bluuebunny/crossref_metadata_2025 huggingface dataset:
The pentose phosphate pathway regulates chronic neuroinflammation and dopaminergic neurodegeneration
Authors: Tu Dezhen , Gao Yun , Yang Ru , Guan Tian , Hong Jau-Shyong , Gao Hui-Ming
Published: December 2019

Background

Metabolic dysfunction and neuroinflammation are increasingly implicated in Parkinson’s disease (PD). The pentose phosphate pathway (PPP, a metabolic pathway parallel to glycolysis) converts glucose-6-phosphate into pentoses and generates ribose-5-phosphate and NADPH thereby governing anabolic biosynthesis and redox homeostasis. Brains and immune cells display high activity of glucose-6-phosphate dehydrogenase (G6PD), the rate-limiting enzyme of the PPP. A postmortem study reveals dysregulation of G6PD enzyme in brains of PD patients. However, spatial and temporal changes in activity/expression of G6PD in PD remain undetermined. More importantly, it is unclear how dysfunction of G6PD and the PPP affects neuroinflammation and neurodegeneration in PD.

Methods

We examined expression/activity of G6PD and its association with microglial activation and dopaminergic neurodegeneration in multiple chronic PD models generated by an intranigral/intraperitoneal injection of LPS, daily subcutaneous injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) for 6 days, or transgenic expression of A53T α-synuclein. Primary microglia were transfected with G6PD siRNAs and treated with lipopolysaccharide (LPS) to examine effects of G6PD knockdown on microglial activation and death of co-cultured neurons. LPS alone or with G6PD inhibitor(s) was administrated to mouse substantia nigra or midbrain neuron-glia cultures. While histological and biochemical analyses were conducted to examine microglial activation and dopaminergic neurodegeneration in vitro and in vivo, rotarod behavior test was performed to evaluate locomotor impairment in mice.

Results

Expression and activity of G6PD were elevated in LPS-treated midbrain neuron-glia cultures (an in vitro PD model) and the substantia nigra of four in vivo PD models. Such elevation was positively associated with microglial activation and dopaminergic neurodegeneration. Furthermore, inhibition of G6PD by 6-aminonicotinamide and dehydroepiandrosterone and knockdown of microglial G6PD attenuated LPS-elicited chronic dopaminergic neurodegeneration. Mechanistically, microglia with elevated G6PD activity/expression produced excessive NADPH and provided abundant substrate to over-activated NADPH oxidase (NOX2) leading to production of excessive reactive oxygen species (ROS). Knockdown and inhibition of G6PD ameliorated LPS-triggered production of ROS and activation of NF-кB thereby dampening microglial activation.

Conclusions

Our findings indicated that G6PD-mediated PPP dysfunction and neuroinflammation exacerbated each other mediating chronic dopaminergic neurodegeneration and locomotor impairment. Insight into metabolic-inflammatory interface suggests that G6PD and NOX2 are potential therapeutic targets for PD.
Response of membrane tension to gravity in an approximate cell model
Authors: Wang Lili , Chen Weiyi , Guo Hongmei , Qian Airong
Published: December 2019

Background

Gravity, especially hypergravity, can affect the morphology of membranes, and further influence most biological processes. Since vesicle structures are relatively simple, the vesicle can be treated as a vital model to study the mechanical properties of membranes in most cases. Basic research on membrane tension has become a vital research topic in cellular biomechanics.

Methods

In this study, a new vesicle model is proposed to quantitatively investigate the response of membrane tension to gravity. In the model, the aqueous lumen inside the vesicle is represented by water, and the vesicle membrane is simplified as a closed, thin, linear elastic shell. Then, the corresponding static equilibrium differential equations of membrane tension are established, and the analytical expression is obtained by the semi-inverse method. The model parameters of the equations are accurately obtained by fitting the reported data, and the values calculated by the model agree well with the reported results.

Results

The results are as follows: First, both the pseudo-ellipsoidal cap and the pseudo-spherical cap can be used to describe the deformed vesicle model; however, the former can better represent the deformation of the vesicle model because the variance of the pseudo-ellipsoidal cap is smaller. Second, the value of membrane tension is no longer a constant for both models. Interestingly, it varies with the vesicle height under the action of gravity. The closer it is to the substrate, the greater the membrane tension. Finally, the inclination between the tangent and the radial lines at a certain point is nearly proportional to the radius of the cross section in both models.

Conclusion

These findings may be helpful to study the vesicle model spreading more accurately by taking into account the influence of gravity because it could affect the distribution of membrane tension. Furthermore, it may also provide some guidance for cell spreading and may have some implications for membrane tension-related mechanobiology studies, especially in the hypergravity conditions.
Low rates of prior HIV testing among HIV-positive adults accessing outpatient services in Eswatini
Authors: Nuwagaba-Biribonwoha Harriet , Wu Yingfeng , Gachuhi Averie Baird , McNairy Margaret L. , Madau Veli , Lamb Mathew , Mazibuko Sikhathele , Mnisi Zandile , Burke Sean , Philip Neena , Sahabo Ruben , El Sadr Wafaa M.
Published: December 2019

Prior HIV testing and awareness of HIV-positive status were assessed among HIV-positive adults at 20 clinics in Eswatini. Of 2196 HIV-positive adults, 1183 (53.8%) reported no prior HIV testing, and 1948 (88.7%) were unaware of their HIV-positive status. Males [adjusted odds ratio, AOR, (95% confidence interval): 0.7 (0.5–0.9)], youth 18–25 years [AOR 0.6 (0.4–0.95)], adults ≥ 50 years [AOR 0.5 (0.3–0.9)], those needing family support [AOR 0.6 (0.5–0.8)], and those living ≥ 45 min from clinic [AOR 0.5 (0.4–0.8)] were less likely to know their HIV-positive status. More HIV testing is needed to achieve 95-95-95 targets, with targeted strategies for those less likely to test for HIV.
A second puncture and injection technique for treating osteoporotic vertebral compression fractures
Authors: Zhang Zhaofei , Jiao Feng , Huang He , Feng Yonghui , Xie Chunliang , Liu Donghua , Qin Fengwei , Zhang Sineng , Wu Peiyu , Tan Weiguang , Tang Wang
Published: December 2019

Objective

To evaluate the clinical effect of the second puncture and injection technique during a percutaneous vertebroplasty (PVP) procedure.

