Rescue Effect Research Articles

The Effect of Rescue and Relief Services on the Severity of Traffic Accidents

INTRODUCTION: According to the official statistics, a high rate of road traffic deaths occurs during the transportation of the injured and in medical centers. The way of transporting the injured to the medical centers, the standard time of dispatch and timely treatment in the hospitals, the land use planning of medical centers and rehabilitation are the three missing links of the post-event phase. Therefore, the researchers aimed to assess the effect of rescue and relief services on the severity of traffic accidents in this study. METHODS: This applied research was conducted based on an exploratory mixed method. The statistical population in the qualitative section included Amin Police Academy professors, experts of both Red Crescent Society (RCS) of Tehran province and Tehran Emergency Medical Service (EMS), who were selected via purposive sampling method. In the qualitative section, the theme analysis method was used and analyzed by using Maxqda-2020 software. The statistical population of the quantitative part included excellent accident experts of 28 districts of Tehran and experts of the RCS and EMS of Tehran province in a stratified random method. In the following, confirmatory factor analysis using structural equation modeling method was used and analyzed with SmartPLS3 software. FINDINGS: The dimensions of the impact of rescue and relief services on the severity of traffic accidents from the perspective of Red Crescent Society and Emergency Medical Services experts included equipment (31.077), land use planning (25.610), expertise (23.598), and education (8.243). The most marked impact was related to equipment, and the least impact pertained to education. Furthermore, the dimensions of the effect of rescue and relief services on the severity of traffic accidents from the perspective of accident experts included equipment (42.009), land use planning (34.419), education (33.770), and expertise (8.951). The greatest impact was related to equipment, and the least effect pertained to expertise. In addition, the factor loadings of all dimensions were more than 0.5 being significant at the 0.95 level. CONCLUSION: Strengthening and developing equipment, land use planning balanced with the geography of missions, and continuous specialized training of rescue and relief of RCS and EMS experts will positively reduce the severity of traffic accidents in the country.

Effect of dienogest on serum anti-Mullerian hormone level after laparoscopic cystectomy of ovarian endometrioma

ObjectiveThis comparative study aimed to evaluate the effect of postoperative dienogest treatment on serum anti-Mullerian hormone (AMH) levels in patients undergoing laparoscopic cystectomy of ovarian endometriomas. Materials and methodsA total of 71 patients with ovarian endometriomas treated at our department were enrolled. After surgery, 54 patients received oral dienogest 2 mg daily continuously for 6 months (dienogest group). The other 17 patients did not receive postoperative medical treatment (control group). Serum AMH levels were measured before surgery, at 3-month period after surgery, and at the end of 6-month follow-up period. Serial changes of AMH levels were compared between the two groups. ResultsThe age, endometrioma size, and serum AMH level before surgery were comparable between the dienogest group and the control group. The AMH levels decreased significantly at 3-month period after surgery in the dienogest group (a decrease of 65.5%; p < 0.001) and the control group (a decrease of 64.8%; p = 0.018). The AMH levels increased gradually from the nadir at 3-month period after surgery and recovered partially at the end of 6-month follow-up period in both groups. There were no statistically significant differences in the rate of reduction of serum AMH levels between the two groups (p = 0.707). ConclusionLaparoscopic cystectomy of ovarian endometrioma causes a significant decrease in serum AMH levels. The rates of reduction of AMH levels are similar regardless of dienogest treatment. Postoperative dienogest treatment for 6 months has no rescue effect on serum AMH levels in these patients.

Consequences of Traceable Mobility in Populations Exhibiting Strong Allee Effect

In this research, we study the impacts of the traceable mobility in a two-patch environment when the population in each patch exhibits strong Allee effects. Traveling individuals are traced across patches by budgeting the average time spent in each patch while keeping their place of residency. Particularly, we focus on the impact that the effective population (residents and visitors) produces on regional dynamics.Our results show that low mobility across regions produces simple dynamics, where orbits converge to single or double extinction, or to a coexistence steady state. We derive mobility conditions under which an endangered population may benefit of the presence of a visitant one and avoid extinction -- the rescue effect. Nonetheless, increments in the visiting population would also lead the resident population to extinction -- the induced extinction effect.

