Aurora A kinase inhibition induces accumulation of SCLC tumor cells in mitosis with restored interferon signaling to increase response to PD-L1.

Despite small cell lung cancers (SCLCs) having a high mutational burden, programmed death-ligand 1 (PD-L1) immunotherapy only modestly increases survival. A subset of SCLCs that lose their ASCL1 neuroendocrine phenotype and restore innate immune signaling (termed the "inflammatory" subtype) have durable responses to PD-L1. Some SCLCs are highly sensitive to Aurora kinase inhibitors, but early-phase trials show short-lived responses, suggesting effective therapeutic combinations are needed to increase their durability. Using immunocompetent SCLC genetically engineered mouse models (GEMMs) and syngeneic xenografts, we show durable efficacy with the combination of a highly specific Aurora A kinase inhibitor (LSN3321213) and PD-L1. LSN3321213 causes accumulation of tumor cells in mitosis with lower ASCL1 expression and higher expression of interferon target genes and antigen-presentation genes mimicking the inflammatory subtype in a cell-cycle-dependent manner. These data demonstrate that inflammatory gene expression is restored in mitosis in SCLC, which can be exploited by Aurora A kinase inhibition.

Regulation of neuroendocrine plasticity by the RNA-binding protein ZFP36L1.

Some small cell lung cancers (SCLCs) are highly sensitive to inhibitors of the histone demethylase LSD1. LSD1 inhibitors are thought to induce their anti-proliferative effects by blocking neuroendocrine differentiation, but the mechanisms by which LSD1 controls the SCLC neuroendocrine phenotype are not well understood. To identify genes required for LSD1 inhibitor sensitivity in SCLC, we performed a positive selection genome-wide CRISPR/Cas9 loss of function screen and found that ZFP36L1, an mRNA-binding protein that destabilizes mRNAs, is required for LSD1 inhibitor sensitivity. LSD1 binds and represses ZFP36L1 and upon LSD1 inhibition, ZFP36L1 expression is restored, which is sufficient to block the SCLC neuroendocrine differentiation phenotype and induce a non-neuroendocrine "inflammatory" phenotype. Mechanistically, ZFP36L1 binds and destabilizes SOX2 and INSM1 mRNAs, two transcription factors that are required for SCLC neuroendocrine differentiation. This work identifies ZFP36L1 as an LSD1 target gene that controls the SCLC neuroendocrine phenotype and demonstrates that modulating mRNA stability of lineage transcription factors controls neuroendocrine to non-neuroendocrine plasticity.

Geneticin reduces mRNA stability.

Messenger RNA (mRNA) translation can lead to higher rates of mRNA decay, suggesting the ribosome plays a role in mRNA destruction. Furthermore, mRNA features, such as codon identities, which are directly probed by the ribosome, correlate with mRNA decay rates. Many amino acids are encoded by synonymous codons, some of which are decoded by more abundant tRNAs leading to more optimal translation and increased mRNA stability. Variable translation rates for synonymous codons can lead to ribosomal collisions as ribosomes transit regions with suboptimal codons, and ribosomal collisions can promote mRNA decay. In addition to different translation rates, the presence of certain codons can also lead to higher or lower rates of amino acid misincorporation which could potentially lead to protein misfolding if a substituted amino acid fails to make critical contacts in a structure. Here, we test whether Geneticin-G418, an aminoglycoside antibiotic known to promote amino acid misincorporation-affects mRNA stability. We observe that G418 decreases firefly luciferase mRNA stability in an in vitro translation system and also reduces mRNA stability in mouse embryonic stem cells (mESCs). G418-sensitive mRNAs are enriched for certain optimal codons that contain G or C in the wobble position, arguing that G418 blunts the stabilizing effects of codon optimality.

Plasticity in the Absence of NOTCH Uncovers a RUNX2-Dependent Pathway in Small Cell Lung Cancer.

Neuroendocrine to nonneuroendocrine plasticity supports small cell lung cancer (SCLC) tumorigenesis and promotes immunogenicity. Approximately 20% to 25% of SCLCs harbor loss-of-function (LOF) NOTCH mutations. Previous studies demonstrated that NOTCH functions as a SCLC tumor suppressor, but can also drive nonneuroendocrine plasticity to support SCLC growth. Given the dual functionality of NOTCH, it is not understood why SCLCs select for LOF NOTCH mutations and how these mutations affect SCLC tumorigenesis. In a CRISPR-based genetically engineered mouse model of SCLC, genetic loss of Notch1 or Notch2 modestly accelerated SCLC tumorigenesis. Interestingly, Notch-mutant SCLCs still formed nonneuroendocrine subpopulations, and these Notch-independent, nonneuroendocrine subpopulations were driven by Runx2-mediated regulation of Rest. Notch2-mutant nonneuroendocrine cells highly express innate immune signaling genes including stimulator of interferon genes (STING) and were sensitive to STING agonists. This work identifies a Notch-independent mechanism to promote nonneuroendocrine plasticity and suggests that therapeutic approaches to activate STING could be selectively beneficial for SCLCs with NOTCH2 mutations. SIGNIFICANCE: A genetically engineered mouse model of NOTCH-mutant SCLC reveals that nonneuroendocrine plasticity persists in the absence of NOTCH, driven by a RUNX2-REST-dependent pathway and innate immune signaling.