Methods

Patients treated with a second puncture and injection (group A) or a single puncture and injection (group B) during PVP at our institution during 2010–2017 were reviewed. Vertebral height loss, visual analogue scale (VAS) score, Oswestry disability index (ODI), adjacent vertebral fractures, and cement leakage were compared between the groups.

Results

A total of 193 patients were enrolled (86 cases in group A, 107 cases in group B). The follow-up period was 15.64 (12–20) months. The loss of anterior (group A 0.01 ± 0.03; group B 0.14 ± 0.17) and middle (group A 0.13 ± 0.12; group B 0.16 ± 0.11) vertebral height in group B was significantly higher than that in group A (P < 0.05). The VAS score and ODI were also significantly higher in group B than in group A at the final follow-up; the VAS score and ODI in group B were 1.65 ± 0.70 and 14.50 ± 4.16, respectively, and those in group A were 1.00 ± 0.74 and 12.81 ± 4.02, respectively (P < 0.05). Three patients in group A and two in group B experienced adjacent vertebral fractures. Regarding mild, moderate, and severe cement leakage, there were 25 (29%), 5 (5%), and 0 cases, respectively, in group A and 28 (26%), 3 (2.8%), and 1 (0.009%) case, respectively, in group B (P > 0.05).

Conclusions

The second puncture and injection technique may effectively increase the dispersion of cement, thus preventing recompression of the cemented vertebral body, and it does not increase the risk of cement leakage or adjacent vertebral fracture.
Evolutionarily novel genes are expressed in transgenic fish tumors and their orthologs are involved in development of progressive traits in humans
Authors: Matyunina E. A. , Emelyanov A. V. , Kurbatova T. V. , Makashov A. A. , Mizgirev I. V. , Kozlov A. P.
Published: December 2019

Earlier we suggested a new hypothesis of the possible evolutionary role of hereditary tumors (Kozlov, Evolution by tumor Neofunctionalization, 2014), and described a new class of genes – tumor specifically expressed, evolutionarily novel (TSEEN) genes - that are predicted by this hypothesis (Kozlov, Infect Agents Cancer 11:34, 2016). In this paper we studied evolutionarily novel genes expressed in fish tumors after regression, as a model of evolving organs. As evolutionarily novel genes may not yet have organismal functions, we studied the acquisition of new gene functions by comparing fish evolutionarily novel genes with their human orthologs. We found that many genes involved in development of progressive traits in humans (lung, mammary gland, placenta, ventricular septum, etc.) originated in fish and are expressed in fish tumors and tumors after regression. These findings support a possible evolutionary role of hereditary tumors, and in particular the hypothesis of evolution by tumor neofunctionalization.

Research highlights

Earlier we described a new class of genes that are tumor-specifically expressed and evolutionarily novel (TSEEN). As the functions of TSEEN genes are often uncertain, we decided to study TSEEN genes of fishes so that we could trace the appearance of their new functions in higher vertebrates. We found that many human genes which are involved in development of progressive traits (placenta development, mammary gland and lung development etc.,) originated in fishes and are expressed in fish tumors.
Simultaneously inactivating Src and AKT by saracatinib/capivasertib co-delivery nanoparticles to improve the efficacy of anti-Src therapy in head and neck squamous cell carcinoma
Authors: Lang Liwei , Shay Chloe , Zhao Xiangdong , Xiong Yuanping , Wang Xuli , Teng Yong
Published: December 2019

Background

Src, an oncoprotein that drives progression of head and neck squamous cell carcinoma (HNSCC), is commonly hyperactivated in this disease. Unfortunately, the clinical benefit of targeting Src is significantly dampened in HNSCC patients, because the cytotoxic effects of anti-Src therapy and tumor resistance to it are less predictable. Thus, understanding the mechanism of tumor resistance to Src inhibition and seeking a way to overcome it are warranted.

Methods

Dual drug-loaded nanoparticles (NPs) were developed to co-deliver Src inhibitor saracatinib (AZD0530) and AKT inhibitor capivasertib (AZD5363) into the same population of tumor cells. An orthotopic tongue tumor model was generated to evaluate the in vivo therapeutic effects. Cell growth was determined by CellTiter-Glo® Luminescent Cell Viability Kit, colony formation, and 3D culture, and tumor growth was determined by bioluminescence and tumor size. The molecular changes induced by the treatments were assessed by Western blotting and immunohistochemistry.

Results

Capivasertib inactivated the AKT-S6 signaling and re-sensitized saracatinib-resistant HNSCC cells to saracatinib. Combination of capivasertib with saracatinib suppressed HNSCC growth more efficiently than either drug alone. Cathepsin B-sensitive NPs for co-delivering saracatinib and capivasertib significantly improved the efficacy of tumor repression without increasing side effects, which were due to highly specific tumor-targeting drug delivery system and synergistic anticancer effects by co-inactivation of AKT and Src in HNSCC cells.

