The effectiveness of decontamination procedures used in forensic hair analysis

Abstract

Hair is a mainstream specimen used in forensic toxicology to determine drug use and exposure. However, the interpretation of an analytical hair result can be complicated by the presence of external drug contamination. Decontamination procedures are included in hair analysis methods to remove external contamination, but the capacity of these washes to completely remove contamination for all drugs is controversial. It is evident that there is no consensus on the most effective decontamination procedure, nor can decontamination procedures consistently remove external drug contamination to less than reportable cut-offs for all analytes. ∆⁹-tetrahydrocannabinol deposited from cannabis smoke is mostly removed by organic solvents, whereas ionizable drugs are more effectively removed by an aqueous wash. Organizations such as the Society of Hair Testing recommend a hair decontamination procedure should include both an organic and aqueous washing step, which is in accordance with the reviewed literature. Studies involving a systematic evaluation of various solvents have shown that the most effective organic solvent was methanol and the most effective aqueous solvent contained sodium dodecyl sulfate detergent. If future systematic studies can demonstrate similar findings, a consensus on the most effective decontamination procedure for forensic hair analysis may be established.

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Imaging of chemokine receptor CXCR4 expression in culprit and nonculprit coronary atherosclerotic plaque using motion-corrected [ 68 Ga]pentixafor PET/CT

Abstract

Purpose

The chemokine receptor CXCR4 is a promising target for molecular imaging of CXCR4⁺ cell types, e.g. inflammatory cells, in cardiovascular diseases. We speculated that a specific CXCR4 ligand, [⁶⁸Ga]pentixafor, along with novel techniques for motion correction, would facilitate the in vivo characterization of CXCR4 expression in small culprit and nonculprit coronary atherosclerotic lesions after acute myocardial infarction by motion-corrected targeted PET/CT.

Methods

CXCR4 expression was analysed ex vivo in separately obtained arterial wall specimens. [⁶⁸Ga]Pentixafor PET/CT was performed in 37 patients after stent-based reperfusion for a first acute ST-segment elevation myocardial infarction. List-mode PET data were reconstructed to five different datasets using cardiac and/or respiratory gating. Guided by CT for localization, the PET signals of culprit and various groups of nonculprit coronary lesions were analysed and compared.

Results

Ex vivo, CXCR4 was upregulated in atherosclerotic lesions, and mainly colocalized with CD68⁺ inflammatory cells. In vivo, elevated CXCR4 expression was detected in culprit and nonculprit lesions, and the strongest CXCR4 PET signal (median SUV_max 1.96; interquartile range, IQR, 1.55–2.31) was observed in culprit coronary artery lesions. Stented nonculprit lesions (median SUV_max 1.45, IQR 1.23–1.88; P = 0.048) and hot spots in naive remote coronary segments (median SUV_max 1.34, IQR 1.23–1.74; P = 0.0005) showed significantly lower levels of CXCR4 expression. Dual cardiac/respiratory gating provided the strongest CXCR4 PET signal and the highest lesion detectability.

Conclusion

We demonstrated the basic feasibility of motion-corrected targeted PET/CT imaging of CXCR4 expression in coronary artery lesions, which was triggered by vessel wall inflammation but also by stent-induced injury. This novel methodology may serve as a platform for future diagnostic and therapeutic clinical studies targeting the biology of coronary atherosclerotic plaque.

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Investing appropriately to alleviate child poverty in Canada [Letters]

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Anticonvulsants in the treatment of low back pain and lumbar radicular pain: a systematic review and meta-analysis [Research]

BACKGROUND:

The use of anticonvulsants (e.g., gabapentin, pregabalin) to treat low back pain has increased substantially in recent years despite limited supporting evidence. We aimed to determine the efficacy and tolerability of anticonvulsants in the treatment of low back pain and lumbar radicular pain compared with placebo.

METHODS:

A search was conducted in 5 databases for studies comparing an anticonvulsant to placebo in patients with nonspecific low back pain, sciatica or neurogenic claudication of any duration. The outcomes were self-reported pain, disability and adverse events. Risk of bias was assessed using the Physiotherapy Evidence Database (PEDro) scale, and quality of evidence was assessed using Grading of Recommendations Assessment, Development and Evaluation (GRADE). Data were pooled and treatment effects were quantified using mean differences for continuous and risk ratios for dichotomous outcomes.

RESULTS:

Nine trials compared topiramate, gabapentin or pregabalin to placebo in 859 unique participants. Fourteen of 15 comparisons found anticonvulsants were not effective to reduce pain or disability in low back pain or lumbar radicular pain; for example, there was high-quality evidence of no effect of gabapentinoids versus placebo on chronic low back pain in the short term (pooled mean difference [MD] –0.0, 95% confidence interval [CI] –0.8 to 0.7) or for lumbar radicular pain in the immediate term (pooled MD –0.1, 95% CI –0.7 to 0.5). The lack of efficacy is accompanied by increased risk of adverse events from use of gabapentinoids, for which the level of evidence is high.

INTERPRETATION:

There is moderate- to high-quality evidence that anticonvulsants are ineffective for treatment of low back pain or lumbar radicular pain. There is high-quality evidence that gabapentinoids have a higher risk for adverse events.

Protocol registration:

PROSPERO-CRD42016046363

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CMA phasing out annual General Council meeting [News]

https://ift.tt/2z6LbGB

Care of community-dwelling older adults with dementia and their caregivers [Review]

https://ift.tt/2lNPv44

Mediterranean diets and metabolic syndrome status in the PREDIMED randomized trial [Expression of Concern]

https://ift.tt/2tX1Kz7

Prepubertal vulvovaginitis [Practice]

https://ift.tt/2lP7bwg

Inorganic phosphate as a potential risk factor for chronic disease [Commentary]

https://ift.tt/2z6KP2J

Perirenal soft tissue infiltration from immunoglobulin G4-related disease [Practice]

https://ift.tt/2lNPlK0

Inequity a root cause of poor health among Indigenous peoples [Letters]

https://ift.tt/2z6KA7P

A day in the life of burnout [Humanities]

https://ift.tt/2lNPf56

Medical school admission requirements lock out many Indigenous students [News]

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Reducing food insecurity and improving health with a basic income guarantee [Letters]

https://ift.tt/2lRm5SU

Physician health charter calls on health systems, organizations to share responsibility for burnout [News]

https://ift.tt/2z6KaON

Genome-Wide Association and Regional Heritability Mapping of Plant Architecture, Lodging and Productivity in Phaseolus vulgaris

The availability of high-density molecular markers in common bean has allowed to explore the genetic basis of important complex agronomic traits with increased resolution. Genome-Wide Association Studies (GWAS) and Regional Heritability Mapping (RHM) are two analytical approaches for the detection of genetic variants. We carried out GWAS and RHM for plant architecture, lodging and productivity across two important growing environments in Brazil in a germplasm of 188 common bean varieties using DArTseq genotyping strategies. The coefficient of determination of G x E interaction (c²_int) was equal to 17, 21 and 41%, respectively for the traits architecture, lodging, and productivity. Trait heritabilities were estimated at 0.81 (architecture), 0.79 (lodging) and 0.43 (productivity), and total genomic heritability accounted for large proportions (72% to 100%) of trait heritability. At the same probability threshold, three marker-trait associations were detected using GWAS, while RHM detected eight QTLs encompassing 145 markers along five chromosomes. The proportion of genomic heritability explained by RHM was considerably higher (35.48 to 58.02) than that explained by GWAS (28.39 to 30.37). In general, RHM accounted for larger fractions of the additive genetic variance being captured by markers effects inside the defined regions. Nevertheless, a considerable proportion of the heritability is still missing (~42% to ~64%), probably due to LD between markers and genes and/or rare allele variants not sampled. RHM in autogamous species had the potential to identify larger-effect QTLs combining allelic variants that could be effectively incorporated into whole-genome prediction models and tracked through breeding generations using marker-assisted selection.

https://ift.tt/2KF3tjv

Genomics-Assisted Identification and Characterization of the Genetic Variants Underlying Differential Nitrogen Use Efficiencies in Allotetraploid Rapeseed Genotypes

Nitrogen (N) is a non-mineral macronutrient essential for plant growth and development. Oilseed rape (A_nA_nC_nC_n, 2n=4x=38) has a high requirement for N nutrients whereas showing the lowest N use efficiency (NUE) among crops. The mechanisms underlying NUE regulation in Brassica napus remain unclear because of genome complexity. In this study, we performed high-depth and -coverage whole-genome re-sequencing (WGS) of an N-efficient (higher NUE) genotype "XY15" and an N-inefficient (lower NUE) genotype "814" of rapeseed. More than 687 million 150-bp paired-end reads were generated, which provided about 93% coverage and 50x depth of the rapeseed genome. Applying stringent parameters, we identified a total of 1,449,157 single-nucleotide polymorphisms (SNPs), 335,228 InDels, 175,602 structure variations (SVs) and 86,280 copy number variations (CNVs) between the N-efficient and -inefficient genotypes. The largest proportion of various DNA polymorphisms occurred in the inter-genic regions. Unlike CNVs, the SNP/InDel and SV polymorphisms showed variation bias of the A_n and C_n subgenomes, respectively. Gene ontology analysis showed the genetic variants were mapped onto the genes involving N compound transport and ATPase complex metabolism, but not including N assimilation-related genes. On basis of identification of N-starvation responsive genes through high-throughput expression profiling, we also mapped these variants onto some key NUE-regulating genes, and validated their significantly differential expression between the N-efficient and -inefficient genotypes through qRT-PCR assays. Our data provide genome-wide high resolution DNA variants underlying NUE divergence in allotetraploid rapeseed genotypes, which would expedite the effective identification and functional validation of key NUE-regulating genes through genomics-assisted improvement of crop nutrient efficiency.

https://ift.tt/2KDsyPi

Spread of blaCTX-M-15-producing Enterobacteriaceae and OXA-23-producing Acinetobacter baumannii ST2 in Tunisian seafood [PublishAheadOfPrint]

Bivalves are filter-feeding animals and markers of bacterial pollution. We report a massive spread of bla_CTX-M-15 through dominant E. coli and K. pneumoniae lineages and/or plasmid subtypes (F31:A4:B1) as well as OXA-23-producing Acinetobacter baumannii ST2 in seafood, highlighting a direct risk for the consumer. These findings should urge authorities to consider hospital effluents, but also farm and urban effluents, as important sources of ESBL/carbapenemase-producers that filter-feeding animals can concentrate and further spread to humans.

https://ift.tt/2KHgPMk

Is it time for systematic voriconazole pharmacogenomic investigation for central nervous system aspergillosis? [PublishAheadOfPrint]

Voriconazole is the standard treatment for invasive aspergillosis but requires therapeutic drug monitoring for optimizing therapy. We report two cases of central nervous system aspergillosis treated with voriconazole. Because of low trough plasma concentrations, we identified gain-of-function mutations of CYP2C19 that were partially responsible for therapeutic failure of voriconazole. We suggest systematic voriconazole pharmacogenomic investigation in cerebral aspergillosis to avoid effective therapy delay in this life-threatening disease.

https://ift.tt/2KsiZU9

Broad-spectrum adaptive antibiotic resistance associated with Pseudomonas aeruginosa mucin-dependent surfing motility [PublishAheadOfPrint]

Surfing motility is a novel form of surface adaptation exhibited by the nosocomial pathogen, Pseudomonas aeruginosa, in the presence of the glycoprotein mucin that is found in high abundance at mucosal surfaces especially the lungs of cystic fibrosis and bronchiectasis patients. Here we investigated the adaptive antibiotic resistance of P. aeruginosa under conditions in which surfing occurs compared to cells undergoing swimming. P. aeruginosa surfing cells were significantly more resistant to several classes of antibiotics including aminoglycosides, carbapenems, polymyxins, and fluroquinolones. This was confirmed by incorporation of antibiotics into growth medium, which revealed a concentration-dependent inhibition of surfing motility that occurred at concentrations much higher than those needed to inhibit swimming. To investigate the basis of resistance, RNA-Seq was performed and revealed that surfing influenced the expression of numerous genes. Included amongst genes dysregulated under surfing conditions were multiple genes from the Pseudomonas resistome, which are known to affect antibiotic resistance when mutated. Screening transposon mutants in these surfing-dysregulated resistome genes revealed that several of these mutants exhibited changes in susceptibility to one or more antibiotics under surfing conditions, consistent with a contribution to the observed adaptive resistance. In particular, several mutants in resistome genes, including armR, recG, atpB, clpS, nuoB, and certain hypothetical genes such as PA5130, PA3576 and PA4292, showed contributions to broad-spectrum resistance under surfing conditions and could be complemented by their respective cloned genes. Therefore, we propose that surfing adaption led to extensive multidrug adaptive resistance as a result of the collective dysregulation of diverse genes.