P-596 Rectal progesterone administration secures high ongoing pregnancy rate in a personalized Hormone Replacement Therapy Frozen Embryo Transfer (HRT-FET) protocol - a prospective interventional study

Abstract Study question Can supplementation with additional rectal administration of progesterone secure high ongoing pregnancy rates (OPR) in patients with low serum progesterone (P4) at blastocyst transfer (ET)? Summary answer Rectally administered progesterone starting on the blastocyst transfer day secures high OPR in patients with serum P4 levels lower than 35 nmol/l (11ng/ml). What is known already Low serum P4 levels at peri-implantation in HRT-FET cycles impact reproductive outcomes negatively. However, studies have shown that patients with low P4 after a standard vaginal progesterone treatment can obtain live birth rates (LBR) comparable to patients with optimal P4 levels if they receive additional subcutaneous progesterone, starting close to the day of blastocyst transfer. In contrast, increasing vaginal progesterone supplementation in low serum P4 patients does not increase LBR. Another route of administration rarely used in ART is the rectal route, although progesterone is well absorbed and serum P4 levels reach a maximum level after approximately two hours. Study design, size, duration This prospective interventional study included a cohort of 488 patients treated with single blastocyst HRT-FET, in which a total of 374 patients had a serum P4 level ≥35nmol/l (11ng/ml) at ET, and 114 patients had a serum P4 level &amp;lt;35nmol/l (11ng/ml). The study was conducted from January 2020 to November 2022. Participants/materials, setting, methods Patients undergoing HRT-FET in a public Fertility Clinic. Endometrial preparation included oral oestradiol (6mg/24hours), followed by vaginal micronized progesterone, 400mg/12hours. Single blastocyst transfer and P4 measurements were performed on the 6th day of progesterone administration and patients with serum P4 &amp;lt;35nmol/l (11ng/ml) additional rectal administration of progesterone (400mg/12hour) was started the same day. In pregnant patients, vaginal and rectal administration continued until week 8, and oestradiol and vaginal progesterone treatment continued until week 10. Main results and the role of chance Among 488 HRT-FET single blastocyst transfers, the mean age of the patients at oocyte retrieval (OR) was 30.9 ±4.6 years and the mean BMI at ET 25.1 ±3.5 kg/m2. The mean serum P4 level after vaginal progesterone administration on the day of blastocyst transfer was 48.9 ±21.0 nmol/l (15.4 ±6.6ng/ml), and a total of 23% (114/488) of the patients had a serum P4 level lower than 35nmol/l (11ng/ml). The overall, positive hCG rate, clinical pregnancy rate, OPR week 12, and total pregnancy loss rate were 66% (320/488), 54% (265/488), 45% (221/488), and 31% (99/320), respectively. There was no significant difference in neither OPR week 12 nor total pregnancy loss rate between patients with P4 ≥35nmol/l (11ng/ml) and patients with P4 &amp;lt;35nmol/l, who received rescue in terms of rectally administered progesterone, 45% vs. 46%, p = 0.77 and 30% vs. 34%, p = 0.53, respectively. OPR did not depend on whether patients had initially low P4 and rectal rescue or were above the P4 cut-off. Logistic regression analysis showed that only age at OR and blastocyst scoring correlated with OPR week 12, independently of other factors like BMI, and vitrification day of blastocysts (day 5 or 6). Limitations, reasons for caution In this study vaginal progesterone (Cyclogest®), a solid pessary with progesterone suspended in vegetable hard fat, was used vaginally as well as rectally. It is unknown whether other vaginal progesterone products could be used rectally as easily with the same rescue effect. Wider implications of the findings A substantial part of HRT-FET patients receiving vaginal progesterone treatment has low serum P4. Adding rectally administered progesterone in these patients increases the reproductive outcome. Importantly, rectal progesterone administration is considered convenient by the majority of patients; moreover, progesterone pessaries are easy to administer rectally and of low cost. Trial registration number EudraCT no.: 2019-001539-29

1628-P: Role of CD38 in the Regulation of Muscle Stem Cell Functions in Aged Mice