Can low serum brain-derived neurotrophic factor levels be associated with lifelong premature ejaculation?; A pilot study.

This study aimed to present the association between the serum level of brain-derived neurotrophic factor (BDNF) and the lifelong pre-mature ejaculation (PE). The data of 40 patients with lifelong PE and 40 healthy controls were evaluated prospectively. PE diagnostic tool and patient-reported outcome measures were performed to the participants. The serum BDNF level measurement was made after the collecting of blood samples in both groups. The mean ± SD age of the PE and control group was 34.43 ± 5.71 and 33.03 ± 3.97 years respectively (p = .228). Only the participant who has been married included in the study, and there was no difference in the mean marriage duration. In both groups, smoking status, alcohol use and body mass index were similar. The median PE diagnostic tool scores were 15 (11-20), and serum BDNF levels were 225.3 (26.1-689.6) ng/ml in the PE group, 5 (0-9) and 540.9 (102.9-769.2) ng/ml in the control group respectively (p < .001 for both). The patients with PE had significantly lower serum BDNF levels. Our study suggests that lower serum BDNF levels may be directly related to lifelong PE.

Features of asthma which provide meaningful insights for understanding the disease heterogeneity.

BACKGROUND: Data-driven methods such as hierarchical clustering (HC) and principal component analysis (PCA) have been used to identify asthma subtypes, with inconsistent results. OBJECTIVE: To develop a framework for the discovery of stable and clinically meaningful asthma subtypes. METHODS: We performed HC in a rich data set from 613 asthmatic children, using 45 clinical variables (Model 1), and after PCA dimensionality reduction (Model 2). Clinical experts then identified a set of asthma features/domains which informed clusters in the two analyses. In Model 3, we reclustered the data using these features to ascertain whether this improved the discovery process. RESULTS: Cluster stability was poor in Models 1 and 2. Clinical experts highlighted four asthma features/domains which differentiated the clusters in two models: age of onset, allergic sensitization, severity, and recent exacerbations. In Model 3 (HC using these four features), cluster stability improved substantially. The cluster assignment changed, providing more clinically interpretable results. In a 5-cluster model, we labelled the clusters as: "Difficult asthma" (n = 132); "Early-onset mild atopic" (n = 210); "Early-onset mild non-atopic: (n = 153); "Late-onset" (n = 105); and "Exacerbation-prone asthma" (n = 13). Multinomial regression demonstrated that lung function was significantly diminished among children with "Difficult asthma"; blood eosinophilia was a significant feature of "Difficult," "Early-onset mild atopic," and "Late-onset asthma." Children with moderate-to-severe asthma were present in each cluster. CONCLUSIONS AND CLINICAL RELEVANCE: An integrative approach of blending the data with clinical expert domain knowledge identified four features, which may be informative for ascertaining asthma endotypes. These findings suggest that variables which are key determinants of asthma presence, severity, or control may not be the most informative for determining asthma subtypes. Our results indicate that exacerbation-prone asthma may be a separate asthma endotype and that severe asthma is not a single entity, but an extreme end of the spectrum of several different asthma endotypes.

Bond strength of resin composite to light activated bleached enamel.

OBJECTIVES: This study evaluated the microtensile bond strength (µTBS) of a resin composite bonded to bleached enamel as a function of bleaching conditions. MATERIALS AND METHODS: The whiteness hydrogen peroxide (HP) bleaching agent containing 35% HP was applied to the central incisors' facial enamel surface and irradiated as follows: No treatment (G1; control); no light (G2); light-emitting diode, the 40s (G3); diode laser, the 20s (G4); and neodymium:yttrium aluminum garnet laser, 20s (G5). A Variolink II resin composite structure was then built up incrementally on the surface. The teeth were sectioned into three 1.2 mm × 1.2 mm wide "I"-shaped sections. The specimens were then subjected to microtensile testing at a crosshead speed of 1 mm/min. Data were analyzed using one-way ANOVA (α =0.05) followed by the Tukey Honestly Significant Difference post-hoc test. The fractured surfaces were observed with a stereomicroscope at × 100 magnification. RESULTS: One-way ANOVA revealed no statistical differences among the groups (P > 0.05). No differences appeared between the groups bonded 14 days after bleaching (P > 0.05). Mean µTBS values (MPa) were as follows: 22.05 ± 5.01 (G1); 19.6 ± 5.6 (G2); 19.3 ± 5.4 (G3); 20.08 ± 2.08 (G4); and 18.1 ± 4.8 (G5). Many adhesive failures occurred at the bleached and irradiated enamel surfaces. CONCLUSION: The various irradiation treatments following the application of the whiteness HP bleaching agent to enamel did not significantly reduce the µTBS within a 14-day period.

