Transannular C-H functionalization of cycloalkane carboxylic acids.

Cyclic organic molecules are common among natural products and pharmaceuticals1,2. In fact, the overwhelming majority of small-molecule pharmaceuticals contain at least one ring system, as they provide control over molecular shape, often increasing oral bioavailability while providing enhanced control over the activity, specificity and physical properties of drug candidates3-5. Consequently, new methods for the direct site and diastereoselective synthesis of functionalized carbocycles are highly desirable. In principle, molecular editing by C-H activation offers an ideal route to these compounds. However, the site-selective C-H functionalization of cycloalkanes remains challenging because of the strain encountered in transannular C-H palladation. Here we report that two classes of ligands-quinuclidine-pyridones (L1, L2) and sulfonamide-pyridones (L3)-enable transannular γ-methylene C-H arylation of small- to medium-sized cycloalkane carboxylic acids, with ring sizes ranging from cyclobutane to cyclooctane. Excellent γ-regioselectivity was observed in the presence of multiple ß-C-H bonds. This advance marks a major step towards achieving molecular editing of saturated carbocycles: a class of scaffolds that are important in synthetic and medicinal chemistry3-5. The utility of this protocol is demonstrated by two-step formal syntheses of a series of patented biologically active small molecules, prior syntheses of which required up to 11 steps6.

Hydrogen-bond-acceptor ligands enable distal C(sp3)-H arylation of free alcohols.

The functionalization of C-H bonds in organic molecules is one of the most direct approaches for chemical synthesis. Recent advances in catalysis have allowed native chemical groups such as carboxylic acids, ketones and amines to control and direct C(sp3)-H activation1-4. However, alcohols, among the most common functionalities in organic chemistry5, have remained intractable because of their low affinity for late transition-metal catalysts6,7. Here we describe ligands that enable alcohol-directed arylation of δ-C(sp3)-H bonds. We use charge balance and a secondary-coordination-sphere hydrogen-bonding interaction-evidenced by structure-activity relationship studies, computational modelling and crystallographic data-to stabilize L-type hydroxyl coordination to palladium, thereby facilitating the assembly of the key C-H cleavage transition state. In contrast to previous studies in C-H activation, in which secondary interactions were used to control selectivity in the context of established reactivity8-13, this report demonstrates the feasibility of using secondary interactions to enable challenging, previously unknown reactivity by enhancing substrate-catalyst affinity.

Association of the Life's Essential 8 cardiovascular health score with periodontitis among US adults.

AIM: To investigate whether cardiovascular health (CVH) is associated with periodontitis. MATERIALS AND METHODS: We used data from the 2009 to 2014 National Health and Nutrition Examination Survey. We quantified CVH using Life's Essential 8, which includes four health behaviours (diet, smoking, physical activity and sleep) and four health factors (body mass index, blood cholesterol, glucose and pressure). We categorized scores as low (0-49), moderate (50-79) and high (80-100). We calculated subscores of health behaviours and factors and categorized them as low, moderate and high. We used logistic regression to assess the association of CVH with periodontitis, adjusting for age, gender, race/ethnicity, education, poverty index, marital status and health insurance. We computed odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: This study included 9296 adults ≥30 years old. Multivariable-adjusted models showed that subjects with moderate (OR, 0.62; 95% CI: 0.52-0.74) or high (OR, 0.43; 95% CI: 0.33-0.57) CVH had significantly lower odds of periodontitis compared to those with low CVH. These results were consistent in the health behaviours model, but the estimates in the health factors model were not significant. CONCLUSIONS: Improving CVH may help prevent periodontitis. Longitudinal studies are needed to confirm our findings.

Investigation of Asymptomatic Infection of Phellinus noxius in Herbaceous Plants.