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Rapid antibiotic combination testing for carbapenem-resistant Gram negative bacteria within 6h using adenosine triphosphate bioluminescence [PublishAheadOfPrint]

To guide the timely selection of antibiotic combinations against carbapenem-resistant Gram negative bacteria (CR-GNB), an in vitro test with a short turn-around time is essential. We developed an in vitro ATP bioluminescent assay to determine effective antibiotic combinations against CR-GNB within 6h. We tested 42 clinical CR-GNB strains (14 Acinetobacter baumannii, 14 Pseudomonas aeruginosa and 14 Klebsiella pneumoniae) against 74 single and two-antibiotic combinations. Approximately 5log₁₀ CFU/ml bacteria were incubated with antibiotic(s) at 35°C, and ATP bioluminescence was measured at 6h and 24h, and compared to 24h viable counts. Receiver operating characteristic (ROC) curves were used to determine the optimal thresholds (T_RLU) for differentiating between inhibitory and non-inhibitory combinations. The area under the 6h and 24h ROC curves were compared using the DeLong method. Prospective validation of the established thresholds was conducted using 18 additional CR-GNB. The predictive accuracy of T_RLU for the 6h ATP bioluminescence assay was 77.5% for when all species were collectively analyzed. Predictive accuracies ranged from 73.7% – 82.7% when each species were individually analyzed. Upon comparison of the area under the 6h and 24h ROC curves, the 6h assay performed significantly better (p <0.01). Predictive accuracy remained high upon prospective validation of the 6h ATP assay (predictive accuracy, 79.8%; 95% CI, 77.6 – 81.9%), confirming the external validity of the assay. Our findings indicate that our 6h ATP bioluminescence assay can provide guidance for prospective combination selection against CR-GNB in a timely manner, and may be useful in the management of CR-GNB infections.

https://ift.tt/2KAL7nh

Characterization of a novel SXT/R391 Integrative and Conjugative Element carrying cfr, blaCTX-M-65, fosA3 and aac(6')-Ib-cr in Proteus mirabilis [PublishAheadOfPrint]

A novel 139,487-bp SXT/R391 Integrative and Conjugative Element, ICEPmiChnBCP11, was characterized in Proteus mirabilis of swine origin in China. ICEPmiChnBCP11 harbors twenty different antimicrobial resistance genes, including clinically important rRNA methyltransferase gene cfr, extended-spectrum β-lactamase gene bla_CTX-M-65, fosfomycin resistance gene fosA3 and fluoroquinolones resistance gene aac(6')-Ib-cr. An ISPpu12-mediated composite transposon containing various resistance genes and ten copies of IS26 is inserted in hot spot 4. ICEPmiChnBCP11 was successfully transferred to Escherichia coli.

https://ift.tt/2KHbQvc

Azithromycin in combination with ceftriaxone reduces systemic inflammation and provides survival benefit in murine model of polymicrobial sepsis [PublishAheadOfPrint]

Sepsis is a life threatening systemic inflammatory condition triggered as a result of excessive host immune response to infection. In the past, immunomodulators have demonstrated protective effect in sepsis. Azithromycin (macrolide antibiotic) having immunomodulatory activity was therefore evaluated in combination with ceftriaxone in a clinically relevant murine model of sepsis induced by caecal ligation and puncture (CLP). First, mice underwent CLP and 3 h later were administered with vehicle or sub-protective dose of ceftriaxone (100 mg/kg, subcutaneous) alone or in combination with immunomodulatory dose of azithromycin (100 mg/kg, intraperitoneal). Survival was monitored for 5 days. In order to assess the immunomodulatory activity, parameters such as plasma and lung cytokines concentrations (interleukin IL-6, IL-1β, tumor necrosis factor-α), plasma glutathione (GSH), plasma and lung myeloperoxidase (MPO), body temperature, blood glucose, total white blood cell count, along with bacterial load in blood, peritoneal fluid and lung homogenate were measured 18 h after CLP challenge. Azithromycin in presence of ceftriaxone significantly improved the survival of CLP challenged mice. Further, the combination attenuated the elevated levels of inflammatory cytokines and MPO in plasma and lung tissue and increased the body temperature, blood glucose and GSH which were otherwise markedly decreased in CLP mice. Ceftriaxone exhibited significant reduction in bacterial load, while co-administration of azithromycin did not show further reduction. Therefore, the survival benefit by azithromycin was due to immunomodulation and not by its antibacterial action. Findings of this study indicate that azithromycin could provide clinical benefits in sepsis in conjunction with appropriate antibacterial agents.

https://ift.tt/2KrQXrQ

Pharmacodynamic target attainment for cefepime, meropenem and piperacillin/tazobactam using a pharmacokinetic/pharmacodynamic-based dosing calculator in critically ill patients [PublishAheadOfPrint]

This was a prospective study to determine if pharmacokinetic/pharmacodynamic (PK/PD)-based antibiotic dosing software aids in achieving concentration targets in critically ill patients receiving cefepime (n=10), meropenem (n=20), or piperacillin/tazobactam (n=19). Antibiotic calculator doses targeting >90% probability of target attainment (PTA) differed from package-insert doses for 22.4% (11/49) patients. Target attainment was achieved for 98% of patients (48/49). A PK/PD based antibiotic dosing calculator provides beta-lactam doses with high likelihood of PTA in critically ill patients.

https://ift.tt/2KEUDSR

Gold nanoparticle conjugated Cinnamic acid exhibit Antiacanthamoebic and antibacterial properties [PublishAheadOfPrint]

Trans-Cinnamic acid (CA) is a natural organic compound. Using amoebicidal assays, for the first time we showed that CA affected viability of protist pathogen, Acanthamoeba castellanii. Conjugation with gold nanoparticles (AuNPs) enhanced antiamoebic effects of CA. Cinnamic acid coated gold nanoparticles (CA-AuNPs) also exhibited significant excystation and encystation activity as compared to CA and AuNPs alone. Pre-treatment of amoebae with CA-AuNPs inhibited A. castellanii-mediated host cell cytotoxicity. Moreover, CA-AuNPs exhibited potent effects against methicillin-resistant Staphylococcus aureus, and neuropathogenic Escherichia coli K1 as well as protected host cells against bacterial-mediated host cell death.

https://ift.tt/2KqYBTo

Prevalence and genetic analysis of mcr-3-positive Aeromonas species from humans, retail meat, and environmental water samples [PublishAheadOfPrint]

The mobile colistin-resistance gene mcr-3 is globally disseminated in both Enterobacteriaceae and Aeromonas species, the latter of which potentially serves as a reservoir for this gene. Here, we investigated the prevalence of mcr-3 from rectal swabs of humans, food-producing animals and their products, and the aquatic environment, and investigated the genetic relationships between the mcr-3-positive isolates. An enriched broth screening method was used to detect mcr-3 in samples, and species identification of isolates from the positive samples was carried out by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and shotgun sequencing. All mcr-3-positive isolates were subjected to antimicrobial susceptibility testing, conjugation, and whole genome sequencing. Ten Aeromonas isolates, two from human rectal swabs, one from pork, three from chicken meat, and four from the aquatic environment, were positive for mcr-3, but only two showed resistance to colistin. Besides the mcr-3-variants identified before– the novel variants were termed mcr-3.13 to mcr-3.18 – all isolates also harbored mcr-3-like genes downstream of the mcr-3 variants. The MCR-3.13 to MCR-3.18 proteins exhibited only 84–85% amino acid identity to the original MCR-3 protein. Whole genome sequence analysis indicated diversity within the genetic environment of mcr-3-positive Aeromonas isolates, and possible transmission between different sources in China, and even worldwide. Close relationships between mcr-3-positive and -negative Aeromonas isolates suggested that mcr-3 might be common in Aeromonas species, which are not inherent hosts of mcr-3 but may act as an important reservoir of this mobile colistin-resistance gene.

https://ift.tt/2KDV6ou

Superior Pyronaridine Single Dose Pharmacodynamics Compared to Artesunate, Chloroquine and Amodiaquine In A Murine Malaria-Luciferase Model [PublishAheadOfPrint]

Many previous in vitro and in vivo preclinical malaria drug studies have relied on low parasite number drug inhibition numerically compared to untreated controls. In contrast, human malaria drug studies measure the high parasite density killing near 100 million/mL. Here we compared the in vivo single dose pharmacodynamic properties of artesunate and 4-aminoquinolines-pyronaridine, chloroquine, and amodiaquine, in a Plasmodium bergheiANKA-GFP-Luciferase-based murine malaria blood stage model. Pyronaridine exhibited dose-dependent killing, achieving parasite reduction near 5-6 logs at 48 hours with complete cure at 10 mg/kg compared to artesunate, which exhibited a 48 hour dose-dependent killing with a 2 log drop at the noncurative 250 mg/kg dose. Chloroquine, noncurative, and amodiaquine, partially curative, have nearly the same initial dose-independent killing with a lag phase of minimal parasite reduction at all doses between 6 and 24 hours, followed by 2.5 log reduction at 48 hours. In drug treated, washed infected blood transfer experiments to naïve mice, chloroquine and amodiaquine showed less viable parasites at the 24 hour compared to 8 hour transfer measured by a prolonged return to parasitemia, despite a similar parasite log reduction at these time points, in contrast to the correlation of parasite log reduction to viable parasites with artesunate and pyronaridine. Artesunate in combination with pyronaridine exhibited a similar initial parasite reduction to pyronaridine, while with chloroquine or amodiaquine, the reduction was similar to artesunate. Single oral dose pyronaridine was much more potent in vivo than artesunate, chloroquine and amodiaquine during the initial decline in parasites and cure.

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Prevalence of Inappropriate Antibiotic Prescribing in Primary Care Clinics within a Veterans Affairs Healthcare System [PublishAheadOfPrint]

Data are needed from outpatient settings to better inform antimicrobial stewardship. In this study, a random sample of outpatient antibiotic prescriptions by primary care providers (PCPs) at our healthcare system was reviewed and compared to consensus guidelines. Over twelve months, 3,880 acute antibiotic prescriptions were written by 76 PCPs caring for 40,734 patients (median panel 600 patients; range 33-1,547). PCPs ordered a median of 84 antibiotic prescriptions per 1,000 patients per year. Azithromycin (25.8%), amoxicillin-clavulanate (13.3%), doxycycline (12.4%), amoxicillin (11%), fluoroquinolones (11%), and trimethoprim-sulfamethoxazole (10.6%) were prescribed most commonly. Medical records corresponding to 300 prescriptions from 59 PCPs were analyzed in depth. The most common indications for these prescriptions were acute respiratory tract infection (28.3%), urinary tract infection (23%), skin and soft tissue infection (15.7%), and COPD exacerbation (6.3%). In 5.7% of cases, no reason for the prescription was listed. No antibiotic was indicated in 49.7% of cases. In 12.3% of cases, an antibiotic was indicated but the prescribed agent was guideline-discordant. In another 14% of cases, a guideline-concordant antibiotic was given for a guideline-discordant duration. Therefore, 76% of reviewed prescriptions were inappropriate. Ciprofloxacin and azithromycin were most likely to be prescribed inappropriately. A non-face-to-face encounter prompted 34% of prescriptions. The condition for which an antibiotic was prescribed was not listed in primary or secondary diagnosis codes in 54.5% of clinic visits. In conclusion, there is an enormous opportunity to reduce inappropriate outpatient antibiotic prescriptions.

