A lactate-induced response to hypoxia.

Organisms must be able to respond to low oxygen in a number of homeostatic and pathological contexts. Regulation of hypoxic responses via the hypoxia-inducible factor (HIF) is well established, but evidence indicates that other, HIF-independent mechanisms are also involved. Here, we report a hypoxic response that depends on the accumulation of lactate, a metabolite whose production increases in hypoxic conditions. We find that the NDRG3 protein is degraded in a PHD2/VHL-dependent manner in normoxia but is protected from destruction by binding to lactate that accumulates under hypoxia. The stabilized NDRG3 protein binds c-Raf to mediate hypoxia-induced activation of Raf-ERK pathway, promoting angiogenesis and cell growth. Inhibiting cellular lactate production abolishes the NDRG3-mediated hypoxia responses. Our study, therefore, elucidates the molecular basis for lactate-induced hypoxia signaling, which can be exploited for the development of therapies targeting hypoxia-induced diseases.

Leptin Receptor Deficiency Impairs Lymph Node Development and Adaptive Immune Response.

Activation and clonal expansion of the Ag-specific adaptive immune response in the draining lymph node is essential to clearing influenza A virus infections. Activation sufficient for virus clearance is dependent on the lymph node's architectural organization that is maintained by stromal cells, chiefly fibroblastic reticular cells. During an analysis of influenza A virus clearance in leptin receptor knockout (DB/DB) mice, we observed that the DB/DB mice have markedly reduced numbers of lymph node fibroblastic reticular cells at the steady state. The reduction in lymph node fibroblastic reticular cells resulted in abnormal lymph node organization and diminished numbers of adaptive immune cells in the lymph nodes under homeostatic conditions. As a consequence, the DB/DB mice were impaired in their ability to generate an effective influenza-specific adaptive immune response, which prevented virus clearance. Using leptin receptor mutant mice with point mutations at distinct signaling sites in the leptin receptor, we were able to link the leptin receptor's signaling domain tyrosine 985, which does not contribute to obesity, to lymph node fibroblastic reticular cell development and function. These results demonstrate a novel role for leptin receptor signaling in regulating lymph node development in a manner that is crucial to the generation of Ag-specific adaptive immune responses.

Bone marrow stromal cell antigen-1 deficiency protects from acute kidney injury.

This study aimed to investigate the role of bone marrow stromal cell antigen-1 (Bst1; also known as CD157) in acute kidney injury (AKI). Bst1 is a cell surface molecule with various enzymatic activities and downstream intracellular signaling pathways that modulate the immune response. Previous research has linked Bst1 to diseases such as ovarian cancer, Parkinson's disease, and rheumatoid arthritis. We used bilateral ischemia-reperfusion injury (IRI) as an AKI model and created bone marrow chimeric mice to evaluate the role of Bst1 in bone marrow-derived cells. We also used flow cytometry to identify Bst1/CD157 expression in hematopoietic cells and evaluate immune cell dynamics in the kidney. The findings showed that Bst1-deficient (Bst1-/-) mice were protected against renal bilateral IRI. Bone marrow chimera experiments revealed that Bst1 expression on hematopoietic cells, but not parenchymal cells, induced renal IRI. Bst1 was mainly found in B cells and neutrophils by flow cytometry of the spleen and bone marrow. In vitro, migration of neutrophils from Bst1-/- mice was suppressed, and adoptive transfer of neutrophils from wild-type Bst1+/+ mice abolished the renal protective effect in Bst1 knockout mice. In conclusion, the study demonstrated that Bst1-/- mice are protected against renal IRI and that Bst1 expression in neutrophils plays a crucial role in inducing renal IRI. These findings suggest that targeting Bst1 in neutrophils could be a potential therapeutic strategy for AKI.NEW & NOTEWORTHY Acute kidney injury (AKI), a serious disease for which there is no effective Federal Drug Administration-approved treatment, is associated with high mortality rates. Bone marrow stromal cell antigen-1 (Bst1) is a cell surface molecule that can cause kidney fibrosis, but its role in AKI is largely unknown. Our study showed that Bst1-/- mice revealed a protective effect against renal bilateral ischemia-reperfusion injury (IRI). Adoptive transfer studies confirmed that Bst1 expression in hematopoietic cells, especially neutrophils, contributed to renal bilateral IRI.

