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Quantitative analysis of the spreading of the mouse trophoblast in vitro: a model for early invasion.

The outgrowth of the mouse blastocyst in culture represents an in vitro model of trophoblastic invasion. In the present study we analysed trophoblast spreading by time lapse video microscopy. Trophoblast spreading consists of (1) the migration and (2) the giant cell transformation of trophoblast cells, (3) the proliferation of ectoplacental cone (EPC) cells and (4) the subsequent transformation of EPC cells into the secondary giant cells. During migration, ruffling of the trophoblast cell membrane is followed by the formation of lamellipodia. The mean surface areas of the spreading trophoblast, measured in more than 100 cultured blastocysts, increased linearly from 48 to 96 h of culture, while the linear migratory speed at the periphery of the outgrowth declined as the time of culture advanced. The EPC cells increased in size approximately eightfold during the giant cell transformation. The apparent nuclear:cytoplasmic ratios, i.e., ratios between the size of nucleus and that of the cytoplasm, measured as the surface areas on the photomicrographs, of EPC cells increased between 40-46 h of culture, but a sharp decline in the ratio occurred between 50 and 51 h of culture, reflecting either the sudden and tremendous increase in the cellular volume and/or spreading of the cytoplasm. The rates of trophoblast spreading varied considerably among the blastocysts of different genetic constitution examined (ICR, C57BL/6, C3H/He and (B6 x C3)F1. It was fastest in blastocysts obtained from matings of males and females of (B6 x C3)F1, and slowest in the C57BL/6 embryos. The differences in the rate of outgrowth observed may not simply be ascribed to difference in the developmental speed of the early embryos, because the rate of outgrowth reached a plateau at about 96-120 h and no "catch-up' was observed by leaving the blastocysts in culture longer. Our results strongly suggest the possible presence of genetic regulatory mechanisms underlying trophoblast outgrowth; further analysis of the phenomenon may provide clues to understand the molecular mechanisms of trophoblastic invasion during the early phase of implantation, hopefully leading to improved success rates of in vitro fertilization-embryo transfer.

Animals↗

Phagocyte spreading and phagocytosis in the compound ascidian Botryllus schlosseri: evidence for an integrin-like, RGD-dependent recognition mechanism.

The involvement of integrins in phagocyte spreading and phagocytosis was investigated in the compound ascidian Botryllus schlosseri. The number of spreading cells was significantly reduced when adhesion occurred in the presence of the tetrapeptide Arg--Gly--Asp--Ser (RGDS), but not of Arg--Gly--Glu--Ser (RGES) indicating the involvement of RGD-mediated adhesion mechanisms in phagocyte spreading. The significant decrease of the fraction of spreading cells in the presence of Botryllus blood plasma suggests the presence of RGD-containing molecules in the blood of our species. The increase in the same index when blood plasma-coated slides as well as fibrinogen- and fibronectin-coated coverslips were used, fits with the above hypothesis. Adhesion in the presence of RGDS leads to a consistent alteration of the actin cytoskeleton, in agreement with the known role of integrin adhesion in microfilament organization. Phagocytosis was greatly reduced by RGDS in the incubation medium, but not by RGES, and was significantly increased by coating yeast cells with fibronectin or blood plasma. Both spreading and phagocytic capability were severely inhibited by wortmannin, suggesting the importance of phosphatidylinositol-3-kinase in integrin-mediated signal transduction in ascidians.

Androstadienes↗

Intramolecular epitope spreading induced by staphylococcal enterotoxin superantigen reactivation of experimental allergic encephalomyelitis.