Skeletal muscle repair and regeneration is a complex process and is known to be delayed with aging, diabetes and obesity. This is in part due to infiltration of inflammatory macrophages which can delay the recovery process. Both macrophages and satellite cells require nicotinamide adenine dinucleotide (NAD+) for their biological functions. Aging-induced decline in NAD+ have been shown to worsen insulin resistance and muscle dysfunction. CD38 is an ectoenzyme that degrades NAD+ and is increased in inflammatory macrophages in aging, obesity and other insulin-resistant states. However, how inhibition of CD38 affects satellite cell functions in aged or obese mice, remains unknown. Here, we utilized aged CD38 knockout mice to assess the role of this protein and NAD+ in the recovery process in injured muscle. CD38 KO mice were found to be resistant to diet-induced obesity and had improved insulin sensitivity. Our data show that genetic deletion of CD38 in aged mice increased NAD+ levels and thus enhanced muscle stem/satellite cells differentiation and promote myogenesis. Deletion of CD38 also resulted in an enhanced number of regenerating myofibers. Furthermore, we found that deletion of CD38 significantly reduces fibrosis of muscle and also improved limb grip strength. Pharmacological inhibition of CD38 in C2C12 myoblasts and ex-vivo primary myoblasts significantly increased NAD+ levels and also enhanced the differentiation of satellite cells into myoblasts and myofibers. We, therefore, examined the rescue effects of 78c (CD38i) on human recombinant CD38 (hCD38) inhibition of muscle stem cells. Administration of hCD38 inhibited expression of MyoD and MyoG in primary myoblasts, while this effect was rescued through 78c treatment. Taken together, these data demonstrate that restoration of NAD+ levels either through genetic deletion/pharmacological inhibition of CD38 can improve muscle stem cell functions and muscle regeneration and might promote muscle regeneration in obesity, type 2 diabetes, and aging. Disclosure A.Nawaz: None. K.Yaku: None. I.Jeelani: None. K.Tobe: Other Relationship; MSD K.K., Novo Nordisk Pharma Ltd., Takeda Pharmaceutical Company Limited, Daiichi Sankyo, Mitsubishi Tanabe Pharma Corporation, Suntory Global Corporation, Ltd.,, Taiho Pharmaceutical Co. Ltd., Japan Diabetes Foundation, Japan Association for Diabetes Education and Care. T.Nakagawa: None.

Wnt5a-YAP signaling axis mediates mechanotransduction in cardiac myocytes and contributes to contractile dysfunction induced by pressure overload

Non-canonical Wnt signaling activated by Wnt5a/Wnt11 is required for the second heart field development in mice. However, the pathophysiological role of non-canonical Wnt signaling in the adult heart has not been fully elucidated. Here we show that cardiomyocyte-specific Wnt5a knockout mice exhibit improved systolic function and reduced expression of mechanosensitive genes including Nppb when subjected to pressure overload. In cultured cardiomyocytes, Wnt5a knockdown reduced Nppb upregulation induced by cyclic cell stretch. Upstream analysis revealed that TEAD1, a transcription factor that acts with Hippo pathway co-activator YAP, was downregulated both invitro and invivo by Wnt5a knockdown/knockout. YAP nuclear translocation was induced by cell stretch and attenuated by Wnt5a knockdown. Wnt5a knockdown-induced Nppb downregulation during cell stretch was rescued by Hippo inhibition, and the rescue effect was canceled by knockdown of YAP. These results collectively suggest that Wnt5a-YAP signaling axis mediates mechanotransduction in cardiomyocytes and contributes to heart failure progression.

The mitochondrial Ca2+ channel MCU is critical for tumor growth by supporting cell cycle progression and proliferation.

Introduction: The mitochondrial uniporter (MCU) Ca2+ ion channel represents the primary means for Ca2+ uptake by mitochondria. Mitochondrial matrix Ca2+ plays critical roles in mitochondrial bioenergetics by impinging upon respiration, energy production and flux of biochemical intermediates through the TCA cycle. Inhibition of MCU in oncogenic cell lines results in an energetic crisis and reduced cell proliferation unless media is supplemented with nucleosides, pyruvate or α-KG. Nevertheless, the roles of MCU-mediated Ca2+ influx in cancer cells remain unclear, in part because of a lack of genetic models. Methods: MCU was genetically deleted in transformed murine fibroblasts for study in vitro and in vivo. Tumor formation and growth were studied in murine xenograft models. Proliferation, cell invasion, spheroid formation and cell cycle progression were measured in vitro. The effects of MCU deletion on survival and cell-death were determined by probing for live/death markers. Mitochondrial bioenergetics were studied by measuring mitochondrial matrix Ca2+ concentration, membrane potential, global dehydrogenase activity, respiration, ROS production and inactivating-phosphorylation of pyruvate dehydrogenase. The effects of MCU rescue on metabolism were examined by tracing of glucose and glutamine utilization for fueling of mitochondrial respiration. Results: Transformation of primary fibroblasts in vitro was associated with increased MCU expression, enhanced MCU-mediated Ca2+ uptake, altered mitochondrial matrix Ca2+ concentration responses to agonist stimulation, suppression of inactivating-phosphorylation of pyruvate dehydrogenase and a modest increase of mitochondrial respiration. Genetic MCU deletion inhibited growth of HEK293T cells and transformed fibroblasts in mouse xenograft models, associated with reduced proliferation and delayed cell-cycle progression. MCU deletion inhibited cancer stem cell-like spheroid formation and cell invasion in vitro, both predictors of metastatic potential. Surprisingly, mitochondrial matrix [Ca2+], membrane potential, global dehydrogenase activity, respiration and ROS production were unaffected. In contrast, MCU deletion elevated glycolysis and glutaminolysis, strongly sensitized cell proliferation to glucose and glutamine limitation, and altered agonist-induced cytoplasmic Ca2+ signals. Conclusion: Our results reveal a dependence of tumorigenesis on MCU, mediated by a reliance on MCU for cell metabolism and Ca2+ dynamics necessary for cell-cycle progression and cell proliferation.

