Comparative outcomes of robot-assisted minimally invasive versus open esophagectomy in patients with esophageal squamous cell carcinoma: a propensity score-weighted analysis.

Robots are increasingly used in minimally invasive surgery. We evaluated the clinical benefits of robot-assisted minimally invasive esophagectomy (RAMIE) in comparison with the conventional open esophageal surgery. From 2012 to 2016, 371 patients with esophageal squamous cell carcinoma underwent an Ivor Lewis or McKeown procedure at our institution. Of these, 130 patients underwent laparoscopic gastric conduit formation followed by RAMIE, whereas 241 patients underwent conventional esophageal surgery, including laparotomy and open esophagectomy (OE). We compared the short- and long-term clinical outcomes of these patients using the propensity score-based inverse probability of treatment weighting technique (IPTW). Among the early outcomes, the OE group showed a higher incidence of pneumonia (P = 0.035) and a higher requirement for vasopressors (P = 0.001). Regarding the long-term outcomes, all-cause mortality was significantly higher (P = 0.001) and disease-free survival was lower (P = 0.006) in the OE group. Wound-related problems also occurred more frequently in the OE group (P = 0.020) during the long-term follow-up. There was no statistical intergroup difference in the recurrence rates (P = 0.191). The Cox proportional-hazard analysis demonstrated that wound problems (HR 0.16, 95% CI 0.02-0.57; P = 0.017), pneumonia (HR 0.23, 95% CI 0.06-0.68; P = 0.019), and use of vasopressors (HR 0.14, 95% CI 0.08-0.25; P = 0.001) were independent predictors of mortality. RAMIE could be a better surgical option for selected patients with esophageal squamous cell carcinoma.

Correlation of cytokine elaboration with mononuclear cell adhesion to platelet storage bag plastic polymers: a pilot study.

The basis for many febrile nonhemolytic transfusion reactions associated with platelet transfusion therapy is cytokine elaboration and accumulation in the storage bag, which correlate with the leukocyte content and the length of platelet storage. We propose that a possible additional variable in the elaboration and accumulation of cytokines is the differential adhesion of mononuclear cells to the plastic substrate of the platelet storage bag. We hypothesize that mononuclear cell adhesion-induced cytokine release is greater in random-donor platelet bags composed of the polyolefin polymer compared to the single-donor apheresis platelet bags composed of the polyvinyl chloride polymer with the tri-(2-ethylhexyl) trimellitate (TEHTM) plasticizer. For four blood donors, we demonstrate preferential mononuclear cell adhesion, in vitro, to discs of polyolefin polymer versus discs of polyvinyl chloride polymer with the TEHTM plasticizer. Scanning electron microscopy corroborates this. In addition, proinflammatory cytokine (interleukin 1beta [IL-1beta] and tumor necrosis factor alpha [TNF-alpha]) levels are greater in culture wells containing discs of polyolefin polymer than in those containing discs of polyvinyl chloride polymer with the TEHTM plasticizer, and even more so in storage bags containing polyolefin polymer versus polyvinyl chloride polymer with the TEHTM plasticizer (IL-1beta, TNF-alpha, IL-6, and IL-8). This study suggests, for the first time, that differential plastic substrate mononuclear cell adhesion may contribute to cytokine release during platelet storage. This may represent an additional variable in the pathophysiology of febrile nonhemolytic transfusion reactions in patients receiving stored platelet units.

Cyclic-strain-induced endothelial cell expression of adhesion molecules and their roles in monocyte-endothelial interaction.

Vascular endothelial cells (ECs) are constantly subjected to hemodynamic forces that may regulate monocyte-endothelial interaction in vivo. To examine the effects of cyclic strain on endothelial expression of monocyte adhesion molecules, E-selectin, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) ECs were exposed to physiologically relevant levels of cyclic strain. When ECs were under 25% maximal strain at 30 cycles/min for 24 h, the expression of E-selectin significantly (p<0.05) increased, by 83%, compared to control ECs under static conditions. Similarly, monocyte adhesion to ECs under strain (maximum of 15 or 25% at 30 and 60 cycles/min for 24 h) also significantly (p<0.05) increased, by >82%. This cyclic-strain-induced monocyte adhesion was substantially inhibited (83.5%) by anti-E-selectin antibody. ICAM-1 expression also significantly increased, by 62%, when ECs were under 25% maximal strain at 30 cycles/min for 3 h whereas VCAM-1 expression by ECs under strain (for 0.5, 3, and 24 h) did not change compared to static ECs. When ECs were treated with anti-ICAM-1 antibody and monocytes with anti-VLA-4 antibody, an increase in monocyte adhesion to ECs under cyclic strain was reduced significantly. These results demonstrate that cyclic strain can induce EC expression of monocyte adhesion molecules E-selectin, ICAM-1, and VCAM-1 in a time-dependent manner and thus can mediate monocyte adhesion.