The staphylococcal enterotoxin superantigens are among the most potent T cell stimulators known. They have been shown to alter the course of disease in experimental allergic encephalomyelitis, an animal model for multiple sclerosis (MS). We have previously demonstrated that two of the staphylococcal enterotoxins, SEA and SEB, are able to reactivate paralysis in PL/J mice which had been immunized with myelin basic protein (MBP) and resolved an initial episode of paralysis. In PL/J mice, Ac1-11 is the dominant encephalitogenic determinant of MBP. We hypothesized that superantigen reactivation of experimental allergic encephalomyelitis (EAE) may result in the spreading of T cell specificities for other epitopes of MBP. PL/J mice which had resolved an initial episode of EAE were treated with SEA and developed a second episode of paralysis. At the onset of symptoms, mice were sacrificed and splenocytes were stimulated in vitro with a panel of MBP peptides. EAE reactivation by SEA resulted in the spreading of T cell specificites to residues 100 to 120 of MBP. While intramolecular spreading did occur, spreading to other antigens did not, as evidenced by the lack of response to a proteolipid protein (PLP) peptide and heat shock protein 60 (hsp 60). To further characterize the epitope MBP 100-120, PL/J mice were immunized with MBP 100-120. No initial development of disease was observed. However, administration of SEA 2 weeks after MBP 100-120 immunization resulted in the onset of paralysis. In addition to a proliferative response to MBP 100-120, these mice also exhibited a proliferative response to the flanking MBP peptides 81-100 and 120-140. Thus, SEA is able to induce intramolecular epitope spreading in PL/J mice after reactivation of EAE.

Animals↗

Protein kinase C regulation of cell spreading in the molluscan Biomphalaria glabrata embryonic (Bge) cell line.

Cellular adhesion and spreading are critical components involved in the processes of cell and tissue development, and immune responses in molluscs, but at present, little is known regarding the signaling pathways involved in these basic cellular functions. In the present study, the molluscan Biomphalaria glabrata embryonic (Bge) cell line was used as an in vitro model to study the signal transduction pathways regulating molluscan cell adhesion and spreading behavior. Western blot analysis using antibodies specific to mitogen-activated protein kinase (MAPK) revealed the presence of an MAPK-like immunoreactive protein in Bge cells, that was phosphorylated upon exposure to phorbol myristate acetate (PMA). Moreover, Bge cell treatment with inhibitors of protein kinase C (PKC), Ras and MAPK kinase (Mek) suppressed PMA-induced expression of activated MAPK, suggesting that PKC-, Ras- and Mek-like molecules may be acting upstream of MAPK. Similarly, in vitro Bge cell-spreading assays were performed in conjunction with the same panel of inhibitors to determine the potential involvement of PKC, Ras and Mek in cellular adhesion/spreading. Results revealed a similar pattern of inhibition of cell-spreading behavior strongly implying that the Bge cell spreading also may be regulated through a MAPK-associated signal transduction pathway(s) involving proteins similar to PKC, Ras and Mek.

Alkaloids↗

Pathogenesis of neurotropic herpesviruses: role of viral glycoproteins in neuroinvasion and transneuronal spread.

Neuroinvasion by herpesviruses requires entry into nerve endings in the periphery, transport to the cell body, replication in the cell body, axonal transport to the synapse and transneuronal viral spread. Entry occurs after receptor binding by fusion of virion envelope and cellular plasma membrane followed by microtubuli-assisted transport of capsids to the nuclear pore. By transneuronal spread, the virus gains access to synaptically linked neuronal circuits. A common set of herpesvirus glycoproteins is involved in entry and direct viral cell-cell spread. However, both processes can be distinguished by involvement of additional viral components. Interestingly, transneuronal spread appears to be functionally linked to intracytoplasmic formation of mature virions. This review will focus on the importance of herpesvirus envelope glycoproteins for infection of neurons and transneuronal spread, and their influence on viral pathogenesis.

Animals↗

Perineural spread of cutaneous squamous and basal cell carcinoma: CT and MR detection and its impact on patient management and prognosis.

