The correlation between altmetric score and traditional measures of article impact for studies pertaining to spine trauma.

PURPOSE: It is becoming increasingly common for researchers to share scientific literature via social media. Traditional bibliometrics have long been utilized to measure a study's academic impact, but they fail to capture the impact generated through social media sharing. Altmetric Attention Score (AAS) is a weighted count of all the online attention garnered by a study, and it is currently unclear whether a relationship with traditional bibliometrics exists. METHODS: We identified the five highest-rated spine-specific and five highest-rated general orthopedic journals by Scopus CiteScore 2020. We then identified all the spine trauma studies across a 5-year span (2016-2020) within these journals and compared AAS with traditional bibliometrics using Independent t-tests and Pearson's correlational analyses. RESULTS: No statistically significant relationships were identified between AAS and traditional bibliometrics for articles pertaining to spine trauma: Level of Evidence (R = - 0.02, p = 0.34), H-Index Primary Author (R = < - 0.01, p = 0.50), H-Index Senior Author (R = - 0.04, p = 0.24), and Number of Citations (R = 0.01, p = 0.40). The top five articles by AAS include those pertaining to motorcycle injuries (AAS = 687), orthosis in thoracolumbar fractures (AAS = 199), golfing injuries (AAS = 166), smartphone-based teleradiology (AAS = 41), and auto racing injuries (AAS = 39). CONCLUSION: The lack of overlap between these types of metrics suggests that AAS or similar alternative metrics should be used to measure an article's social impact. The social impact of an article should likewise be a factor in determining an article's overall impact along with its academic impact as measured by bibliometrics.

Inhibition of LPS-mediated activation in rat Kupffer cells by N-acetylcysteine occurs subsequent to NF-kappaB translocation and requires protein synthesis.

Activation of the resident hepatic macrophage population, Kupffer cells, leads to production of mediators that initiate, potentiate, and modulate hepatic injury. Recent studies have shown that activation of the pluripotent transcription factor nuclear factor-kappaB (NF-kappaB) is an important step in the induction of inflammatory cytokines, chemokines, growth factors, cell adhesion proteins, and cytokine receptors, thus efforts have been focused to modulate its activity. A common observation in diverse experimental systems is that oxidant stress activates NF-kappaB and antioxidant drugs prevent activation and subsequent inflammatory gene transcription. However, we have recently shown that the inhibitory effect of N-acetylcysteine (NAC) is independent of its role as a substrate of glutathione synthesis and NAC can inhibit Kupffer cell activation at points beyond the initiation of activation. The goal of this study was to characterize the mechanism for NAC-mediated inhibition of Kupffer cell activation. We show for the first time that this process requires a cellular synthetic response to prevent both NF-kappaB and tumor necrosis factor alpha (TNF-alpha) mRNA activation. Furthermore, NAC-mediated inhibition occurs after degradation of IkappaB-alpha and nuclear translocation of NF-kappaB. These data suggest that inhibition of Kupffer cell activation by NAC is a nuclear event and offers a potential approach to modulate Kupffer cell activation during hepatic injury.

CD14 mediate endotoxin induction of nitric oxide synthase in cultured brain glial cells.

Here we show that lipopolysaccharide (LPS) induction of glial inducible nitric oxide synthase (iNOS) requires membrane (m) and soluble (s) forms of CD14. In glial cell cultures, an anti-rat CD14 monoclonal antibody detected CD14 protein in whole cells and cell lysates, and reduced LPS-dependent iNOS expression. Glial cells and normal brain tissue expressed CD14 mRNA, as revealed by isolation of a rat CD14 clone (rCD14) from an astrocyte cDNA library and RT-PCR analysis. Finally, serum the ED(50) of LPS required for glial iNOS expression, and antibodies against sCD14 blocked the potentiating effect of serum.

Alterations in tumor necrosis factor-alpha expression by hepatic macrophages following acute cholestatic liver injury.