The white-rot fungus Phellinus noxius is known to cause brown root rot disease (BRRD) in woody trees and shrubs. To understand the pathogenicity of P. noxius in herbaceous plants, we investigated 23 herbaceous weed and turfgrass species in 32 BRRD-infested sites in Taiwan and/or tested them by artificial inoculation. In the field survey, P. noxius was isolated from seven symptomless herbaceous species (i.e., Typhonium blumei, Paspalum conjugatum, Paspalum distichum, Oplismenus compositus, Bidens pilosa, Digitaria ciliaris, and Zoysia matrella). Potted plant inoculation assays suggested that P. noxius is able to infect Artemisia princeps, O. compositus, and Z. matrella but not Axonopus compressus, Eremochloa ophiuroides, Ophiopogon japonicus, or Cynodon dactylon. A. princeps plants wilted within 2 weeks postinoculation, but inoculated O. compositus and Z. matrella were asymptomatic, and P. noxius could be isolated from only inoculated sites. The colonization of P. noxius in the cortex and vascular cylinder of roots was visualized by paraffin sectioning and trypan blue staining of juvenile seedlings grown on water agar. To evaluate the effect of replantation for the remediation of BRRD-infested sites, P. noxius-inoculated wood strips were buried in soil with or without vegetation. After 4 weeks, P. noxius could be detected only in the bare soil group. For the control of BRRD, the herbaceous hosts should be removed around the diseased trees/stumps and non-host turfgrasses (e.g., A. compressus, E. ophiuroides, O. japonicus, or C. dactylon) planted to accelerate the degradation of P. noxius.

Decreased junctional adhesion molecule 3 expression induces reactive oxygen species production and apoptosis in trophoblasts†.

Junctional adhesion molecule 3 (JAM3) is involved in epithelial cell junction, cell polarity, and motility. The molecular mechanisms underlying the role of JAM3 in placental dysfunction remain unclear. We hypothesized that JAM3 expression regulates trophoblast fusion, differentiation, proliferation, and apoptosis. Our results revealed that JAM3 was expressed in the cytotrophoblasts and syncytiotrophoblasts of first-trimester and term placental villi. JAM3 expression in cell-cell junctions decreased with the formation of syncytiotrophoblasts. Using trophoblasts as an in vitro model, we observed that forskolin and JAM3 knockdown significantly reduced JAM3 expression and increased syncytium formation. JAM3 knockdown additionally inhibited trophoblast proliferation and increased the number of trophoblasts in the sub-G1 and G2/M phases, indicating cell-cycle disturbance and apoptosis. Cell-cycle arrest was associated with the engagement of checkpoint kinase 2-cell division cycle 25C-cyclin-dependent kinase 1/cyclin B1 signaling. Increased expression of BIM, NOXA, XAF1, cytochrome c, and cleaved caspase-3 further indicated trophoblast apoptosis. Overexpression of JAM3 or recombinant JAM3 protein enhanced trophoblast adhesion and migration, which were inhibited by JAM3 knockdown. JAM3 knockdown induced reactive oxygen species and syncytin 2 expression in trophoblasts. Furthermore, H2O2-induced oxidative stress reduced JAM3 expression in trophoblasts and cell culture supernatants. H2O2 simultaneously induced trophoblast apoptosis. JAM3 expression was significantly decreased in the plasmas and placentas of patients with early-onset severe preeclampsia. Thus, our results show that JAM3 may not only be a structural component of trophoblast cell junctions but also regulates trophoblast fusion, differentiation, proliferation, apoptosis, and motility. Dysregulated trophoblast JAM3 expression is crucial in preeclampsia development.

Placental multipotent mesenchymal stromal cell-derived Slit2 may regulate macrophage motility during placental infection.