PURPOSE: To analyze the pretreatment imaging findings and outcome of patients with perineural spread of squamous or basal cell carcinoma of the face and scalp treated with radiotherapy, to determine whether CT (computed tomography) or MR (magnetic resonance) imaging can be effectively used to identify patients who would benefit from aggressive treatment, and to characterize the imaging features associated with cure. METHODS: Thirty-five patients had perineural spread of squamous and basal cell carcinoma along the divisions of the trigeminal and/or facial nerves based on clinical findings and/or histopathological proof. Perineural extension seen on imaging was divided into three zones of involvement. The volume of perineural disease was graded semiquanitatively. All patients received radiotherapy with curative intent. RESULTS: Eighteen of the 35 patients had imaging evidence of perineural spread of tumor, and the remaining 17 were imaging negative for perineural spread. The absolute 5-year survival of the imaging positive group was 50% compared with 86% in the imaging-negative group (p = 0.048). CONCLUSIONS: Imaging can be used to identify patients with advanced perineural spread who warrant aggressive radiotherapy. Imaging evidence of perineural invasion worsens prognosis; however, low-volume and peripheral perineural disease is radiocurable. Greater perineural tumor volume with more central disease was associated with an unfavorable outcome.

Adult↗

Prognostic significance of microscopic and macroscopic extracapsular spread from metastatic tumor in the cervical lymph nodes.

It has been established that the presence or absence of cervical node metastases in patients with head and neck squamous cell carcinoma (HNSCC) is a powerful prognostic indicator. This report reviews the evolution of thinking over the past 70 years with regard to the import and detection of cervical nodal metastases which exhibit spread of tumor beyond the confines of the original encompassing nodal capsule. In the process, this discussion touches upon clinical examination, gross and microscopic pathologic examination, and radiographic imaging studies. In particular, the distinction between gross nodal extracapsular spread of tumor and microscopic nodal extracapsular spread of tumor has been drawn in recent reports; this raises the possibility that identification of microscopic breaching of the nodc capsule by tumor might provide clinically significant information which is not provided by the gross observation of an intact lymph node capsule. While it remains to be seen whether microscopic extracapsular spread alone will prove to be an important prognostic factor, it is recommended that selective neck dissection continue to be offered even in those patients with clinically negative necks; further studies should aid in defining the import of microscopic extracapsular tumor spread in patients with positive cervical nodes.

Carcinoma, Squamous Cell↗

Studies on the 1967-8 foot-and-mouth disease epidemic. The relation of weather to the spread of disease.

An analysis of the 1967-8 foot-and-mouth disease epidemic with reference to the initial spread, the origin of outbreaks more than 60 km. from the main epidemic area, the series of outbreaks near Worcester, a specific case history and the daily rate of spread of the epidemic, strongly suggests that the weather played a major part in the spread of disease. The two main factors involved in this type of spread are wind and precipitation. It is noted that after the epidemic had been checked, following anticyclonic weather, the association between the weather and the spread of disease was less apparent.

Air Microbiology↗

Possible spread of African horse sickness on the wind.

Analyses of outbreaks of African horse sickness showed that movement of infected Culicoides midges on the wind was most likely responsible for the spread of the disease over the sea from Morocco to Spain in 1966, from Turkey to Cyprus in 1960, and from Senegal to the Cape Verde Islands in 1943. The pattern of spread of the epidemic in the Middle East in 1960 could have been laid down by the infected midges carried on spells of south-east winds, and analyses of outbreaks in Algeria in 1965 and India in 1960 also suggested windborne spread of the disease. Each spread occurred when the presence of virus, host and vector coincided either with a spell of winds unusual for a particular time of year (Spain, Cyprus, Cape Verde Islands and Algeria) or with a series of disturbances usual at that time of the year (Middle East and India). Inferred flight endurance of the midge varied up to at least 20 h and flight range from 40 to 700 km. Flight occurred when temperatures were likely to have been in the range of 15-25 degrees C if it was at night or 20 to about 40 degrees C if it was by day.It is suggested that likely movements of midges on the wind can be estimated from synoptic weather charts, and should be taken into account when planning control of the disease in the face of an outbreak. Such control includes a ban on movement of horses, vaccination and spraying of insecticide.The risk of spread to countries outside the endemic areas should be assessed by reference to possible wind dispersal of infected midges.