CCR2 monocytes as therapeutic targets for acute disc herniation and radiculopathy in mouse models.

OBJECTIVE: Back pain and radiculopathy caused by disc herniation are major health issues worldwide. While macrophages are key players in disc herniation induced inflammation, their roles and origins in disease progression remain unclear. We aim to study the roles of monocytes and derivatives in a mouse model of disc herniation. METHODS: Using a CCR2-CreER; R26R-EGFP (Ai6) transgenic mouse strain, we fate-mapped C-C chemokine receptor type 2 (CCR2) expressing monocytes and derivatives at disc herniation sites, and employed a CCR2RFP/RFP mouse strain and a CCR2-specific antagonist to study the effects of CCR2+ monocytes on local inflammatory responses, pain level, and disc degeneration by immunostaining, flow cytometry, and histology. RESULTS: CCR2+ monocytes (GFP+) increased at the sites of disc hernia over postoperative day 4, 6, and 9 in CCR2-CreER; Ai6 mice. F4/80+ cells increased, and meanwhile, CD11b+ cells trended downward. Co-localization analysis revealed that both GFP+CD11b+ and GFP+F4/80+ constituted the majority of CD11b+ and F4/80+ cells at disc hernia sites. Fluorescence activated cell sorter purified GFP+ cells exhibited higher cytokine expressions than GFP- cells. Inhibition of CCR2 signaling reduced infiltration of monocytes and macrophages, alleviated pain, maintained disc height, and reduced osteoclast activity in adjacent cortical bone for up to 1 month. CONCLUSION: Our findings suggest that circulating CCR2+ monocytes play important roles in initiating and promoting the local inflammatory responses, pain sensitization, and degenerative changes after disc herniation, and thus may serve as therapeutic targets for disc herniation induced back and leg pain.

IL-33 Coordinates Innate Defense to Systemic Candida albicans Infection by Regulating IL-23 and IL-10 in an Opposite Way.

Invasive candidiasis has high mortality rates in immunocompromised patients, causing serious health problems. In mouse models, innate immunity protects the host by rapidly mobilizing a variety of resistance and tolerance mechanisms to systemic Candida albicans infection. We have previously demonstrated that exogenous IL-33 regulates multiple steps of innate immunity involving resistance and tolerance processes. In this study, we systematically analyzed the in vivo functions of endogenous IL-33 using Il33 -/- mice and in vitro immune cell culture. Tubular epithelial cells mainly secreted IL-33 in response to systemic C. albicans infection. Il33 -/- mice showed increased mortality and morbidity, which were due to impaired fungal clearance. IL-33 initiated an innate defense mechanism by costimulating dendritic cells to produce IL-23 after systemic C. albicans infection, which in turn promoted the phagocytosis of neutrophils through secretion of GM-CSF by NK cells. The susceptibility of Il33 -/- mice was also associated with increased levels of IL-10, and neutralization of IL-10 resulted in enhanced fungal clearance in Il33 -/- mice. However, depletion of IL-10 overrode the effect of IL-33 on fungal clearance. In Il10 -/- mouse kidneys, MHC class II+F4/80+ macrophages were massively differentiated after C. albicans infection, and these cells were superior to MHC class II-F4/80+ macrophages that were preferentially differentiated in wild-type mouse kidneys in killing of extracellular hyphal C. albicans Taken together, our results identify IL-33 as critical early regulator controlling a serial downstream signaling events of innate defense to C. albicans infection.

Testosterone promotes the migration, invasion and EMT process of papillary thyroid carcinoma by up-regulating Tnnt1.

