Farnesyl diphosphate synthase exacerbates nonalcoholic steatohepatitis via the activation of AHR-CD36 axis.

Nonalcoholic steatohepatitis (NASH) has become a major concern that threatens human health worldwide. The underlying pathogenesis was crucial but remained poorly understood. Here, we found that the expression of hepatic farnesyl diphosphate synthase (FDPS) was increased in mice and patients with NASH. Elevated FDPS levels were positively correlated with NASH severity. Overexpression of FDPS in mice provoked increased lipid accumulation, inflammation, and fibrosis, while hepatic FDPS deficiency protected mice from NASH progression. Importantly, pharmacological inhibition of FDPS with clinically used alendronate remarkably attenuated NASH-associated phenotypes in mice. Mechanistically, we demonstrated that FDPS increased its downstream product farnesyl pyrophosphate levels, which could function as an aryl hydrocarbon receptor (AHR) agonist to upregulate the expression of fatty acid translocase CD36, to accelerate the development of NASH. Collectively, these findings suggest that FDPS exacerbates NASH via AHR-CD36 axis and identify FDPS as a promising target for NASH therapy.

Farnesyl Diphosphate Synthase Gene Associated with Loss of Bone Mass Density and Alendronate Treatment Failure in Patients with Primary Osteoporosis.

Aminobisphosphonates (NBPs) are the first-choice medication for osteoporosis (OP); NBP treatment aims at increasing bone mineral density (BMD) by inhibiting the activity of farnesyl diphosphate synthase (FDPS) enzyme in osteoclasts. Despite its efficacy, inadequate response to the drug and side effects have been reported. The A allele of the rs2297480 (A > C) SNP, found in the regulatory region of the FDPS gene, is associated with reduced gene transcription. This study evaluates the FDPS variant rs2297480 (A > C) association with OP patients' response to alendronate sodium treatment. A total of 304 OP patients and 112 controls were enrolled; patients treated with alendronate sodium for two years were classified, according to BMD variations at specific regions (lumbar spine (L1-L4), femoral neck (FN) and total hip (TH), as responders (OP-R) (n = 20) and non-responders (OP-NR) (n = 40). We observed an association of CC genotype with treatment failure (p = 0.045), followed by a BMD decrease in the regions L1-L4 (CC = -2.21% ± 2.56; p = 0.026) and TH (CC = -2.06% ± 1.84; p = 0.015) after two years of alendronate sodium treatment. Relative expression of the FDPS gene was also evaluated in OP-R and OP-NR patients. Higher expression of the FDPS gene was also observed in OP-NR group (FC = 1.84 ± 0.77; p = 0.006) when compared to OP-R. In conclusion, the influence observed of FDPS expression and the rs2897480 variant on alendronate treatment highlights the importance of a genetic approach to improve the efficacy of treatment for primary osteoporosis.

miR-128-3p inhibits intramuscular adipocytes differentiation in chickens by downregulating FDPS.

BACKGROUND: Intramuscular fat (IMF) content is the major indicator for evaluating chicken meat quality due to its positive correlation with tenderness, juiciness, and flavor. An increasing number of studies are focusing on the functions of microRNAs (miRNAs) in intramuscular adipocyte differentiation. However, little is known about the association of miR-128-3p with intramuscular adipocyte differentiation. Our previous RNA-seq results indicated that miR-128-3p was differentially expressed at different periods in chicken intramuscular adipocytes, revealing a possible association with intramuscular adipogenesis. The purpose of this research was to investigate the biological functions and regulatory mechanism of miR-128-3p in chicken intramuscular adipogenesis. RESULTS: The results of a series of assays confirmed that miR-128-3p could promote the proliferation and inhibit the differentiation of intramuscular adipocytes. A total of 223 and 1,050 differentially expressed genes (DEGs) were identified in the mimic treatment group and inhibitor treatment group, respectively, compared with the control group. Functional enrichment analysis revealed that the DEGs were involved in lipid metabolism-related pathways, such as the MAPK and TGF-ß signaling pathways. Furthermore, target gene prediction analysis showed that miR-128-3p can target many of the DEGs, such as FDPS, GGT5, TMEM37, and ASL2. The luciferase assay results showed that miR-128-3p targeted the 3' UTR of FDPS. The results of subsequent functional assays demonstrated that miR-128-3p acted as an inhibitor of intramuscular adipocyte differentiation by targeting FDPS. CONCLUSION: miR-128-3p inhibits chicken intramuscular adipocyte differentiation by downregulating FDPS. Our findings provide a theoretical basis for the study of lipid metabolism and reveal a potential target for molecular breeding to improve meat quality.

