAPL2 protein complexes using interaction proteomics. This approach identified the ER-associated protein and lipid droplet (LD) biogenesis factor ACSL3 as a stabilizing GABARAPL2-binding partner. GABARAPL2 bound ACSL3 in a manner dependent on its LC3-interacting regions, whose binding site in GABARAPL2 was required to recruit the latter to the ER. Through this interaction, the UFM1-activating enzyme UBA5 became anchored at the ER. Furthermore, ACSL3 depletion and LD induction affected the abundance of several ufmylation components and ER-phagy. Together these data allow us to define ACSL3 as a novel regulator of the enigmatic UFM1 conjugation pathway.
Microtubule polyglutamylation is important for regulating cytoskeletal architecture and motility in Trypanosoma brucei
Authors: Jentzsch Jana , Sabri Adal , Speckner Konstantin , Lallinger-Kube Gertrud , Weiss Matthias , Ersfeld Klaus
Published: August 2020

The shape of kinetoplastids, such as Trypanosoma brucei, is precisely defined during the stages of the life cycle and governed by a stable subpellicular microtubule cytoskeleton. During the cell cycle and transitions between life cycle stages, this stability has to transiently give way to a dynamic behaviour to enable cell division and morphological rearrangements. How these opposing requirements of the cytoskeleton are regulated is poorly understood. Two possible levels of regulation are activities of cytoskeleton-associated proteins and microtubule post-translational modifications (PTMs). Here, we investigate the functions of two putative tubulin polyglutamylases in T. brucei, TTLL6A and TTLL12B. Depletion of both proteins leads to a reduction in tubulin polyglutamylation in situ and is associated with disintegration of the posterior cell pole, loss of the microtubule plus-end-binding protein EB1 and alterations of microtubule dynamics. We also observe a reduced polyglutamylation of the flagellar axoneme. Quantitative motility analysis reveals that the PTM imbalance correlates with a transition from directional to diffusive cell movement. These data show that microtubule polyglutamylation has an important role in regulating cytoskeletal architecture and motility in the parasite T. brucei.

This article has an associated First Person interview with the first author of the paper.
Intraflagellar transport during assembly of flagella of different length in Trypanosoma brucei isolated from tsetse flies
Authors: Bertiaux Eloïse , Mallet Adeline , Rotureau Brice , Bastin Philippe
Published: August 2020

Multicellular organisms assemble cilia and flagella of precise lengths differing from one cell to another, yet little is known about the mechanisms governing these differences. Similarly, protists assemble flagella of different lengths according to the stage of their life cycle. Trypanosoma brucei assembles flagella of 3 to 30 µm during its development in the tsetse fly. This provides an opportunity to examine how cells naturally modulate organelle length. Flagella are constructed by addition of new blocks at their distal end via intraflagellar transport (IFT). Immunofluorescence assays, 3D electron microscopy and live-cell imaging revealed that IFT was present in all T. brucei life cycle stages. IFT proteins are concentrated at the base, and IFT trains are located along doublets 3–4 and 7–8 and travel bidirectionally in the flagellum. Quantitative analysis demonstrated that the total amount of flagellar IFT proteins correlates with the length of the flagellum. Surprisingly, the shortest flagellum exhibited a supplementary large amount of dynamic IFT material at its distal end. The contribution of IFT and other factors to the regulation of flagellum length is discussed.
Mammalian copper homeostasis requires retromer-dependent recycling of the high-affinity copper transporter 1
Authors: Curnock Rachel , Cullen Peter J.
Published: August 2020

The concentration of essential micronutrients, such as copper (used here to describe both Cu+ and Cu2+), within the cell is tightly regulated to avoid their adverse deficiency and toxicity effects. Retromer-mediated sorting and recycling of nutrient transporters within the endo-lysosomal network is an essential process in regulating nutrient balance. Cellular copper homeostasis is regulated primarily by two transporters: the copper influx transporter copper transporter 1 (CTR1; also known as SLC31A1), which controls the uptake of copper, and the copper-extruding ATPase ATP7A, a recognised retromer cargo. Here, we show that in response to fluctuating extracellular copper, retromer controls the delivery of CTR1 to the cell surface. Following copper exposure, CTR1 is endocytosed to prevent excessive copper uptake. We reveal that internalised CTR1 localises on retromer-positive endosomes and, in response to decreased extracellular copper, retromer controls the recycling of CTR1 back to the cell surface to maintain copper homeostasis. In addition to copper, CTR1 plays a central role in the trafficking of platinum. The efficacy of platinum-based cancer drugs has been correlated with CTR1 expression. Consistent with this, we demonstrate that retromer-deficient cells show reduced sensitivity to the platinum-based drug cisplatin.
Humming hummingbirds, insect flight tones, and a model of animal flight sound
Authors: Clark Christopher J. , Mistick Emily A.
Published: August 2020

Why do hummingbirds hum and insects whine when their wings flap in flight? Gutin proposed that a spinning propeller produces tonal sound because the location of the center of aerodynamic pressure on each blade oscillates relative to an external receiver. Animal wings also move, and in addition, aerodynamic force produced by animal wings fluctuates in magnitude and direction over the course of the wingbeat. Here we model animal wing tone as the equal, opposite reaction to aerodynamic forces on the wing, using Lowson's equation for the sound field produced by a moving point force. Two assumptions of Lowson's equation are met: animal flight is low (< 0.3) Mach and animals from albatrosses to mosquitoes are acoustically compact, meaning they have a small spatial extent relative to the wavelength of their wingbeat frequency. This model predicts the acoustic waveform of a hovering Costa's Hummingbird (Calypte costae), which varies in the X, Y, and Z directions around the animal. We modeled the wing forces of a hovering animal as a sinusoid with an amplitude equal to body weight. This model predicts wing sound pressure levels below a hovering hummingbird and mosquito to within 2 dB; and that far-field mosquito wing tone attenuates to 20 dB within about 0.2 m of the animal, while hummingbird humming attenuates to 20 dB at about 10 m. Wing tones play a role in communication of certain insects, such as mosquitoes, and influences predator-prey interactions, because it potentially reveals the predator's presence to its intended prey.
Both maternal and embryonic exposure to mild hypoxia influence embryonic development of the intertidal gastropod Littorina littorea (Linnaeus, 1758)
Authors: McCoy James C. S. , Spicer John I. , Tills Oliver , Rundle Simon D.
Published: August 2020