Empirical Evidence for the Rescue Effect from a Natural Microcosm.

Ecological theory predicts that populations which receive immigrants are less vulnerable to extinction than those that do not receive immigrants (the "rescue effect"). A parallel but opposite process may also exist, where emigration increases the risk of local extinction (the "abandon-ship effect"). Using a natural microcosm of plant-specialist frogs from Madagascar, empirical evidence for both processes is provided. Populations receiving immigrants were less extinction-prone than those without immigration, and those populations losing individuals through emigration were more extinction-prone than those in which no emigration occurred. The number of immigrants and emigrants was also elevated and depressed (respectively) in patches that did not go extinct. These data provide some of the first definitive empirical evidence for the rescue effect and provide suggestive initial data on the abandon-ship effect. Both of these processes may be important to understanding the dynamics of populations.

C11orf54 promotes DNA repair via blocking CMA-mediated degradation of HIF1A

C11orf54 is an ester hydrolase highly conserved across different species. C11orf54 has been identified as a biomarker protein of renal cancers, but its exact function remains poorly understood. Here we demonstrate that C11orf54 knockdown decreases cell proliferation and enhances cisplatin-induced DNA damage and apoptosis. On the one hand, loss of C11orf54 reduces Rad51 expression and nuclear accumulation, which results in suppression of homologous recombination repair. On the other hand, C11orf54 and HIF1A competitively interact with HSC70, knockdown of C11orf54 promotes HSC70 binding to HIF1A to target it for degradation via chaperone-mediated autophagy (CMA). C11orf54 knockdown-mediated HIF1A degradation reduces the transcription of ribonucleotide reductase regulatory subunit M2 (RRM2), which is a rate-limiting RNR enzyme for DNA synthesis and DNA repair by producing dNTPs. Supplement of dNTPs can partially rescue C11orf54 knockdown-mediated DNA damage and cell death. Furthermore, we find that Bafilomycin A1, an inhibitor of both macroautophagy and chaperone-mediated autophagy, shows similar rescue effects as dNTP treatment. In summary, we uncover a role of C11orf54 in regulating DNA damage and repair through CMA-mediated decreasing of HIF1A/RRM2 axis.

Brain responses to energy intake and its association with cognition in Alzheimer’s Disease

AbstractBackgroundTo evaluate the brain response to energy intake using cerebral blood flow(CBF) in Alzheimer’s disease (AD), mild cognitive impairment (MCI) and healthy controls (HC) and its relationship with cognition.MethodA self‐comparison design was used to test the effects of energy intake on brain activity in participants with AD (n = 30), MCI participants (n = 32), and HC (n = 30) and its relationship with cognitive function. Brain perfusion was measured using arterial spin labeling (ASL) functional brain imaging. Participants arrived at the site in the morning and underwent the ASL‐MRI with at least 10 hours fasting overnight. Then the second ASL‐MRI at 30 minutes after eating a standard breakfast was performed. Energy response ΔCBF was computed by subtracting the fasting CBF from the postprandial CBF obtained from ASL‐MRI. Whole brain grey matter voxel‐wise paired T test and linear regression analyses were used for statistical analysis.ResultAD and MCI group had different energy response ΔCBF pattern with HC group. In AD group, energy intake induced a significantly decreased CBF in temporal lobe including lateral inferior and superior temporal gyrus, right superior temporal gyrus, parietal lobe including bilateral superior and inferior parietal lobe, postcentral gyrus and precuneus, most of occipital lobe and frontal lobe including right rolandic operculum, right precentral gyrus and left middle frontal gyrus. (q &lt; 0.05, false discovery rate corrected). MCI had similar energy intake response pattern with AD. HC group had no significantly changed CBF response to energy intake. Energy response ΔCBF in the vicinity of the superior temporal gyrus showed significant negative correlation with Minimum Mental State Examination (MMSE) in all AD, MCI and HC groups.ConclusionThe different energy response ΔCBF pattern may help to characterize the energy disorder in AD and may indicate the different brain energy rescue effect in the three groups. Energy response ΔCBF in the superior temporal gyrus is an indicator of cognitive function in AD.