Effects of fermented dairy products on inflammatory biomarkers: A meta-analysis.

AIM: Fermented dairy products (FDPs) are made from raw milk under the action of specific microorganisms by lactic acid bacteria fermentation or co-fermentation of lactic acid bacteria, bifidobacteria, and yeast. The aim of this study was to explore the effects of FDPs on inflammatory biomarkers. DATA SYNTHESIS: A comprehensive search was conducted on four electronic databases, including PubMed, Web of Science, Embase, and the Cochrane Library. Finally, fourteen trials (15 arms) were included in this meta-analysis: yogurt (n = 9), fermented milk (n = 4), and kefir (n = 2). Additionally, the random effects model or fixed-effects model was used to pool the study results. Firstly, the analysis indicated that FDPs' supplementation decreased the levels of C-reactive protein (CRP) (SMD = -0.21; 95% CI: -0.40, -0.02; P = 0.033) and increased interferon-gamma (IFN-γ) levels (SMD = 0.12; 95% CI: 0.01, 0.23; P = 0.033). Furthermore, we obtained some statistically significant results in the following subgroups: CRP decreased in participants with metabolic diseases. IFN-γ increased in the intervention that lasted ≥12 weeks, Asian, yogurt, and healthy population. Finally, there was no significant effect on tumor necrosis factor-alpha, interleukin (IL)-6, IL-10, and IL-2. CONCLUSIONS: FDPs reduced CRP and increased IFN-γ, but they had no effect on other inflammatory markers. The results showed that the consumption of FDPs was slightly associated with reduced inflammation, but because of the limited literature, these results should be interpreted with caution.

The Effect of Preceramic Soldering on Fracture Resistance of 4-Unit Zirconia Fixed Dental Prostheses.

PURPOSE: Preceramic soldering of zirconia may deliver better fitting restorations. However, there is not sufficient evidence regarding the influence of preceramic soldering of zirconia restorations on mechanical strength. The aim of this study was to evaluate the effect of preceramic soldering on the fracture load of 4-unit zirconia fixed dental prostheses (FDPs). MATERIALS AND METHODS: Eighty samples of 4-unit FDPs between maxillary right first premolar and maxillary right second molar were prepared and two restorative materials were used as a framework (Z) and monolithic restoration (M). The samples were divided into two subgroups as control (C) and study (S). The restorations of study groups (S) were divided into two pieces and soldered with a bonding material (DCM HotBond Zirkon). The groups were divided into two subgroups for thermal cycle (T) application. After soldering and thermal cycling application, 4-point bending test was applied to the samples at a cross-head speed of 1 mm/min in a universal testing machine and the fracture load was recorded. The data was analyzed statistically, and the level of significance was set at α = 0.001. RESULTS: Statistically significant differences were found among the groups, based on the results of maximum failure loads (p < 0.001). The highest mean failure load was observed in the ZCT(-) group (1094.1 + 139.77 N), while the lowest mean failure load was obtained in the ZST(+) group (627.7 + 82.14 N). No significant difference was found among the groups MC and MS, MC, and ZC groups (p > 0.001). Thermal aging application caused lower fracture resistance in control and soldering groups (p < 0.001). CONCLUSIONS: The preceramic soldering applications affected zirconia group negatively. However, the values were above the clinically acceptable static load bearing capacity for posterior teeth. Soldering did not cause a statistically significant difference for the fracture strengths of monolithic zirconia groups. Thermal cycling affected the fracture strength of zirconia and monolithic zirconia restorations negatively.