Cyclic strain effects on human monocyte interactions with endothelial cells and extracellular matrix proteins.

Human vascular endothelial cells (ECs) are exposed to various levels of hemodynamic forces, cyclic strain, and shear stress in vivo. Here, we examined the in vitro effects of the various levels (0-6%, 7-16%, and 17-25%) of strain at 60, 30, and 15 cycles per minute (cpm) on human monocyte adherence to endothelial cells and extracellular matrix protein preabsorbed surfaces. Monocyte adhesion to endothelial cells under cyclic strain significantly increased. At both 30 and 60 cpm, ECs under strains of 7-16% and 17-25% showed >52% and >117% higher monocyte adhesion than endothelial cells under static condition when monocytes were added for 0.5 h. This increase in monocyte adhesion to ECs under cyclic strain remained significantly higher even after 24 h of incubation. Human monocyte adhesion to extracellular matrix protein preabsorbed surfaces differed depending on the specific extracellular matrix protein. Monocytes adhered to collagen type I and fibronectin preabsorbed surfaces >50% under 0-6% strain, >23% under 7-16% strain, and >52% under 17-25% strain at 15 and 30 cpm compared to the collagen type V preabsorbed surface. However, when extracellular protein preabsorbed surfaces under cyclic strain were compared to the control static condition, monocyte adhesion did not significantly change on most of other surfaces. These results suggest that cyclic strain may play a role in the regulation of monocyte-endothelial cells/extracellular matrix interactions in vivo.

Propensity for macrophage apoptosis is related to the pattern of expression and function of integrin extracellular matrix receptors.

Ligation of integrins to an extracellular matrix activates signal transduction systems which produce multiple responses in different cell types. Adhesion often provides a survival signal to cells; disruption of adhesion frequently results in apoptosis. Our laboratory has utilized apoptosis-sensitive and -resistant cell lines to investigate the role of integrin expression and function in regulation of apoptosis in macrophages. Chronic exposure of murine macrophage-like RAW264.7 cells to apoptosis-inducing agents (bacterial lipopolysaccharide and interferon-gamma) resulted in the generation of a derivative cell line (RES) resistant to apoptosis. Observation of RAW and RES cultures indicated a difference in adhesion between the two cell types. The two cell lines also exhibit significant differences in expression of integrins previously characterized to be important in apoptosis.

Cellular adaptive responses to low oxygen tension: apoptosis and resistance.

Oxygen plays such a critical role in the central nervous system that a specialized mechanism of oxygen delivery to neurons is required. Reduced oxygen tension, or hypoxia, may have severe detrimental effects on neuronal cells. Several studies suggest that hypoxia can induce cellular adaptive responses that overcome apoptotic signals in order to minimize hypoxic injury or damage. Adaptive responses of neuronal cells to hypoxia may involve activation of various ion channels, as well as induction of specific gene expression. For example, ATP sensitive K+ channels are activated by hypoxia in selective neuronal cells, and may play a role in cell survival during hypoxia/anoxia. Additionally, hypoxia-induced c-Jun, bFGF and NGF expression appear to be associated with prevention (or delay) of neuronal cell apoptosis. In this paper, these adaptive responses to hypoxia in neuronal cells are discussed to examine the possible role of hypoxia in pathophysiology of diseases.

Inflammatory mediators are perpetuated in macrophages resistant to apoptosis induced by hypoxia.