Conclusions

Addition of AKT blockade improves anti-HNSCC efficacy of anti-Src therapy, and co-delivery of capivasertib and saracatinib by tumor-targeting NPs has the potential to achieve better treatment outcomes than the free drug combination.
NOS1 inhibits the interferon response of cancer cells by S-nitrosylation of HDAC2
Authors: Xu Pengfei , Ye Shuangyan , Li Keyi , Huang Mengqiu , Wang Qianli , Zeng Sisi , Chen Xi , Gao Wenwen , Chen Jianping , Zhang Qianbing , Zhong Zhuo , Lin Ying , Rong Zhili , Xu Yang , Hao Bingtao , Peng Anghui , Ouyang Manzhao , Liu Qiuzhen
Published: December 2019

Background

The dysfunction of type I interferon (IFN) signaling is an important mechanism of immune escape and metastasis in tumors. Increased NOS1 expression has been detected in melanoma, which correlated with dysfunctional IFN signaling and poor response to immunotherapy, but the specific mechanism has not been determined. In this study, we investigated the regulation of NOS1 on the interferon response and clarified the relevant molecular mechanisms.

Methods

After stable transfection of A375 cells with NOS1 expression plasmids, the transcription and expression of IFNα-stimulated genes (ISGs) were assessed using pISRE luciferase reporter gene analysis, RT-PCR, and western blotting, respectively. The effect of NOS1 on lung metastasis was assessed in melanoma mouse models. A biotin-switch assay was performed to detect the S-nitrosylation of HDAC2 by NOS1. ChIP-qPCR was conducted to measure the binding of HDAC2, H4K16ac, H4K5ac, H3ac, and RNA polymerase II in the promoters of ISGs after IFNα stimulation. This effect was further evaluated by altering the expression level of HDAC2 or by transfecting the HDAC2-C262A/C274A site mutant plasmids into cells. The coimmunoprecipitation assay was performed to detect the interaction of HDAC2 with STAT1 and STAT2. Loss-of-function and gain-of-function approaches were used to examine the effect of HDAC2-C262A/C274A on lung metastasis. Tumor infiltrating lymphocytes were analyzed by flow cytometry.

Results

HDAC2 is recruited to the promoter of ISGs and deacetylates H4K16 for the optimal expression of ISGs in response to IFNα treatment. Overexpression of NOS1 in melanoma cells decreases IFNα-responsiveness and induces the S-nitrosylation of HDAC2-C262/C274. This modification decreases the binding of HDAC2 with STAT1, thereby reducing the recruitment of HDAC2 to the ISG promoter and the deacetylation of H4K16. Moreover, expression of a mutant form of HDAC2, which cannot be nitrosylated, reverses the inhibition of ISG expression by NOS1 in vitro and decreases NOS1-induced lung metastasis and inhibition of tumor infiltrating lymphocytes in a melanoma mouse model.

Conclusions

This study provides evidence that NOS1 induces dysfunctional IFN signaling to promote lung metastasis in melanoma, highlighting NOS1-induced S-nitrosylation of HDAC2 in the regulation of IFN signaling via histone modification.
PTPRD-inactivation-induced CXCL8 promotes angiogenesis and metastasis in gastric cancer and is inhibited by metformin
Authors: Bae Won Jung , Ahn Ji Mi , Byeon Hye Eun , Kim Seokwhi , Lee Dakeun
Published: December 2019

Background

Protein tyrosine phosphatase receptor delta (PTPRD) is frequently inactivated in various types of cancers. Here, we explored the underlying mechanism of PTPRD-loss-induced cancer metastasis and investigated an efficient treatment option for PTPRD-inactivated gastric cancers (GCs).

Methods

PTPRD expression was evaluated by immunohistochemistry. Microarray analysis was used to identify differentially expressed genes in PTPRD-inactivated cancer cells. Quantitative reverse transcription (qRT-PCR), western blotting, and/or enzyme-linked immunosorbent assays were used to investigate the PTPRD-CXCL8 axis and the expression of other related genes. An in vitro tube formation assay was performed using HUVECs. The efficacy of metformin was assessed by MTS assay.

Results

PTPRD was frequently downregulated in GCs and the loss of PTPRD expression was associated with advanced stage, worse overall survival, and a higher risk of distant metastasis. Microarray analysis revealed a significant increase in CXCL8 expression upon loss of PTPRD. This was validated in various GC cell lines using transient and stable PTPRD knockdown. PTPRD-loss-induced angiogenesis was mediated by CXCL8, and the increase in CXCL8 expression was mediated by both ERK and STAT3 signaling. Thus, specific inhibitors targeting ERK or STAT3 abrogated the corresponding signaling nodes and inhibited PTPRD-loss-induced angiogenesis. Additionally, metformin was found to efficiently inhibit PTPRD-loss-induced angiogenesis, decrease cell viability in PTPRD-inactivated cancers, and reverse the decrease in PTPRD expression.

Conclusions

Thus, the PTPRD-CXCL8 axis may serve as a potential therapeutic target, particularly for the suppression of metastasis in PTPRD-inactivated GCs. Hence, we propose that the therapeutic efficacy of metformin in PTPRD-inactivated cancers should be further investigated.
Nonoperative management of splenic injury in closely monitored patients with reduced consciousness is safe and feasible
Authors: Teuben Michel , Spijkerman Roy , Blokhuis Taco , Pfeifer Roman , Teuber Henrik , Pape Hans-Christoph , Leenen Luke
Published: December 2019

Background

Treatment of blunt splenic injury has changed over the past decades. Nonoperative management (NOM) is the treatment of choice. Adequate patient selection is a prerequisite for successful NOM. Impaired mental status is considered as a relative contra indication for NOM. However, the impact of altered consciousness in well-equipped trauma institutes is unclear. We hypothesized that impaired mental status does not affect outcome in patients with splenic trauma.

