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Supplemental Table 3: Differential gene expression analysis between B16 KO and WT tumors in vivo at day 6. Showing genes with q < 0.05 and log2FC > 1 or < -1.

Supplementary Figure 1: Impact of PAK4 deletion on the expression of chemokines.

Supplementary Figure 8: PAK4 KO in human melanoma cells impairs B-catenin/WNT signalling.

Supplementary Figure 4: PAK4 deletion does not affect MHC-I and II surface expression.

Supplemental Table 6: List of extracellular matrix genes enriched in PAK4 KO anti-PD-1 treated samples.

Supplementary Figure 2: PAK4 deletion does not affect CCL21 secretion.

Supplementary Figure 3: No difference in signalling through different stimuli between PAK4 KO and WT cells.

Supplemental Table 4: Differential gene expression analysis between B16 PAK4 KO anti-PD-1 vs PAK4 KO isotype tumors in vivo at day 10. Showing genes with q < 0.05 and log2FC > 2 or < -2.

Supplemental Table 2: Differential gene expression analysis between B16 KO and WT cells in vitro. Showing genes with q < 0.05 and log2FC > 1 or < -1.

Supplementary Table 1: Panel of immune markers for dendritic cell characterization.

Supplementary Figure 6: Spatial colocalization of CD8 and CD31 positive cells.

Supplemental Table 5: Differential gene expression analysis between B16 PAK4 KO anti-PD-1 vs WT anti-PD-1 tumors in vivo at day 10. Showing genes with q < 0.05 and log2FC > 1 or < -1.

Supplementary Figure 9: PAK4 inhibitor, A0317859, also synergizes with PD-1 blockade in YUMM2.1 melanoma cells.

Supplementary Figure 7: PAK4 protein expression levels in the different cell lines.

Supplementary Figure 5: PAK4 deletion increases CXCL10 expression in vivo.

PAK4 inhibition can sensitize tumors to immune checkpoint blockade (ICB) therapy; however, the underlying mechanisms remain unclear. We report that PAK4 inhibition reverses immune cell exclusion by increasing the infiltration of CD8 T cells and CD103+ dendritic cells (DC), a specific type of DCs that excel at cross-presenting tumor antigens and constitute a source of CXCL10. Interestingly, in melanoma clinical datasets, PAK4 expression levels negatively correlate with the presence of CCL21, the ligand for CCR7 expressed in CD103+ DCs. Furthermore, we extensively characterized the transcriptome of PAK4 knockout (KO) tumors, in vitro and in vivo, and established the importance of PAK4 expression in the regulation of the extracellular matrix, which can facilitate immune cell infiltration. Comparison between PAK4 wild type and KO anti-PD-1 treated tumors revealed how PAK4 deletion sensitizes tumors to ICB from a transcriptomic perspective. In addition, we validated genetically and pharmacologically that inhibition of PAK4 kinase activity is sufficient to improve antitumor efficacy of anti-PD-1 blockade in multiple melanoma mouse models. Therefore, this study provides novel insights into the mechanism of action of PAK4 inhibition and provides the foundation for a new treatment strategy that aims to overcome resistance to PD-1 blockade by combining anti-PD-1 with a small-molecule PAK4 kinase inhibitor. Significance: Our findings provide new insights into PAK4 inhibition mechanism of action as well as the scientific foundation for specifically blocking PAK4 kinase activity using a novel and specific PAK4 kinase inhibitor to overcome resistance to PD-1 blockade.

We describe a case of mild lower urinary tract symptoms and microscopic hematuria in a 53-year-old-male with hypertension found to have urethral stricture disease suspicious for urothelial carcinoma. During the investigation, cystoscopy and biopsy demonstrated eosinophilic amyloid proteins consistent with primary localized urethral amyloidosis. No systemic evidence of amyloidosis was demonstrated. Following a trial of conservative management with serial dilatations, the patient elected to proceed with surgical management by anterior urethroplasty using an excision and primary anastomosis technique. The patient has done well with resolution of his symptoms and no further recurrence of urethral amyloid disease at ongoing follow up.