RITA selectively inhibits proliferation of BAP1-deficient cutaneous melanoma cells in vitro

To screen for small molecular compounds with selective inhibitory activity against cutaneous melanoma cells with BAP1 deletion. Cutaneous melanoma cells expressing wild-type BAP1 were selected to construct a BAP1 knockout cell model using CRISPR-Cas9 system, and small molecules with selective inhibitory activity against BAP1 knockout cells were screened from a compound library using MTT assay. Rescue experiment was carried out to determine whether the sensitivity of BAP1 knockout cells to the candidate compounds was directly related to BAP1 deletion. The effects of the candidate compounds on cell cycle and apoptosis were detected with flow cytometry, and the protein expressions in the cells were analyzed with Western blotting. The p53 activator RITA from the compound library was shown to selectively inhibit the viability of BAP1 knockout cells. Overexpression of wild-type BAP1 reversed the sensitivity of BAP1 knockout cells to RITA, while overexpression of the mutant BAP1 (C91S) with inactivated ubiquitinase did not produce any rescue effect. Compared with the control cells expressing wild-type BAP1, BAP1 knockout cells were more sensitive to RITA-induced cell cycle arrest and apoptosis (P < 0.0001) and showed an increased expression of p53 protein, which was further increased by RITA treatment (P < 0.0001). Loss of BAP1 results in the sensitivity of cutaneous melanoma cells to p53 activator RITA. In melanoma cells, the activity of ubiquitinase in BAP1 is directly related to their sensitivity to RITA. An increased expression of p53 protein induced by BAP1 knockout is probably a key reason for RITA sensitivity of melanoma cells, suggesting the potential of RITA as a targeted therapeutic agent for cutaneous melanoma carrying BAP1-inactivating mutations.

Unexpected phenotypic and molecular changes of combined glucocerebrosidase and acid sphingomyelinase deficiency.

Heterozygous variants in GBA1, encoding glucocerebrosidase (GCase), are the most common genetic risk factor for Parkinson's disease (PD). Moreover, sporadic PD patients also have a substantial reduction of GCase activity. Genetic variants of SMPD1 are also overrepresented in PD cohorts, whereas a reduction of its encoded enzyme (acid sphingomyelinase or ASM) activity is linked to an earlier age of PD onset. Despite both converging on the ceramide pathway, how the combined deficiencies of both enzymes might interact to modulate PD has yet to be explored. Therefore, we created a double-knockout (DKO) zebrafish line for both gba1 (or gba) and smpd1 to test for an interaction in vivo, hypothesising an exacerbation of phenotypes in the DKO line compared to those for single mutants. Unexpectedly, DKO zebrafish maintained conventional swimming behaviour and had normalised neuronal gene expression signatures compared to those of single mutants. We further identified rescue of mitochondrial Complexes I and IV in DKO zebrafish. Despite having an unexpected rescue effect, our results confirm ASM as a modifier of GBA1 deficiency in vivo. Our study highlights the need for validating how genetic variants and enzymatic deficiencies may interact in vivo.