Slit proteins have been reported to act as axonal repellents in Drosophila; however, their role in the placental microenvironment has not been explored. In this study, we found that human placental multipotent mesenchymal stromal cells (hPMSCs) constitutively express Slit2. Therefore, we hypothesized that Slit2 expressed by hPMSCs could be involved in macrophage migration during placental inflammation through membrane cognate Roundabout (Robo) receptor signaling. In order to develop a preclinical in vitro mouse model of hPMSCs in treatment of perinatal infection, RAW 264.7 cells were used in this study. Slit2 interacted with Robo4 that was highly expressed in RAW 264.7 macrophages: their interaction increased the adhesive ability of RAW 264.7 cells and inhibited migration. Lipopolysaccharide (LPS)-induced CD11bCD18 expression could be inhibited by Slit2 and by hPMSC-conditioned medium (CM). LPS-induced activation of p38 and Rap1 was also attenuated by Slit2 and by hPMSC-CM. Noticeably, these inhibitory effects of hPMSC-CM decreased after depletion of Slit2 from the CM. Furthermore, we found that p38 siRNA inhibited LPS-induced Rap1 expression in RAW 264.7 cells, indicating that Rap1 functions downstream of p38 signaling. p38 siRNA increased cell adhesion and inhibited migration through reducing LPS-stimulated CD11bCD18 expression in RAW 264.7 cells. Thus, hPMSC-derived Slit2 may inhibit LPS-induced CD11bCD18 expression to decrease cell migration and increase adhesion through modulating the activity and motility of inflammatory macrophages in placenta. This may represent a novel mechanism for LPS-induced placental infection.

The effects of using erbium, chromium-doped:yttrium-scandium-gallium-garnet laser on the surface modification, bacterial decontamination, and cell adhesion on zirconia discs: an in vitro study.

The use of zirconia for implants and abutments has become more prevalent in implant dentistry as an alternative to the commonly used titanium implants, and peri-implant disease can still affect them. The erbium, chromium-doped:yttrium-scandium-gallium-garnet (Er, Cr:YSGG) laser has emerged as a promising treatment modality. The purposes of this in vitro study were to (1) determine the effects of the laser on the surface roughness of zirconia discs; (2) determine the extent of removal of a single species biofilm, E. coli, on the zirconia discs after applying the laser; (3) determine the amount of cell adhesion and proliferation utilizing fibroblasts on zirconia discs after treatment with the laser. All treatments will be compared with the commonly used ultrasonic instrumentation and hand scalers. For the first aim, gross examination revealed noticeable surface damage on the discs when using ultrasonic and scalers but not for the laser group. For surface roughness, the mean roughness was Pa= 0.623±0.185 µm, 0.762±0.421 µm, 0.740±0.214 µm, and 0.724±0.168 µm for control discs, and discs treated with either the Er,Cr:YSGG laser, ultrasonic instrumentation, and hand scalers respectively. There was no statistical significance among the groups (p=0.628). For bacteria decontamination, there was a statistical significance among the groups (p< 0.0001). Statistical significance was seen between the control group and each of the three treatment groups, favoring the treatment groups (p< 0.0001). Statistical significance was seen when comparing ultrasonic instrumentation and hand scalers (p= 0.000) as well as when comparing the Er,Cr:YSGG laser to hand scalers (p= 0.007), favoring both the ultrasonic instrumentation and Er,Cr:YSGG laser. No significance between the Er,Cr:YSGG laser group and the ultrasonic instrumentation group was noted (p =0.374). When comparing the cell attachment following treatment in each of the three groups and also without treatment (control), there was a statistical significance among the groups (p<0.0001) in terms of total cell count, favoring the control and the laser groups. Further evaluations with SEM showed differences in cell morphology indicating more adherent cells on Er,Cr:YSGG laser-treated surfaces. In conclusion, gross examination of the discs show clear surface changes when using ultrasonic instrumentation and hand scalers compared to the Er,Cr:YSGG laser group. The Er,Cr:YSGG laser was able to effectively ablate bacteria from zirconia disc. Fibroblast attachment on the surfaces of the zirconia discs shows more adherence when treated with Er,Cr:YSGG laser.

The In Vitro Evaluation of Preosteoblast Migration From 3-D-printed Scaffolds to Decontaminated Smooth and Minimally Rough Titanium Surfaces: A Pilot Study.