PURPOSE: To explore the key genes and molecular pathways in the progression of thyroid papillary carcinoma (PTC) promoted by testosterone using RNA-sequencing technology, and to provide new drug targets for improving the therapeutic effect of PTC. METHODS: Orchiectomy (ORX) was carried out to construct ORX mouse models. TPC-1 cells were subcutaneously injected for PTC formation in mice, and the tumor tissues were collected for RNA-seq. The key genes were screened by bioinformatics technology. Tnnt1 expression in PTC cells was knocked down or overexpressed by transfection. Cell counting kit-8 (CCK-8), colony formation assay, scratch assay and transwell assay were adopted, respectively, for the detection of cell proliferation, colony formation, migration and invasion. Besides, quantification real-time polymerase chain reaction (qRT-PCR) and western blot were utilized to determine the mRNA and protein expression levels of genes in tissues or cells. RESULTS: Both estradiol and testosterone promoted the growth of PTC xenografts. The key gene Tnnt1 was screened and obtained by bioinformatics technology. Functional analysis revealed that overexpression of Tnnt1 could markedly promote the proliferation, colony formation, migration, invasion, and epithelial-to-mesenchymal transition (EMT) process of PTC cells, as well as could activate p38/JNK pathway. In addition, si-Tnt1 was able to inhibit the cancer-promoting effect of testosterone. CONCLUSION: Based on the outcomes of bioinformatics and basic experiments, it is found that testosterone can promote malignant behaviors such as growth, migration, invasion and EMT process of PTC by up-regulating Tnnt1 expression. In addition, the function of testosterone may be achieved by activating p38/JNK signaling pathway.

Computer guided root tip extraction and implant placement: A clinical report.

Extraction of a residual root tip and implant placement can be challenging because of the complexity and invasiveness of the procedure. Improvised application of a guided implant surgery may avoid such challenges. This clinical report presents an innovative technique combining a 3-dimensionally printed surgical guide with conventional instrumentation for a residual root tip extraction in a minimally invasive and predictable way.

Retrieval of cement-retained zirconia implant-supported crowns with an Er,Cr:YSGG laser.

STATEMENT OF PROBLEM: Cement-retained implant-supported crowns can be challenging to retrieve from the abutment once technical or biological issues arise. Removal traditionally requires sectioning the crown with rotary instruments, which causes irreversible damage to the crown and potential damage to the periodontal apparatus stabilizing the implant. PURPOSE: The purpose of this in vitro study was to evaluate an erbium, chromium-doped yttrium, scandium, gallium, and garnet (Er,Cr:YSGG) laser as a minimally invasive alternative for the retrieval of zirconia implant-supported crowns from titanium abutments. Time, temperature, and structural changes to the crown after retrieval were assessed. Appropriate laser parameters were established for this method of crown retrieval. MATERIAL AND METHODS: Twenty zirconia crowns were milled for a maxillary left second premolar based on a CAD-CAM implant analog cast. Ten of these crowns were cemented with a noneugenol zinc oxide dental cement (group Temp) (n=10). The remaining 10 were cemented with a self-adhesive universal resin cement (group Resin) (n=10). Er,Cr:YSGG laser irradiation was performed with the Waterlase iPlus for 1-minute cycles. An attempt was made to remove the crown with a mechanical instrument after each cycle. A type K thermocouple continuously recorded temperature at the level of the abutment. For statistical comparison of decementation time and temperature, the Mann-Whitney test was used (α=.05). Scanning electron microscopy of the nonirradiated and the irradiated crowns was used for analysis of structural and dimensional changes. RESULTS: A significant difference (P<.001) was found in the time ±standard deviation required to retrieve the crowns between group Temp (02:40 ±00:18 minutes:seconds) and group Resin (05:26 ±00:36 minutes:seconds). A significant difference (P<.001) was found in the mean ±standard deviation temperature recorded between group Temp (24.0 ±1.19 °C) and group Resin (25.7 ±0.66 °C). No structural changes to crowns were observed after irradiation. CONCLUSIONS: Retrieval of cement-retained zirconia implant-supported crowns with an Er,Cr:YSGG laser is safe and efficient. Crowns luted with zinc oxide dental cement were retrieved significantly faster while maintaining a significantly lower average temperature than those luted with resin cement. Laser irradiation for decementation did not cause structural changes to zirconia implant-supported crowns.

Enhancing scanning accuracy of digital implant scans: A systematic review on application methods of scan bodies.