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Predictors of Peripherally Inserted Central Catheter Occlusion in the Outpatient Parenteral Antimicrobial Therapy Setting [PublishAheadOfPrint]

In this retrospective study of 285 patients receiving outpatient parenteral antibiotic therapy (OPAT), duration of antibiotic, use of double-lumen catheters, and receipt of penicillin G and cloxacillin appeared to increase the risk of PICC occlusion. Physicians should consider these factors when prescribing long-term antibiotic therapy. Further studies are needed to evaluate methods to reduce PICC occlusion, particularly when double-lumen PICCs are necessary, and when cloxacillin or penicillin G are the preferred treatment.

https://ift.tt/2KoxOa2

Clinical regimens of favipiravir inhibit Zika virus (ZIKV) replication in the Hollow Fiber Infection Model (HFIM) [PublishAheadOfPrint]

Zika virus (ZIKV) infection is associated with serious, long-term neurological manifestations. There are currently no approved therapies for the treatment or prevention of ZIKV. Favipiravir (FAV) is a viral polymerase inhibitor with broad-spectrum activity. Our prior studies used static FAV concentrations and demonstrated promising activity. However, the anti-ZIKV activity of dynamic FAV concentrations has never been evaluated in a human cell line. Here we employed the hollow fiber infection model (HFIM) to simulate human pharmacokinetic (PK) profiles associated with clinically-utilized influenza and Ebola FAV dosage regimens and assess the viral burden profiles. Clinically achievable FAV concentrations inhibited ZIKV replication in HUH-7 cells in a dose-dependent fashion (EC₅₀ = 236.5 uM). The viral burden profiles under dynamic FAV concentrations were predicted by mechanism-based mathematical modeling (MBM) and subsequently successfully validated in the HFIM. This validated, translational MBM can now be used to predict the anti-ZIKV activity for other FAV dosage regimens in presence of between patient variability in pharmacokinetics. This approach can be extended to rationally optimize FAV combination dosage regimens which hold promise to treat ZIKV infections in non-pregnant patients.

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Pembrolizumab OK'd for Cervical Cancer [News in Brief]

PD-1 inhibitor becomes the first approved for a gynecologic cancer.

https://ift.tt/2tW6xkk

Pediatric Hospitalization for Gun Injuries Higher in Urban Areas

MONDAY, July 2, 2018 -- Urban areas have higher overall pediatric hospitalization rates for firearm injuries, with the highest rates for urban 15- to 19-year-olds, according to a study published online July 2 in Pediatrics. Bradley R. Herrin, M.D.,...

https://ift.tt/2Kr3NXc

Irrigation Water Likely Cause of Romaine Lettuce E. Coli Outbreak

MONDAY, July 2, 2018 -- Tainted irrigation water is likely to blame for a 36-state Escherichia coli outbreak linked to romaine lettuce that sickened 200 people and caused five deaths, U.S. health officials say. The illnesses were previously...

https://ift.tt/2MGq8wA

Ophthalmologists Warn About Eye Injury Risk With Fireworks

MONDAY, July 2, 2018 -- With the Fourth of July approaching, ophthalmologists are issuing a warning on the dangers of fireworks and the risk they pose for eye injuries. According to the American Academy of Ophthalmology (AAO), about 10,000 people...

https://ift.tt/2Kr3Mm6

Pembrolizumab Not Better Than PTX for Advanced Gastric Cancer

MONDAY, July 2, 2018 -- For patients with previously treated advanced gastric cancer or gastro-esophageal junction cancer, pembrolizumab does not result in a significant improvement in overall survival compared with paclitaxel, according to a study...

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High-throughput functional evaluation of variants of unknown significance in ERBB2

Purpose: The advent of next-generation sequencing technologies has enabled the identification of several activating mutations of Erb-B2 receptor tyrosine kinase 2 (ERBB2) among various cancers. However, the significance of infrequent mutations has not been fully investigated. Herein, we comprehensively assessed the functional significance of the ERBB2 mutations in a high-throughput manner. Experimental Design: We evaluated the transforming activities and drug sensitivities of 55 nonsynonymous ERBB2 mutations using the mixed-all-nominated-in-one (MANO) method. Results: G776V, G778_S779insG, and L841V were newly revealed to be activating mutations. Although afatinib, neratinib and osimertinib were shown to be effective against most of the ERBB2 mutations, only osimertinib demonstrated good efficacy against L755P and L755S mutations, the most common mutations in breast cancer. In contrast, afatinib and neratinib were predicted to be more effective than other inhibitors for the A775_776insYVMA mutation, the most frequent ERBB2 mutation in lung cancer. We surveyed the prevalence of concurrent ERBB2 mutation with gene amplification and found that approximately 30% of ERBB2-amplified urothelial carcinomas simultaneously carried ERBB2 mutations, altering their sensitivity to trastuzumab, a monoclonal antibody against ERBB2. Furthermore, the MANO method was applied to evaluate the functional significance of 17 compound mutations within ERBB2 reported in the COSMIC database, revealing that compound mutations involving L755S were sensitive to osimertinib but insensitive to afatinib and neratinib. Conclusions: Several ERBB2 mutations showed varying sensitivities to ERBB2-targeted inhibitors. Our comprehensive assessment of ERBB2 mutations offers a fundamental database to help customize therapy for ERBB2-driven cancers.

https://ift.tt/2tY8xIN

Oligosaccharyltransferase inhibition Reduces Receptor Tyrosine Kinase Activation and Enhances Glioma Radiosensitivity

Purpose: Parallel signaling reduces the effects of receptor tyrosine kinase (RTK) targeted therapies in glioma. We hypothesized that inhibition of protein N-linked glycosylation, an endoplasmic reticulum co- and post-translational modification crucial for RTK maturation and activation, could provide a new therapeutic approach for glioma radiosensitization. Experimental design: We investigated the effects of a small molecule inhibitor of the oligosaccharyltransferase (NGI-1) on EGFR family receptors, MET, PDGFR, and FGFR1. The influence of glycosylation state on tumor cell radiosensitivity, chemotherapy induced cell toxicity, DNA damage, and cell cycle arrest were determined and correlated with glioma cell receptor expression profiles. The effects of NGI-1 on xenograft tumor growth were tested using a nanoparticle formulation validated by in vivo molecular imaging. A mechanistic role for RTK signaling was evaluated through the expression of a glycosylation independent CD8-EGFR chimera. Results: NGI-1 reduced glycosylation, protein levels, and activation of most RTKs. NGI-1 also enhanced the radiosensitivity and cytotoxic effects of chemotherapy in those glioma cells with elevated ErbB family activation, but not in cells without high levels of RTK activation. NGI-1 radiosensitization was associated with increases in both DNA damage and G1 cell cycle arrest. Combined treatment of glioma xenografts with fractionated radiation and NGI-1 significantly reduced tumor growth compared to controls. Expression of the CD8-EGFR eliminated NGI-1's effects on G1 arrest, DNA damage, and cellular radiosensitivity, identifying RTK inhibition as the principal mechanism for the NGI-1 effect. Conclusion: This study suggests that OST inhibition with NGI-1 is a novel approach to radiosensitize malignant gliomas with enhanced RTK signaling.

https://ift.tt/2lLYPWe

CD8+ T-cell density imaging with 64Cu-labeled cys-diabody informs immunotherapy protocols

Purpose: Noninvasive and quantitative tracking of CD8+ T-cells by positron emission tomography (PET) has emerged as a potential technique to gauge response to immunotherapy. We apply an anti-CD8 cys-diabody, labeled with 64Cu, to assess the sensitivity of PET imaging of normal and diseased tissue. Experimental Design Radiolabeling of an anti-CD8 cys-diabody (169cDb) with 64Cu was developed. The accumulation of 64Cu-169cDb was evaluated with PET/CT imaging (0, 5, and 24 hours) and biodistribution (24 hours) in wild-type mouse strains (n = 8 per group studied with imaging and immunohistochemistry or flow cytometry) after intravenous administration. Tumor-infiltrating CD8+ T-cells in tumor bearing mice treated with CpG and aPD-1 were quantified and mapped (n = 6-8 per group studied with imaging and immunohistochemistry or flow cytometry). Results We demonstrate the ability of immunoPET to detect small differences in CD8+ T-cell distribution between mouse strains and across lymphoid tissues, including the intestinal tract of normal mice. In FVB mice bearing a syngeneic HER2-driven model of mammary adenocarcinoma (NDL), 64Cu-169cDb PET imaging accurately visualized and quantified changes in tumor-infiltrating CD8+ T-cells in response to immunotherapy. A reduction in the circulation time of the imaging probe followed the development of treatment-related liver and splenic hypertrophy and provided an indication of off-target effects associated with immunotherapy protocols. Conclusion 64Cu-169cDb imaging can spatially map the distribution of CD8+ T-cells in normal organs and tumors. ImmunoPET imaging of tumor-infiltrating cytotoxic CD8+ T-cells detected changes in T-cell density resulting from adjuvant and checkpoint immunotherapy protocols in our pre-clinical evaluation.

https://ift.tt/2z1uhZW

Dynamics of genome alterations in Crohn's disease associated colorectal carcinogenesis

Purpose: Patients with inflammatory bowel diseases, i.e., ulcerative colitis (UC) and Crohn's disease (CD), face an increased risk of developing colorectal cancer (CRC). Evidence, mainly from UC, suggests that TP53 mutations represent an initial step in the progression from inflamed colonic epithelium to CRC. However, the pathways involved in the evolution of CRC in CD patients are poorly characterized. Experimental Design: Here, we analyzed 73 tissue samples from 28 patients with CD-CRC, including precursor lesions, by targeted next generation sequencing of 563 cancer-related genes and array-based comparative genomic hybridization. The results were compared to 24 sporadic CRCs with similar histomorphology (i.e., mucinous adenocarcinomas), and to The Cancer Genome Atlas data. Results: CD-CRCs showed somatic copy number alterations (SCNAs) similar to sporadic CRCs with one notable exception: the gain of 5p was significantly more prevalent in CD-CRCs. CD-CRCs had a distinct mutation signature: TP53 (76% in CD-CRCs versus 33% in sporadic mucinous CRCs), KRAS (24% versus 50%), APC (17% versus 75%), and SMAD3 (3% versus 29%). TP53 mutations and SCNAs were early and frequent events in CD progression, while APC, KRAS and SMAD2/4 mutations occurred later. In four CD-CRC patients, at least one mutation and/or SCNAs were already present in non-dysplastic colonic mucosa, indicating occult tumor evolution. Conclusions: Molecular profiling of CD-CRCs and precursor lesions revealed an inflammation-associated landscape of genome alterations: 5p gains and TP53 mutations occurred early in tumor development. Detection of these aberrations in precursor lesions may help predicting disease progression and distinguishes CD-associated from sporadic colorectal neoplasia.

https://ift.tt/2lNIBfb

A Combination Of Approved Antibodies Overcomes Resistance Of Lung Cancer To Osimertinib By Blocking Bypass Pathways

Purpose: Because of emergence of resistance to osimertinib, a third-generation EGFR tyrosine kinase inhibitor (TKI), no targeted treatments are available for patients with lung cancer who lose sensitivity due to new mutations or bypass mechanisms. We examined in animals and in vitro an alternative therapeutic approach making use of antibodies. Experimental Design: An osimertinib-sensitive animal model of lung cancer, which rapidly develops drug resistance, has been employed. To overcome compensatory hyper-activation of ERK, which we previously reported, an anti-EGFR antibody (cetuximab) was combined with other antibodies, as well as with a sub-therapeutic dose of osimertinib, and cancer cell apoptosis was assayed. Results: Our animal studies identified a combination of three clinically approved drugs, cetuximab, trastuzumab (an anti-HER2 mAb) and osimertinib (low dose), as an effective and long-lasting treatment, able to prevent onset of resistance to osimertinib. A continuous schedule of concurrent treatment was sufficient for effective tumor inhibition and for prevention of relapses. Studies employing cultured cells and analyses of tumor extracts indicated that the combination of two mAbs and a sub-therapeutic TKI dose sorted EGFR and HER2 for degradation, cooperatively enhanced apoptosis, inhibited activation of ERK, and reduced abundance of several bypass proteins, namely MET, AXL and HER3. Conclusions: Our in vitro assays and animal studies identified an effective combination of clinically approved drugs, which might overcome resistance to irreversible TKIs in clinical settings. The results we present attribute the long-lasting effect of the drug combination to simultaneous blockade of several well-characterized mechanisms of drug resistance.