Genome-wide identification and functional characterization of the PheE2F/DP gene family in Moso bamboo.

BACKGROUND: E2F/DP proteins have been shown to regulate genes implicated in cell cycle control and DNA repair. However, to date, research into the potential role of the Moso bamboo E2F/DP family has been limited. RESULTS: Here, we identified 23 E2F/DPs in the Moso bamboo genome, including nine E2F genes, six DP genes, eight DEL genes and one gene with a partial E2F domain. An estimation of the divergence time of the paralogous gene pairs suggested that the E2F/DP family expansion primarily occurred through a whole-genome duplication event. A regulatory element and coexpression network analysis indicated that E2F/DP regulated the expression of cell cycle-related genes. A yeast two-hybrid assay and expression analysis based on transcriptome data and in situ hybridization indicated that the PheE2F-PheDP complex played important roles in winter Moso bamboo shoot growth. The qRT-PCR results showed that the PheE2F/DPs exhibited diverse expression patterns in response to drought and salt treatment and diurnal cycles. CONCLUSION: Our findings provide novel insights into the Moso bamboo E2F/DP family and partial experimental evidence for further functional verification of the PheE2F/DPs.

Impact of timing of dental implant placement and loading: Summary and consensus statements of group 1-The 6th EAO Consensus Conference 2021.

OBJECTIVES: This publication reports on the EAO workshop group 1 summaries, discussions and consensus statements based on four systematic reviews evaluating the impact of timing of dental implant placement and loading. MATERIALS AND METHODS: The first of the systematic reviews was on the influence of the timing of implant placement and loading in the biological outcomes of implant-supported fixed partial dentures. The second systematic review evaluated the influence of the timing of implant placement and loading on the aesthetic outcomes in single-tooth implants. The third systematic review was on the long-term outcomes of maxillary single-tooth implants in relation to timing protocols of implant placement and loading and the fourth on patient's perception of timing concepts in implant dentistry. The group evaluated these systematic reviews, provided comments and additions as required and agreed on the relevant consensus statements as well as on clinical and research recommendations. RESULTS: Different timings of implant placement/loading presented with high implant survival rates. The systematic reviews evaluated from this working group provided a number of conclusions based on the available/current literature. However, the specific topic of timing is an area that further research is required in order to provide detailed guidelines for the different protocols to be employed.

FDPS promotes glioma growth and macrophage recruitment by regulating CCL20 via Wnt/ß-catenin signalling pathway.

Glioma is one of the most lethal tumours and common malignant in the central nervous system (CNS), which exhibits diffuse invasion and aggressive growth. Several studies have reported the association of FDPS to tumour development and progression. However, the role of FDPS in progression of glioma and macrophage recruitment is not well-elucidated. In the current study, a remarkable enhancement in FDPS level was observed in glioma tissues and associated with poor prognosis, contributed to tumour growth. FDPS was correlated with macrophage infiltration in glioma and pharmacological deletion of macrophages largely abrogated the oncogenic functions of FDPS in glioma. Mechanistically, FDPS activated Wnt/ß-catenin signalling pathway and ultimately facilitates macrophage infiltration by inducing CCL20 expression. In conclusion, overexpressed FDPS exhibits an immunomodulatory role in glioma. Therefore, targeting FDPS may be an effective therapeutic strategy for glioma.

Biological and mechanical complications of angulated abutments connected to fixed dental prostheses: A systematic review with meta-analysis.