Methods

Our prospectively composed trauma database was used and adult patients with blunt splenic injury were included during a 14-year time period. Treatment guidelines remained unaltered over time. Patients were grouped based on the presence (Group GCS: < 14) or absence (Group GCS: 14–15) of impaired mental status. Outcome was compared.

Results

A total of 161 patients were included, of whom 82 were selected for NOM. 36% of patients had a GCS-score < 14 (N = 20). The median GCS-score in patients with reduced consciousness was 9 (range 6–12). Groups were comparable except for significantly higher injury severity scores in the impaired mental status group (19 vs. 17,p = 0.007). Length of stay (28 vs. 9 days,p < 0.001) and ICU-stay (8 vs. 0 days,p = 0.005) were longer in patients with decreased GCS-scores. Failure of NOM, total splenectomy rates, complications and mortality did not differ between both study groups.

Conclusion

This study shows that NOM for blunt splenic trauma is a viable treatment modality in well-equipped institutions, regardless of the patients mental status. However, the presence of neurologic impairment is associated with prolonged ICU-stay and hospitalization. We recommend, in institutions with adequate monitoring facilities, to attempt nonoperative management for blunt splenic injury, in all hemodynamically stable patients without hollow organ injuries, also in the case of reduced consciousness.
Cortisol total/CRP ratio for the prediction of hospital-acquired pneumonia and initiation of corticosteroid therapy in traumatic brain-injured patients
Authors: Bouras Marwan , Roquilly Antoine , Mahé Pierre-Joachim , Cinotti Raphaël , Vourc’h Mickaël , Perrot Bastien , Bach-Ngohou Kalyane , Masson Damien , Asehnoune Karim
Published: December 2019

Background

To propose a combination of blood biomarkers for the prediction of hospital-acquired pneumonia (HAP) and for the selection of traumatic brain-injured (TBI) patients eligible for corticosteroid therapy for the prevention of HAP.

Methods

This was a sub-study of the CORTI-TC trial, a multicenter, randomized, double-blind, controlled trial evaluating the risk of HAP at day 28 in 336 TBI patients treated or not with corticosteroid therapy. Patients were between 15 and 65 years with severe traumatic brain injury (Glasgow coma scale score ≤ 8 and trauma-associated lesion on brain CT scan) and were enrolled within 24 h of trauma. The blood levels of CRP and cortisoltotal&free, as a surrogate marker of the pro/anti-inflammatory response balance, were measured in samples collected before the treatment initiation. Endpoint was HAP on day 28.

Results

Of the 179 patients with available samples, 89 (49.7%) developed an HAP. Cortisoltotal&free and CRP blood levels upon ICU admission were not significantly different between patients with or without HAP. The cortisoltotal/CRP ratio upon admission was 2.30 [1.25–3.91] in patients without HAP and 3.36 [1.74–5.09] in patients with HAP (p = 0.021). In multivariate analysis, a cortisoltotal/CRP ratio > 3, selected upon the best Youden index on the ROC curve, was independently associated with HAP (OR 2.50, CI95% [1.34–4.64] p = 0.004). The HR for HAP with corticosteroid treatment was 0.59 (CI95% [0.34–1.00], p = 0.005) in patients with a cortisoltotal/CRP ratio > 3, and 0.89 (CI95% [0.49–1.64], p = 0.85) in patients with a ratio < 3.

Conclusion

A cortisoltotal/CRP ratio > 3 upon admission may predict the development of HAP in severe TBI. Among these patients, corticosteroids reduce the occurrence HAP. We suggest that this ratio may select the patients who may benefit from corticosteroid therapy for the prevention of HAP.
Improved spectrophotometric assay for lytic polysaccharide monooxygenase
Authors: Breslmayr Erik , Daly Sarah , Požgajčić Alen , Chang Hucheng , Rezić Tonči , Oostenbrink Chris , Ludwig Roland
Published: December 2019

Background

The availability of a sensitive and robust activity assay is a prerequisite for efficient enzyme production, purification, and characterization. Here we report on a spectrophotometric assay for lytic polysaccharide monooxygenase (LPMO), which is an advancement of the previously published 2,6-dimethoxyphenol (2,6-DMP)-based LPMO assay. The new assay is based on hydrocoerulignone as substrate and hydrogen peroxide as cosubstrate and aims toward a higher sensitivity at acidic pH and a more reliable detection of LPMO in complex matrices like culture media.

Results

An LPMO activity assay following the colorimetric oxidation of hydrocoerulignone to coerulignone was developed. This peroxidase activity of LPMO in the presence of hydrogen peroxide can be detected in various buffers between pH 4–8. By reducing the substrate and cosubstrate concentration, the assay has been optimized for minimal autoxidation and enzyme deactivation while maintaining sensitivity. Finally, the optimized and validated LPMO assay was used to follow the recombinant expression of an LPMO in Pichia pastoris and to screen for interfering substances in fermentation media suppressing the assayed reaction.