https://ift.tt/2zeMXWz

The anti-cancer activity of a first-in-class small molecule targeting PCNA

Purpose: Proliferating cell nuclear antigen (PCNA) plays an essential role in regulating DNA synthesis and repair and is indispensable to cancer cell growth and survival. We previously reported a novel cancer associated PCNA isoform (dubbed caPCNA), which was ubiquitously expressed in a broad range of cancer cells and tumor tissues, but not significantly in non-malignant cells. We found the L126-Y133 region of caPCNA is structurally altered and more accessible to protein-protein interaction. A cell permeable peptide harboring the L126-Y133 sequence blocked PCNA interaction in cancer cells and selectively kills cancer cells and xenograft tumors. Based on these findings, we sought small molecules targeting this peptide region as potential broad-spectrum anti-cancer agents. Experimental Design: By computer modeling and medicinal chemistry targeting a surface pocket partly delineated by the L126-Y133 region of PCNA, we identified a potent PCNA inhibitor (AOH1160) and characterized its therapeutic properties and potential toxicity. Results: AOH1160 selectively kills many types of cancer cells at below micromolar concentrations without causing significant toxicity to a broad range of non-malignant cells. Mechanistically, AOH1160 interferes with DNA replication, blocks homologous recombination-mediated DNA repair, and causes cell cycle arrest. It induces apoptosis in cancer cells and sensitizes them to cisplatin treatment. AOH1160 is orally available to animals and suppresses tumor growth in a dosage form compatible to clinical applications. Importantly, it doesn't cause significant toxicity at 2.5 times of an effective dose. Conclusion: These results demonstrated the favorable therapeutic properties and the potential of AOH1160 as a broad-spectrum therapeutic agent for cancer treatment.

https://ift.tt/2lNNKnF

Inhibition of Rspo-Lgr4 facilitates checkpoint blockade therapy by switching macrophage polarization

Therapies targeting immune checkpoints have shown great clinical potential in a subset of cancer patients but may be hampered by a failiure to reverse the immunosuppressive tumor microenvironment (TME). As the most abundant immune cells in TME, tumor-associated macrophages (TAM) play non-redundant roles in restricting anti-tumor immunity. The leucine-rich repeat-containing G-protein-coupled receptor 4 (Lgr4, also known as Gpr48) has been associated with multiple physiological and pathological functions. Lgr4 and its ligands R-spondin 1-4 have been shown to promote the growth and metastasis of tumor cells. However, whether Lgr4 can promote tumor progression by regulating the function of immune cells in the tumor microenvironment remains largely unknown. Here we demonstrate that Lgr4 promotes macrophage M2 polarization through Rspo/Lgr4/Erk/Stat3 signaling. Notably, urethane-induced lung carcinogenesis, Lewis Lung carcinoma (LLC,) and B16F10 melanoma tumors were all markedly reduced in Lgr4fl/flLyz2cre/+ mice, characterized by fewer protumoral M2 TAM and increased CD8+ T lymphocyte infiltration in the TME. Furthermore, LLC tumor growth was greatly depressed when Rspo/Lgr4/Erk/Stat3 signaling was blocked with either the LGR4 extracellular domain or an anti-Rspo1 antibody. Importantly, blocking Rspo-Lgr4 signaling overcame LLC resistance to anti-PD-1 therapy and improved the efficacy of PD-1 immunotherapy against B16F10 melanoma, indicating vital roles of Rspo-Lgr4 in host anti-tumor immunity and a potential therapeutic target in cancer immunotherapy.

https://ift.tt/2lRH1cu

High-throughput screening of combinatorial immunotherapies with patient-specific in silico models of metastatic colorectal cancer

Solid tumors are rich ecosystems of numerous different cell types whose interactions lead to immune escape and resistance to immunotherapy in virtually all patients with metastatic cancer. Here we have developed a 3D model of human solid tumor tissue that includes tumor cells, fibroblasts, and myeloid and lymphoid immune cells and can represent over a million cells over clinically relevant timeframes. This model accurately reproduced key features of the tissue architecture of human colorectal cancer (CRC) and could be informed by individual patient data, yielding in silico tumor explants. Stratification of growth kinetics of these explants corresponded to significantly different overall survival in a cohort of metastatic CRC patients. We used the model to simulate the effect of chemotherapy, immunotherapies, and cell migration inhibitors alone and in combination. We classified tumors according to tumor and host characteristics, showing that optimal treatment strategies markedly differed between these classes. This platform can complement other patient-specific ex vivo models and can be used for high-throughput screening of combinatorial immunotherapies.

https://ift.tt/2u0ZNSl

CTGF mediates tumor-stroma interactions between hepatoma cells and hepatic stellate cells to accelerate HCC progression.

Connective tissue growth factor (CTGF) is a matricellular protein related to hepatic fibrosis. This study aims to clarify the roles of CTGF in hepatocellular carcinoma (HCC), which usually develops from fibrotic liver. CTGF was overexpressed in 93 human HCC compared with non-tumorous tissues, primarily in tumor cells. Increased CTGF expression was associated with clinicopathological malignancy of HCC. CTGF was upregulated in hepatoma cells in hepatocyte-specific Kras-mutated mice (Alb-Cre KrasLSL-G12D/+). Hepatocyte-specific knockout of CTGF in these mice (Alb-Cre KrasLSL-G12D/+ CTGFfl/fl) decreased liver tumor number and size. Hepatic stellate cells (HSC) were present in both human and murine liver tumors, and α-SMA expression, a marker of HSC activation, positively correlated with CTGF expression. Forced expression of CTGF did not affect growth of PLC/PRF/5 cells, a hepatoma cell line with little CTGF expression, but facilitated their growth in the presence of LX-2 cells, a hepatic stellate cell line. The growth of HepG2 cells, which express high levels of CTGF, was promoted by co-culture with LX-2 cells compared with monoculture. Growth promotion by LX-2 cells was negated by an anti-CTGF antibody in both culture and xenografts. Co-culturing LX-2 cells with HepG2 cells drove LX-2-derived production of IL-6, which led to STAT-3 activation and proliferation of HepG2 cells. An anti-CTGF antibody reduced IL-6 production in LX-2 cells and suppressed STAT-3 activation in HepG2 cells. In conclusion, our data identify tumor cell-derived CTGF as a keystone in the HCC microenvironment, activating nearby HSC which transmit pro-growth signals to HCC cells, this interaction is susceptible to inhibition by an anti-CTGF antibody.

https://ift.tt/2lOHnAs

CircNT5E acts as a sponge of microRNA-422a to promote glioblastoma tumorigenesis

Circular RNA and long noncoding RNA function as efficient microRNA sponges that regulate gene expression in eukaryotes. However, the sponges of functional miRNAs in glioblastoma remain largely unknown. Here we identify a subset of circRNAs and lncRNAs that are specifically increased in miR-422a-downregulated glioblastoma tissues. We characterized a novel circRNA derived from NT5E, named circNT5E, that is regulated by ADARB2 binding to sites flanking circRNA-forming introns. We hypothesized that circNT5E may serve as a sponge against miR-422a in glioblastoma tumorigenesis. circNT5E controlled multiple pathological processes, including cell proliferation, migration, and invasion. CircNT5E directly bound miR-422a and inhibited miR-422a activity. Furthermore, circNT5E was observed to sponge other miRNAs, exhibiting tumor suppressor-like features in glioblastoma. Taken together, these findings highlight a novel oncogenic function of circRNA in glioblastoma tumorigenesis.

https://ift.tt/2z3i1It

CRISPR/Cas9-mediated knockout of DGK improves anti-tumor activities of human T cells

The efficacy of T cell therapy is inhibited by various tumor-associated immunosuppressive ligands and soluble factors. Such inhibitory signals turn specific T cell signaling pathways on or off, impeding the anti-cancer functions of T cells. Many studies have focused on PD-1 or CTLA-4 blockade to invigorate T cell functions through CD28/B7 signaling, but obtaining robust clinical outcomes remains challenging. In this study, we use CRISPR/Cas9 to potentiate T cell function by increasing CD3 signaling via knockout of diacylglycerol kinase (DGK), an enzyme that metabolizes diacylglycerol to phosphatidic acid (PA). Knockout of DGK augmented the effector functions of CAR-T cells in vitro via increased TCR signaling. DGK knockout from CAR-T cells rendered them resistant to soluble immunosuppressive factors such as TGF-β and prostaglandin E2 and sustained effector functions under conditions of repeated tumor stimulation. Moreover, DGK knockout caused significant regression of U87MGvIII glioblastoma tumors through enhanced effector functions in a xenograft mouse model. Collectively, our study shows that knockout of DGK effectively enhances the effector functions of CAR-T cells, suggesting that CRISPR/Cas9-mediated knockout of DGK could be applicable as part of a multifaceted clinical strategy to treat solid cancers.

https://ift.tt/2lMYD9f

Combined blockade of IL-6 and PD-1/PD-L1 signaling abrogates mutual regulation of their immunosuppressive effects in the tumor microenvironment

Recently emerging cancer immunotherapies combine the applications of therapeutics to disrupt the immunosuppressive conditions in tumor-bearing hosts. In this study, we found that targeting the pro-inflammatory cytokine interleukin (IL)-6 enhances tumor-specific Th1 responses and subsequent anti-tumor effects in tumor-bearing mice. IL-6 blockade upregulated expression of the immune checkpoint molecule programmed death-ligand 1 (PD-L1) on melanoma cells. This PD-L1 induction was canceled in IFN-gamma-deficient mice or CD4+ T cell-depleted mice, suggesting that CD4+ T cell-derived IFN-gamma is important for PD-L1 induction in tumor-bearing hosts. In some patients with melanoma, however, treatment with the anti-PD-1 antibody Nivolumab increased systemic levels of IL-6, which was associated with poor clinical responses. This PD-L1 blockade-evoked induction of IL-6 was reproducible in melanoma-bearing mice. We found that PD-1/PD-L1 blockade prompted PD-1+ macrophages to produce IL-6 in the tumor microenvironment. Depletion of macrophages in melanoma-bearing mice reduced the levels of IL-6 during PD-L1 blockade, suggesting macrophages are responsible for the defective CD4+ Th1 response. Combined blockade of the mutually regulated immunosuppressive activities of IL-6 and PD-1/PD-L1 signals enhanced expression of T cell-attracting chemokines and promoted infiltration of IFN-gamma-producing CD4+ T cells in tumor tissues, exerting a synergistic anti-tumor effect, while PD-L1 blockade alone did not promote Th1 response. Collectively, these findings suggest that IL-6 is a rational immunosuppressive target for overcoming the narrow therapeutic window of anti-PD-1/PD-L1 therapy.

https://ift.tt/2z6KlK6

Microenvironmental Cues Determine Tumor Cell Susceptibility to Neutrophil Cytotoxicity

We have recently shown that neutrophil anti-tumor cytotoxicity is Ca2+-dependent and is mediated by TRPM2, an H2O2-dependent Ca2+ channel. However, neutrophil anti-tumor activity is dependent on context and is manifested in the premetastatic niche, but not at the primary site. We therefore hypothesized that expression of TRPM2 and the consequent susceptibility to neutrophil cytotoxicity may be associated with the epithelial/mesenchymal cellular state. We found that TRPM2 expression was upregulated during epithelial-to-mesenchymal transition (EMT), and mesenchymal cells were more susceptible to neutrophil cytotoxicity. Conversely, cells undergoing mesenchymal-to-epithelial transition (MET) expressed reduced levels of TRPM2, rendering them resistant to neutrophil cytotoxicity. Cells expressing reduced levels of TRPM2 were protected from neutrophil cytotoxicity and seeded more efficiently in the premetastatic lung. These data identify TRPM2 as the link between environmental cues at the primary tumor site, tumor cell susceptibility to neutrophil cytotoxicity, and disease progression. Furthermore, these data identify EMT as a process enhancing tumor-cell immune susceptibility and, by contrast, MET as a novel mode of immune evasion.