OBJECTIVES: To evaluate the biological and mechanical complications of angulated abutments on full-arch and partial jaw rehabilitations with a follow-up for at least 1 year. METHODS: Electronic search was carried out in MEDLINE, EMBASE and Web of Science. Studies published between January 2000 and January 2019 were included. The quality of the included studies was assessed. The data extraction was focused on implant loss, marginal bone loss and mechanical complications, and meta-analyses were performed for marginal bone loss, mechanical complications and implant failure. RESULTS: Nine studies, three prospective and six retrospective cohort studies were included. They reported on 797 patients that received 4127 implants. The total number of abutments was 4079 of which 1673 were angulated, and 2406 were straight. All abutments were prefabricated. Angulated abutments were associated with increased implant failure rates (two studies; RR = 7.30; 95% CI = 2.79-19.08) and an effect that was both statistically significant (P < .001) and clinically relevant. Three studies reported differentiated data for mechanical and technical complications at 1 year of follow-up, being mostly related to the retention screw while screw fracture. Angulated abutments were associated with a statistically significant increase in MBL 1 year after insertion compared to straight abutments (three studies; MD = 0.08 mm; 95% CI = 0.01-0.14 mm; P = .02), which might be, however, clinically negligible. CONCLUSIONS: The prosthetic complications such as screw loosening and abutment loosening were frequent. After 1 year of follow-up, implants supporting angulated abutments yielded significantly more marginal bone loss than those supporting straight abutments.

Fracture behavior of all-ceramic, implant-supported, and tooth-implant-supported fixed dental prostheses.

OBJECTIVES: In vitro investigation of the effects of fixed dental prosthesis (FDP) support and loading conditions on the fracture behavior of all-ceramic, zirconia-based FDP veneered with computer-aided design/computer-aided manufacturing (CAD/CAM)-manufactured lithium disilicate ceramic. MATERIALS AND METHODS: Based on a model for a 3-unit FDP in the molar region (tooth in region 15, implant in region 17), 16 identical zirconia frameworks were fabricated and veneered with milled lithium disilicate ceramic. Another 16 FDPs were manufactured similarly, using a model in which the tooth was replaced by an implant. The specimens underwent 10,000 thermal cycles between 6.5 and 60 °C and 1,200,000 chewing cycles with a force magnitude of 100 N. All were then subsequently loaded until fracture in a universal testing device. Half of the FDPs were subjected to centric and axial loading on the pontic, the others to eccentric and oblique loading on one cusp of the pontic. RESULTS: No failures were observed after artificial aging. Fracture loads of tooth-implant-supported restorations were 1636 ± 158 and 1086 ± 156 N for axial and oblique loading, respectively; implant-supported FDPs fractured at 1789 ± 202 and 1200 ± 68 N, respectively. Differences were significant for load application (P < 0.001) and support type (P = 0.020). For the two types of load application, fracture mode differed substantially: complete fracture was observed for centric and axial loading whereas mixed cohesive/adhesive failure was observed for many FDPs loaded eccentrically and obliquely. CONCLUSIONS: The high incidence of chipping of manually veneered implant-supported all-ceramics restorations might be reduced by use of CAD/CAM-manufactured lithium disilicate veneers. CLINICAL RELEVANCE: FDPs veneered with lithium disilicate resist occlusal forces of 500 N, irrespective of load application and support type. The fracture resistance of implant-supported FDPs was, however, higher than that of combined tooth-implant-supported FDPs. Their clinical use seems to be justified.

Fracture Load Before and After Veneering Zirconia Posterior Fixed Dental Prostheses.

PURPOSE: To evaluate the fracture load of 3-unit zirconia-based posterior fixed dental prostheses (FDPs) before and after veneering the frameworks. MATERIALS AND METHODS: Forty standardized stainless-steel master dies were fabricated (height: 5 mm, convergence: 6º, chamfer: 1 mm) and randomly screwed in pairs onto metal bases. The bases were randomly divided into two groups (n = 20 each) according to the zirconia CAD/CAM system used for constructing 3-unit structures for FDPs: group 1 (L): Lava All-ceramic, group 2 (Z): IPS e.max ZirCAD. Half of the zirconia structures per group were randomly selected and veneered, while the remaining half was left unveneered. The specimens were luted in standard fashion onto the stainless steel master dies using conventional glass ionomer cement. All specimens were tested for fracture load (FL). Specimens were subjected to a three-point bending test until fracture by applying an axial compressive load at the central fossa of the pontics with a universal testing machine at a 0.5 mm/min crosshead speed. Wilcoxon's rank-sum test and Weibull statistics were used for statistical analysis (α = 0.05). RESULTS: L structures recorded significantly higher values of load to fracture than the Z group both before and after veneering. Within each ceramic group, no differences were found between unveneered and veneered frameworks. CONCLUSIONS: Although further studies are necessary to corroborate these findings, both zirconia systems could be recommended for restoring posterior teeth on the basis of the fracture load values recorded in this experiment (>1000 N). The veneering procedure did not affect the overall load to fracture in any group.