STATEMENT OF PROBLEM: Scan bodies play a crucial role in the accuracy of digital implant scans by serving as implant-positioning transfer devices. Previous literature has demonstrated the effects of scan body characteristics on the accuracy of digital implant scans. However, the optimal application methods of scan bodies to enhance scanning accuracy remain unclear. PURPOSE: The purpose of this systematic review was to determine the optimal application methods of scan bodies to enhance the accuracy of digital implant scans. MATERIAL AND METHODS: An electronic search was conducted by using the PubMed (MEDLINE), Web of Science, Cochrane Library, and Embase databases from November 2018 to 2023. Relevant references from the included studies were further screened manually for eligibility. Following the population, intervention, comparison, and outcome (PICO) criteria, a research question focused on identifying the optimal application method for effectively using scan bodies to enhance scanning accuracy was developed. Specific inclusion criteria involved in vitro and in vivo studies. The Checklist for Reporting In Vitro Studies (CRIS) guidelines were followed and the assessment of the risk of bias in the included studies was conducted. RESULTS: Sixteen articles that met the eligibility criteria were included in this systematic review. Two studies investigated the effect of scan body bevel orientation on the accuracy of digital implant scans, and 3 examined the impact of tightening torque on scan bodies. Among the studies focusing on completely edentulous arches, 5 recommended the use of auxiliary geometric devices on the dental arch to enhance scanning accuracy. However, 2 studies reported no improvements in accuracy after splinting scan bodies with thread. CONCLUSIONS: Different techniques for applying scan bodies, such as configuring bevel orientation, adjusting tightening torque, and attaching auxiliary geometric devices, influence the accuracy of digital implant scans. For scanning completely edentulous arches, attaching auxiliary devices to scan bodies to cover the edentulous ridge effectively enhances scanning accuracy.

Analyzing the accuracy of a cross-mounting technique utilizing digitized interocclusal records.

PURPOSE: To compare the accuracy of a partially digital cross-mounting workflow of direct scans of interocclusal records to a conventional workflow by analyzing the deviations of sequentially cross-mounted casts. MATERIALS AND METHODS: A set of reference casts, comprising maxillary and mandibular full-arch prepared casts and interim prostheses, was articulated, mounted, and scanned to generate four reference casts for cross-mounting. In the conventional approach, 15 sets of these four casts were printed. Polyvinylsiloxane (PVS) records were made using the reference casts and utilized for sequential cross-mounting. In the partially digital group, the same PVS interocclusal records were scanned and used for digital cross-mounting via design software. The mean deviations of both groups from the reference cast were analyzed using a 3D inspection software program. Statistical tests, including paired t-test and analysis of variance (ANOVA), were conducted to compare the average discrepancies between the two groups and to evaluate discrepancies in the anterior and posterior regions (α = 0.05). RESULTS: The range of discrepancies was similar in both the conventional and partially digital groups. The final set of related casts had a mean deviation of 201.58 ± 136.98 mm in the conventional workflow and 248.69 ± 164.71 mm in the partially digital workflow. No statistically significant difference was found between conventional and partially digital groups (p = 0.091). Error propagation was examined by comparing discrepancies at each step within the cross-mounting process. In the conventional group, no significant difference was found (p = 0.148), but a significant difference was found among groups in the partially digital group at each step of sequential mounting (p < 0.001). A significant difference was observed between anterior and posterior deviations in the partially digital group (p < 0.001), but not in the conventional group (p = 0.143). CONCLUSIONS: The study reveals that there is no statistically significant difference between conventional and partially digital cross-mounting workflows. However, within the partially digital group, a significant difference in deviation emerges across cross-mounting steps, with increased deviation in the anterior region.

Ultrasensitive and selective colorimetric detection of uric acid using peroxidase mimetic activity of biogenic palladium nanoparticles.

Uric acid (2,6,8-trihydroxypurine) is a metabolic product of purine, which is one of the important markers of human health. The development of a rapid, facile, highly sensitive, and selective method for uric acid detection is critical for the diagnosis of related diseases and is still a strategic challenge. In this study, we developed a highly sensitive and selective colorimetric assay for the detection of uric acid using biogenic palladium nanoparticles (Pd NPs). The synthesized nanoparticles were shown to acquire peroxidase mimetic activity that oxidized 3,3',5,5'-tetramethylbenzidine and produced a blue colour in an assay. The developed colorimetric assay is instrument-free detection of uric acid with a limit of detection of 0.05 µM and a 1.11 µM limit of quantification (LOQ). This is the first report determining the LOQ for a colorimetric assay that gives the lowest quantity of analyte that can be evaluated with more precision under the specified conditions of the analysis. The developed assay had a linear response at low uric acid concentrations of 0.05 to 1 µM and a 0.99841 linear regression correlation coefficient. This colorimetric detection provides a rapid, cost-effective, and easy-to-use platform for the clinical diagnosis of uric acid biomarkers.