https://ift.tt/2lOHf3W

Determination of Tumor Margins with Surgical Specimen Mapping Using Near-Infrared Fluorescence

For many solid tumors, surgical resection remains the gold standard and tumor-involved margins are associated with poor clinical outcomes. Near-infrared (NIR) fluorescence imaging using molecular agents has shown promise for in situ imaging during resection. However, for cancers with difficult imaging conditions, surgical value may lie in tumor-mapping of surgical specimens. We thus evaluated a novel approach for real-time, intraoperative tumor margin assessment. 21 adult patients with biopsy-confirmed squamous cell carcinoma arising from the head and neck (HNSCC) scheduled for standard-of-care surgery were enrolled. Cohort 1 (n=3) received panitumumab-IRDye800CW at an intravenous microdose of 0.06 mg/kg, cohort 2A (n=5) received 0.5mg/kg, cohort 2B (n=7) received 1mg/kg, and cohort 3 (n=6) received 50 mg. Patients were followed 30 days post-infusion and adverse events were recorded. Imaging was performed using several closed- and wide-field devices. Fluorescence was histologically correlated to determine sensitivity and specificity. In situ imaging demonstrated tumor-to-background ratio (TBR) of 2-3, compared to ex vivo specimen imaging TBR of 5-6. We obtained clear differentiation between tumor and normal tissue, with a three-fold signal difference between positive and negative specimens (p<0.05). We achieved high correlation of fluorescence intensity with tumor location with sensitivities and specificities >89%; fluorescence predicted distance of tumor tissue to the cut surface of the specimen. This novel method of detecting tumor-involved margins in surgical specimens using a cancer-specific agent provides highly sensitive and specific, real-time, intraoperative surgical navigation in resections with complex anatomy which are otherwise less amenable to image guidance.

https://ift.tt/2u0IdxF

PDSS2 deficiency induces hepatocarcinogenesis by decreasing mitochondrial respiration and reprogramming glucose metabolism

Glucose metabolic reprogramming from oxidative phosphorylation to glycolysis is one of the hallmarks of cancer development. Coenzyme Q10 (CoQ10) is essential for electron transport in the mitochondrial respiratory chain and antioxidant defense. Here we investigated the role of a key factor in CoQ10 synthesis, prenyldiphosphate synthase subunit 2 (PDSS2), in hepatocellular carcinoma (HCC) tumorigenesis. PDSS2 was frequently downregulated in HCC tissues and was significantly associated with poorer HCC prognosis (P=0.027). PDSS2 downregulation was an independent prognostic factor independent of T status and staging (P=0.028). Downregulation of CoQ10 significantly correlated with downregulation of PDSS2 in HCC tumor tissues (R=0.414, P<0.001). Six different splicing isoforms of PDSS2 and five variants, except full-length PDSS2, showed loss of function in HCC. Reintroduction of full-length PDSS2 into HCC cells increased CoQ10 and mitochondrial electron transport complex I activity and subsequently induced a metabolic shift from aerobic glycolysis to mitochondrial respiration in cells. Reintroduction of PDSS2 also inhibited foci formation, colony formation in soft agar, and tumor formation in nude mice. Knockdown of PDSS2 induced chromosomal instability in the immortalized human liver cell line MIHA. Furthermore, knockdown of PDSS2 in MIHA induced malignant transformation. Overall, our findings indicate that PDSS2 deficiency might be a novel driving factor in HCC development.

https://ift.tt/2z4RniF

Expression of long non-coding RNA YIYA promotes glycolysis in breast cancer

Long non-coding RNA (lncRNA) have yet to be linked to cancer metabolism. Here we report that upregulation of the lncRNA LINC00538 (YIYA) promotes glycolysis, cell proliferation and tumor growth in in breast cancer. YIYA associated with the cytosolic cyclin-dependent kinase CDK6 and regulated CDK6-dependent phosphorylation of the fructose bisphosphatase PFK2 (PFKFB3) in a cell cycle-independent manner. In breast cancer cells, these events promoted catalysis of glucose 6-phosphate to fructose-2,6-bisphosphate/fructose-1,6-bisphosphate. CRISPR/Cas9-mediated deletion of YIYA or CDK6 silencing impaired glycolysis and tumor growth in vivo. In clinical specimens of breast cancer, YIYA was expressed in ~40% of cases where it correlated with CDK6 expression and unfavorable survival outcomes. Our results define a functional role for lncRNA in metabolic reprogramming in cancer, with potential clinical implications for its therapeutic targeting.

https://ift.tt/2lR4Tgk

Four Japanese Patients with Congenital Nephrogenic Diabetes Insipidus due to the AVPR2 Mutations

Almost 90% of nephrogenic diabetes insipidus (NDI) is caused by mutations in the arginine vasopressin receptor 2 gene (AVPR2) on the X chromosome. Herein, we reported clinical and biochemical parameters in four cases of three unrelated Japanese families and analyzed the status of the AVPR2. Two of the four patients had poor weight gain. However, in the male and female sibling cases, neither had poor weight gain while toddlers, but in the male sibling, episodes of recurrent fever, polyuria, and polydipsia led to the diagnosis of NDI at 4 years of age. Analysis of AVPR2 identified two nonsense mutations (c.299_300insA; p.K100KfsX91 and c.296G > A; p.W99X) and one missense mutation (c.316C > T; p.R106C). These mutations were previously reported. The patient with c.316C > T; p.R106C had milder symptoms consistent with previous reports. Of the familial cases, the sister was diagnosed as having NDI, but a skewed X-inactivation pattern in her peripheral blood lymphocytes was not identified. In conclusion, our study expands the spectrum of phenotypes and characterized mutations in AVPR2 in NDI.

https://ift.tt/2lR2UbP

Musculoskeletal Ultrasonography Assessment of Functional Magnetic Stimulation on the Effect of Glenohumeral Subluxation in Acute Poststroke Hemiplegic Patients

Background. Glenohumeral subluxation (GHS) is common in patients with acute hemiplegia caused by stroke. GHS and upper limb function are closely related. Objective. Using musculoskeletal ultrasonography (MSUS) to objectively evaluate the efficacy of functional magnetic stimulation (FMS) in the treatment of GHS in acute hemiplegic patients after stroke. Methods. The study used prospective case control study. Stroke patients with GHS were recruited and assigned to control group and FMS group. Control group received electrode stimulation at the supraspinatus and deltoid muscles of the hemiplegic side, while FMS group was stimulated at the same locations. Before and after treatment, the distances of the acromion-greater tuberosity (AGT), acromion-lesser tuberosity (ALT), acromiohumeral distance (AHD), supraspinatus thickness (SST), and deltoid muscle thickness (DMT) in patients' bilateral shoulder joint were measured by MSUS, respectively. Meanwhile, Fugl-Meyer Assessment (FMA) was used to evaluate the improvement of upper limb function. Results. 30 patients were recruited. After FMS treatment, there was a significant decrease in the difference value between ipsilateral side and contralateral side of AGT [, ], ALT [, ], AHD [, ], SST [, ], and DMT [, ], and FMA score increased [, ]. Compared with control group, FMS group decreased more significantly in the difference value between ipsilateral side and contralateral side of AGT [, ], ALT [, ], AHD [, ], SST [, ], and the DMT [, ], and FMA score increased more significantly in FMS group [, ]. Conclusion. The study preliminarily shows that the MSUS can objectively and dynamically evaluate the treatment effect of GHS in hemiplegic patients. Meanwhile, compared with control group, the FMS is more effective and has fewer side effects, and the long-term effect of FMS is worth further study. This trial is registered with ChiCTR1800015352.

https://ift.tt/2lNTzRZ

Management of Mucinous Appendiceal Tumors

https://ift.tt/2MJrWou

Long-Term Outcome After Surgery for a Localized Retroperitoneal Soft Tissue Sarcoma in Elderly Patients: Results from a Retrospective, Single-Center Study

Abstract

Background

To evaluate short- and long-term results after curative surgery for a retroperitoneal sarcoma (RPS) in elderly patients.

Methods

We retrospectively analyzed data of all patients operated in our single, tertiary care center for a nonmetastatic RPS and identified patients aged 70 years and older.

Results

Among 296 patients with an RPS treated between 1994 and 2015, 60 (20%) were aged 70 years and older (median age 74 years; range 70–85). The median tumor size was 24 cm (range 6–46). Forty-six patients (77%) had mass-related symptoms at the time of diagnosis. The most frequent histological subtypes were de-differentiated liposarcoma (53%, n = 32) and well-differentiated liposarcoma (35%, n = 21). Twenty-two patients (37%) had perioperative radiotherapy and/or chemotherapy. Fifty-eight patients (97%) had macroscopically complete resection. The postoperative mortality was 8% and severe morbidity (Dindo/Clavien ≥ 3) was 32%. A reoperation was required for ten patients (17%). After a median follow-up of 20 months (range 1–121), the 5-year overall survival (OS) rate was 90% (95% confidence interval [CI] 79–100%), and median OS was not reached. The cancer-specific death rate was 88%. No prognostic factor for disease-specific survival was detected. The 5-year disease-free survival (DFS) rate was 52% (95% CI 33–84%) and 5-year locoregional recurrence-free survival rate was 52% (95% CI 33–84%). Median DFS was 94 months (95% CI 35–NA). Reoperation after inappropriate surgery and postoperative morbidity were independent predictive factors of locoregional relapse. No predictive factors of distant metastasis were found.

Conclusions

Curative surgery is feasible in selected elderly patients but with higher mortality and morbidity rates than in younger patients. It enables a prolonged survival. Future studies should focus on selection process to minimize postoperative mortality and morbidity.

https://ift.tt/2KFoI8d

Routine Axillary Ultrasound for Patients with T1–T2 Breast Cancer Does Not Increase the Rate of Axillary Lymph Node Dissection Based on Predictive Modeling

Abstract

Background

Since publication of the American College of Surgeons Oncology Group Z0011 trial results, demonstrating that many patients with nonpalpable axillary lymph nodes and one or two positive sentinel nodes do not require axillary lymph node dissection (ALND), preoperative axillary ultrasound (AUS) has become controversial. Clinicians are concerned that AUS may lead to unnecessary ALND. The authors developed an algorithm (Algorithm 1) in which the number of AUS-suspicious nodes and tumor biology direct management. For estrogen receptor-positive (ER+)/human epidermal growth factor receptor 2-negative (HER2–) breast cancer with a single AUS-suspicious node and a positive lymph node needle biopsy (LNNB), sentinel lymph node biopsy (SLNB) is performed with a specimen X-ray documenting retrieval of the clipped node. Other patients with positive LNNB receive neoadjuvant chemotherapy. The authors hypothesized that routine AUS and this algorithm could decrease ALND compared with a strategy of no preoperative AUS.

Methods

Decision-tree analysis and Monte Carlo simulation were used to assess the expected number of ALNDs under two strategies (routine AUS vs no AUS). Probabilities were drawn from a literature review and an institutional database. The authors assumed nodal pathologic complete response rates as reported in the literature. Four additional algorithms were created to assess whether any other treatment model could decrease the rate of ALND.

Results

Using the routine AUS and the authors' algorithm, the predicted ALND rate was 9%, versus 10% for a strategy of no AUS, with overlapping uncertainty intervals. The remaining treatment algorithms showed similar results.