The liver is unique for its large resident macrophage (HM phi) population as a potential source of immunoregulatory cytokines. The present study was designed to determine HM phi function in a rat model of cholestasis (CBDL). Northern blot analysis of TNF-alpha mRNA showed a profound difference in the dose response to bacterial lipopolysaccharide (LPS) between sham and CBDL HM phi. Sham HM phi demonstrated an 8-fold difference in induction of TNF-alpha mRNA versus CBDL HM phi. TNF-alpha secretion, determined by enzyme-linked immunosorbent assay, was significantly higher from LPS-activated sham HM phi versus the same cells activated with Gram-positive bacterial peptidoglycan while CBDL HM phi were more responsive to peptidoglycan than to LPS. These results demonstrate stimulus- and response-specific functional alterations in the HM phi population during acute cholestatic injury. We speculate that these functional alterations are phenotypically induced in acute liver injury resulting in responses that are not characteristic of normal HM phi.

Lipopolysaccharide and tumor necrosis factor-alpha synergy potentiate serum-dependent responses of rat macrophages.

Tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) are major mediators of sepsis and multiple organ failure. Serum-mediated macrophage activation requires lipopolysaccharide (LPS) and its serum binding protein, lipopolysaccharide binding protein as a ligand for the receptor CD14. This study was designed to determine whether cytokines participate in regulation of serum-mediated LPS activation. Rat macrophages were stimulated with LPS with and-without TNF-alpha or IL-1 beta and activation was determined by detection of TNF-alpha by specific enzyme-linked immunosorbent assay or TNF-alpha mRNA by Northern blot analysis. The addition of TNF-alpha but not IL-1 beta, in the presence of serum, leads to potentiation of macrophage activation after LPS stimulation. This effect could be specifically inhibited by neutralization of LPS with polymyxin B or an antibody against TNF-alpha. This study shows that LPS and TNF-alpha synergize to potentiate serum-mediated macrophage activation. These results demonstrate another element of the control mechanism of cytokine secretion following macrophage activation in sepsis.

CD14-lipopolysaccharide receptor activity in hepatic macrophages after cholestatic liver injury.

BACKGROUND: Increased septic complications and altered cytokine responses to bacterial endotoxins are lethal consequences of chronic liver disease. Kupffer cells (HM phi) control the clearance of lipopolysaccharide (LPS) and subsequent cytokine responses. The purpose of this study is to determine the phenotype and function of HM phi after cholestatic liver injury. METHODS: Male Sprague-Dawley rats underwent 4 days of common bile duct division and ligation or sham laparotomy. HM phi were isolated by collagenase-pronase perfusion and purified by centrifugal elutriation. RESULTS: Indirect immunofluorescence and Western blot analysis with monoclonal antibodies OX42 and ED9 showed that HM phi from cholestatic rats expressed MAC-1 (CD11b) and the LPS receptor CD14, respectively. HM phi from sham animals were negative for CD14 and CD11b by use of immunofluorescence; however, Western blot detected low levels of CD14 in controls. Functional analysis for tumor necrosis factor-alpha release after LPS stimulation showed that HM phi isolated from cholestatic livers exhibited serum (normal rat serum) dependent stimulation (1 ng/ml LPS + 1% normal rat serum; n = 12; p < 0.05 at 8 hours, t test) consistent with the presence of CD14 on their cell surface. CONCLUSIONS: This study shows that HM phi isolated after common bile duct division and ligation are phenotypically and functionally different from normal HM phi. The appearance of serum-dependent LPS responses within the injured liver may play an important role in the immunologic alterations associated with hepatic disease.

Molecular and cellular control points in pediatric liver injury and repair.

Several exciting areas of cellular and molecular biology of the liver have led to a better understanding of the mechanisms of pediatric liver injury and repair. Soon these advances will lead to treatment options for specialized areas of pediatric hepatology. Most of the current goals of surgical therapy lead to either successful hepatic resection for tumors or biliary decompression for atresia. We have come to accept the unfortunate fact that ongoing chronic liver disease ultimately leads to replacement by transplantation. The purpose of this article is to demonstrate areas of recent basic science advancement, directly related to pediatric liver disease, that may provide opportunities and new strategies to obviate the progression from early injury to end-stage liver disease or to augment repair of the injured liver.