Association of vitamin D binding protein polymorphisms with bronchopulmonary dysplasia: a case-control study of gc globulin and bronchopulmonary dysplasia.

OBJECTIVE: The pathophysiologies of bronchopulmonary dysplasia (BPD) are inflammation, infection, tissue damage, angiogenesis defects and genetic susceptibility. Because of the role of the vitamin D binding protein (Gc globulin) on these factors, we investigated the relationship between Gc globulin polymorphisms and BPD. STUDY DESIGN: This case-control study was performed with 160 neonates (⩽32 gestational ages, ⩽1500 g). PCR DNA sequence analyses were used for GC gene rs4588 and rs7041 single-nucleotide polymorphisms. RESULT: In the univariate analyses, it was observed that Gc2 was the only variant that was protective against BPD (Odd ratio (OR)=0.47, 95% coinfidence interval (CI)=0.24 to 0.89, P=0.020). In the multivariate analyses, Gc2 decreased the risk of disease (OR=0.15, 95% CI=0.029 to 0.79, P=0.026) independent of gestational age, birth weight, 5-min Appearance, Pulse, Grimace, Activity, and Respiration scores, respiratory distress syndrome and sepsis. CONCLUSION: The Gc2 variant was, after adjusting for confounders, associated with a decrease in the frequency of BPD. Our study adds Gc globulin to the list of candidate genes that potentially contribute to the etiology of the disease.

Evaluation of the permeability of five desensitizing agents using computerized fluid filtration.

OBJECTIVE: The aim of this study was to evaluate the permeability of five desensitizing agents using computerized fluid filtration (CFF) test method. MATERIALS AND METHODS: Sixty dentin discs of 500 ± 200-mm-thick were prepared from middle dentin of bovine incisors without exposed the pulp and then randomly divided into five groups (n = 12). The permeability of the discs was measured using the CFF test method before and after application of the following desensitizers: Admira Protect (Voco, Cuxhaven, Germany), Seal and Protect (Dentsply, Konstanz, Germany), Sensi Kill (DFL, Brazil), Systemp Desensitizer (Ivoclar Vivadent, Liechtenstein), BisBlock (Bisco, USA). Fluid movement measurements were made at 2-min intervals for 8 min, and a mean of the values obtained was calculated for each specimen. The results were analyzed using Kruskal-Wallis test and Wilcoxon signed ranks tests with a significance threshold of P < 0.05. RESULTS: There were no significant differences in permeability among desensitizing agents (P > 0.05); however dentin permeability was reduced in all groups (P < 0.05). CONCLUSION: The in vitro fluid conductance of dentin discs were reduced by treating with these five desensitizing agents.

Polymorphisms of endotoxin pathway and endotoxin exposure: in vitro IgE synthesis and replication in a birth cohort.

BACKGROUND: Genetic variants in endotoxin signaling pathway are important in modulating the effect of environmental endotoxin on asthma and atopic phenotypes. Our objective was to determine the single nucleotide polymorphisms (SNPs) in the endotoxin signaling pathway that may influence in vitro IgE synthesis and to investigate the relationship between these variants and endotoxin exposure in relation to the development of asthma and atopy in a birth cohort. METHODS: Peripheral blood mononuclear cells from 45 children with asthma were stimulated with 2 and 200 ng/ml lipopolysaccharide in vitro and IgE was measured in the culture supernatants. Children were genotyped for 121 SNPs from 30 genes in the endotoxin signaling pathway. Variants with a dose-response IgE production in relation to lipopolysaccharide (LPS) were selected for replication in a population-based birth cohort, in which we investigated the interaction between these SNPs and endotoxin exposure in relation to airway hyper-responsiveness, wheeze, and atopic sensitization. RESULTS: Twenty-one SNPs in nine genes (CD14, TLR4, IRF3, TRAF-6, TIRAP, TRIF, IKK-1, ST-2, SOCS1) were found to modulate the effect of endotoxin on in vitro IgE synthesis, with six displaying high linkage disequilibrium. Of the remaining 15 SNPs, for seven we found significant relationships between genotype and endotoxin exposure in the genetic association study in relation to symptomatic airway hyper-responsiveness (CD14-rs2915863 and rs2569191, TRIF-rs4807000), current wheeze (ST-2-rs17639215, IKK-1-rs2230804, and TRIF-rs4807000), and atopy (CD14-rs2915863 and rs2569192, TRAF-6-rs5030411, and IKK-1-rs2230804). CONCLUSIONS: Variants in the endotoxin signaling pathway are important determinants of asthma and atopy. The genotype effect is a function of the environmental endotoxin exposure.