A hypoxic/anoxic microenvironment has been proposed to exist within a vascular lesion due to intimal or medial cell proliferation in vascular diseases. Here, we examined whether hypoxia alters macrophage function by exposing murine macrophage-like RAW 264.7 (RAW) cells to hypoxia (2% O2). When cells were exposed to hypoxia, a significant number of RAW cells underwent apoptosis. Additionally, small subpopulations of RAW cells were resistant to hypoxia-induced apoptosis. Through repeated cycles of hypoxia exposure, hypoxia-induced apoptosis-resistant macrophages (HARMs) were selected; HARM cells demonstrate >70% resistance to hypoxia-induced apoptosis, as compared with the parental RAW cells. When heat shock protein (HSP) expression was examined after hypoxia, we observed a significant decrease in constitutive heat shock protein 70 (HSC 70) in RAW cells, but not in HARMs, as compared with the control normoxic condition (21% O2). In contrast, the expression level of glucose-regulated protein 78 (GRP 78) in RAW and HARM cells after hypoxia treatment was not altered, suggesting that HSC 70 and not GRP 78 may play a role in protection against hypoxia-induced apoptosis. When tumor necrosis factor alpha (TNF-alpha) production was examined after hypoxic treatment, a significant increase in TNF-alpha production in HARM but decrease in RAW was observed, as compared with cells cultured in normoxic conditions. HARM cells also exhibit a much lower level of modified-LDL uptake than do RAW cells, suggesting that HARMs may not transform into foam cells. These results suggest that a selective population of macrophages may adapt to potentially pathological hypoxic conditions by overcoming the apoptotic signal.

In situ changes in the relative abundance of human epidermal cytokine messenger RNA levels following exposure to the poison ivy/oak contact allergen urushiol.

Abstract: Epidermal keratinocytes in culture have been shown to produce many cytokines, and their proteins have been identified in skin tissue samples. It has therefore been assumed that these cytokines are transcribed in vivo by the epidermis in response to contact allergens. In this report, in situ hybridization was used to detect the messenger RNAs for interleukin-1 alpha (IL-1 alpha), interleukin-1 beta (IL-1 beta) and tumour necrosis factor-alpha (TNF-alpha) in samples of human skin prior to and at various times after application of urushiol, the immunogenic component of poison ivy/oak. In sensitive subjects, IL-1 alpha and TNF-alpha mRNAs showed a progressive increase in transcript levels that paralleled the clinical and histological features of the inflammatory process. The time-course of the IL-1 beta response differed from that of IL-1 alpha and TNF-alpha, in that there was an early (by 6 h after urushiol administration) elevation in IL-1 beta mRNA that occurred before there was evidence of inflammation and had returned to background levels by 72 h when the reaction had reached its peak. In contrast to urushiol-sensitive subjects, urushiol-anergic individuals did not exhibit an increase in IL-1 alpha, IL-1 beta or TNF-alpha mRNA levels. The data provide evidence for an in vivo role for epidermal IL-1 alpha, IL-1 beta and TNF-alpha transcription in the regulation of IL-1 beta and TNF-alpha polypeptide levels in the epidermis in response to this common contact allergen.

Messenger RNAs for the multifunctional cytokines interleukin-1 alpha, interleukin-1 beta and tumor necrosis factor-alpha are present in adnexal tissues and in dermis of normal human skin.

Interleukin-1 alpha (IL-1 alpha), interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) are 3 cytokines that play a key rôle in cutaneous homeostasis and in the immunopathogenesis of a number of dermatologic diseases. Most studies have focused on their production by keratinocytes and Langerhans cells. To determine whether there are non-epidermal sites of cytokine transcription, biopsy specimens of normal human skin were probed for IL-1 alpha, IL-1 beta and TNF-alpha messenger RNAs using the method of in situ hybridization. The results demonstrate that each cytokine mRNA is present at multiple sites within the skin, including epidermal appendages and adnexal structures (hair follicles, sebaceous glands), the dermal microvasculature, arrectores pilorum smooth muscle, and the dermal connective tissue. These data provide evidence that in vivo there are multiple sites other than the epidermis of constitutive IL-1 alpha, IL-1 beta, and TNF-alpha gene transcription in normal human skin.

Adhesion and cytokine production by monocytes on poly(2-methacryloyloxyethyl phosphorylcholine-co-alkyl methacrylate)-coated polymers.