Conclusions

The biphenol hydrocoerulignone is a better substrate for LPMO than the monophenol 2,6-DMP, because of a ~ 30 times lower apparent KM value and a 160 mV lower oxidation potential. This greatly increases the measured LPMO activity when using hydrocoerulignone instead of 2,6-DMP under otherwise similar assay conditions. The improved activity allows the adaptation of the LPMO assay toward a higher sensitivity, different buffers and pH values, more stable assay conditions or to overcome low concentrations of inhibiting substances. The developed assay protocol and optimization guidelines increase the adaptability and applicability of the hydrocoerulignone assay for the production, purification, and characterization of LPMOs.
The SMART App: an interactive web application for comprehensive DNA methylation analysis and visualization
Authors: Li Yin , Ge Di , Lu Chunlai
Published: December 2019

Background

Data mining of The Cancer Genome Atlas (TCGA) data has significantly facilitated cancer genome research and provided unprecedented opportunities for cancer researchers. However, existing web applications for DNA methylation analysis does not adequately address the need of experimental biologists, and many additional functions are often required.

Results

To facilitate DNA methylation analysis, we present the SMART (Shiny Methylation Analysis Resource Tool) App, a user-friendly and easy-to-use web application for comprehensively analyzing the DNA methylation data of TCGA project. The SMART App integrates multi-omics and clinical data with DNA methylation and provides key interactive and customized functions including CpG visualization, pan-cancer methylation profile, differential methylation analysis, correlation analysis and survival analysis for users to analyze the DNA methylation in diverse cancer types in a multi-dimensional manner.

Conclusion

The SMART App serves as a new approach for users, especially wet-bench scientists with no programming background, to analyze the scientific big data and facilitate data mining. The SMART App is available at http://www.bioinfo-zs.com/smartapp.
Safety, tolerability, pharmacokinetics, and pharmacodynamics of PF-06650833, a selective interleukin-1 receptor-associated kinase 4 (IRAK4) inhibitor, in single and multiple ascending dose randomized phase 1 studies in healthy subjects
Authors: Danto Spencer I. , Shojaee Negin , Singh Ravi Shankar P. , Li Cheryl , Gilbert Steven A. , Manukyan Zorayr , Kilty Iain
Published: December 2019

Background

PF-06650833 is a potent, selective inhibitor of interleukin-1 receptor-associated kinase 4 (IRAK4). Two randomized, double-blind, sponsor-open phase 1 studies evaluated the safety, pharmacokinetics, and pharmacodynamics of single (SAD) and multiple ascending doses (MAD) of PF-06650833 immediate-release (IR) and modified-release (MR) oral formulations in healthy adult subjects.

Methods

Study 1 (NCT02224651) was a 96-day, placebo-substitution, SAD study of once-daily (QD) oral PF-06650833 IR 1 to 6000 mg and MR 30 to 300 mg in fasted and fed states. Study 2 (NCT02485769) was a 14-day, placebo-controlled, MAD study of PF-06650833 IR 25 to 750 mg twice daily, IR 1000 mg four times per day, IR 330 mg three times per day, and MR 300 mg QD.

Results

PF-06650833 was generally well tolerated, with no dose-limiting treatment-emergent adverse events (TEAEs) identified in either study. TEAEs were generally mild in severity, with headache, gastrointestinal disorders, and acne most commonly reported. No serious AEs or deaths were reported. A maximum tolerated dose was not established in either study. In the SAD study, food intake delayed absorption of IR 30 mg and increased total exposure by 33%. Delayed absorption was achieved with the MR formulation (Tmax of 1 h versus 8 h for IR 100 mg and MR 100 mg formulations, respectively). Food had no effect on total exposure for MR 30 mg, but reduced half-life 1.8-fold and increased Cmax by 62%. In the MAD study, accumulation ranged from 0.9-fold to 1.4-fold for AUCtau and 0.9-fold to 1.3-fold for Cmax. Less than 1% of the dose was recovered unchanged in urine for all dose groups, with renal clearance ranging from 14 to 23 mL/min for IR < 750 mg and MR 300 mg. There was a sustained decrease in serum high-sensitivity C-reactive protein for IR ≥ 250 mg and MR 300 mg. Based on the cholesterol/hydroxycholesterol ratio, no apparent CYP3A induction or inhibition was observed.

Conclusions

PF-06650833, the first IRAK4 inhibitor to enter clinical development, has a favorable safety and pharmacokinetic profile and has shown evidence of pharmacological effect. The data support continued evaluation in human clinical trials for the treatment of rheumatic and autoimmune diseases.

Trial registration

Clinicaltrials.gov, NCT02224651, registered 25 August 2014; NCT02485769, registered 30 June 2015
Point of care testing evaluation of lateral flow immunoassay for diagnosis of cryptococcus meningitis in HIV-positive patients at an urban hospital in Nairobi, Kenya, 2017
Authors: Gitonga Lawrence Kirimi , Boru Waqo Gufu , Kwena Arthur , Maritim Marybeth , Wamicwe Joyce , Ransom James
Published: December 2019

Objectives

The objective of this study was to evaluate the performance of lateral flow immunoassay (LFA) against latex agglutination (LA), India ink and culture in point-of-care diagnosis of cryptococcus meningitis (CM). We conducted cross-sectional study among HIV-positive patients with suspected CM at Mbagathi Hospital, Nairobi, April–July 2017.