In vitro evaluations are essential to gaining a better understanding of re-osseointegration, while reducing animal use and the overall costs of peri-implantitis studies. This pilot study evaluated preosteoblast migration from 3-D-printed scaffolds to decontaminated titanium microimplants, creating a system that tries to mimic the bone-implant interface. Smooth (S) and minimally rough (R) titanium microimplants were incubated in Escherichia coli cultures and divided into six groups according to the decontamination protocol applied: EDTA gel (EDTA); chlorhexidine (CHL); chlorhexidine-soaked gauze (GCHL); scaling (SC); titanium brush (TiB); and implantoplasty (IP). Pristine S and R microimplants were used as the controls (C). After the decontamination procedures, the microimplants were inserted in 3-D-printed polyurethane-based scaffolds previously inoculated with preosteoblast cell cultures. Cellular migration was assessed after 24, 72 and 120 hours by ATP quantification. At the 120-hour time point, there was no statistically significant difference between S-C, S-EDTA, S-CHL, S-GCHL and S-SC (p > 0.05), and between R-C, R-EDTA and R-GCHL (p > 0.05). The in vitro model developed in this pilot study successfully demonstrated cell migration on the different decontaminated surfaces. This methodology suggests that on smooth microimplants, EDTA, GCHL, SC and TiB decontamination may have a reduced impact on preosteoblast migration, while on minimally rough microimplants, EDTA and GCHL decontamination affected cell migration the least. However, when selecting a decontamination protocol, the effectiveness of the decontamination per se must also be considered.

Establishment of perpendicular protrusion of type I collagen on TiO2 nanotube surface as a priming site of peri-implant connective fibers.

Natural teeth are supported by connective tissue collagen fibers that insert perpendicularly in the tooth cementum. Perpendicular insertion plays an important role in the maintenance of the junction between the oral epithelium and the periodontal connective tissue. Most titanium dental implant surfaces have no micro or macro structure to support perpendicularly oriented collagen attachment. Without this tight biologic seal to resist bacterial invasion and epithelial downgrowth, progressive bone loss in peri-implantitis is seen around dental implants. The purpose of this study was to establish the perpendicularly oriented collagen attachment to titanium oxide nanotube (TNT), and to assess its binding stability. TNT was prepared on the titanium-surface by anodization. Scanning electron microscopy (SEM) showed a regularly aligned TNT with an average 67 nm-diameter when anodized at 30 V for 3 h. Subsequently, collagen type I (CoI) was electrophoretically fused to anodic TNT in native polyacrylamide gel system where negatively charged CoI-C term was perpendicularly navigated to TNT. SEM and atomic force microscopy (AFM) were used to analyze CoI on the TiO2 and TNT surface. Several tens of nanometers of CoI protrusion were recorded by AFM. These protrusions may be long enough to be priming sites for cell-secreted CoI. CoI laid parallel to the titanium surface when fused by a chemical linker. Binding resistance of CoI against drastic ultrasonication was measured by Fourier-transform infrared spectroscopy attenuated total reflection (FTIR-ATR). The electrophoretically fused CoI in the titanium nanotube (TNT-CoIEPF) showed the significantly greatest binding resistance than the other groups (P < 0.01, a 1-way ANOVA and Tukey HSD post hoc test). Furthermore, TNT-CoIEPF surface rejected epithelial cell stretching and epithelial sheet formation. Chemically linked horizontal CoI on titanium oxide (TiO2) facilitated epithelial cell stretching and sheet formation.

The compatibility of Bacillus thuringiensis Cry protein-solubilizing buffers with the droplet feeding method in fall armyworm larvae.

The volumes of alkaline (pH > 10), Bacillus thuringiensis Cry protein-solubilizing buffers imbibed by fall armyworm larvae in droplet feeding assays were determined. The buffers differed in the presence or concentration of key ingredients, including buffering agents, chelating agents, reducing agents, and protease inhibitors. For both first and second instar larvae, the buffer used had a significant effect on the volume imbibed. The study showed that the droplet feeding method is compatible with Cry protein-solubilizing buffers, but that it is important to determine the volume imbibed for every buffer used in dose-dependent bioassays in order to reduce dose errors.