Structural changes and carbon reduction due to habitat loss in Atlantic Forest

Habitat loss is a significant threat to biodiversity in tropical forests, leading to changes in composition, structure, and function. In this study, we aimed to evaluate the influence of the forest amount at the landscape scale on the structure and the stored carbon of tree communities. Eleven landscape samples of 36 km2 (6 × 6 km), with different amounts of forest cover at the landscape scale (between 5 and 60%), were randomly selected within a wide latitudinal range of Atlantic Forests in Brazil. We also used a sample design to control and avoid some biases in landscape-scale studies during the landscape selection, such as the rescue effect from the surrounding landscapes, differences in matrix permeability in each landscape, and the control for the same vegetation type or successional stage. In each selected landscape, we surveyed all woody individuals with a diameter at breast height (DBH) ≥ 2.5 cm in eight plots of 250 m2, randomly placed in the landscape’s forested areas. The structural variables evaluated were: total abundance, the abundance of prominent families, the abundance of large trees (DBH ≥ 30 cm), mean DBH, and average height, besides carbon storage. From the set of 8,179 individuals, we observed a positive correlation between forest amount and the total abundance, the abundance of large trees, and carbon stock. The abundance of the prominent hardwood tree families were positively correlated with forest amount. When comparing the two landscapes with the lowest proportion of habitats (5–15%) and the highest proportion (55−60%), almost one thousand trees and more than 150 T of carbon per hectare are lost. This reduction in the overall abundance and carbon indicates no compensatory effects on the tree community due to a turnover in composition or substitution of large trees for smaller ones. The Atlantic Forest is undergoing a process of homogenization and functional collapse, derived directly or indirectly from habitat loss, leading to a possible regime shift.

C1q/TNF-related protein 4 mediates proliferation and migration of vascular smooth muscle cells during vascular remodelling.

Vascular remodelling is an essential pathophysiological state in many circulatory diseases. Abnormal vascular smooth muscle cell (VSMC) behaviour leads to neointimal formation and may eventually results in major adverse cardiovascular events. The C1q/TNF-related protein (C1QTNF) family is closely associated with cardiovascular disease. Notably, C1QTNF4 has unique two C1q domains. However, the role of C1QTNF4 in vascular diseases remains unclear. C1QTNF4 expression was detected in human serum and artery tissues using ELISA and multiplex immunofluorescence (mIF) staining. Scratch assay, transwell assay and confocal microscopy were used to investigate C1QTNF4 effects on VSMC migration. EdU incorporation, MTT assay and cell counting experiment revealed C1QTNF4 effects on VSMC proliferation. C1QTNF4-transgenic, C1QTNF4-/- and AAV9-mediated VSMC-specific C1QTNF4 restoration C1QTNF4-/- mouse and rat disease models were generated. RNA-seq, quantitative real-time PCR, western blot, mIF, proliferation and migration assays were used to investigate the phenotypic characteristics and underlying mechanisms. Serum C1QTNF4 levels were decreased in patients with arterial stenosis. C1QTNF4 shows colocalisation with VSMC in human renal arteries. In vitro, C1QTNF4 inhibits VSMC proliferation and migration and alters VSMC phenotype. In vivo, an adenovirus-infected rat balloon injury model, C1QTNF4-transgenic and C1QTNF4-/- mouse wire-injury models with or without VSMC-specific C1QTNF4 restoration were established to mimic the VSMC repair and remodelling. The results show that C1QTNF4 decreases intimal hyperplasia. Especially, we displayed the rescue effect of C1QTNF4 in vascular remodelling using AAV vectors. Next, transcriptome analysis of artery tissue identified the potential mechanism. In vitro and in vivo experiments confirm that C1QTNF4 ameliorates neointimal formation and maintains vascular morphology by downregulating the FAK/PI3K/AKT pathway. Our study demonstrated that C1QTNF4 is a novel inhibitor of VSMC proliferation and migration that acts by downregulating the FAK/PI3K/AKT pathway, thus protecting blood vessels from abnormal neointima formation. These results provide new insights into promising potent treatments for vascular stenosis diseases.

Targeting the elevated IFN-γ in vitiligo patients by human anti- IFN-γ monoclonal antibody hampers direct cytotoxicity in melanocyte

BackgroundVitiligo is an autoimmune disease that progressively destroys melanocytes in the skin, resulting in patchy disfiguring depigmentation. The direct pathological effect of IFN-γ, CXCL10 to the melanocytes in vitiligo has been reported, but there are contradictory results to which cytokine exerts the critical cytotoxic effect on melanocytes. ObjectiveThe overarching goal was to study the direct toxicity of highly expressed cytokine in vitiligo skin lesions to melanocytes. MethodsWe obtained the interstitial fluid analyte from lesion and non-lesion skin of vitiligo patients and healthy control and sent for high sensitivity multiplex cytokine panel. We further performed functional study to identify the direct toxicity effect of the highly expressed cytokines. ResultsWe found a significant elevation of IFN-γ, CXCL9, CXCL10, CXCL11 in the vitiligo skin. Ex vivo melanocyte studies support the direct role of IFN-γ per se in melanocyte cell loss, increased oxidative stress and melanogenesis disruption. Interestingly, we found that IFN-γ regulated cell death through oxidative stress-related ferroptosis cell death, which may initiate autoimmunity in vitiligo. In contrast to blocking selected cell death pathway, our in vitro study supports the rescue effect of human anti-IFN-γ monoclonal antibody 2A6Q to IFN-γ induced cell death, oxidative stress, and loss of function in melanocytes by interrupting IFN-γ signaling, which may be a potential therapeutic option for vitiligo. ConclusionThis study further confirms the direct of toxicity effect of IFN-γ per se towards melanocyte in vitiligo skin and the potential utility of human anti-IFN-γ monoclonal antibody in treating vitiligo.