Human monocytes isolated from peripheral venous blood were assayed for their ability to adhere to various polymers. The culture supernatants were also assayed for the cytokines, interleukin-1 beta (IL-beta), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha). The polymers evaluated for adherence and cytokine production included Pellethane, polyethylene and poly[n-butyl methacrylate (BMA)] coated with poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-alkyl methacrylate] copolymers. In some experiments the test polymers were adsorbed with fibrinogen or IgG prior to the addition of monocytes. MPC copolymer-coated materials inhibited monocyte and macrophage adhesion after 1 and 8 days of culture relative to corresponding uncoated polymers and tissue culture polystyrene (TCPS). The degree of inhibition by coated Pellethane compared to uncoated Pellethane was the greatest, while inhibition of adhesion by coated poly(BMA) was the least compared to uncoated poly(BMA). However, adhesion was significantly decreased on both coated and uncoated poly(BMA) by day 8. While IL-1 beta, IL-6, and TNF-alpha release was variably influenced by polymer coating, release was consistently inhibited relative to TCPS on day 1. However, cytokine production was not inhibited compared to corresponding uncoated polymers on day 1. With or without protein preadsorption, IL-1 beta release was not detectable in the supernatants of any polymer on day 8, IL-6 production was diminished on day 8, and TNF-alpha production was sustained on day 8. Overall, MPC copolymer-coated and uncoated poly(BMA) were the least stimulating, while TCPS was the most stimulating.(ABSTRACT TRUNCATED AT 250 WORDS)

Human monocyte/macrophage adhesion and cytokine production on surface-modified poly(tetrafluoroethylene/hexafluoropropylene) polymers with and without protein preadsorption.

To study surface property-dependent human monocyte adhesion and cytokine (IL-1 beta, IL-6, TNF-alpha) production, poly(tetrafluoroethylene/hexafluoropropylene) (FEP) polymer was modified to exhibit neutral, anionic, or cationic properties by incorporating amide (CONH2) and/or carboxyl (COOH) or aminoethyl amide [CONH(CH2CH2NH)nCH2CH2NH2] groups on the surface. Monocyte adhesion on surface-modified FEP polymers and cytokines released by monocytes/macrophages (MC/MO) into the culture medium were compared to control tissue culture polystyrene (TCPS) at days 1 and 8. On day 1, the neutral surface FEP polymer with incorporated amide (NH2) groups showed the greatest inhibition of adhesion, 89% (P < .01), and cytokine production (IL-1 beta with 58%, IL-6 with 70%, and TNF-alpha with 39%) compared to control TCPS. In contrast, the highly cationic [CONH(CH2CH2NH)nCH2CH2NH2] surface did not show significant (P > .01) inhibition of monocyte adhesion and cytokine production. When fibrinogen or IgG was preadsorbed to the surface, the inhibitory effects of the neutral surface FEP polymer on monocyte adhesion and cytokine production were not altered. In addition, other surface-modified FEP polymers showed similar inhibition of monocyte adhesion and cytokine production compared to TCPS. Specifically, as the incorporation of carboxyl (COOH) group content increased on FEP polymer surfaces, monocyte adhesion and cytokine production were also increased on day 1 with IgG preadsorption. On day 8, all surface-modified FEP polymers showed significant (P < .01) inhibition of monocyte adhesion when fibrinogen or IgG was preadsorbed. However, without protein (fibrinogen or IgG) preadsorption, monocyte adhesion was not significantly inhibited compared to control TCPS. In addition, cytokine production detected by ELISAs on day 8 showed no detectable levels of IL-1 beta and significantly decreased levels of IL-6 compared to day 1 for all tested polymers, with or without protein preadsorption. Interestingly, the level of TNF-alpha production on day 8 remained high although not as high as on day 1. Based on these results, we suggest that FEP polymers with neutral hydrophilic surface properties may adhere and activate the least number of monocytes, which are important mediators of biocompatibility.

In situ detection of cytokine messenger RNAs in the eccrine sweat gland of normal human skin.

Human sweat and eccrine sweat glands contained the multifunctional polypeptide cytokines, interleukin-1 alpha and beta (IL-1 alpha and beta). To determine whether the sweat gland itself is an actual site of transcription for cytokines we performed in situ hybridization on histologic sections of normal human skin. In this report we show that the mRNAs encoding the cytokines IL-1 alpha, IL-1 beta, and tumor necrosis factor-alpha (TNF-alpha) are present in normal human skin eccrine sweat gland duct and secretory coil epithelium. These results suggest that in vivo the sweat gland is a production site for cytokine polypeptides and may contribute to the exceedingly large quantities of these cytokine proteins found in the epidermis.