Results

Of 124 capillary blood and serum and 99 cerebrospinal fluid (CSF) samples, LFA and LA had a concurrence on serum of 94.4%, kappa (0.88), sensitivity (100%) and specificity (91%). LFA and LA on CSF, was 97.9%, kappa (0.96), sensitivity (100%) and specificity (96%). LFA and India ink was 96.9%, kappa (0.94), sensitivity (100%) and specificity (94.1%). On CSF culture, concurrence was 72.7%, kappa (0.43), sensitivity (100%) and specificity (64%) and of LFA on capillary blood, serum and CSF was 100% with kappa (1.00), sensitivity and specificity of 100%.
Progression to dementia in memory clinic patients with mild cognitive impairment and normal β-amyloid
Authors: Rosenberg Anna , Solomon Alina , Jelic Vesna , Hagman Göran , Bogdanovic Nenad , Kivipelto Miia
Published: December 2019

Background

Determination of β-amyloid (Aβ) positivity and likelihood of underlying Alzheimer’s disease (AD) relies on dichotomous biomarker cut-off values. Individuals with mild cognitive impairment (MCI) and Aβ within the normal range may still have a substantial risk of developing dementia, primarily of Alzheimer type. Their prognosis, as well as predictors of clinical progression, are not fully understood. The aim of this study was to explore the associations of cerebrospinal fluid (CSF) biomarkers (Aβ42, total tau, phosphorylated tau) and other characteristics, including modifiable vascular factors, with the risk of progression to dementia among patients with MCI and normal CSF Aβ42.

Methods

Three hundred eighteen memory clinic patients with CSF and clinical data, and at least 1-year follow-up, were included. Patients had normal CSF Aβ42 levels based on clinical cut-offs. Cox proportional hazard models with age as time scale and adjusted for sex, education, and cognition (Mini-Mental State Examination) were used to investigate predictors of progression to dementia and Alzheimer-type dementia. Potential predictors included CSF biomarkers, cognitive performance (verbal learning and memory), apolipoprotein E (APOE) ε4 genotype, medial temporal lobe atrophy, family history of dementia, depressive symptoms, and vascular factors, including the Cardiovascular Risk Factors, Aging and Dementia (CAIDE) risk score. Predictive performance of patient characteristics was further explored with Harrell C statistic.

Results

Lower normal Aβ42 and higher total tau and phosphorylated tau were associated with higher dementia risk, and the association was not driven by Aβ42 values close to cut-off. Additional predictors included poorer cognition, APOE ε4 genotype, higher systolic blood pressure, and lower body mass index, but not the CAIDE dementia risk score. Aβ42 individually and in combination with other CSF biomarkers improved the risk prediction compared to age and cognition alone. Medial temporal lobe atrophy or vascular factors did not increase the predictive performance.

Conclusions

Possibility of underlying AD pathology and increased dementia risk should not be ruled out among MCI patients with CSF Aβ42 within the normal range. While cut-offs may be useful in clinical practice to identify high-risk individuals, personalized risk prediction tools incorporating continuous biomarkers may be preferable among individuals with intermediate risk. The role of modifiable vascular factors could be explored in this context.
The shift in the balance between osteoblastogenesis and adipogenesis of mesenchymal stem cells mediated by glucocorticoid receptor
Authors: Han Lizhi , Wang Bo , Wang Ruoyu , Gong Song , Chen Guo , Xu Weihua
Published: December 2019

Mesenchymal stem cells (MSCs) are multipotent cells capable of differentiating into several tissues, such as bone, cartilage, and fat. Glucocorticoids affect a variety of biological processes such as proliferation, differentiation, and apoptosis of various cell types, including osteoblasts, adipocytes, or chondrocytes. Glucocorticoids exert their function by binding to the glucocorticoid receptor (GR). Physiological concentrations of glucocorticoids stimulate osteoblast proliferation and promote osteogenic differentiation of MSCs. However, pharmacological concentrations of glucocorticoids can not only induce apoptosis of osteoblasts and osteocytes but can also reduce proliferation and inhibit the differentiation of osteoprogenitor cells. Several signaling pathways, including the Wnt, TGFβ/BMP superfamily and Notch signaling pathways, transcription factors, post-transcriptional regulators, and other regulators, regulate osteoblastogenesis and adipogenesis of MSCs mediated by GR. These signaling pathways target key transcription factors, such as Runx2 and TAZ for osteogenesis and PPARγ and C/EBPs for adipogenesis. Glucocorticoid-induced osteonecrosis and osteoporosis are caused by various factors including dysfunction of bone marrow MSCs. Transplantation of MSCs is valuable in regenerative medicine for the treatment of osteonecrosis of the femoral head, osteoporosis, osteogenesis imperfecta, and other skeletal disorders. However, the mechanism of inducing MSCs to differentiate toward the osteogenic lineage is the key to an efficient treatment. Thus, a better understanding of the molecular mechanisms behind the imbalance between GR-mediated osteoblastogenesis and adipogenesis of MSCs would not only help us to identify the pathogenic causes of glucocorticoid-induced osteonecrosis and osteoporosis but also promote future clinical applications for stem cell-based tissue engineering and regenerative medicine. Here, we primarily review the signaling mechanisms involved in adipogenesis and osteogenesis mediated by GR and discuss the factors that control the adipo-osteogenic balance.
Aspirin inhibits RANKL-induced osteoclast differentiation in dendritic cells by suppressing NF-κB and NFATc1 activation
Authors: Wu Lili , Luo Zhenhua , Liu Yitong , Jia Lu , Jiang Yiyang , Du Juan , Guo Lijia , Bai Yuxing , Liu Yi
Published: December 2019

Background

Aspirin has been demonstrated to promote osteoblast-mediated bone formation and inhibit osteoclast (OC)-mediated bone resorption. However, it remains unclear whether aspirin influences other immune cells during bone resorption. Dendritic cells (DCs), the most potent antigen-presenting cells, can also transdifferentiate into active OCs in the presence of receptor activator of nuclear factor-κB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). The effects of aspirin on DC-derived OCs (DDOCs) were investigated in the current study.