Carbon dioxide (CO2) emissions can be reduced through newer powertrain concepts with electrification, efficiency optimization, and through alternative fuels. The focus of this work is on alternative fuels and optimizing engine efficiency for selected fuels on a large bore engine (> 170 mm). We surveyed alternative fuels with a lower carbon to hydrogen ratio than diesel fuel, or fuels without carbon. Examples of such fuels are: natural gas (including liquified natural gas (LNG) and its variability), methanol, ethanol, liquefied petroleum gas, ammonia, and hydrogen. The knock limited loads and possible efficiency targets were estimated using a closed cycle simulation with chemical kinetics. The achievable power densities and emitted CO2 levels are presented. In addition to simulation, single cylinder test results are presented for natural gas fuels. The total cost of ownership (TCO) analysis focused on a high fuel use application, in this case, stationary electric power generation. Natural gas is a good alternative fuel for heavy duty applications based on cost and availability.

Urethrocutaneous fistula (UCF) development following primary hypospadias repair is a common complication with high rates of recurrence despite attempts at repair. A novel technique for the management of these fistulae, the PATIO (preserve the tract and turn it inside out) repair, has been described and has shown encouraging outcomes in previous reports.The aim of this study was to evaluate fistula repair outcomes in patients undergoing the PATIO technique compared with standard repair.A retrospective chart-based review was performed for pediatric patients undergoing UCF repair from January 2005 to July 2018. Data including: age, follow-up, meatal location, meatal stenosis, number of fistulae and repairs, UCF location, complications, and outcomes was obtained. Cases were categorized into PATIO repair, standard repair, and PATIO repair following prior standard repair. Surgical outcome with respect to freedom from fistula recurrence was determined.In total, 586 patients underwent hypospadias surgery with 44 patients developing 52 UCF cases that required repair during the study period for a fistula rate of 8.9%. Mean age at repair was 19 months. Median follow-up time was 28 months. For PATIO repair alone, 21/26 (81%) had success. For standard repair alone, 8/18 (44%) had success and for standard repair followed by PATIO repair, 8/8 (100%) were successful. A statistically significant difference was noted for success when comparing standard repair with PATIO repair (p = 0.023, p 0.05) and PATIO repair following standard repair (p = 0.010, p 0.05). There was a statistically non-significant difference between PATIO repair and PATIO repair following standard repair (p = 0.309, p 0.05). Failure following PATIO repair was found in cases where the procedure was early in implementation and experience was limited.UCF repair using the PATIO technique has shown encouraging results in the short-term, with a majority of patients achieving a successful outcome compared with standards techniques. As this procedure continues to be used and experience develops, a larger sample of cases will become available for analysis and longer follow-up will prove necessary in examining the long-term outcomes of this procedure. The outcomes examined have demonstrated consistency with previously reported outcomes in the literature. Limitations include small sample size, short-term follow up, and the retrospective nature of the review.The findings of this study have provided further support to the use of this technique in conventional UCF repair as a means to decrease the risk of recurrence and provide durable results in the short-term. Ongoing follow up will prove necessary to examine success in the long-term.

National joint replacement registries outside North America have been effective in reducing revision risk. However, there is little information on the role of smaller regional registries similar to those found in Canada or the United States. We sought to understand trends in total hip (THA) and knee (TKA) arthroplasty revision patterns after implementation of a regional registry.We reviewed our regional joint replacement registry containing all 30 252 cases of primary and revision THA and TKA performed between Jan. 1, 2005, and Dec. 31, 2013. Each revision case was stratified into early (2 yr), mid (2-10 yr) or late (10 yr), and we determined the primary reason for revision.The early revision rate for TKA dropped from 3.0% in 2005 to 1.3% in 2011 (R(2) = 0.84, p = 0.003). Similarly, the early revision rate for THA dropped from 4.2% to 2.1% (R(2) = 0.78, p = 0.008). Despite primary TKA and THA volumes increasing by 35.5% and 39.5%, respectively, there was no concomitant rise in revision volumes. The leading reasons for TKA revision were infection, instability, aseptic loosening and stiffness. The leading reasons for THA revision were infection, instability, aseptic loosening and periprosthetic fracture. There were no discernible trends over time in reasons for early, mid-term or late revision for either TKA or THA.After implementation of a regional joint replacement registry we observed a significant reduction in early revision rates. Further work investigating the mechanism by which registry reporting reduces early revision risk is warranted.Ailleurs qu'en Amérique du Nord, les registres nationaux des remplacements articulaires ont été efficaces pour réduire le risque de révision. Cependant, il y a peu d'information sur le rôle des plus petits registres régionaux comme ceux qu'on trouve au Canada et aux États-Unis. Nous avons donc cherché à comprendre les tendances en matière de révision des arthroplasties totales de la hanche (ATH) et du genou (ATG) après la création d'un registre régional.Nous avons passé en revue notre registre régional des remplacements articulaires, qui contient les 30 252 ATH et ATG primaires et de révision effectuées entre le 1er janvier 2005 et le 31 décembre 2013. Chaque cas de révision a été classé précoce (2 ans), moyen (de 2 à 10 ans) ou tardif (10 ans), et nous avons déterminé la raison principale de la révision.Le taux de révision précoce pour l'ATG a diminué de 3,0 % en 2005 à 1,3 % en 2011 (Nous avons observé une baisse significative des taux de révision précoce après la mise en œuvre d'un registre régional des remplacements articulaires. Il serait pertinent d'étudier plus en profondeur le mécanisme par lequel le signalement dans un registre réduit le risque de révision précoce.