There is growing evidence that maternal exposure to environmental stressors can alter offspring phenotype and increase fitness. Here, we investigate the relative and combined effects of maternal and developmental exposure to mild hypoxia (65% and 74% air saturation respectively) on the growth and development of embryos of the marine gastropod Littorina littorea. Differences in embryo morphological traits were driven by the developmental environment, whereas the maternal environment and interactive effects of maternal and developmental environment were the main driver of differences in the timing of developmental events. While developmental exposure to mild hypoxia significantly increased the area of an important respiratory organ, the velum, it significantly delayed hatching of veliger larvae and reduced their size at hatching and overall survival. Maternal exposure had a significant effect on these traits, and interacted with developmental exposure to influence the time of appearance of morphological characters, suggesting that both are important in affecting developmental trajectories. A comparison between embryos that successfully hatched and those that died in mild hypoxia revealed that survivors exhibited hypertrophy in the velum and associated pre-oral cilia suggesting these traits are linked with survival in low oxygen environments. We conclude that both maternal and developmental environments shape offspring phenotype in a species with a complex, developmental life history, and that plasticity in embryo morphology arising from exposure to even small reductions in oxygen tensions impacts the hatching success of these embryos.
Biosonar spatial resolution along the distance axis: revisiting the clutter interference zone
Authors: Wagenhäuser Peter A. , Wiegrebe Lutz , Baier A. Leonie
Published: August 2020

Many echolocating bats forage close to vegetation - a chaotic arrangement of prey and foliage where multiple targets are positioned behind one another. Bats excel at determining distance: they measure the delay between outgoing call and returning echo. In their auditory cortex, delay-sensitive neurons form a topographic map, suggesting that bats can resolve echoes of multiple targets along the distance axis - a skill crucial for the forage-amongst-foliage scenario. We tested this hypothesis combining an auditory virtual reality with formal psychophysics: We simulated a prey item embedded in two foliage elements, one in front of and one behind the prey. The simulated spacing between “prey” (target) and “foliage” (maskers) was defined by the inter-masker delay (IMD). We trained Phyllostomus discolor bats to detect the target in the presence of the maskers, systematically varying both loudness and spacing of the maskers. We show that target detection is impaired when maskers are closely spaced (IMD<1 ms), but remarkably improves when the spacing is increased: the release from masking is about 5 dB for intermediate IMDs (1-3 ms) and increases to over 15 dB for large IMDs (≥ 9 ms). These results are well comparable to earlier work on bats’ clutter interference zone (Simmons et al., 1988). They suggest that prey would enjoy considerable acoustic protection from closely spaced foliage, but also that the range resolution of bats would let them “peek into gaps”. Our study puts target ranging into a meaningful context and highlights the limitations of computational topographic maps.
A rare bacterial RNA motif is implicated in the regulation of thepurFgene whose encoded enzyme synthesizes phosphoribosylamine
Authors: Malkowski Sarah N. , Atilho Ruben M. , Greenlee Etienne B. , Weinberg Christina E. , Breaker Ronald R.
Published: August 2020

The Fibro-purFmotif is a putative structured noncoding RNA domain that was discovered previously in species ofFibrobacterby using comparative sequence analysis methods. An updated bioinformatics search yielded a total of only 30 unique-sequence representatives, exclusively found upstream of thepurFgene that codes for the enzyme amidophosphoribosyltransferase. This enzyme synthesizes the compound 5-phospho-D-ribosylamine (PRA), which is the first committed step in purine biosynthesis. The consensus model for Fibro-purFmotif RNAs includes a predicted three-stem junction that carries numerous conserved nucleotide positions within the regions joining the stems. This architecture appears to be of sufficient size and complexity for the formation of the ligand-binding aptamer portion of a riboswitch. In this study, we conducted biochemical analyses of a representative Fibro-purFmotif RNA to confirm that the RNA generally folds according to the predicted consensus model. However, due to the instability of PRA, binding of this ligand candidate by the RNA could not be directly assessed. Genetic analyses were used to demonstrate that Fibro-purFmotif RNAs regulate gene expression in accordance with predicted PRA concentrations. These findings indicate that Fibro-purFmotif RNAs are genetic regulation elements that likely suppress PRA biosynthesis when sufficient levels of this purine precursor are present.
Case Report: Peripartum periventricular leukomalacia resulting in cerebral palsy associated with placenta previa in Japan
Authors: Suzuki Shunji
Published: August 2020

Intrapartum fetal heart rate monitoring abnormalities had been reported to correlate with decreased umbilical artery base excess associated with neonatal seizures. However, we present an infant born at 35 weeks of gestation diagnosed with cerebral palsy associated with periventricular leukomalacia (PVL) without fetal heart rate (FHR) monitoring abnormalities, According to the summary reports of PVL cases published on the home page of the Japan Obstetric Compensation System for Cerebral Palsy (JOCSC)), the percentage of placenta previa without FHR monitoring abnormalities in the cases of PVL was 5.7% (12/209), which seemed to be higher than the total percentage of placenta previa reported in Japan (0.3-0.5%).