The inflammatory response in pediatric biliary disease: macrophage phenotype and distribution.

PURPOSE: Extrahepatic biliary obstruction in infants and children leads to ductal hyperplasia and portal fibrosis. Inflammatory mediators responsible for increased cellular proliferation and matrix deposition are hypothesized to result from the intrahepatic recruitment and activation of lymphocytes and macrophages (M phi). The authors previously showed components of this mechanism in studies that demonstrated increased adhesion molecule expression in biliary atresia, as well as evidence of altered hepatic M phi function during the course of experimental cholestatic liver injury. Therefore they sought determine the expression of macrophage receptor markers CD68 and CD14 in pediatric biliary disease. METHODS: Sixteen liver specimens were snap-frozen and cryosectioned onto polylysine-coated slides. Sections were stained with murine monoclonal antibodies to CD68 (resident M phi) and CD14 (monocyte-M phi lipopolysaccharide [LPS] receptor) glycoproteins. The sections were analyzed using a semiquantitative scale of proliferation and were position-graded from 0 to 3 (maximal). RESULTS: Blinded analysis showed that marked proliferation of CD68-positive cells occurred in five of the six patients with biliary atresia (BA) and in one patient who had severe cholestasis. Normal perisinusoidal liver M phi were found in specimens from patients with hepatitis (2), choledochal cyst (1), and congenital hepatic fibrosis (1). Similarly, expression of CD14 periportal M phi was found only in patients with BA or cholestasis (1.9 +/- 0.3 [mean +/- SEM]) and was absent in other diseases. Strong sinusoidal expression of CD14 was evident in all patients who had extrahepatic biliary obstruction. An early biopsy specimen from a premature infant with BA did not show cholestasis, fibrosis, CD68 Mø proliferation, or CD14 expression; however, another biopsy specimen, obtained further in the course of jaundice showed the progressive development of all features. CONCLUSION: These findings suggest proliferation of resident M phi in association with cholestasis. The presence of the LPS receptor on periportal cells during cholestatic liver injury points to a potential source of cytokines responsible for the inflammatory reaction of biliary atresia.

Uptake and modification of 125I-lipopolysaccharide by isolated rat Kupffer cells.

While it is generally believed that hepatic clearance of lipopolysaccharide involves Kupffer cells, the mechanism involved has not been fully elucidated. This study assesses this phenomenon in terms of in vitro uptake and post-uptake modification experiments with an 125I-labeled Salmonella minnesota lipopolysaccharide. 125I-Lipopolysaccharide was added to Kupffer cells in suspension cultures under a variety of conditions. In vitro uptake of 125I-Lipopolysaccharide was not saturable up to concentrations of 33.33 micrograms per ml. Kinetics experiments performed at 16.67 micrograms per ml demonstrated that Kupffer cells were unsaturable after 60 min of incubation. The kinetics of uptake could be inhibited, however, by incubation in the presence of a 10-fold excess of unlabeled lipopolysaccharide, indicating that a component of the uptake process may be limited. Energy dependence in this process was demonstrated by incubation in the presence of 1 mM 2-deoxyglucose which inhibited 125I-lipopolysaccharide uptake by approximately 30%. Pretreatment with 7.5 x 10(-5) M colchicine had no effect on kinetics, implying no role for the cell cytoskeleton in lipopolysaccharide uptake. These results are inconsistent with a receptor-mediated process as previously suggested. Modification of internalized label has been demonstrated by changes in buoyant density in CsCl isopyknic density gradients following overnight incubation with Kupffer cells. These results indicate that Kupffer cells clear bacterial endotoxin in vitro and post-uptake degradation occurs within 20 hr of incubation.