Preservation of Hypermobile Teeth by Establishing Posterior Occlusal Support Using Implant Prostheses: A 5-Year Follow-Up.

For patients with periodontally compromised, hypermobile teeth, implant-supported fixed dental prostheses (FDPs) or removable dentures are often used after extracting mobile teeth. The loss of native teeth may carry social consequences, depending upon the patient's age, state of health, and degree of social functioning. This report represents successful stabilization and preservation of questionable, hypermobile teeth that have been damaged by traumatic occlusion due to the loss of posterior support with a cross-arch splinted FDP, as well as the implementation of posterior support using implant-supported prostheses.

Two Novel and Three Recurrent Mutations in the Mevalonate Pathway Genes in Chinese Patients with Porokeratosis.

Purpose: Porokeratosis (PK) is a chronic autosomal-dominant cutaneous keratinization disorder exhibiting clinical and genetic heterogeneity. Mevalonate decarboxylase (MVD), farnesyl diphosphate synthase (FDPS), phosphomevalonate kinase(PMVK), and mevalonate kinase genes(MVK), which encode the mevalonate pathway, are disease-causing genes in PK. Patients and Methods: Data and blood samples were collected from two Chinese families and five sporadic patients with porokeratosis. Whole-exome and Sanger sequencing were performed to detect pathogenic gene mutation in the patients. Results: Five heterozygous mutations were identified, including a novel FDPS stop-gain mutation c.438T>G (p.Tyr146Ter), a novel MVD missense mutation c.683G>C (p.R228P), and three previously reported MVD mutations: c.746T>C (p.F249S), c.875A>G (p.N292S), and c.1111_1113del (p.371_371del). The novel FDPS c.438T>G mutation was predicted as "disease-causing" (p = 1) by Mutation Taster. The other novel MVD c.683G>C was also predicted as "deleterious" (score = 0.00) by Sorting Intolerant From Tolerant (SIFT), "probably damaging" (score = 1) by PolyPhen2, and "disease-causing" (p = 0.999) by Mutation Taster. Conclusion: Our results extended the mutation spectrum of mevalonate pathway genes in porokeratosis and provided useful strategies for a more accurate diagnosis and genetic counseling.

The influence of pontic distribution on the marginal and internal gaps of CAD/CAM five-unit anterior zirconia framework.

Background/purpose: Nowadays, zirconia-based framework has been used for longspan or full-arch fixed dental prostheses (FDPs). This study aimed to evaluate the effect of pontic distribution on marginal and internal gaps of five-unit anterior zirconiabased DPs. Materials and methods: Right maxillary central incisor and second premolar were selected as terminal abutments and three different edentulous conditions with one nonterminal abutment were simulated. Marginal and internal gaps in each zirconia-based samples(n = 10) were examined by computer-aided replica technique. Five regions, including marginal gaps at mesial or distal finishing line, internal gaps at the mesial or distal axial wall, and occlusal surface, were statistically analyzed (α = .05). Results: Most of marginal gaps and internal gaps at axial wall were clinically acceptable, but larger at occlusal surface. For the three experimental groups, clinically accepted percentage with qualified gaps were less than 30%.There were statistical differences at axial wall over pontic side and marginal gaps over non-pontic side between groups (P<0.05). For sum of gaps of all abutments in each group, statistical differences were found at marginal and axial wall (P < 0.05). As for those on terminal and non-terminal abutments, statistical differences were found on second premolar (P < 0.05). Conclusion: Except for occlusal surface, the overall marginal gaps and internal gaps at axial wall of five-unit anterior zirconia-based FDPs with different pontic distribution were clinically acceptable. However, the percentage with qualified gaps were low (<30%). Greater gaps were noted when adjacent pontic existed. Different pontic size and distribution with curvature had an influence on the gaps.