PurposeThe purpose of this paper is to provide details on implementation of inexpensive and accurate automation solution for performing brisket cutting while the sheep/lamb is hanging by forelegs.Design/methodology/approachThe system developed uses 6 Degree of freedom (DOF) industrial manipulator, custom build tool, sterilizer, and other off the shelf products to create a machine, all the parameters in the system are designed to ensure that the highest levels of safety and hygiene standards are met. The primary system has been developed for sheep/lamb.FindingsIt is shown that the system developed enhanced both the productivity and quality of the process, while adhering to the hygiene standard in the meat processing plants.Research limitations/implicationsThe process of precise brisket can be very challenging; the system designed uses customised tools to provide a good model for the trajectory planning.Originality/valueThe system developed uses statistical tools to calculate the correct trajectories.

Although energy efficiency implementation is increasingly being recognized by policymakers worldwide as one of the most effective means to mitigating rising energy prices, tackling potential environmental risks, and enhancing energy security, mainstreaming its financing in developing country markets continues to be a challenge. Experience shows that converting cost-effective energy savings potential, particularly the demand-side improvement opportunities across sectors, into investments face many barriers and unforeseen transaction costs. This paper draws upon selected experiences with financing energy efficiency in developing countries to explore the key factors of various programmatic approaches and financing instruments that have been applied successfully for delivering energy efficiency solutions. Through case studies, a diverse range of institutional issues are examined related to the identification, packaging, designing, and monitoring approaches that have been used to catalyze traditional and innovative financing of energy efficiency projects. With adequate liquidity in major developing country markets and availability of modern energy savings technologies, it is often the institutional issues that become a key challenge to address in order to finance and implement robust programs. As further operational experience is gained, increased knowledge sharing can lead to scaling-up of such energy efficiency investments. The paper concludes with some ideas for accelerating implementation.

The maize Golden2-like or GLK genes encode proteins belonging to a class of GARP domain transcriptional activators. GLKs have been observed to be involved in chloroplast development and stress adaptation. In spite of the importance of GLKs in plant processes, the in planta targets of GLK regulated transcription, however, have not been identified. We use GLK1 overexpression (OE) to study reprogramming of gene expression networks by GLK1 in Arabidopsis and to identify an associated phenotype. GLK1 OE in Arabidopsis confers resistance to Fusarium graminearum, a broad host pathogen responsible for major losses in cereal crops. Affymetrix Gene Chip and RT-PCR analyses indicated that GLK1 OE in Arabidopsis reprogrammed gene expression networks to enhance a high constitutive expression of genes encoding proteins involved in basal defences but downregulated genes that are associated with programmed cell death and hypersensitive responses. The nature of the role of GLK1 overexpression in selectively reprogramming defence gene networks could explain its effectiveness in enhancing the development of resistance to the necrotrophic phase of F. graminearum pathogenesis.

In silico methods and array technologies have identified genes differentially expressed in prostate cancer. Biological functions of the identified genes are often unclear. Considering the biological significance of androgens in prostate cancer, we profiled the prostate transcripts of congenital androgen-deficient mice with or without androgen replacement in vivo using murine gene expression array. In parallel genes differentially expressed in human prostate cancer were identified by Digital Differential Display and the Serial Analysis of Gene Expression. Androgen dependence of the identified genes was then determined by the steady-state mRNA levels of the murine orthologs in response to androgen treatment. The annotation is supported by the finding that some of the androgen target genes have been reported previously with independent experiments.