Influence of varying titanium base abutment heights on retention of zirconia restorations: An in vitro study.

STATEMENT OF PROBLEM: Implant manufacturers have introduced titanium base (Ti-Base) abutments with increased abutment heights, ostensibly, to increase the retention of the bonded restoration and to improve overall strength. However, evidence regarding the effects of increasing Ti-Base height on improving retention is lacking. PURPOSE: The purpose of this in vitro study was to evaluate the effect of different Ti-Base abutment heights on the retention of zirconia implant-supported crowns. MATERIAL AND METHODS: Thirty Ti-Base abutments of the same diameter and heights of 3.5 mm (n=10), 4.5 mm (n=10), and 5.5 mm (n=10), were used for testing. Zirconia restorations were cemented onto the Ti-Base abutments with a resin cement after treatment with a 10-methacryloyloxydecyl dihydrogen phosphate primer by a single operator using a positioning device. The zirconia-Ti-Base restorations were tightened to an implant analog embedded in an autopolymerizing resin block. The specimens were placed and tested in a universal testing machine for pull-out testing. Retention was measured by recording the force at load drop. Statistical analysis was performed using 1-way analysis of variance with the Tukey method for pairwise comparisons. RESULTS: The abutment height had a significant effect on retention (P=.010). Ti-Base abutments of 4.5 and 5.5 mm had significantly greater retention than Ti-Base abutments of 3.5 mm (P=.020, P=.040, respectively). However, Ti-Base abutments of 4.5 and 5.5 mm in height were statistically similar (P=.890). CONCLUSIONS: An increase in the height of Ti-Base abutments above the standard 3.5 mm height significantly improved the retention of the overlying restoration.

Accuracy of impression scan bodies for complete arch fixed implant-supported restorations.

STATEMENT OF PROBLEM: Despite the extensive studies on and comparisons of different implant impression techniques for completely edentulous patients, studies on novel techniques that combine conventional impression making with digital scanning are lacking. PURPOSE: The primary aim of this study was to compare the accuracy of the impression scan body technique with conventional impression making and digital scanning for a completely edentulous mandibular arch. The secondary aim was to assess the relationship between different implant angulations and interimplant distances in relation to the recording accuracy. MATERIAL AND METHODS: An edentulous mandibular definitive cast (control) was designed with 5 implants placed at different angles and interimplant distances. Three recording techniques were tested: a conventional impression with splinted copings (conventional) (n=15), an impression scan body technique where impression scan bodies were attached to the splinted impression copings in a conventional elastomeric impression and then digitally scanned with an extraoral scanner (n=15), and an intraoral digital scanning technique (digital) (n=15). For comparison, the definitive cast and the conventional impression stone casts were digitized into standard tessellation language (STL) datasets using the extraoral scanner. The 3-dimensional (3D) deviations between the 3 test groups and the control were calculated by superimposing the STL datasets. The 3D deviations from the control were compared by using the Kruskal-Wallis test followed by the Dunn post hoc test (α=.05). The Mann-Whitney test was used to investigate the effect of implant angulation and interimplant distance on impression accuracy (α=.05). RESULTS: The conventional splinted-coping impression technique showed a mean 3D deviation of 0.408 mm. The impression scan body and intraoral digital scan showed similar mean 3D deviations, 0.219 mm and 0.257 mm, respectively (P=.334). Both techniques showed significantly lower 3D deviations than the conventional technique (P<.001). Implants at an angle of 5 degrees and 10 degrees showed a statistically significant difference (P=.010) with mean 3D deviations of 0.340 mm and 0.396 mm, respectively. Implants with 5-mm and 10-mm interimplant distance showed a significant difference (P<.001) with mean 3D deviations of 0.301 mm and 0.423 mm, respectively. CONCLUSIONS: The impression scan body technique is comparable with intraoral digital scanning for a completely edentulous arch. Increased implant angulation and increased interimplant distance significantly reduced the accuracy of implant impression making or scanning.