Long-term clinical outcome of three-unit fixed-fixed posterior zirconia ceramic inlay-retained FDPs with a modified design.

OBJECTIVES: To assess the long-term clinical outcome of posterior inlay-retained fixed dental prostheses (IRFDPs) with a modified design made from zirconia ceramic. METHODS: In 30 patients thirty 3-unit IRFDPs were placed to replace 7 premolars (4 in the maxilla, 3 in the mandible), and 23 first molars (15 in the maxilla, 8 in the mandible). Preparations were accomplished in agreement with the general principles for ceramic inlay restorations and modified with a short retainer-wing bevel preparation within the enamel at the buccal and oral sides. The frameworks were milled from 3Y-TZP ceramic, and the pontics were veneered with feldspathic ceramic. All IRFDPs were bonded with a phosphate monomer containing luting resin after air-abrasion of the intaglio surfaces. The patients were recalled 6-12 months after placement, and then annually. Kaplan-Meier analysis was used to calculate the survival and complication rates of the IRFDPs. RESULTS: The mean observation time was 10.6 ± 1.5 years. The 10-year cumulative survival rate was 89% with 4 failures, two of them were due to deep secondary caries with loss of retention, one due to repeated debonding with enamel fractures, and one due to generalized progressive periodontitis. The most common complication was chipping of the veneering ceramic (20.1%). Eighteen IRFDPs were free of any type of complication up to 15.4 years, which corresponds to a 10-year cumulative success rate of 70.4%. CONCLUSION: The long-term clinical performance of modified IRFDPs made of veneered zirconia ceramics was favorable after 10 years, therefore, they represent a treatment alternative to replace posterior single missing teeth. CLINICAL SIGNIFICANCE: Zirconia-based IRFDPs fabricated in the modified design may represent a substance-preserving alternative to conventional posterior FDPs to replace posterior single missing teeth, particularly in cases where implants cannot be placed, and when the adjacent teeth already have small restorations or defects.

Zoledronic Acid Targeting of the Mevalonate Pathway Causes Reduced Cell Recruitment and Attenuates Pulmonary Fibrosis.

Background and aim: Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease causing irreparable scarring of lung tissue, with most patients succumbing rapidly after diagnosis. The mevalonate pathway, which is involved in the regulation of cell proliferation, survival, and motility, is targeted by the bisphosphonate zoledronic acid (ZA). The aim of this study was to assess the antifibrotic effects of ZA and to elucidate the mechanisms by which potential IPF treatment occurs. Methods: A series of in vitro and in vivo models were employed to identify the therapeutic potential of ZA in treating IPF. In vitro transwell assays were used to assess the ability of ZA to reduce fibrotic-related immune cell recruitment. Farnesyl diphosphate synthase (FDPS) was screened as a potential antifibrotic target using a bleomycin mouse model. FDPS-targeting siRNA and ZA were administered to mice following the onset of experimentally-induced lung fibrosis. Downstream analyses were conducted on murine lung tissues and lung fluids including 23-plex cytokine array, flow cytometry, histology, Western blotting, immunofluorescent staining, and PCR analysis. Results: In vitro administration of ZA reduced myofibroblast transition and blocked NF-κB signaling in macrophages leading to impaired immune cell recruitment in a transwell assay. FDPS-targeting siRNA administration significantly attenuated profibrotic cytokine production and lung damage in a murine lung fibrosis model. Furthermore, ZA treatment of mice with bleomycin-induced lung damage displayed decreased cytokine levels in the BALF, plasma, and lung tissue, resulting in less histologically visible fibrotic scarring. Bleomycin-induced upregulation of the ZA target, FDPS, was reduced in lung tissue and fibroblasts upon ZA treatment. Confirmatory increases in FDPS immunoreactivity was seen in human IPF resected lung samples compared to control tissue indicating potential translational value of the approach. Additionally, ZA polarized macrophages towards a less profibrotic phenotype contributing to decreased IPF pathogenesis. Conclusion: This study highlights ZA as an expedient and efficacious treatment option against IPF in a clinical setting.