Targeted inactivation of murine Ddx3x: essential roles of Ddx3x in placentation and embryogenesis.

The X-linked DEAD-box RNA helicase DDX3 (DDX3X) is a multifunctional protein that has been implicated in gene regulation, cell cycle control, apoptosis, and tumorigenesis. However, the precise physiological function of Ddx3x during development remains unknown. Here, we show that loss of Ddx3x results in an early post-implantation lethality in male mice. The size of the epiblast marked by Oct3/4 is dramatically reduced in embryonic day 6.5 (E6.5) Ddx3x-/Y embryos. Preferential paternal X chromosome inactivation (XCI) in extraembryonic tissues of Ddx3x heterozygous (Ddx3x-/+) female mice with a maternally inherited null allele leads to placental abnormalities and embryonic lethality during development. In the embryonic tissues, Ddx3x exhibits developmental- and tissue-specific differences in escape from XCI. Targeted Ddx3x ablation in the epiblast leads to widespread apoptosis and abnormal growth, which causes embryonic lethality in the Sox2-cre/+;Ddx3xflox/Y mutant around E11.5. The observation of significant increases in γH2AX and p-p53Ser15 indicates DNA damage, which suggests that loss of Ddx3x leads to higher levels of genome damage. Significant upregulation of p21WAF1/Cip1 and p15Ink4b results in cell cycle arrest and apoptosis in Ddx3x-deficient cells. These results have uncovered that mouse Ddx3x is essential for both embryo and extraembryonic development.

Omega-3 polyunsaturated fatty acids reduce preterm labor by inhibiting trophoblast cathepsin S and inflammasome activation.

Preterm labor is associated with inflammation and infection. The mechanisms underlying the role of omega-3 fatty acid in inflammasome activation and prevention of preterm labor remain unknown. We hypothesized that omega-3 fatty acid can reduce the rate of preterm birth induced by infection and trophoblast inflammation. In the present study, we found that inflammasome-related molecules and IL-1ß in trophoblasts were activated by TNF-α derived from lipopolysaccharide (LPS)-stimulated THP-1 cell-conditioned medium (CM) and recombinant TNF-α protein. The results demonstrated that stimulation with TNF-α caused lysosomal rupture in trophoblasts, which accelerated cathepsin S (CTSS) diffusion from lysosomes to the cytosol and activated NLRP1 (nacht domain-leucine-rich repeat, and pyd-containing protein 1) and absent in melanoma 2 (AIM2) inflammasomes, thereby increasing IL-1ß secretion. Moreover, in response to LPS challenge, TNF-α increased trophoblast cell death and decreased cell viability through inflammasome and CTSS activation. Stearidonic acid (SDA; 18:4n-3) and docosahexaenoic acid (DHA; 22:6n-3) inhibited inflammasome-related molecule synthesis and CTSS and caspase-1 activation, which further reduced the preterm delivery rate of pregnant mice induced by LPS (92.9 compared with 69.7% (DHA); 92.9 compared with 53.5% (SDA)). Higher expression of TNF-α, IL-1ß, prostaglandin E2, and CTSS, but lower resolvin D1 expression, was observed in preterm pregnant mice than in controls. Similarly, resolvin D1 was highly expressed in women with term delivery compared with women with preterm delivery. Thus, SDA and DHA may attenuate macrophage-derived TNF-α inducing CTSS and inflammasome activation, IL-1ß secretion, and placental trophoblast cell death. These functions are implicated in the preventive effects of SDA and DHA on preterm labor.

Association of dysfunctional synapse defective 1 (SYDE1) with restricted fetal growth - SYDE1 regulates placental cell migration and invasion.