Methods

Flow cytometry and mixed lymphocyte reaction (MLR) assays were used for DC identification. The proliferative capacity of DCs was determined by BrdU assays. Apoptosis was examined by flow cytometry. The osteoclastic potential of DCs was tested using tartrate-resistant acid phosphatase (TRAP) staining, western blotting, and reverse transcription polymerase chain reaction (RT-PCR). Western blotting was also used to examine signaling pathways. A mandibular bone defect model was established to assess the effect of aspirin on bone resorption.

Results

Aspirin had no influence on the surface phenotype, proliferation, or apoptosis of DCs, though aspirin significantly inhibited osteoclast differentiation in RANKL-stimulated DCs. DC osteoclast differentiation was modulated by aspirin via the nuclear factor kappa B (NF-κB)/nuclear factor of activated T cell, cytoplasmic 1 (NFATc1) signaling pathway. Aspirin treatment also had favorable therapeutic effects on bone regeneration in the bone defect model, and the number of osteoclasts was decreased.

Conclusions

Aspirin inhibited RANKL-induced OC differentiation in DCs via the NF-κB pathway, downregulating expression of NFATc1. Aspirin treatment promoted bone regeneration by inhibiting DDOC activation in the early stages of inflammation in a rat mandibular bone defect model.
HastaLaVista, a web-based user interface for NMR-based untargeted metabolic profiling analysis in biomedical sciences: towards a new publication standard
Authors: Wist Julien
Published: December 2019

Metabolic profiling has been shown to be useful to improve our understanding of complex metabolic processes. Shared data are key to the analysis and validation of metabolic profiling and untargeted spectral analysis and may increase the pace of new discovery. Improving the existing portfolio of open software may increase the fraction of shared data by decreasing the amount of effort required to publish them in a manner that is useful to others. However, a weakness of open software, when compared to commercial ones, is the lack of user-friendly graphical interface that may discourage inexperienced researchers. Here, a web-browser-oriented solution is presented and demonstrated for metabolic profiling analysis that combines the power of R for back-end statistical analyses and of JavaScript for front-end visualisations and user interactivity. This unique combination of statistical programming and web-browser visualisation brings enhanced data interoperability and interactivity into the open source realm. It is exemplified by characterizing the extent to which bariatric surgery perturbs the metabolisms of rats, showing the value of the approach in iterative analysis by the end-user to establish a deeper understanding of the system perturbation. HastaLaVista is available at: (https://github.com/jwist/hastaLaVista, 10.5281/zenodo.3544800) under MIT license. The approach described in this manuscript can be extended to connect the interface to other scripting languages such as Python, and to create interfaces for other types of data analysis.
Cytokine production and phenotype of Histomonas meleagridis-specific T cells in the chicken
Authors: Lagler Julia , Mitra Taniya , Schmidt Selma , Pierron Alix , Vatzia Eleni , Stadler Maria , Hammer Sabine E. , Mair Kerstin H. , Grafl Beatrice , Wernsdorf Patricia , Rauw Fabienne , Lambrecht Bénédicte , Liebhart Dieter , Gerner Wilhelm
Published: December 2019

The protozoan parasiteHistomonas meleagridisis the causative agent of the re-emerging disease histomonosis of chickens and turkeys. Due to the parasite’s extracellular occurrence, a type-2 differentiation ofH. meleagridis-specific T cells has been hypothesized. In contrast, a recent study suggested that IFN-γ mRNA+cells are involved in protection against histomonosis. However, the phenotype and cytokine production profile ofH. meleagridis-specific T cells still awaits elucidation. In this work, clonal cultures of a virulent monoxenic strain ofH. meleagridiswere used for infecting chickens to detect IFN-γ protein and IL-13 mRNA by intracellular cytokine staining and PrimeFlow™ RNA Assays, respectively, in CD4+and CD8β+T cells. Infection was confirmed by characteristic pathological changes in the cecum corresponding withH. meleagridisdetection by immunohistochemistry andH. meleagridis-specific antibodies in serum. In splenocytes stimulated either withH. meleagridisantigen or PMA/ionomycin, IFN-γ-producing CD4+T cells from infected chickens increased in comparison to cells from non-infected birds 2 weeks and 5 weeks post-infection. Additionally, an increase of IFN-γ-producing CD4−CD8β−cells uponH. meleagridisantigen and PMA/ionomycin stimulation was detected. Contrariwise, frequencies of IL-13 mRNA-expressing cells were low even after PMA/ionomycin stimulation and mainly had a CD4−CD8β−phenotype. No clear increase of IL-13+cells related toH. meleagridisinfection could be found. In summary, these data suggest thatH. meleagridisinfection induces a type-1 differentiation of CD4+T cells but also of non-CD4+cells. This phenotype could include γδ T cells, which will be addressed in future studies.
Development of loop-mediated isothermal amplification coupled with nanoparticle-based lateral flow biosensor assay for Mycoplasma pneumoniae detection
Authors: Wang Yacui , Wang Yi , Jiao Weiwei , Li Jieqiong , Quan Shuting , Sun Lin , Wang Yonghong , Qi Xue , Wang Xingyun , Shen Adong
Published: December 2019