Clearance of gut-derived endotoxins by the liver. Release and modification of 3H, 14C-lipopolysaccharide by isolated rat Kupffer cells.

This paper describes experiments that were designed to study postuptake modification by isolated rat Kupffer cells of a 3H,14C-biosynthetically labeled endotoxin purified from Escherichia coli J5 as assessed by cesium chloride isopyknic density gradients and gel permeation chromatography. Pulse-chase experiments demonstrated that half as much of the endotoxin's lipid, relative to polysaccharide, was released by the cells. Density gradients revealed that native endotoxin equilibrated at a density of 1.412 g/ml, whereas endotoxin retained by Kupffer cells equilibrated at densities of 1.274 and 1.295 g/ml. Gel permeation chromatography indicated that endotoxin retained by Kupffer cells formed a larger micelle than either exocytosed or native endotoxin. Endotoxin exocytosed by Kupffer cells fractionated into two peaks, one with a smaller and one with a larger apparent micelle size than native endotoxin but both smaller than the retained lipopolysaccharide. Both systems indicated that the Kupffer cell modified endotoxin by enriching the lipid content of the molecule and shortening the length of the O-antigen. Thus, the Kuffer cell, in its mode of action on the endotoxin molecule, appears to play a prominent role in the initial phase of a biochemical process for endotoxin clearance and detoxification.

Comparative studies of endotoxin uptake by isolated rat Kupffer and peritoneal cells.

The process of uptake of endotoxin by cells of the reticuloendothelial system is of current interest. Rabbit peritoneal macrophages have been used to study macrophage-endotoxin interactions and have suggested a receptor-mediated process. It is generally believed that the site of in vivo endotoxin clearance is the liver and that this clearance involves the Kupffer cell population. In the current report, the uptake characteristics of iodine-125-labeled Salmonella minnesota lipopolysaccharide (LPS) were compared in both isolated rat Kupffer cells and elicited rat peritoneal cells. Both types of cells were isolated from male Sprague-Dawley rats fed a semisynthetic AIN-76 5% saturated-fat diet either by peritoneal lavage for peritoneal cells or by collagenase perfusion followed by purification on a 17.5% metrizamide gradient for Kupffer cells. Hot phenol water-extracted S. minnesota LPS was labeled with iodine by the chloramine-T method following a reaction with methyl-p-hydroxybenzimidate. The in vitro uptake of [125I]LPS by Kupffer cells was unsaturable up to concentrations of 33.33 micrograms/ml, while peritoneal cells became saturated at between 16.67 and 25 micrograms of LPS per ml. Uptake by both types of cells could be inhibited by a 10-fold excess of unlabeled LPS. Kinetic experiments demonstrated that Kupffer cells were unsaturable after 60 min of incubation, while peritoneal cells were saturable after 40 min of incubation. Pretreatment with 75 mM colchicine inhibited uptake by peritoneal cells but not Kupffer cells, while pretreatment with 12 mM 2-deoxyglucose inhibited uptake by Kupffer cells but not peritoneal cells. These results are consistent with a process of receptor-mediated endocytosis for peritoneal cells, while Kupffer cells may internalize endotoxins by absorptive pinocytosis. These results suggest that studies of peritoneal cell-endotoxin interactions do not accurately describe the physiologic process within the liver, the major site for the clearance of gut-derived endotoxins.

NF-kappaB activation and modulation in hepatic macrophages during cholestatic injury.