The transcription factor glial cells missing 1 (GCM1) regulates trophoblast differentiation and function during placentation. Decreased GCM1 expression is associated with pre-eclampsia, suggesting that abnormal expression of GCM1 target genes may contribute to the pathogenesis of pregnancy complications. Here we identified a novel GCM1 target gene, synapse defective 1 (SYDE1), which encodes a RhoGAP that is highly expressed in human placenta, and demonstrated that SYDE1 promotes cytoskeletal remodelling and cell migration and invasion. Importantly, genetic ablation of murine Syde1 results in small fetuses and placentas with aberrant phenotypes in the placental-yolk sac barrier, maternal-trophoblast interface, and placental vascularization. Microarray analysis revealed altered expression of renin-1, angiotensin I converting enzyme 2, angiotensin II type 1a receptor, and membrane metalloendopeptidase of the renin-angiotensin system in Syde1-knockout placenta, which may compensate for the vascular defects to maintain normal blood pressure. As pregnancy proceeds, growth restriction of the Syde1-/- fetuses and placentas continues, with elevated expression of the Syde1 homologue Syde2 in placenta. Syde2 may compensate for the loss of Syde1 function because SYDE2, but not the GAP-dead SYDE2 mutant, reverses migration and invasion activities of SYDE1-knockdown JAR trophoblast cells. Clinically, we further detected decreased SYDE1 expression in preterm and term IUGR placentas compared with gestational age-matched controls. Our study suggests a novel mechanism for GCM1 and SYDE1 in regulation of trophoblast cell migration and invasion during placental development and that decreased SYDE1 expression is associated with IUGR. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

The Prognostic Role of STEAP1 Expression Determined via Immunohistochemistry Staining in Predicting Prognosis of Primary Colorectal Cancer: A Survival Analysis.

STEAP1 (six transmembrane epithelial antigen of the prostate 1) is a transmembrane protein that functions as a potential channel or transporter protein. It is overexpressed in certain cancers and is viewed as a promising therapeutic target. However, the prognostic role of STEAP1 is still controversial, and no role for STEAP1 has yet been indicated in colorectal cancer. The aim of this study was to investigate the possible association of STEAP1 expression with colorectal cancer prognosis. STEAP1 expression was analyzed by immunohistochemical staining of a tissue array of 165 cancer specimens from primary colorectal cancer patients. The mean and medium follow-up times after surgery were 5.1 and 3.9 years, respectively. A total of 139 patients died during the 13 years of follow-up in the survey period. The prognostic value of STEAP1 with respect to overall survival was analyzed by Kaplan-Meier analysis and Cox proportional hazard models. In total, 164 samples displayed detectable STEAP1 expression in the cytoplasm and membrane. Low STEAP1 expression was correlated with poor overall survival (five-year survival: 33.7% vs. 57.0%, low expression vs. high expression, p = 0.020). Accordingly, multivariate analysis identified low STEAP1 expression as an independent risk factor (hazard ratio = 1.500, p = 0.018), especially in elderly patients or those with late stage cancers, late T values, and early N values. We suggest that analysis of STEAP1 expression by immunohistochemical staining could serve as an independent prognostic marker for colorectal patients. This finding should be validated by other investigative groups.

Human placenta-derived multipotent mesenchymal stromal cells involved in placental angiogenesis via the PDGF-BB and STAT3 pathways.