BACKGROUND: To report the clinicopathologic features of a fibrocellular membrane in a pseudophakic eye with retained silicone oil in the absence of any capsular contraction syndrome, necessitating repeat anterior capsulorhexis. METHODS: Clinicopathologic report of a case. RESULTS: Histopathologic study of the membrane showed absence of any true periodic acid Schiff-positive capsule within the specimen. Clinically, there was no evidence of any anterior capsular contraction. The fibrocellular membrane may have occurred as a result of inflammatory and mechanical effects of silicone oil in the anterior chamber or as a result of the intraocular lens design. CONCLUSION: The authors report the clinicopathologic features of a fibrocellular membrane occluding the anterior capsular opening in a pseudophakic eye with retained silicone oil, in the absence of any capsular contraction. The stimulus for its formation may have been the inflammatory and mechanical effects of retained silicone oil in the anterior chamber.

Four orthologues of the Arabidopsis CBF/Dreb transcriptional activator genes were isolated from the winter Brassica napus, cv. Jet neuf. All four BNCBF clones encode a putative DRE/CRT (LTRE)-binding protein with an AP2 DNA-binding domain, a putative nuclear localization signal and a possible acidic activation domain. Deduced amino acid sequences suggested that BNCBFs 5, 7and 16 are very similar to the Arabidopsis CBFI whereas BNCBF17 is different in that it contains two extra regions of 16 and 21 amino acids in the acidic domain. Transcripts hybridizing specifically to BNCBF17 and to one or more of the other BNCBFs accumulated in leaves within 30 min of cold exposure of the Brassica seedlings and preceded transcript accumulation of the cold-inducible BN28 gene, a Brassica orthologue of the cor6.6 or KIN gene from Arabidopsis. Cold-induced accumulation of BNCBF17 mRNA was rapid but was short-lived compared to transcripts hybridizing to BNCBF5/7/16. Transcripts hybridizing to one or more of BNCBF5/7/16 accumulated at low levels after the plants were subjected to prolonged exposure to salt stress. BNCBF17 was not responsive to salt stress. BNCBF transcript accumulation was similar in both spring and winter Brassica but the persistence of the transcripts in the cold were generally shorter in the spring than in the winter type. BNCBF5 and 17 proteins bind in vitro to the LTRE domains of the cold-inducible BN115 (cor15a orthologue) or BN28 promoters. Differential binding preferences, however, to LTREs between BNI 15 and BN28 were observed. Mutation of the core CCGAC sequence of the LTRE indicated that BNCBF17 had a lower sequence binding specificity than BNCBF5. Furthermore, experiments indicated that the LTREs were able to drive BNCBF5 and 17 trans-activation of the Lac-Z reporter gene in yeast. We conclude that the BNCBFs reported here could function as trans-acting factors in low-temperature responses in Brassica, controlling the expression of cold-induced genes through an ABA-independent pathway.

PURPOSE The purpose of this study was to describe a patient with an untreated rhegmatogenous retinal detachment, first diagnosed during pregnancy, that underwent spontaneous resolution after normal labor and childbirth. METHODS Descriptive case report. RESULTS AND DISCUSSION It has been extensively documented that normal labor and childbirth are safe in the context of rhegmatogenous retinal detachment. Although exudative retinal detachment is a well-known complication of preeclampsia, spontaneous reattachment of a rhegmatogenous retinal detachment is a rare phenomenon. In this case, labor-induced physiologic changes may have contributed to the resolution of the detachment.

A robust phenotypic plasticity to ward off adverse environmental conditions determines performance and productivity in crop plants. Flax (linseed), is an important cash crop produced for natural textile fiber (linen) or oilseed with many health promoting products. This crop is prone to drought stress and yield losses in many parts of the world. Despite recent advances in drought research in a number of important crops, related progress in flax is very limited. Since, response of this plant to drought stress has not been addressed at the molecular level; we conducted microarray analysis to capture transcriptome associated with induced drought in flax. This study identified 183 differentially expressed genes (DEGs) associated with diverse cellular, biophysical and metabolic programs in flax. The analysis also revealed especially the altered regulation of cellular and metabolic pathways governing photosynthesis. Additionally, comparative transcriptome analysis identified a plethora of genes that displayed differential regulation both spatially and temporally. These results revealed co-regulated expression of 26 genes in both shoot and root tissues with implications for drought stress response. Furthermore, the data also showed that more genes are upregulated in roots compared to shoots, suggesting that roots may play important and additional roles in response to drought in flax. With prolonged drought treatment, the number of DEGs increased in both tissue types. Differential expression of selected genes was confirmed by qRT-PCR, thus supporting the suggested functional association of these intrinsic genes in maintaining growth and homeostasis in response to imminent drought stress in flax. Together the present study has developed foundational and new transcriptome data sets for drought stress in flax.