Mycoplasma pneumoniae (MP) is one of the most common pathogens causing respiratory tract infection, especially for community-acquired pneumonia (CAP) in school-age children. There was considerable amount of studies on loop-mediated isothermal amplification (LAMP) assay for MP detection. However, the result interpretation of these developed LAMP assays was sophisticated and subjective. Therefore, we developed and evaluated a LAMP coupled with nanoparticle-based lateral flow biosensor (LFB) assay (LAMP-LFB) for simple, reliable, and objective identification of MP (MP-LAMP-LFB). Six primers specific to P1 gene of MP were designed, and the preferred temperature for this assay was confirmed to be 65 °C. The amplification products could be visually interpreted by LFB within 2 min. The MP-LAMP-LFB assay specifically identified DNA templates of MP, and no cross-reactivity with other pathogens was obtained. The limit of the detection for this assay was 600 fg of DNA templates in pure cultures, which was in complete accordance with colorimetric indicator detection and agarose gel electrophoresis analysis. This assay was applied to 209 oropharyngeal swab specimens collected from children with acute respiratory tract infection for clinical evaluation, and compared to real-time PCR detection. Using the LAMP-LFB and real-time PCR assay, the positive rates of MP were 47.8% and 31.6%, respectively. Results suggested that the LAMP-LFB assay displayed high sensitivity compared to real-time PCR method. In summary, LAMP-LFB assay established here was a simple, objective, and sensitive assay for MP detection, which can be widely applied in clinical settings, especially in rural areas.
Introducing phonetic information to speaker embedding for speaker verification
Authors: Liu Yi , He Liang , Liu Jia , Johnson Michael T.
Published: December 2019

Phonetic information is one of the most essential components of a speech signal, playing an important role for many speech processing tasks. However, it is difficult to integrate phonetic information into speaker verification systems since it occurs primarily at the frame level while speaker characteristics typically reside at the segment level. In deep neural network-based speaker verification, existing methods only apply phonetic information to the frame-wise trained speaker embeddings. To improve this weakness, this paper proposes phonetic adaptation and hybrid multi-task learning and further combines these into c-vector and simplified c-vector architectures. Experiments on National Institute of Standards and Technology (NIST) speaker recognition evaluation (SRE) 2010 show that the four proposed speaker embeddings achieve better performance than the baseline. The c-vector system performs the best, providing over 30% and 15% relative improvements in equal error rate (EER) for the core-extended and 10 s–10 s conditions, respectively. On the NIST SRE 2016, 2018, and VoxCeleb datasets, the proposed c-vector approach improves the performance even when there is a language mismatch within the training sets or between the training and evaluation sets. Extensive experimental results demonstrate the effectiveness and robustness of the proposed methods.
Associations between serum levels of insulin-like growth factor-I and bone mineral acquisition in pubertal children: a 3-year follow-up study in Hamamatsu, Japan
Authors: Kouda Katsuyasu , Iki Masayuki , Ohara Kumiko , Nakamura Harunobu , Fujita Yuki , Nishiyama Toshimasa
Published: December 2019

Background

Epidemiological data regarding the association between serum levels of IGF-I and bone mineral acquisition during childhood are scarce. Here, we investigated the association between serum levels of IGF-I and bone status during puberty.

Methods

We analyzed prospective 3-year follow-up data of 254 community-dwelling children who completed both baselines (at age 11.2 years) and follow-up (at age 14.2 years) surveys in Hamamatsu, Japan. Total body (TB) bone area and bone mineral parameters were assessed using dual-energy X-ray absorptiometry.

Results

During the 3-year follow-up period, there were significant (P< 0.05) increases in total body less head (TBLH) areal bone mineral density (aBMD), TBLH bone mineral content (BMC), and TB bone area, and a significant decrease in TB bone mineral apparent density (BMAD, volumetric bone mineral density, vBMD). IGF-I levels showed significant positive relationships with TBLH BMC and TBLH aBMD at both baseline and follow-up. TBLH aBMD in boys and TB BMAD in girls at follow-up showed significant increases from the lowest to highest quartiles of baseline IGF-I levels after adjusting for confounding factors. Similarly, changes in TBLH aBMD in boys and TB BMAD in girls during the 3-year follow-up period showed significant increases from the lowest to highest quartiles of baseline IGF-I levels after adjusting for confounding factors.

Conclusions

These results suggest that pubertal children with high levels of serum IGF-I tended to have high bone mineral acquisition later on.
Whitebark pine encroachment into lower-elevation sagebrush grasslands in southwest Montana, USA
Authors: Flanary Sarah J. , Keane Robert E.
Published: December 2019

Background

Projections for the future health and abundance of whitebark pine (Pinus albicaulis Engelm.) populations in western North America are dire. Not only has the species been declining due to the combined effects of fire exclusion policies, mountain pine beetle (Dendroctonus ponderosae Hopkins 1902) outbreaks, and white pine blister rust (Cronartium ribicola J.C. Fisch), many are predicting that climate change will amplify these effects and whitebark pine habitat may be lost in many areas. One method to evaluate the validity of future predictions is to examine current migration patterns of whitebark pine into surrounding non-forested areas. In this paper, we documented the findings of a case study surveying the encroachment of whitebark pine into adjacent lower-elevation sagebrush grasslands and higher-elevation subalpine meadows in southwestern Montana, USA.

Results

We found abundant evidence of whitebark pine trees (>40 trees ha−1) in sagebrush grasslands below the mature seed source, but there was no evidence of any whitebark pine regeneration in subalpine meadows above the seed-source stand.

Conclusions

These results are in stark contrast to the findings of statistical modeling studies that predict that whitebark pine will expand into higher-elevation areas as a response to climate change. We believe that the reason for this low-elevation encroachment may be in response to fire exclusion rather than climate change. Although observations in this case study are for a limited geographic area, they do serve to illustrate the complexity of predicting ecological responses of tree species to climate change.