We studied the smooth muscle cell differentiation capability of human placental multipotent mesenchymal stromal cells (hPMSCs) and identified how endothelial cells recruit hPMSCs participating in vessel formation. hPMSCs from term placentas were induced to differentiate into smooth muscle cells under induction conditions and different matrix substrates. We assessed endothelial cells from umbilical veins for platelet-derived growth factor (PDGF)-BB expression and to induce hPMSC PDGFR-beta and STAT3 activation. Endothelial cells were co-cultured with hPMSCs for in vitro angiogenesis. Cell differentiation ability was then further assessed by mouse placenta transplantation assay. hPMSCs can differentiate into smooth muscle cells; collagen type I and IV or laminin support this differentiation. Endothelial cells expressed significant levels of PDGF-BB and activated STAT3 transcriptional activity in hPMSCs. Endothelial cell-conditioned medium induced hPMSC migration, which was inhibited by STAT3 small interfering RNA transfection or by pretreatement with PDGFR-beta-blocking antibody but not by PDGFR-alpha-blocking antibody or isotype immunoglobulin G (IgG; P < 0.001). hPMSCs can incorporate into endothelial cells with tube formation and promote endothelial cells, forming capillary-like networks than endothelial cells alone (tube lengths: 12 024.1 ± 960.1 vs. 9404.2 ± 584.7 pixels; P < 0.001). Capillary-like networks were significantly reduced by hPMSCs pretreated with PDGFR-beta-blocking antibody but not by PDGFR-alpha-blocking antibody or isotype IgG (P < 0.001). Transplantation of hPMSCs into mouse placentas revealed incorporation of the hPMSCs into vessel walls, which expressed alpha-smooth muscle actin, calponin, and smooth muscle myosin (heavy chain) in vivo. In conclusion, endothelial cell-hPMSC interactions occur during vessel development of placenta. Placental endothelial cell-derived PDGF-BB recruits hPMSCs involved in vascular development via PDGFR-beta/STAT3 activation.

High nuclear/cytoplasmic ratio of Cdk1 expression predicts poor prognosis in colorectal cancer patients.

BACKGROUND: Cdk1 (cyclin-dependent kinase 1) is critical regulator of the G2-M checkpoint. Cyclin-dependent kinase pathways are considered possible targets for cancer treatment; however, the prognostic role of Cdk1 in colorectal cancer is still controversial. Therefore, we attempted to determine the impact of Cdk1 on the clinical outcome of colorectal cancer patients to further identify its role in colorectal cancer. METHODS: Cdk1 immunoreactivity was analyzed by immunohistochemistry (IHC) in 164 cancer specimens from primary colorectal cancer patients. The medium follow-up time after surgery was 3.7 years (range: 0.01 to 13.10 years). The prognostic value of Cdk1 on overall survival was determined by Kaplan-Meier analysis and Cox proportional hazard models. RESULTS: All samples displayed detectable Cdk1 expression with predominant location in the cytoplasm and nucleus. A high Cdk1 nuclear/cytoplasmic (N/C) expression ratio was correlated with poor overall survival (5-year survival rate: 26.3% vs 46.9%, N/C ratio ≥1.5 vs N/C ratio <1.5, log-rank p = 0.027). Accordingly, a Cdk1 N/C expression ratio ≥1.5 was identified as an independent risk factor by multivariate analysis (hazard ratio = 1.712, P = 0.039). CONCLUSIONS: We suggest that Cdk1 N/C expression ratio determined by IHC staining could be an independent prognostic marker for colorectal cancer.

Difluorophenylglycinols as new modulators of proteolytic processing of amyloid precursor proteins.

Synthesis and evaluation of difluorophenylglycinols as new modulators of proteolytic processing of the amyloid-ß precursor proteins for Alzheimer's therapies were described. A range of N-substituted (R)- and (S)-difluorophenylglycinols, structured on the amino alcohol framework, were explored by incorporating the arylsulfonyl moieties and various N-substituents. Evans' chiral auxiliary strategy was employed for the asymmetric synthesis of these enantiomeric difluorophenylglycinols. Compounds with effects on the γ-secretase inhibition and ERK-mediated signaling pathways were evaluated on cell-based assays. Among them, N-cyclopropylmethyl derivatives R-12c and R-13c showed modest γ-secretase inhibition as well as ERK-dependent activation.

A rare malignancy of the parotid gland in a 13-year-old Taiwanese boy: case report of a mammary analogue secretory carcinoma of the salivary gland with molecular study.