Disruption of FDPS/Rac1 axis radiosensitizes pancreatic ductal adenocarcinoma by attenuating DNA damage response and immunosuppressive signalling.

BACKGROUND: Radiation therapy (RT) has a suboptimal effect in patients with pancreatic ductal adenocarcinoma (PDAC) due to intrinsic and acquired radioresistance (RR). Comprehensive bioinformatics and microarray analysis revealed that cholesterol biosynthesis (CBS) is involved in the RR of PDAC. We now tested the inhibition of the CBS pathway enzyme, farnesyl diphosphate synthase (FDPS), by zoledronic acid (Zol) to enhance radiation and activate immune cells. METHODS: We investigated the role of FDPS in PDAC RR using the following methods: in vitro cell-based assay, immunohistochemistry, immunofluorescence, immunoblot, cell-based cholesterol assay, RNA sequencing, tumouroids (KPC-murine and PDAC patient-derived), orthotopic models, and PDAC patient's clinical study. FINDINGS: FDPS overexpression in PDAC tissues and cells (P < 0.01 and P < 0.05) is associated with poor RT response and survival (P = 0.024). CRISPR/Cas9 and pharmacological inhibition (Zol) of FDPS in human and mouse syngeneic PDAC cells in conjunction with RT conferred higher PDAC radiosensitivity in vitro (P < 0.05, P < 0.01, and P < 0.001) and in vivo (P < 0.05). Interestingly, murine (P = 0.01) and human (P = 0.0159) tumouroids treated with Zol+RT showed a significant growth reduction. Mechanistically, RNA-Seq analysis of the PDAC xenografts and patients-PBMCs revealed that Zol exerts radiosensitization by affecting Rac1 and Rho prenylation, thereby modulating DNA damage and radiation response signalling along with improved systemic immune cells activation. An ongoing phase I/II trial (NCT03073785) showed improved failure-free survival (FFS), enhanced immune cell activation, and decreased microenvironment-related genes upon Zol+RT treatment. INTERPRETATION: Our findings suggest that FDPS is a novel radiosensitization target for PDAC therapy. This study also provides a rationale to utilize Zol as a potential radiosensitizer and as an immunomodulator in PDAC and other cancers. FUNDING: National Institutes of Health (P50, P01, and R01).

Fibrinolysis in Dogs with Intracavitary Effusion: A Review.

Physiologic fibrinolysis is a localized process in which stable fibrin strands are broken down by plasmin in response to thrombosis. Plasmin activation can also take place separately from the coagulation process, resulting in pathologic fibrinolysis. When plasmin activation exceeds the neutralizing capacity of plasmin inhibitors, severe bleeding can potentially take place. Although the processes which regulate coagulation and fibrinolysis in the blood are well known, it is less clear as to what extent the same processes take place in the body cavities and whether they influence systemic hemostasis. The results of the studies herein cited demonstrate that coagulation followed by fibrinogenolytic/fibrinolytic activity takes place in all kinds of canine ascitic and pleural fluids. Moreover, systemic clotting abnormalities suggesting primary fibrinolysis/primary hyperfibrinolysis (i.e., elevated plasma fibrin/fibrinogen degradation products [FDPs] and normal D-dimer concentrations with fibrinogen concentrations ≤ 100 mg/dL or above this cut-off, respectively) occur in dogs with intracavitary effusion. Enhanced fibrinolytic activity in dogs with intracavitary effusion can also be detected using rotational thromboelastometry (ROTEM), although the degree of agreement between ROTEM and FDPs, D-dimer and fibrinogen concentrations is poor. Finally, contrary to the thrombotic events commonly documented in some humans and cats with cardiac diseases, bleeding tendencies due to primary fibrinolysis/primary hyperfibrinolysis have been documented in dogs with cardiogenic ascites.