A clinical comparison of sagittal condylar inclination and Bennett angle derived from a conventional electronic tracking device and an optical jaw tracking device.

STATEMENT OF PROBLEM: The challenges of conventional methods to measure the sagittal condylar inclination and the Bennett angle include patient discomfort and an output of averaged values calculated on the arc of mandibular movement. Programming these average values into a mechanical or mathematically simulated articulator may introduce occlusal errors. PURPOSE: The purpose of this clinical study was to compare the sagittal condylar inclination and Bennett angle measurements derived from a conventional electronic tracking device and an optical tracking device. MATERIAL AND METHODS: Fifteen dentate participants with at least 12 occluding units, 25 years old and above, participated in the study. Each participant's mandibular movements were recorded by the 2 systems for different condylar guidance values. A conventional tracking device, the Cadiax Compact 2, and an optical tracking device, the MODJAW, were used for this study. Sagittal condylar inclination was measured at 3 mm and 5 mm using protrusive records and the values of the Bennett angle at 3 mm were measured using excursive records along the path traced by mandibular condyles. The Wilcoxon signed-rank test was used to analyze the paired data from the 2 methods. RESULTS: A significant difference was found between right (P=.007) and left sagittal condylar inclinations (P=.010) measured at 3 mm with the conventional and optical tracking devices. A significant difference was found between right (P=.015) and left sagittal condylar inclinations (P=.004) measured at 5 mm with the conventional and optical tracking devices. The right and left Bennett angles measured at 3 mm with the conventional and optical tracking devices were statistically different (P=.043 and P=.035 respectively). CONCLUSIONS: The sagittal condylar inclination and Bennett angle measured at different positions along the path of movement exhibited significant differences. The sagittal condylar inclination and Bennett angle values obtained from the conventional tracking device were generally significantly less than those derived from the optical tracking device.

Accuracy comparison of bilateral versus complete arch interocclusal registration scans for virtual articulation.

STATEMENT OF PROBLEM: Alternatives to the bilateral interocclusal registration scanning technique to improve virtual articulation have not been fully investigated. PURPOSE: The purpose of this in vitro study was to compare the accuracy of virtually articulating digital casts by using bilateral interocclusal registration scans versus a complete arch interocclusal scan. MATERIAL AND METHODS: A set of maxillary and mandibular reference casts were hand-articulated and mounted on an articulator. The mounted reference casts were scanned, and the maxillomandibular relationship record was scanned 15 times using 2 different scanning techniques, the bilateral interocclusal registration scan (BIRS) and complete arch interocclusal registration scan (CIRS), with an intraoral scanner. The generated files were transferred to a virtual articulator, and each set of scanned casts was articulated using BIRS and CIRS. The virtually articulated casts were saved as a set and transferred to a 3-dimensional (3D) analysis program. The scanned casts were set in the same coordinate system as the reference cast and overlaid on top of the reference cast for analysis. Two anterior and 2 posterior points were selected to determine points of comparison between the reference cast and test casts virtually articulated with BIRS and CIRS. The mean discrepancy between the 2 test groups and the anterior and posterior mean discrepancy within each group were tested for significance by using the Mann-Whitney U test (α=.05). RESULTS: A significant difference was found between the virtual articulation accuracy of BIRS and CIRS (P<.001). The mean deviation for BIRS was 0.053 ±0.051 mm and that for CIRS was 0.265 ±0.241 mm. Furthermore, significant differences were found between the anterior and posterior deviations in both BIRS (P=.020) and CIRS (P<.001). The mean deviation for BIRS was 0.034 ±0.026 mm in the anterior and 0.073 ±0.062 mm in the posterior. The mean deviation for CIRS was 0.146 ±0.108 mm anteriorly and 0.385 ±0.277 mm posteriorly. CONCLUSIONS: BIRS was more accurate than CIRS for virtual articulation. Moreover, the alignment accuracy of anterior and posterior sites for both BIRS and CIRS exhibited significant differences, with the anterior alignment exhibiting better accuracy in relation to the reference cast.