Mammary analogue secretory carcinoma (MASC) is a recently described malignancy of the salivary glands characterized by an ETV6-NTRK3 (EN) fusion gene. Morphologically, MASC is sometimes difficult to distinguish from acinic cell carcinoma. Consequently, identifying the chromosomal translocation is essential for diagnosis. We present a case of parotid gland MASC in a 13-year-old boy. To the best of our knowledge, this is the youngest case reported in the literature. Histologic evaluation showed a tumor composed of microcysts, tubular structures, solid nests, or papillary architecture, with secretions within the lumens of the cysts or tubules. Immunohistochemically, tumor cells showed diffuse positive staining of S-100 protein, cytokeratin 19, and vimentin. ETV6 rearrangement was detected by fluorescence in situ hybridization and EN fusion transcripts were verified by reverse transcription (RT-PCR) assay.

The effect of larval exposure to acids and detergents on the life history of the major malaria vector Anopheles arabiensis Patton (Diptera: Culicidae).

BACKGROUND: Anopheles arabiensis, a highly adaptable member of the Anopheles gambiae complex, poses a challenge for control efforts due to its outdoor biting and resting behaviour. Consequently, indoor insecticide-based control methods are ineffective against An. arabiensis. Furthermore, An. arabiensis are adapting to breeding in polluted waters, and may be contributing to residual malaria and malaria in urban areas. There have been some advances in understanding the effect of rural pollutants on Anopheles mosquitoes, but the effect of urban pollutants is poorly understood. Thus, in this study, the effect of acidic pollutants [nitric acid (HNO3) and hydrochloric acid (HCl)] and alkaline pollutants (phosphate-free and phosphate-containing detergent) on two laboratory-reared An. arabiensis strains - an insecticide susceptible strain (SENN) and an insecticide-resistant strain selected from SENN (SENN-DDT) - were determined. RESULTS: The median lethal concentration (LC50) and larval exposure on larval development, adult longevity and insecticide tolerance were evaluated. Nitric acid and phosphate-containing detergent were found to be more toxic than HCl and phosphate-free detergent in terms of LC50 values. Detergent exposure (both phosphate-containing and phosphate-free) increased adult longevity of both strains. Nitric acid reduced larval development time in both SENN and SENN-DDT, whereas HCl reduced larval development time in SENN only. By contrast, both phosphate-containing and phosphate-free detergents increased larval development time of both strains. Furthermore, HNO3 and phosphate-containing detergent increased insecticide tolerance the most. CONCLUSION: The two An. arabiensis strains responded to urban pollutants differently. Thus, this study provides insight into the adaptation of An. arabiensis to acidic and alkaline urban pollutants. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Characterization of the Tissue and Strain-Specific Microbiota of Anopheles funestus Giles (Diptera: Culicidae).

The mosquito microbiota is a critical determinant of mosquito life history. It is therefore a target for novel vector control strategies like paratransgenesis. However, the microbiota in Anopheles funestus, a major African malaria vector, is poorly characterized. Thus, the study aimed to investigate the overall bacterial landscape in the salivary glands, ovaries and midguts of three laboratory strains of An. funestus differing in insecticide-resistant phenotype by sequencing the V3-V4 hypervariable region of bacterial 16S rRNA genes. When examining alpha diversity, the salivary glands harbored significantly more bacteria in terms of species richness and evenness compared to ovaries and midguts. On the strain level, the insecticide-susceptible FANG strain had significantly lower bacterial diversity than the insecticide-resistant FUMOZ and FUMOZ-R strains. When looking at beta diversity, the compositions of microbiota between the three tissues as well as between the strains were statistically different. While there were common bacteria across all three tissues and strains of interest, each tissue and strain did exhibit differentially abundant bacterial genera. However, overall, the top five most abundant genera across all tissues and strains were Elizabethkingia, Acinetobacter, Aeromonas, Cedecea and Yersinia. The presence of shared microbiota suggests a core microbiota that could be exploited for paratransgenesis efforts.