Discussion

Use of AUS may help to tailor patient care without leading to overutilization of ALND, as long as neoadjuvant chemotherapy is administered when appropriate.

https://ift.tt/2MGqS4Q

Surgical Management of Lobular Carcinoma In Situ: Analysis of the National Cancer Database

Abstract

Background

Current guidelines recommend counseling on risk-reduction strategies, including lifestyle modification, endocrine therapy, and bilateral mastectomy, for patients with classic-type lobular carcinoma in situ (LCIS) detected on core biopsy or surgical excision. Importantly, current diagnosis and treatment guidelines for classic-type LCIS do not include unilateral mastectomy for primary treatment or risk reduction. Prior studies reporting national practice patterns suggest increasing use of mastectomy for management of LCIS, with considerable variation by geographic region. However, these studies did not distinguish between uni- and bilateral mastectomies. This study aimed to investigate national practice patterns and factors associated with unilateral mastectomy.

Methods

The study used the National Cancer Database to identify women with a diagnosis of LCIS from 2004 to 2013. Descriptive statistics were used to describe surgical treatment, and multinomial logistic regression was used to identify temporal, patient, and facility-level factors associated with receipt of uni- and bilateral mastectomy.

Results

The study identified 30,105 women with LCIS. Of these woman, 5.4% received no surgery, 84.8% had surgical excision, 4% underwent unilateral mastectomy, and 5.1% underwent bilateral mastectomy. Adjusted analysis showed that young age, white race, insurance coverage, greater comorbidity, and geographic region (p < 0.001) were associated with receipt of both uni- and bilateral mastectomy. Additionally, more recent year of diagnosis was associated with receipt of bilateral mastectomy. Unilateral mastectomy rates within geographic regions ranged from 2.7% in New England to 8% in the South.

Conclusions

Nearly as many patients underwent unilateral (4%) as bilateral mastectomy (5.1%), representing inappropriate care. These findings highlight an opportunity to reduce unnecessary care through improved provider and patient education regarding optimal management of LCIS.

https://ift.tt/2KENtBq

Heterogeneity in Outcomes of Pathologic T1-2N1 Breast Cancer After Mastectomy: Looking Beyond Locoregional Failure Rates

Abstract

Purpose

A meta-analysis of 22 randomized trials accrued from 1964 to 1986 demonstrated significantly higher rates of locoregional failure (LRF) and breast-cancer mortality in women with 1–3 positive nodes without postmastectomy radiotherapy (PMRT) after mastectomy (mast.). Recent data demonstrate that PMRT reduces distant metastases (DM) in women with pN1 disease. The challenge today is whether all patients with pathologic T1-2pN1 disease have similar substantial LRF/DM risk that routinely warrants PMRT.

Methods

We reviewed patients with pT1-2N1 breast cancer treated with mast. ± adjuvant systemic therapy without PMRT from 2000 to 2013. The endpoints were LRF and DM rates, estimated by cumulative incidence method.

Results

We identified 468 patients with median follow-up of 6.3 years. Most (71%) were estrogen receptor/progesterone receptor + human epidermal growth factor receptor 2 (HER2). There were 269 patients with 1+ node, 140 patients with 2+ nodes, and 59 patients with 3+ nodes. The 6-year LRF/DM rates were 4.1%/8.4%. Patients with 1+, 2+, and 3+ nodes had 6-year LRF of 2.3, 5.1 and 8.9%, respectively (p = 0.13). The 6-year DM rate was higher in patients with 3+ nodes versus 1–2+ nodes: 15.7% versus 7.4% (p = 0.02). Several subgroups had low 6-year LRF and DM rates, including T1/1+ node (0.8%/4.1% LRF/DM) and micrometastases (0%/5.8% LRF/DM).

Conclusions

Patients with pT1-2pN1 represent a heterogeneous group with a wide range of LRF/DM rates. In particular, patients with pT1 tumors and 1 + LN, and patients with micrometastases, had low event rates. These groups would derive small absolute reductions in LRF and DM with addition of PMRT, underscoring the importance of patient selection for PMRT in pT1-2pN1 breast cancer.

https://ift.tt/2MGqOSE

Neoadjuvant Chemotherapy Use in Breast Cancer is Greatest in Excellent Responders: Triple-Negative and HER2+ Subtypes

Abstract

Background

While breast cancer has historically been treated with surgery followed by adjuvant chemotherapy (AC) and radiation when indicated, neoadjuvant chemotherapy (NAC) use is thought to be increasing; however, the trends of its use in various biological subtypes have not been evaluated. We sought to evaluate the trend of NAC use over time by biological subtype.

Methods

We identified all patients with invasive breast cancer who underwent curative intent surgery and were treated with chemotherapy from 2010 to 2015 from the National Cancer Database. An unadjusted analysis of trends in proportions over time was performed using Cochran–Armitage trend tests stratified by hormone receptor (HR) and human epidermal growth factor receptor 2 (HER2) status.

Results

Of 315,264 patients who received chemotherapy, 251,726 (79.8%) received AC and 63,538 (20.2%) received NAC. From 2010 to 2015, significant increases in NAC use were seen in all biologic subtypes (all p < 0.001). The highest proportions and greatest increases in proportions of NAC were seen among triple-negative breast cancers (TNBC; 19.5–33.7%) and HER2+ (HR−/HER2+, 21.5–39.8%; HR+/HER2+, 17.0–33.7%) tumors. HR+/HER2− tumors also had a statistically significant increase in use but this increase was less dramatic (13.0–16.8%) and NAC use in recent years was significantly lower than in other subtypes (p < 0.001).

Conclusion

Within patients receiving chemotherapy for breast cancer, its receipt in the neoadjuvant setting has been increasing among all biologic subtypes. The highest use of NAC is in TNBC and HER2+ disease, with use in these subgroups being twice as frequent as in HR+/HER2− disease.

https://ift.tt/2KENsxm

Crystal Ball or Magic8 Ball? Reply Hazy, Try Again

https://ift.tt/2MFUuiY

Comparison of Outcome of Esophagectomy Versus Nonsurgical Treatment for Resectable Esophageal Cancer with Clinical Complete Response to Neoadjuvant Therapy

Abstract

Background

Treatment for patients who have achieved clinical complete response (cCR) after neoadjuvant therapy has not been established, and there is no consensus regarding the indications for either esophagectomy or nonsurgical treatment.

Methods

Among 1,545 patients with esophageal cancer at Toranomon Hospital between January 2006 and August 2017, 39 who achieved cCR after neoadjuvant treatment were divided into two groups according to treatment: esophagectomy group (n = 18) and nonsurgical treatment group (n = 21) for comparison.

Results

No significant intergroup difference was observed in baseline characteristics. Pathological complete response was confirmed in 13 (72.2%) of the 18 patients who underwent esophagectomy, whereas residual tumor was detected at the location of primary tumor in 2 (11.1%) patients, and lymph node metastasis was found in 3 (16.7%) patients. Recurrence-free survival (RFS) was significantly longer in the esophagectomy group than in the nonsurgical group (p = 0.002). Disease-specific survival (DSS) was significantly longer in the esophagectomy group (p = 0.007). However, no significant intergroup difference was observed in overall survival estimated based on all deaths, including respiratory failure and aspiration pneumonia (p = 0.451).

Conclusions

With improved diagnostic accuracy, nonsurgical treatment can be an option for patients estimated as cCR after treatment administered in a neoadjuvant setting. However, surgical resection is considered more appropriate because of residual tumor in some patients with cCR and because of superior DSS and RFS following esophagectomy compared with nonsurgical treatment. Future studies must focus on ameliorating late postoperative complications, such as respiratory failure and aspiration pneumonia.

https://ift.tt/2KvaDv3

RNA-Seq of Circulating Tumor Cells in Stage II–III Breast Cancer

Abstract

Background

We characterized the whole transcriptome of circulating tumor cells (CTCs) in stage II–III breast cancer to evaluate correlations with primary tumor biology.

Methods

CTCs were isolated from peripheral blood (PB) via immunomagnetic enrichment followed by fluorescence-activated cell sorting (IE/FACS). CTCs, PB, and fresh tumors were profiled using RNA-seq. Formalin-fixed, paraffin-embedded (FFPE) tumors were subjected to RNA-seq and NanoString PAM50 assays with risk of recurrence (ROR) scores.

Results

CTCs were detected in 29/33 (88%) patients. We selected 21 cases to attempt RNA-seq (median number of CTCs = 9). Sixteen CTC samples yielded results that passed quality-control metrics, and these samples had a median of 4,311,255 uniquely mapped reads (less than PB or tumors). Intrinsic subtype predicted by comparing estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) versus PAM50 for FFPE tumors was 85% concordant. However, CTC RNA-seq subtype assessed by the PAM50 classification genes was highly discordant, both with the subtype predicted by ER/PR/HER2 and by PAM50 tumors. Two patients died of metastatic disease, both of whom had high ROR scores and high CTC counts. We identified significant genes, canonical pathways, upstream regulators, and molecular interaction networks comparing CTCs by various clinical factors. We also identified a 75-gene signature with highest expression in CTCs and tumors taken together that was prognostic in The Cancer Genome Atlas and Molecular Taxonomy of Breast Cancer International Consortium datasets.

Conclusion

It is feasible to use RNA-seq of CTCs in non-metastatic patients to discover novel tumor biology characteristics.

https://ift.tt/2MJfvsK

Compliance with an Enhanced Recovery After a Surgery Program for Patients Undergoing Gastrectomy for Gastric Carcinoma: A Phase 2 Study

Abstract

Background

Enhanced recovery after surgery (ERAS) programs have gained widespread acceptance in different fields of major surgery. However, most elements of perioperative care in ERAS are based on practices that originated from colorectal surgery. This study investigated compliance with the main elements of ERAS for patients undergoing gastrectomy for gastric carcinoma.

Methods

This phase 2 study enrolled 168 patients undergoing elective gastrectomy for gastric carcinoma. An ERAS program consisting of 18 main elements was implemented, and compliance with each element was evaluated (ClinicalTrials.gov, NCT01653496).

Results

Distal gastrectomy was performed for 142 patients (84.5%) and total gastrectomy for 26 patients (10.1%). Laparoscopic surgery was performed for 141 patients (86%). The postoperative morbidity rate was 9.5%, and the mortality rate was 0%. The rates of compliance with the 18 main elements of ERAS ranged from 88.1 to 100%. The lowest compliance rate was observed in the restriction of intravenous fluid element (88.1%). Overall, all ERAS elements were successfully applied for 122 patients (72.6%). In the multivariate analysis, the significant factors that adversely affected compliance with ERAS were surgery during the early study period [odds ratio (OR) 0.39; p = 0.038], open surgery (OR 0.15; p <0.001), and postoperative morbidity (OR 0.16; p = 0.003).

Conclusions

Most elements of ERAS can be successfully applied for patients undergoing gastrectomy for gastric carcinoma. Multimodal collaboration between providers is essential to achieve proper application of ERAS.

https://ift.tt/2KvaAPT

Inhibition Mechanism of Acellular Dermal Matrix on Capsule Formation in Expander–Implant Breast Reconstruction After Postmastectomy Radiotherapy

Abstract

Background

Capsular contracture is one of the most common complications of expander–implant breast reconstruction. Recently, clinical reports have shown that use of an acellular dermal matrix (ADM) to cover breast implants decreases incidence of capsular contracture, but the underlying mechanism is unclear. Here, we examine how ADM reduces capsular formation in expander–implant breast reconstruction and identify cellular and molecular mechanisms of ADM-mediated reduction of capsular contracture in nonirradiated and irradiated patients.

Methods

Thirty patients who underwent immediate two-stage implant-based breast reconstruction were included; 15 received radiotherapy. While the tissue expander was changed to permanent silicone implant, biopsies of the subpectoral capsule and ADM capsule were performed. Capsule thickness, immunohistochemistry of α-smooth muscle actin (αSMA), vimentin, CD31, F4/80 expression, αSMA and CD31 coexpression, and relative gene expression levels of transforming growth factor (TGF)-β1 and platelet-derived growth factor (PDGF)-B were investigated.