An analog-digital hybrid technique for immediate dentures with idealized border extensions.

In patients with severe tooth mobility, conventional impression making can be challenging because of the risk of accidental tooth extraction. Digital intraoral scanning avoids such a complication but does not capture optimal border extensions for a complete denture. This clinical report presents a combined digital and analog recording technique which allows the recording of optimal vestibular border extensions without the risk of tooth extraction.

Tfh cells with NLRP3 inflammasome activation are essential for high-affinity antibody generation, germinal centre formation and autoimmunity.

OBJECTIVE: NLRP3 inflammasome regulates T cell responses. This study examined the roles of NLRP3 inflammasome activation in the regulation of T follicular helper (Tfh) cells during humoral response to T dependent antigens and in systemic lupus erythematosus (SLE). METHODS: NLRP3 inflammasome activation of Tfh cells was studied in B6, MRL/lpr and NZM2328 mice and in SLE patients and healthy controls using a fluorescence-labelled caspase-1 inhibitor probe. MCC950, a selective inhibitor of NLRP3, was used to investigate the relation between NLRP3 inflammasome activation and germinal centre (GC) reaction, Ab responses to immunisation, and autoantibody production. RESULTS: NLRP3 inflammasome activation in Tfh cells after immunisation was identified in B6 mice. MCC950 inhibited humoral responses to sheep red blood cell and NP-CGG with reduction of the GC reaction. B6 mice with lymphoid cell-specific deletion of NLRP3 or Casp1 mounted suboptimal humoral responses with impaired GC formation and defective affinity maturation. In MRL/lpr and NZM2328 mice, inhibition of NLRP3 activation suppressed NLRP3 activated Tfh cell expansion as well as attenuated lupus-like phenotypes. Tfh cells with activated NLRP3 inflammasome exhibited increased expression of molecules for Tfh cell function and differentiation, and had greater ability to activate B cells. In SLE patients, disease activity was positively correlated with an increase in the activated NLRP3+ Tfh population and this population was markedly reduced in response to therapy. CONCLUSIONS: The activation of NLRP3 inflammasome in Tfh cells is an integral part of responses to immunisation. The activated NLRP3+ Tfh population is essential for optimal humoral responses, GC formation and autoimmunity.

Interscalene Brachial Plexus Block with Liposomal Bupivacaine versus Standard Bupivacaine with Perineural Dexamethasone: A Noninferiority Trial.

BACKGROUND: The interscalene nerve block provides analgesia for shoulder surgery. To extend block duration, provide adequate analgesia, and minimize opioid consumption, the use of adjuvants such as dexamethasone as well as the application of perineural liposomal bupivacaine have been proposed. This randomized, double-blinded, noninferiority trial hypothesized that perineural liposomal bupivacaine is noninferior to standard bupivacaine with perineural dexamethasone in respect to average pain scores in the first 72 h after surgery. METHODS: A total of 112 patients undergoing ambulatory shoulder surgery were randomized into two groups. The liposomal bupivacaine group received a 15-ml premixed admixture of 10 ml of 133 mg liposomal bupivacaine and 5 ml of 0.5% bupivacaine (n = 55), while the bupivacaine with dexamethasone group received an admixture of 15 ml of 0.5% standard bupivacaine with 4 mg dexamethasone (n = 56), respectively. The primary outcome was the average numerical rating scale pain scores at rest over 72 h. The mean difference between the two groups was compared against a noninferiority margin of 1.3. Secondary outcomes were analgesic block duration, motor and sensory resolution, opioid consumption, numerical rating scale pain scores at rest and movement on postoperative days 1 to 4 and again on postoperative day 7, patient satisfaction, readiness for postanesthesia care unit discharge, and adverse events. RESULTS: A liposomal bupivacaine group average numerical rating scale pain score over 72 h was not inferior to the bupivacaine with dexamethasone group (mean [SD], 2.4 [1.9] vs. 3.4 [1.9]; mean difference [95% CI], -1.1 [-1.8, -0.4]; P < 0.001 for noninferiority). There was no significant difference in duration of analgesia between the groups (26 [20, 42] h vs. 27 [20, 39] h; P = 0.851). Motor and sensory resolutions were similar in both groups: 27 (21, 48) h versus 27 (19, 40) h (P = 0.436) and 27 [21, 44] h versus 31 (20, 42) h (P = 0.862), respectively. There was no difference in opioid consumption, readiness for postanesthesia care unit discharge, or adverse events. CONCLUSIONS: Interscalene nerve blocks with perineural liposomal bupivacaine provided effective analgesia similar to the perineural standard bupivacaine with dexamethasone. The results show that bupivacaine with dexamethasone can be used interchangeably with liposomal bupivacaine for analgesia after shoulder surgery.