Single-cell analysis of immune cells on gingiva-derived mesenchymal stem cells in experimental autoimmune uveitis

Gingiva-derived mesenchymal stem cells (GMSCs) have shown astonishing efficacy in the treatment of various autoimmune diseases. However, the mechanisms underlying these immunosuppressive properties remain poorly understood. Here, we generated a lymph node single-cell transcriptomic atlas of GMSC-treated experimental autoimmune uveitis mice. GMSC exerted profound rescue effects on Tcells, B cells, dendritic cells, and monocytes. GMSCs rescued the proportion of T helper 17 (Th17) cells and increased the proportion of regulatory Tcells. In addition to globally altered transcriptional factors (Fosb and Jund), we observed cell type-dependent gene regulation (e.g., Il17a and Rac1 in Th17 cells), highlighting the GMSCs' cell type-dependent immunomodulatory capacity. GMSCs strongly influenced the phenotypes of Th17 cells, suppressing the formation of the highly inflammatory CCR6-CCR2+ phenotype and enhancing the production of interleukin (IL) -10 in the CCR6+CCR2+ phenotype. Integration of the glucocorticoid-treated transcriptome suggests a more specific immunosuppressive effect of GMSCs on lymphocytes.

Correlation of Electrophysiological and Fluorescence-Based Measurements of Modulator Efficacy in Nasal Epithelial Cultures Derived from People with Cystic Fibrosis.

It has been suggested that in vitro studies of the rescue effect of CFTR modulator drugs in nasal epithelial cultures derived from people with cystic fibrosis have the potential to predict clinical responses to the same drugs. Hence, there is an interest in evaluating different methods for measuring in vitro modulator responses in patient-derived nasal cultures. Commonly, the functional response to CFTR modulator combinations in these cultures is assessed by bioelectric measurements, using the Ussing chamber. While this method is highly informative, it is time-consuming. A fluorescence-based, multi-transwell method for assaying regulated apical chloride conductance (Fl-ACC) promises to provide a complementary approach to theratyping in patient-derived nasal cultures. In the present work, we compared Ussing chamber measurements and fluorescence-based measurements of CFTR-mediated apical conductance in matching, fully differentiated nasal cultures derived from CF patients, homozygous for F508del (n = 31) or W1282X (n = 3), or heterozygous for Class III mutations G551D or G178R (n = 5). These cultures were obtained through a bioresource called the Cystic Fibrosis Canada-Sick Kids Program in Individual CF Therapy (CFIT). We found that the Fl-ACC method was effective in detecting positive responses to interventions for all genotypes. There was a correlation between patient-specific drug responses measured in cultures harbouring F508del, as measured using the Ussing chamber technique and the fluorescence-based assay (Fl-ACC). Finally, the fluorescence-based assay has the potential for greater sensitivity for detecting responses to pharmacological rescue strategies targeting W1282X.

Abstract LB256: Type 1 calreticulin mutations in myeloproliferative neoplasms promote glycolysis via intracellular calcium mediated upregulation of GLUT1