Cholestatic liver injury induces an inflammatory response that follows the activation of hepatic macrophages. Constitutive activation of the transcription factor, NF-kappaB, was found in these macrophages over the course of hepatic injury. Since NF-kappaB activation has been shown to have a key role in the inflammatory process, the modulatory effects of the antioxidant, alpha-tocopherol succinate, and the glucocorticoid, dexamethasone, on NF-kappaB activation were examined in this study. Male Sprague Dawley rats underwent 2-7 days of common bile duct division and ligation (CBDL) or sham laparotomy. Hepatic macrophages were isolated by collagenase Pronase perfusion and purified by centrifugal elutriation. Activation was determined by electrophoretic mobility shift assay and ELISA. We determined that NF-kappaB activation in injured hepatic macrophages could only be inhibited by dexamethasone. Dexamethasone-mediated inhibition of NF-kappaB activation required the synthesis of a regulatory protein since cycloheximide-treated cells were resistant to its effects. Furthermore, dexamethasone-treated hepatic macrophages showed elevated steady-state levels of IkappaB-alpha mRNA, suggesting the role of IkappaB-alpha as a potential regulatory mediator. Consistent with constitutive transcriptional activation we showed constitutive secretion of TNF-alpha from injured hepatic macrophages which could be inhibited by dexamethasone. These data show for the first time, in a biologically significant model of hepatic injury, constitutive activation of the key inflammatory transcription factor NF-kappaB and cytokine TNF-alpha. These results support an approach focused on the NF-kappaB/IkappaB-alpha pathway as a critical target for therapeutic intervention during hepatic injury, and the consideration of possible steroid-based therapies.

Inhibition of the Kupffer cell inflammatory response by acute ethanol: NF-kappa B activation and subsequent cytokine production.

Suppression of host defense mechanisms plays a critical role in the response to infectious agents in patients with alcohol-induced liver disease. Kupffer cells, the resident hepatic macrophage population, are an essential component of host defense mechanisms against infection. Thus, regulation of the Kupffer cell inflammatory response by ethanol may be a key component of that immunosuppression. The aim of this study is to test the hypothesis that ethanol directly and specifically inhibits Kupffer cell activation. Kupffer cells were incubated in 100 mM ethanol for 90 minutes, whereupon cells were washed and incubated without ethanol for LPS activation. Such treatments lead to inhibition of LPS-mediated NF-kappa B activation. Consistent with these data, steady state levels of TNF-alpha and TNF-alpha secretion were depressed throughout a range of LPS concentrations. The inhibition induced by ethanol was time dependent and completely reversible. These data show that the suppressive effects of ethanol affect the the earliest steps of the LPS signal transduction cascade as it is currently understood.

Polyenylphosphatidylcholine inhibits PDGF-induced proliferation in rat hepatic stellate cells.

Polyenylphosphatidylcholine (PPC), a polyunsaturated phospholipid extract from soy beans, prevents the development of liver cirrhosis in animal models. Its mechanism of action is unknown. Based on the hypothesis that PPC might act by decreasing hepatic stellate cell proliferation, we studied the effect of PPC and its main components, dilinoleoylphosphatidylcholine (DLPC) and palmitoyl-linoleoylphosphatidylcholine (PLPC), on PDGF-induced stellate cell proliferation and intracellular signal transduction. Normal rat hepatic stellate cells in tissue culture were serumstarved, and incubated with 10ng/ml PDGF in the absence or presence of phospholipids. Cell proliferation was measured by 3H-thymidine incorporation. P44MAPK activation was determined by kinase assay, and AP-1 binding by electrophoretic mobility shift assay. PPC (200 ng/ml) significantly inhibited PDGF-induced proliferation (p < 0.05; ANOVA, n = 3) and antagonized PDGF-induced P44MAPK activation and AP-1 binding. This effect was mimicked by DLPC but not by PLPC. Neither DLPC nor PLPC prevented PDGF receptor activation. We conclude that PPC exerts a previously unrecognized effect on mitogen-induced stellate cell proliferation which may be mediated by DLPC. Inhibition of this cascade represents a potential mechanism for the inhibitory effect of PPC on hepatic fibrogenesis.

Spontaneous insertion of an IS2 element into the promoter region of the lac operon.

A spontaneous mutation in pUC18 has revealed the insertion of a chromosomal insertion sequence (IS)2 element into the promoter region of the lac operon. The IS2 insertion site, at the pentanucleotide sequence TCGAG, is unlike previously described junctional sequences.