Reducing farnesyl diphosphate synthase levels activates Vγ9Vδ2 T cells and improves tumor suppression in murine xenograft cancer models.

Human Vγ9Vδ2 T cells are attractive candidates for cancer immunotherapy due to their potent capacity for tumor recognition and cytolysis of many tumor cell types. However, efforts to deploy clinical strategies for Vγ9Vδ2 T cell cancer therapy are hampered by insufficient potency. We are pursuing an alternate strategy of modifying tumors to increase the capacity for Vγ9Vδ2 T cell activation, as a means for strengthening the anti-tumor response by resident or ex vivo manufactured Vγ9Vδ2 T cells. Vγ9Vδ2 T cells are activated in vitro by non-peptidic antigens including isopentenyl pyrophosphate (IPP), a substrate of farnesyl diphosphate synthase (FDPS) in the pathway for biosynthesis of isoprenoids. In an effort to improve in vivo potency of Vγ9Vδ2 T cells, we reduced FDPS expression in tumor cells using a lentivirus vector encoding a short-hairpin RNA that targets FDPS mRNA (LV-shFDPS). Prostate (PC3) or hepatocellular carcinoma (Huh-7) cells transduced with LV-shFDPS induced Vγ9Vδ2 T cell stimulation in vitro, resulting in increased cytokine expression and tumor cell cytotoxicity. Immune deficient mice implanted with LV-shFDPS transduced tumor cells showed dramatic responses to intraperitoneal injection of Vγ9Vδ2 T cells with strong suppression of tumor growth. In vivo potency was increased by transducing tumor cells with a vector expressing both shFDPS and human IL-2. Tumor suppression by Vγ9Vδ2 T cells was dose-dependent with greater effects observed in mice injected with 100% LV-shFDPS transduced cells compared to mice injected with a mixture of 50% LV-shFDPS transduced cells and 50% control (no vector) tumor cells. Delivery of LV-shFDPS by intratumoral injection was insufficient to knockdown FDPS in the majority of tumor cells, resulting in insignificant tumor suppression by Vγ9Vδ2 T cells. Thus, Vγ9Vδ2 T cells efficiently targeted and suppressed tumors expressing shFDPS in mouse xenotransplant models. This proof-of-concept study demonstrates the potential for suppression of genetically modified tumors by human Vγ9Vδ2 T cells and indicates that co-expression of cytokines may boost the anti-tumor effect.

Effect of Different CAD/CAM Milling and 3D Printing Digital Fabrication Techniques on the Accuracy of PMMA Working Models and Vertical Marginal Fit of PMMA Provisional Dental Prosthesis: An In Vitro Study.

Background: Minimal evidence exists on the efficacy of different digital manufacturing techniques in the fabrication of precise dental working models and provisional prosthesis. Aim of study: The objective was to evaluate the effect of two digital fabrication techniques (CAD/CAM milling and 3D printing) on the accuracy of PMMA working models and marginal fit of PMMA provisional prosthesis. Materials and methods: Two abutment teeth of modified typodont were prepared. A reference stone model was fabricated, and an optical impression was performed to obtain a CAD reference model. Four CAM milled working models and four printed working models were fabricated. CAD software was used to design the provisional prostheses. Group A tested four milled provisional prosthesis, and group B tested four 3D printed prosthesis. The 3D accuracy of working models was assessed by superimposition of the control reference working model on the CAD test working model. A stereo-optical microscope was used to assess vertical marginal fit of the provisional dental prosthesis. Student's t and Mann-Whitney U tests were utilized to compare the two groups. Results: Results showed no statistically significant difference between the two tested groups. Conclusion: The two digital working model fabrication techniques recorded comparable accuracy. Similarly, 3D printed provisional prosthesis showed comparable marginal fit to the CAD/CAM milled ones.