Abstract Myeloproliferative neoplasms (MPNs) comprise a group of cancers of the bone marrow that can transform into a post-MPN acute myeloid leukemia (AML), at which point the overall survival rate is less than 6 months. JAK2 inhibitors in MPN failed to alter disease progression, emphasizing the need for new treatments. Calreticulin (CALR) is one of the MPN driver mutations and encodes an endoplasmic reticulum (ER) calcium (Ca2+) binding protein. CALR mutations are classified as either type 1 or type 2. Only type 1 proteins exhibit loss of the C-terminal Ca2+ binding sites. Additionally, type 1 and 2 CALR mutations engender significant prognostic differences. Previously published work from our lab showed that type 1 CALR exhibits a loss of Ca2+ binding sites, rendering them unable to bind ER Ca2+ and leading to depletion of ER Ca2+ stores. We next wanted to evaluate where this Ca2+ is localized. Through imaging studies, we found that CALR type 1 expressing cells have significantly increased cytosolic Ca2+ and significantly lower mitochondrial Ca2+ compared to both CALR wildtype and CALR type 2 expressing cells. It has been well established in various cancer types that increased glycolytic activity is prompted in part by elevated cytosolic Ca2+. To test if this was the case in our model, we performed a vast array of metabolic assays in UT-7-MPL cells, a human megakaryocytic cell line, including but not limited to lactate secretion, Seahorse XF Glycolysis Stress Test assay, and metabolite tracing with 13-C labelled glucose. These all confirmed a highly glycolytic profile in UT-7-MPL cells expressing the CALR type 1 mutation. In addition, through evaluation of mitochondrial mass, membrane potential and several other assays, mitochondrial metabolism in CALR type 1 mutant cells appears severely dysfunctional. All the glycolytic and mitochondrial phenotypes are almost fully rescued upon the reintroduction of the Ca2+ binding sites of the CALRwt protein (P and C domains; P+C), indicating that perturbed Ca2+ homeostasis is the driver of these changes. Pharmacological treatment with cytosolic Ca2+ chelator BAPTA-AM or with kaempferol, an activator of the mitochondrial Ca2+ uniporter mimic the P+C rescue effect for both the glycolytic and the mitochondrial phenotypes. In our in vitro system CALR type 1 mutant cells express along with other glycolytic factors significantly higher cell surface levels of the glucose transporter GLUT1, and demonstrate enhanced sensitivity to pharmacological GLUT1 inhibition with BAY-876. Preliminary data in a transgenic mouse model as well as in primary patient samples show that GLUT1 expression is higher in CALR type 1 mutant mice and patient cells, and that the mitochondrial mass in this subset of patients is lower compared to CALR type 2 mutant patients. Finally, we validated that all these phenotypes are JAK/STAT pathway independent and that they are worsened by ruxolitinib treatment, providing a strong rationale for a combination treatment. Together, these data both expand the current knowledge of the role that Ca2+ dynamics play in metabolic reprogramming and can lead to improved targeted therapies for MPN patients. Citation Format: Michele Ciboddo, Jonathan Dowgielewicz, George Yan, Deborah Rodriguez, Hunter Blaylock, Nicole Arellano, Chad Coen, Chufan Cai, Nia Hammond, Alex Rosencrance, Elisa Rumi, Daniela Pietra, Daniele Vanni, Silvia Catricalà, Ilaria Carola Casetti, Oscar Borsani, Brandon Faubert, Shannon Elf. Type 1 calreticulin mutations in myeloproliferative neoplasms promote glycolysis via intracellular calcium mediated upregulation of GLUT1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB256.

Demand Grading of Emergency Medical Materials Based on Factor Analysis

Reasonable and scientific classification and urgency classification of emergency materials is the premise and basis of emergency management, which affects the effect and efficiency of emergency rescue. Emergency medical materials are related to people's life, health, and safety. In this paper, based on the early stage of COVID-19, the demand for emergency medical materials is combined to determine the demand grading evaluation indicator system, and factor analysis is used to reduce the dimensionality of data and to determine the weights of each indicator. Therefore, a comprehensive evaluation model is established and the demand for emergency medical materials is graded by clustering.

Not All Friends Are Alike: A Categorization of Friendly Acquisitions Integrating the Acquired Firm Profile with the Acquirer's Approach

AbstractAlthough previous studies have examined the distinct motivations of acquirers and contrasted the differences between friendly and hostile acquisitions, the combination of the acquirer and target firm perspectives, particularly within friendly acquisitions, has not yet been addressed. By adopting a multidisciplinary approach and integrating the acquiring and acquired firms’ perspectives, this study identifies different types of friendly deals and investigates their impact on post‐acquisition performance. A unique dataset of 668 acquisitions was constructed by merging the Zephyr and Orbis databases. A regression analysis revealed that, in general, rescue and appealing targets obtain better post‐acquisition performance than average targets. The effect of rescue and appealing targets is accentuated by greater accumulated knowledge. Conversely, in partial acquisitions, the effect of rescue and appealing targets is diminished for high‐accumulated knowledge. This signal change of the knowledge‐moderating parameter implies that under general conditions, rescue and appealing targets outperform average targets. However, if the acquirer seeks to follow a collaborative approach to extract the accumulated knowledge of the target firm, then it is better to signal a more cooperative stance through a partial acquisition.