The effect of CD14 C159T polymorphism on in vitro IgE synthesis and cytokine production by PBMC from children with asthma.

BACKGROUND: Even though the genotype at the promoter region of the CD14 molecule is known to affect the atopic phenotypes, the cellular and molecular basis of this association is largely unknown. OBJECTIVE: To investigate the effect of lipopolysaccharide (LPS) on IgE production and cytokine profile by peripheral blood mononuclear cells (PBMC) obtained from asthmatic children with the TT and the CC genotypes at position -159 of the CD14 gene. METHODS: Peripheral blood mononuclear cells from asthmatic children with alternative genotypes at CD14 C159T locus were stimulated with 2 and 200 ng/ml LPS in vitro. The IgE, IgG and, IgM response was determined by ELISA and Ig έ-germline, IgG, and IgM transcription by real-time PCR. A cluster of cytokines was measured by cytometric bead array. RESULTS: Asthmatic children with the TT genotype but not those with the CC genotype responded with increased IgE synthesis and germline transcription to LPS stimulation. There were no genotype-related differences in IgG and IgM. TT but not the CC genotype was associated with significantly increased interleukin (IL)-4/IL-12 and IL-4/interferon-gamma (IFN-γ) ratios in the culture supernatant. There were no genotype-related differences in IL-1ß, IL-7, IL-10, IL-13, IL-17A, granulocyte colony stimulating factor, granulocyte macrophage colony stimulating factor, monocyte chemotactic protein, and tumor necrosis factor alpha. CONCLUSION: Peripheral blood mononuclear cells from asthmatic children with the TT genotype at position -159 of the CD14 gene make more IgE than those with the CC genotype following LPS stimulation because of increased germline transcription and have an augmented Th2 cytokine profile.

Antenatal and delivery risk factors and prevalence of cerebral palsy in Duzce (Turkey).

This cross-sectional study aimed at investigating the prevalence and the etiological factors of cerebral palsy (CP) and comparing them with normal population within the rural and urban areas of Duzce province. Of the 102 children with cerebral palsy, 98 were associated with antenatal and delivery risk factors. The mean crude prevalence of cerebral palsy was 1.1 per 1000 live births. The children with CP were compared with 530 control subjects. The mothers of the children with cerebral palsy were significantly younger than the mothers of children in control group, and they had less parity and abortion. Preeclampsia, premature rupture of membranes, home births, prolonged labor, and twin pregnancies were significantly more common in the mothers of children with cerebral palsy, where no significant differences were found between the groups in terms of breech delivery, rate of cesarean births, gestational diabetes, and hemorrhage in late pregnancy. Birth asphyxia, liqueur with meconium stained, prolonged jaundice and neonatal seizure were also significantly more common in the group with cerebral palsy. Of the children with cerebral palsy, 78% were born at term, 20% were born with gestational ages of 32-36 weeks, 2% were born with gestational ages of 30-31 weeks. Nine percent of those children had a birth weight of >or= 3000 g, 12.2% had a birth weight of 2500-2999 g, 33.7% had a birth weight of 1500-2499 g, and 5.1% had a birth weight of

Prevalence of oral lesions in 13- to 16-year-old students in Duzce, Turkey.

OBJECTIVE: The aim of this study was to determine the prevalence of oral lesions in 13- to 16-year-old students. METHODS: A cross-sectional survey was carried out on students in Duzce, a province in the western Black Sea region of Turkey. A total of 993 children aged between 13 and 16 from eight secondary schools were examined. Oral lesions with recurrent behavior, if observed, were recorded at the time of examination. Venous blood samples were obtained for detecting hemoglobin levels. RESULTS: Two hundred sixty adolescents (26.2%) were diagnosed with at least one oral mucosal lesion at the time of the examination. Thirteen different mucosal alterations were diagnosed, and the most common lesions were angular cheilitis (9%), linea alba (5.3%), and aphthous ulceration (3.6%). The correlation between occurrence of mucosal lesions and sex was not statistically significant (P > 0.05). Statistical evaluation of the data revealed a significant relationship only between the presence of angular cheilitis and anemia (P < 0.05). CONCLUSION: This study is the first epidemiologic study of oral mucosal lesions in adolescents in Turkey. Angular cheilitis was the only oral mucosal lesion that had a significant correlation with anemia.