Results

Irradiated submuscular capsules were thicker than nonirradiated submuscular capsules, but the thickness of ADM capsules did not significantly differ between nonirradiated and irradiated groups. Levels of myofibroblasts, fibroblasts, vascularity, EndoMT, and macrophages were significantly lower in ADM capsules than in submuscular capsules. With the exception of EndoMT, all others were increased in irradiated submuscular capsules compared with nonirradiated submuscular capsule, while none significantly differed between nonirradiated and irradiated ADM capsules.

Conclusions

Use of ADM reduced myofibroblasts, vascularity, fibroblasts, and EndoMT in capsule tissues. Moreover, ADM use decreased macrophages, a key regulator of tissue fibrosis, as well as TGF-β1 and PDGF-B expression. We hope that these results provide basic concepts important for prevention of capsular contracture.

https://ift.tt/2MHgJEV

Minimally Invasive Surgery for Retroperitoneal Sarcoma: Just Because We Can Does Not Mean We Should

https://ift.tt/2KrF6dk

Metastatic Melanoma to the Colon, Rectum, and Anus: A 50-Year Experience

Abstract

Background

Melanoma metastatic to the large bowel (colon, rectum, and anus) is rarely diagnosed, with more than 95% of large bowel metastases identified post-mortem. The incidence, natural history, and survival rates of patients with large bowel melanoma metastases are poorly documented in the literature.

Objective

This study aimed to identify the incidence, clinical characteristics, and survival of patients with large bowel melanoma metastases.

Methods

A review was undertaken of all patients with melanoma treated over a 50-year period (1964–2014) at a tertiary referral center. Cases selected for study were those diagnosed with melanoma metastases in the colon, rectum, and anus. Primary colorectal and anal melanomas were excluded. Data were retrieved relating to patient demographics, clinical features, and survival.

Results

Of 38,279 patients with primary melanoma, 106 patients (0.3%, mean age 51.0 years [standard deviation 16.3], 64 males) developed large bowel metastases. The median interval between diagnosis of primary melanoma and large bowel metastasis was 62.8 months (range 1–476). The most common symptom was rectal bleeding (29.2%), and the large bowel was the sole site of metastasis in 47.2% of patients. Median survival from diagnosis of large bowel metastasis was 31.7 months (range 1–315), and overall survival at 1, 2, and 5 years was 68.1, 45.9, and 26.5%, respectively.

Conclusion

Our study provides insights into melanoma metastatic to the colon, rectum, and anus, which had an incidence of 0.3%. There are potentially long intervals between diagnosis of primary melanoma and large bowel metastasis. The most common symptom was rectal bleeding, although some patients were asymptomatic.

https://ift.tt/2ME4IjJ

Multicenter Study of Presentation, Management, and Postoperative and Long-Term Outcomes of Septegenerians and Octogenerians Undergoing Gastrectomy for Gastric Cancer

Abstract

Background

The optimal treatment strategy for elderly patients with gastric cancer is still controversial. This study aimed to assess the impact of age on short- and long-term outcomes after treatment for primary gastric cancer.

Methods

From January 2004 to December 2014, a total of 507 patients underwent gastrectomy for gastric adenocarcinoma at two high-volume upper gastrointestinal (GI) centers. The patients were classified into three groups as follows: group A (patients ≤ 69 years old, n = 266), group B (patients 70–79 years old, n = 166), and group C (patients ≥ 80 years old, n = 75). Clinicopathologic characteristics as well as, short- and long-term outcomes were compared between the groups.

Results

The patients in groups B and C had more comorbidities, whereas the younger subjects (group A) had more advanced tumor stages. Less extensive surgery was performed in the groups B and C. Older patients (age ≥ 70 years) had more postoperative medical complications. Moreover, group C had a higher postoperative mortality rate (8.1%) than group A (1.8%) or group B (1.9%). In the multivariable analysis, age older than 80 years (group C) was a negative independent factor for overall survival (OS) (hazard ratio [HR], 2.36) compared with group A, whereas group B seemed to have a comparable risk (HR, 1.37). Notably, the three groups did not show significant differences in disease-related survival (DRS).

Conclusion

The data suggest that patients 70–79 years of age show a risk of postoperative death comparable with that of younger subjects. However, patients older than 80 years should be carefully selected for surgical treatment due to the increased risk of postoperative mortality.

https://ift.tt/2KrF3OG

Cost Effectiveness of Cytoreductive Surgery and Hyperthermic Intraperitoneal Chemotherapy for Management of Colorectal Peritoneal Carcinomatosis

Abstract

Background

Peritoneal carcinomatosis from colorectal cancer is a stage 4 disease for which palliative chemotherapy has traditionally been considered the mainstay of treatment. Since the development of cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) by Sugarbaker, this combined method treatment has resulted in improved survival outcomes with acceptable morbidity for selected patients with peritoneal carcinomatosis. This study examined the cost effectiveness of CRS and HIPEC compared with palliative chemotherapy for patients with peritoneal carcinomatosis from colorectal cancer within the context of the Singaporean health care system.

Methods

A retrospective review of patients with peritoneal carcinomatosis from histologically proven colorectal cancer treated at the National Cancer Centre Singapore (NCCS) was conducted.

Results

The average cost of CRS and HIPEC per patient was S$83,680.26, and the median overall survival period was 47 months. The calculated cost per life year attained for a patient who underwent CRS and HIPEC was S$21,365.19 per life year. In comparison, the average cost of palliative chemotherapy was S$44,478.87, with a median overall survival of 9 months, and the calculated cost per life year attained for a patient in this treatment group was S$59,305.16 per life year.

Conclusion

The findings show that CRS and HIPEC results in prolonged survival for selected patients with colorectal peritoneal carcinomatosis and a lower cost per life year attained than for the traditionally used palliative chemotherapy. It should logically be the preferred treatment of choice for selected patients with colorectal peritoneal metastasis.

https://ift.tt/2ME4GID

Strategic Delay: Histology- and Biology-Driven Decision-Making in Recurrent Retroperitoneal Sarcoma

https://ift.tt/2KqPNwO

Evaluation of the Relationship Between Flap Tension and Tissue Perfusion in Implant-Based Breast Reconstruction Using Laser-Assisted Indocyanine Green Angiography

Abstract

Background

For implant-based breast reconstruction, inadequate tissue perfusion may cause devastating complications. Tissue tension by inadequate implant volume may reduce tissue perfusion by stretching and collapsing the capillaries. The SPY system is used to perform intraoperative fluorescence angiography with indocyanine green to assess visually the blood flow and evaluate tissue perfusion. However, there is no report yet about how mastectomy flap perfusion changes with the expander-filling volume. Therefore, to analyse the changes of tissue perfusion of the mastectomy flap according to the tension level, we used the SPY system and adjusted the filling volume of the tissue expander to change the tension on the skin flap.

Methods

Ten breasts of ten patients who underwent immediate two-stage, implant-based breast reconstructions were included. The expander-filling volume just before mastectomy flap blanching was set as 100%. Based on this, the expander-filling volume was reduced to 50% and increased to 150%. Ingress and egress rates were evaluated using the SPY system at each condition and analysed by a linear mixed model using least square means.

Results

The mean ingression rates were 138, 100, and 65%, and the mean egression rates were 145, 100, and 66% at 50, 100, and 150% inflation, respectively.

Conclusions

It was objectively proven that tissue perfusion deteriorates as the tension applied on the flap increases. On the basis of this finding, we can control the amount of inflation volume of the expander or remove the skin in the pre-ischaemic condition to reduce complications of implant-based breast reconstruction.

https://ift.tt/2MFrIyS

Walking the Fine Line of Axillary Management in Mastectomy Patients

https://ift.tt/2KExWla

Techniques for Cytoreductive Surgery with Hyperthermic Intraperitoneal Chemotherapy

https://ift.tt/2MJrTJk

Dynamic bimodal changes in CpG and non-CpG methylation genome-wide upon CGGBP1 loss-of-function

Although CpG methylation is well studied, mechanisms of non-CpG methylation in mammals remains elusive. Studying proteins with non-CpG cytosine methylation-sensitive DNA-binding, such as human CGGBP1, can unve...

https://ift.tt/2z77Y5o

Establishing a New Clinical Role for Medical Physicists: A Prospective Phase II Trial

During the course of radiation therapy, patients are inundated with challenging and complicated information related to their care, which often results in anxiety and confusion. In this study, medical physicists routinely met with patients, where they explained the treatment planning and delivery process, reviewed the patient's treatment plan, and answered all technical questions. The results from this clinical trial indicate that these physicist-patient consults have the potential to decrease patient anxiety and increase patient satisfaction.

https://ift.tt/2KHPART

Radiation-induced lung density changes on CT scan for NSCLC: no impact of dose-escalation level or volume

Understanding the regional dose volume effects of lung parenchyma damage induction is crucial to dissect the causes of the multifactorial radiation-induced pulmonary toxicity. The relationship between dose and lung density changes on CT 3 months after treatment was therefore analyzed in the prospective dataset of a dose-escalation study in NSCLC. Lung Hounsfield Unit increase saturated above 60 Gy EQD2 and was independent of dose-escalation level and volume. Mild responses were observed peripherally in the lung.

https://ift.tt/2IQt73q

A Bayesian approach for prediction of patient radiosensitivity

Using Bayesian statistical analysis of appearance and repair of DNA damage in peripheral blood lymphocytes following ex-vivo irradiation, we developed a model that reliably discriminated between radiosensitive and non-radiosensitive ex-radiotherapy patients. This new approach to data analysis improves the prospects of developing an assay to identify radiosensitive patients before they are scheduled for radiotherapy.

https://ift.tt/2KCul7b

Focal Salvage High Dose-Rate Brachytherapy for Locally Recurrent Prostate Cancer after Primary Radiotherapy Failure: Results from a Prospective Clinical Trial

No prospective published data exist on focal salvage high-dose-rate (HDR) brachytherapy for recurrent prostate cancer. In the present prospective trial, we evaluated toxicity, quality of life and efficacy of such an approach in patients with multiparametric MRI-visible, biopsy-confirmed local recurrence after previous definitive external beam radiotherapy. We found that it had a favorable toxicity profile, was associated with promising biochemical control and yielded MRI response in the majority of patients.

https://ift.tt/2Kstu9R

Circulating Tumor Cell Assessment in Presumed Early Stage Non-small Cell Lung Cancer Patients Treated with Stereotactic Body Radiation Therapy: A Prospective Pilot Study

In patients treated with SBRT for presumed early stage non-small cell lung cancer (NSCLC), detection and monitoring of circulating tumor cells (CTCs) may be useful for assessing treatment response safely and non-invasively. No published reports of CTC trends in this patient population exist to date.

https://ift.tt/2KCuqrv

Tox and Hound – The KULTS of Toxicology

by Meghan Spyres       When someone asks me 'why toxicology?', I don't think of the chance to use colorful antidotes like hydroxocobalamin for cyanide or Prussian blue for thallium toxicity. It's not even the chance to manage a blue ringed octopus envenomation. Exotic cases are exciting, but for most of us, we choose […]

EMCrit Project by Tox & Hound.

https://ift.tt/2z4kMt9

The E3 Ubiquitin Ligase MIB-1 Is Necessary To Form the Nuclear Halo in Caenorhabditis elegans Sperm

Unlike the classical nuclear envelope with two membranes found in other eukaryotic cells, most nematode sperm nuclei are not encapsulated by membranes. Instead, they are surrounded by a nuclear halo of unknown composition. How the halo is formed and regulated is unknown. We used forward genetics to identify molecular lesions behind three classical fer (fertilization defective) mutations that disrupt the ultrastructure of the Caenorhabditis elegans sperm nuclear halo. We found fer-2 and fer-4 alleles to be nonsense mutations in mib-1. fer-3 was caused by a nonsense mutation in eri-3. GFP::MIB-1 was expressed in the germline during early spermatogenesis, but not in mature sperm. mib-1 encodes a conserved E3 ubiquitin ligase homologous to vertebrate Mib1 and Mib2, which function in Notch signaling. Here, we show that mib-1 is important for male sterility and is involved in the regulation or formation of the nuclear halo during nematode spermatogenesis.

https://ift.tt/2tO0UFJ

The domino SWI2/SNF2 Gene Product Represses Cell Death in Drosophila melanogaster