Protective role of NO in hepatic microcirculatory dysfunction during endotoxemia.

Nitric oxide (NO) has been reported to have a protective function in attenuating hepatic injury during endotoxemia or sepsis. As a result, the role of NO in attenuating the hepatic microcirculatory alterations associated with endotoxemia was investigated in mice by in vivo microscopy. The livers were examined 2 h after intravenous injection of Escherichia coli 0111:B4 lipopolysaccharide (LPS) alone or in combination with inhibitors of the synthesis of NO, NG-nitro-L-arginine methyl ester or NG-monomethyl-L-arginine. In the animals treated with the combination of NO synthase inhibitors and LPS, leukocyte adherence was increased threefold above that in animals treated with LPS alone. This was accompanied by a 33% reduction in sinusoidal blood flow. Simultaneous administration of L-arginine, but not D-arginine, eliminated these microcirculatory disturbances. The results demonstrate that inhibition of LPS-stimulated NO production results in an early hepatic microvascular inflammatory response to a dose of endotoxin which by itself is scarcely inflammatory. This suggests that NO plays a significant role in stabilizing the hepatic microcirculation during endotoxemia, thereby helping to protect the liver from ischemia and leukocyte-induced oxidative injury.

N-acetylcysteine and alpha-tocopherol reverse the inflammatory response in activated rat Kupffer cells.

Activation of the resident macrophage populations of the reticuloendothelial system is a key component of the complex pathophysiology of sepsis. Macrophage activation leads to production and secretion of inflammatory mediators such as cytokines, vasoactive substances, free radicals, and chemokines, which have been associated with high morbidity and mortality in the septic patient. The goal of the present study was to determine whether antioxidants could suppress Kupffer cell activation at points beyond the initiation of activation. Kupffer cells were studied since they are central to the clearance of bacteria and endotoxins, and have been associated with hepatocellular dysfunction in sepsis. Cells were activated with 10 ng/ml LPS for various times whereupon N-acetylcysteine (30 mM) and alpha-tocopherol (50 microM) were added. Steady state levels of cytokine mRNA, activation of nuclear factor-kappaB, and TNF-alpha secretion were determined when expression was maximal in control cells. The results of this study show that antioxidants can be used to suppress Kupffer cell activation at points beyond the initiation of activation. Furthermore, we show that N-acetylcysteine-mediated inhibition of activation requires secondary protein synthesis, but does not modulate IkappaB-alpha mRNA expression. The inhibitory effect of these drugs occurs at the very earliest steps of the LPS signal transduction cascade as it is currently understood. The results of the present study suggest that the inflammatory response to sepsis may be controlled through appropriate antioxidant therapy.

LPS-mediated NF-kappa beta activation in rat Kupffer cells can be induced independently of CD14.

Lipopolysaccharide (LPS) activation of macrophages occurs after LPS complexed with serum LPS-binding protein (LBP) binds CD14. Activation of the nuclear transcription factor NF-kappa B is directly related to this event. Since the role of CD14 in LPS signaling has not been evaluated in Kupffer cells, the resident hepatic macrophage, the purpose of this study was to characterize LPS-mediated NF-kappa B activation under CD14-dependent (1% serum, as a source of LBP) and CD14-independent (serum-free) conditions. Classic CD14-dependent signaling was seen in peritoneal macrophages where serum potentiated NF-kappa B activation. However, in Kupffer cells, NF-kappa B was activated by LPS under CD14-independent conditions, and this response was not potentiated by serum. The activation of NF-kappa B in Kupffer cells, by 1 ng/ml LPS, reached a maximum within 60 min of stimulation. However, peritoneal macrophage NF-kappa B activation occurred only in serum and increased progressively through 240 min of stimulation. These results suggest a novel mechanism of LPS-mediated activation in Kupffer cells that may represent an adaptation to their role in clearance and detoxification of gut-derived endotoxin.