Right atrial and tricuspid hypoplasia.

Cor triatriatum dexter is a rare cardiac anomaly with division between the sinus and primitive atrial portions of the right atrium. It is supposed that the failure of the venous valves to regress appropriately may create abnormalities in fetal circulation predisposing the fetus to maldevelopment of the right heart structures. The underdevelopment of the right ventricle with hypoplasia or atresia of the tricuspid valve is a well known abnormality, but underdevelopment of the right atrium with tricuspid hypoplasia due to multiple venous valves is extremely rare. This is a case of complex type of cor triatriatum dexter in which the right atrium is hypoplastic and like a tube formed by the superior and inferior vena cava and is divided into 3 portions by 2 membranes.

In vivo evaluation of teicoplanin- and calcium sulfate-loaded PMMA bone cement in preventing implant-related osteomyelitis in rats.

The objective of this study was to evaluate the efficacy of teicoplanin- and calcium sulphate-loaded polymethylmethacrylate (PMMA) bone cements in preventing experimental implant-related osteomyelitis in rats. Four groups of antibiotic-loaded rods were prepared and were implanted into the lateral condylus of the rat femur after inoculation of Staphylococcus aureus. The effectiveness of these were assessed microbiologically, radiographically, and histopathologically. Radiographic evaluation revealed a significant reduction of periostal reaction and osteolysis in rats that received calcium sulphate- and teicoplanin-loaded rods. Histopathological evaluation confirmed these results. Acute infection and bone necrosis were found to be significantly lower in rats that had received calcium sulphate- and teicoplanin-loaded rods. The addition of calcium sulfate to teicoplanin-loaded PMMA bone cement appeared satisfactory as an antibiotic-carrying system for prophylaxis of experimental implant-related osteomyelitis, but further investigations are needed to reach definitive statements for clinical applications.

Immunogenicity study of a combined diphtheria, tetanus, acellular pertussis, inactivated poliomyelitis vaccine used to reconstitute a freeze-dried Haemophilus influenzae type b vaccine (DTaP-IPV//PRP-T) administered simultaneously with a hepatitis B vaccine at two, three and four months of life.

This study was designed to assess the immunogenicity of a vaccine combining diphtheria and tetanus toxoids, acellular pertussis vaccine, and inactivated poliovirus vaccine reconstituting Haemophilus influenzae type b polysaccharide conjugated to tetanus protein (DTaP-IPV//PRP-T; Pasteur Mérieux Connaught, Lyon, France) administered simultaneously in association with hepatitis B vaccine (RECOMBIVAX (¿trade mark omitted¿) Merck, Sharp & Dohme, West Point, PA, USA) for the primary immunization of infants. The vaccines were administered at two, three and four months of age. One hundred and sixty-two healthy infants, aged 8-10 weeks, were enrolled in the study. Blood samples were taken before the first dose and 4 weeks after the third dose. The infants were observed for 15 minutes after vaccination for any immediate reaction. Adverse events requiring a medical consultation were recorded by the parents in a diary over the 7 days following vaccination. Four weeks after the third immunization, the percentages of infants fulfilling seroconversion criteria were 98.9% for pertussis toxin, 95.9% for filamentous haemagglutinin, 100.0% for tetanus, 100.0% for diphtheria, 99.3% for poliovirus type 1, 100.0% for both poliovirus types 2 and 3, 98.0% for Haemophilus influenzae type b, and 100% for hepatitis B surface antigen. No vaccine-related serious adverse event was reported. The simultaneous administration of DTaP-IPV//PRP-T and hepatitis B vaccines at two, three and four months of age yielded clinically satisfactory immune responses to all antigens compared with historical controls and gave a good safety profile.

Swaddling and acute respiratory infections.

In Turkey and China the ancient practice of swaddling is still commonly practiced. Both countries have extremely high rates of pneumonia, especially during the neonatal period. Preliminary evidence on the possibility that swaddling may interfere with normal respiratory function and thereby predispose to pneumonia was gathered in a teaching health center in Ankara. Babies who had been swaddled for at least three months were four times more likely to have developed pneumonia (confirmed radiologically) and upper respiratory infections than babies who were unswaddled. These preliminary findings were highly significant and are being followed up by further studies.