The Drosophila domino locus encodes DNA-dependent ATPases of the SWI2/SNF2 class. This class of chromatin remodeler is associated with an array of cellular activities encompassing transcription, replication, repair and recombination. Moreover, domino was observed initially to maintain a repressive chromatin state via genetic interaction studies with homeotic genes. Although domino mutations were also characterized with a cell death phenotype, its association with a death pathway has not been investigated. Here we have used targeted RNA interference to depress domino function in the wing. Resultant wing damage phenotypes were found to be enhanced through overexpression of pro-apoptotic loci, and suppressed through loss of function of these loci. Loss of wing margin and blade tissue was correlated with activation of the effector Caspase Dcp-1, a marker for apoptosis. The affected wing regions also exhibited lower levels of the DIAP1 protein, an inhibitor of apoptosis. The lower level of DIAP1 protein was not correlated with an effect on the activity of a DIAP1 gene transgenic reporter (thread-LacZ), suggesting that loss of DIAP1 occurred post transcriptionally. In some cases excessive cell proliferation within the targeted tissue, measured through BrdU incorporation, was also observed. Finally, we used a transgenic reporter construct to monitor the chromatin state upstream of the proapoptotic reaper locus. In genotypes exhibiting targeted domino loss and wing phenotypes, we observed increased reporter activity only in the affected areas. These data support the conclusion that domino normally functions to maintain pro-apoptotic genes in a repressed state.

https://ift.tt/2z3QyX5

A High Quality Genome for Mus spicilegus, a Close Relative of House Mice with Unique Social and Ecological Adaptations

Genomic data for the closest relatives of house mice (Mus musculus species complex) are surprisingly limited. Here, we present the first complete genome for a behaviorally and ecologically unique member of the sister clade to house mice, the mound-building mouse, Mus spicilegus. Using read cloud sequencing and de novo assembly we produced a 2.50 Gbp genome with a scaffold N50 of 2.27 Mbp. We constructed >25 000 gene models, of which the majority had high homology to other Mus species. To evaluate the utility of the M. spicilegus genome for behavioral and ecological genomics, we extracted 196 vomeronasal receptor (VR) sequences from our genome and analyzed phylogenetic relationships between M. spicilegus VRs and orthologs from M. musculus and the Algerian mouse, M. spretus. While most M. spicilegus VRs clustered with orthologs in M. musculus and M. spretus, 10 VRs with evidence of rapid divergence in M. spicilegus are strong candidate modulators of species-specific chemical communication. A high quality assembly and genome for M. spicilegus will help to resolve discordant ancestry patterns in house mouse genomes, and will provide an essential foundation for genetic dissection of phenotypes that distinguish commensal from non-commensal species, and the social and ecological characteristics that make M. spicilegus unique.

https://ift.tt/2Niqjze

Assessment of an Organ-Specific de Novo Transcriptome of the Nematode Trap-Crop, Solanum sisymbriifolium

Solanum sisymbriifolium, also known as "Litchi Tomato" or "Sticky Nightshade," is an undomesticated and poorly researched plant related to potato and tomato. Unlike the latter species, S. sisymbriifolium induces eggs of the cyst nematode, Globodera pallida, to hatch and migrate into its roots, but then arrests further nematode maturation. In order to provide researchers with a partial blueprint of its genetic make-up so that the mechanism of this response might be identified, we used single molecule real time (SMRT) sequencing to compile a high quality de novo transcriptome of 41,189 unigenes drawn from individually sequenced bud, root, stem, and leaf RNA populations. Functional annotation and BUSCO analysis showed that this transcriptome was surprisingly complete, even though it represented genes expressed at a single time point. By sequencing the 4 organ libraries separately, we found we could get a reliable snapshot of transcript distributions in each organ. A divergent site analysis of the merged transcriptome indicated that this species might have undergone a recent genome duplication and re-diploidization. Further analysis indicated that the plant then retained a disproportionate number of genes associated with photosynthesis and amino acid metabolism in comparison to genes with characteristics of R-proteins or involved in secondary metabolism. The former processes may have given S. sisymbriifolium a bigger competitive advantage than the latter did.

https://ift.tt/2u0GKr6

Genomic Variation Among and Within Six Juglans Species

Genomic analysis in Juglans (walnuts) is expected to transform the breeding and agricultural production of both nuts and lumber. To that end, we report here the determination of reference sequences for six additional relatives of Juglans regia: Juglans sigillata (also from section Dioscaryon), Juglans nigra, Juglans microcarpa, Juglans hindsii (from section Rhysocaryon), Juglans cathayensis (from section Cardiocaryon), and the closely related Pterocarya stenoptera. While these are 'draft' genomes, ranging in size between 640Mbp and 990Mbp, their contiguities and accuracies can support powerful annotations of genomic variation that are often the foundation of new avenues of research and breeding. We annotated nucleotide divergence and synteny by creating complete pairwise alignments of each reference genome to the remaining six. In addition, we have re-sequenced a sample of accessions from four Juglans species (including regia). The variation discovered in these surveys comprises a critical resource for experimentation and breeding, as well as a solid complementary annotation. To demonstrate the potential of these resources the structural and sequence variation in and around the polyphenol oxidase loci, PPO1 and PPO2 were investigated. As reported for other seed crops variation in this gene is implicated in the domestication of walnuts. The apparently Juglandaceae specific PPO1 duplicate shows accelerated divergence and an excess of amino acid replacement on the lineage leading to accessions of the domesticated nut crop species, Juglans regia and sigillata.

https://ift.tt/2tNtS8u

A Dense Linkage Map of Lake Victoria Cichlids Improved the Pundamilia Genome Assembly and Revealed a Major QTL for Sex-Determination

Genetic linkage maps are essential for comparative genomics, high quality genome sequence assembly and fine scale quantitative trait locus (QTL) mapping. In the present study we identified and genotyped markers via restriction-site associated DNA (RAD) sequencing and constructed a genetic linkage map based on 1,597 SNP markers of an interspecific F2 cross of two closely related Lake Victoria cichlids (Pundamilia pundamilia and P. sp. 'red head'). The SNP markers were distributed on 22 linkage groups and the total map size was 1,594 cM with an average marker distance of 1.01 cM. This high-resolution genetic linkage map was used to anchor the scaffolds of the Pundamilia genome and estimate recombination rates along the genome. Via QTL mapping we identified a major QTL for sex in a ~1.9 Mb region on Pun-LG10, which is homologous to Oreochromis niloticus LG 23 (Ore-LG23) and includes a well-known vertebrate sex-determination gene (amh).

https://ift.tt/2z4uGej

BLAST-XYPlot Viewer: A Tool for Performing BLAST in Whole-Genome Sequenced Bacteria/Archaea and Visualize Whole Results Simultaneously

One of the most commonly used tools to compare protein or DNA sequences against databases is BLAST. We introduce a web tool that allows the performance of BLAST-searches of protein/DNA sequences in whole-genome sequenced bacteria/archaea, and displays a large amount of BLAST-results simultaneously. The circular bacterial replicons are projected as horizontal lines with fixed length of 360, representing the degrees of a circle. A coordinate system is created with length of the replicon along the x-axis and the number of replicon used on the y-axis. When a query sequence matches with a gene/protein of a particular replicon, the BLAST-results are depicted as an "x,y" position in a specially adapted plot. This tool allows the visualization of the results from the whole data to a particular gene/protein in real time with low computational resources.

https://ift.tt/2NkJloW

Phenotypic Data from Inbred Parents Can Improve Genomic Prediction in Pearl Millet Hybrids

Pearl millet is a non-model grain and fodder crop adapted to extremely hot and dry environments globally. In India, a great deal of public and private sectors' investment has focused on developing pearl millet single cross hybrids based on the cytoplasmic-genetic male sterility (CMS) system, while in Africa most pearl millet production relies on open pollinated varieties. Pearl millet lines were phenotyped for both the inbred parents and hybrids stage. Many breeding efforts focus on phenotypic selection of inbred parents to generate improved parental lines and hybrids. This study evaluated two genotyping techniques and four genomic selection schemes in pearl millet. Despite the fact that 6x more sequencing data were generated per sample for RAD-seq than for tGBS, tGBS yielded more than 2x as many informative SNPs (defined as those having MAF > 0.05) than RAD-seq. A genomic prediction scheme utilizing only data from hybrids generated prediction accuracies (median) ranging from 0.73-0.74 (1000-grain weight), 0.87-0.89 (days to flowering time), 0.48-0.51 (grain yield) and 0.72-0.73 (plant height). For traits with little to no heterosis, hybrid only and hybrid/inbred prediction schemes performed almost equivalently. For traits with significant mid-parent heterosis, the direct inclusion of phenotypic data from inbred lines significantly (P < 0.05) reduced prediction accuracy when all lines were analyzed together. However, when inbreds and hybrid trait values were both scored relative to the mean trait values for the respective populations, the inclusion of inbred phenotypic datasets moderately improved genomic predictions of the hybrid genomic estimated breeding values. Here we show that modern approaches to genotyping by sequencing can enable genomic selection in pearl millet. While historical pearl millet breeding records include a wealth of phenotypic data from inbred lines, we demonstrate that the naive incorporation of this data into a hybrid breeding program can reduce prediction accuracy, while controlling for the effects of heterosis per se allowed inbred genotype and trait data to improve the accuracy of genomic estimated breeding values for pearl millet hybrids.

https://ift.tt/2u0GwQM

Development of Diagnostic SNP Markers To Monitor Hybridization Between Sika Deer (Cervus nippon) and Wapiti (Cervus elaphus)

Sika deer (Cervus Nippon) and wapiti (Cervus elaphus) are closely related species and their hybridization can result in significant allele-shift of their gene pool. Additive genetic effects and putative heterotic effects of their hybridization on growth performance could confer considerable economic advantage in deer farming. Here, we used double-digest restriction site-associated DNA sequencing technology (ddRAD-seq) and detected ~320,000 genome-wide SNPs from 30 captive individuals: 7 sika deer, 6 wapiti and 17 F1 hybrids (reciprocal cross). By screening observed heterozygosity of each SNP across four taxonomic groups, we report for the first time a resource of 2,015 putative diagnostic SNP markers (species-specific SNPs for sika deer and wapiti), which can be used to design tools for assessing or monitoring the degree of hybridization between sika deer and wapiti. These ddRAD-seq data and SNP datasets are also valuable resources for genome-wide studies, including trait discovery for breeders of domestic deer.

https://ift.tt/2NjiWYv

Dynamic Changes in Yeast Phosphatase Families Allow for Specialization in Phosphate and Thiamine Starvation

Convergent evolution is often due to selective pressures generating a similar phenotype. We observe relatively recent duplications in a spectrum of Saccharomycetaceae yeast species resulting in multiple phosphatases that are regulated by different nutrient conditions – thiamine and phosphate starvation. This specialization is both transcriptional and at the level of phosphatase substrate specificity. In Candida glabrata, loss of the ancestral phosphatase family was compensated by the co-option of a different histidine phosphatase family with three paralogs. Using RNA-seq and functional assays, we identify one of these paralogs, CgPMU3, as a thiamine phosphatase. We further determine that the 81% identical paralog CgPMU2 does not encode thiamine phosphatase activity; however, both are capable of cleaving the phosphatase substrate, 1-napthyl-phosphate. We functionally demonstrate that members of this family evolved novel enzymatic functions for phosphate and thiamine starvation, and are regulated transcriptionally by either nutrient condition, and observe similar trends in other yeast species. This independent, parallel evolution involving two different families of histidine phosphatases suggests that there were likely similar selective pressures on multiple yeast species to recycle thiamine and phosphate. In this work, we focused on duplication and specialization, but there is also repeated loss of phosphatases, indicating that the expansion and contraction of the phosphatase family is dynamic in many Ascomycetes. The dynamic evolution of the phosphatase gene families is perhaps just one example of how gene duplication, co-option, and transcriptional and functional specialization together allow species to adapt to their environment with existing genetic resources.

https://ift.tt/2u0Gp7O