The Optimal Cell Salvage Settings to Maximize Hematocrit and Minimize Potassium Using the Cobe BRAT2 Autologous Blood Recovery Unit.

OBJECTIVE: The objective was to determine the optimal cell saver device settings (infusion rate and wash rate) at which hematocrit is preserved and potassium and lactate are removed from banked red blood cells (RBC). DESIGN: Red cells were washed using the Cobe BRAT 2 Autologous Blood Recovery Unit and sampled for electrolyte composition and hematocrit pre- and postwash. SETTING: This was a single-center study. INTERVENTIONS: Red cells were washed using six infusion rates (100-1,000 mL/min) and six wash rates (100-1,000 mL/min) for a total of 36 combinations. Hematocrit, potassium, glucose, and lactate were evaluated before and after washing. MEASUREMENTS AND MAIN RESULTS: At wash rates ≤400 mL/min, hematocrit increased independent of infusion rate. At wash rates ≥400 mL/min, slower infusion rates were associated with higher hematocrit compared to faster infusion rates (p < 0.0001 for a wash rate 400-800 mL/min, p < 0.0005 for a wash rate 1,000 mL/min). Maximal wash speeds were associated with decreasing hematocrit. Infusion and wash rate were both independent predictors of potassium change; slower rates were associated with a larger decrease in potassium. Glucose decreased proportionally as infusion and wash rate decreased. Lactate did not show an association with either infusion or wash rate. CONCLUSION: Red-cell washing produces higher hematocrit and lower potassium as infusion rate and wash rate decrease. A 340-mL unit of RBC can be processed in 4.26 minutes without loss of hematocrit or an increase in potassium when both infusion and wash rates are set to 400 mL/min.

Photoprotective effects of sphingomyelin-containing milk phospholipids in ultraviolet B-irradiated hairless mice by suppressing nuclear factor-κB expression.

Ceramide-containing phospholipids improve skin hydration and barrier function and are ideal for use in skin care products. In this study, we evaluated the photoprotective effect of milk phospholipids on the skin condition of UVB-irradiated hairless mice. Skin parameters were assessed following oral administration of milk phospholipids. The UVB irradiation induced photoaging in mice. The animals were divided into 5 groups: a control group (oral administration of saline with no UBV irradiation), UVB group (oral administration of saline with UVB irradiation), and 3 UVB irradiation groups receiving the milk phospholipids at 3 different concentrations of oral administration, 50 mg/kg (ML group), 100 mg/kg (MM group), and 150 mg/kg (MH group), for 8 wk. An increase in skin hydration and transepidermal water loss were improved in the 150 mg/kg of milk phospholipid-administered group. Hematoxylin and eosin staining revealed a decrease in epidermal thickness in the milk phospholipid-administered groups (50, 100, and 150 mg/kg of body weight). In particular, the 100 and 150 mg/kg groups showed significant changes in the area, length, and depth of the wrinkles compared with the UVB group. Moreover, the gene expression of matrix metalloproteins was attenuated, and that of proinflammatory cytokines, especially tumor necrosis factor-α, was significantly reduced in the milk phospholipid-administered groups than in the UVB group. The reduced ceramide and increased sphingosine-1-phosphate levels in the skin tissue due to UVB exposure were restored to levels similar to those of the control group following milk phospholipid administration. These results were confirmed to be due to the downregulation of protein expression of nuclear factor kappa-B (NF-κB) and phosphorylated IκB-α (inhibitor of κB α). Collectively, oral administration of milk phospholipids improves skin health through a synergistic effect on photoprotective activity.