Africa, Northern↗

Structural, topographical, and shear characteristics of milk protein and monoglyceride monolayers spread at the air-water interface.

In this contribution we are concerned with the study of structure, topography, and surface rheological characteristics under shear conditions of monoglyceride (monopalmitin and monoolein) and milk protein (beta-casein, kappa-casein, caseinate, and WPI) spread monolayers at the air-water interface. Combined surface chemistry (surface film balance and surface shear rheometry) and microscopy (Brewster angle microscopy: BAM) techniques have been applied in this study to pure emulsifiers (proteins and monoglycerides) spread at the air-water interface. To study the shear characteristics of spread films, a homemade canal viscometer was used. The experiments have demonstrated the sensitivity of the surface shear viscosity (eta(s)) of protein and monoglyceride films at the air-water interface, as a function of surface pressure (or surface density). The surface shear viscosity was higher for proteins than for monoglycerides. In addition, eta(s) was higher for the globular WPI than for disordered beta-casein and caseinate due to the strong forces acting on spread globular proteins. This technique makes it possible to distinguish between beta-casein and caseinate spread films, with the higher eta(s) values for the later due to the presence of kappa-casein. The eta(s) value varies greatly with the surface pressure (or surface density). In general, the greater the surface pressure, the greater the values of eta(s). Finally, the eta(s) value is also sensitive to the monolayer structure, as was observed for monoglycerides with a rich structural polymorphism (i.e., monopalmitin).

Air↗

Spreading dynamics of chain-like monolayers: a molecular dynamics study.

Using large-scale molecular dynamics simulations, we have shown previously that the spreading dynamics of sessile drops on solid surfaces can be described in detail using the molecular-kinetic theory of dynamic wetting. Here we present our first steps in extending this approach to investigate the spreading dynamics of Langmuir-Blodgett monolayers. We make use of a monolayer model originally developed by Karaborni and Toxvaerd, but somewhat simplified to facilitate large-scale simulations. Our preliminary results are in good agreement with recent experimental observations and also support a molecular-kinetic interpretation in which the driving force for spreading is the lateral pressure in the monolayer. Away from equilibrium, initial spreading rates are constant and logarithmically dependent on pressure. However, near equilibrium, spreading is pseudo-diffusive and follows the square root of time. In both regimes the controlling factor is the equilibrium frequency of molecular displacements within the monolayer.

Journal Article↗

Spreading of viscous liquids at high temperature: silicate glasses on molybdenum.

The spreading of Si-Ca-Al-Ti-O glasses on molybdenum has been investigated. By controlling the oxygen activity in the furnace, spreading can take place under reactive or nonreactive conditions. As the nucleation of the reaction product under reactive conditions is slow in comparison to the spreading kinetics, in both cases the glass front moves on the metal surface with similar spreading velocities. Spreading can be described using a molecular dynamics model where the main contribution to the wetting activation energy comes from the viscous interactions in the liquid. Enhanced interfacial diffusions in low-oxygen activities (reactive cases) form triple-line ridges that can pin the wetting front and cause a stick-slip motion.

Journal Article↗

Diverse spreading behavior of binary polymer nanodroplets.

Molecular dynamics simulations are used to study the spreading of binary polymer nanodroplets in a cylindrical geometry. The polymers, described by the bead-spring model, spread on a flat surface with a surface-coupled Langevin thermostat to mimic the effects of a corrugated surface. Each droplet consists of chains of length 10 or 100 monomers with approximately 350,000 monomers total. The qualitative features of the spreading dynamics are presented for differences in chain length, surface interaction strength, and composition. When the components of the droplet differ only in the surface interaction strength, the more strongly wetting component forms a monolayer film on the surface even when both materials are above or below the wetting transition. In the case where the only difference is the polymer chain length, the monolayer film beneath the droplet is composed of an equal amount of short chain and long chain monomers even when one component (the shorter chain length) is above the wetting transition and the other is not. The fraction of short and long chains in the precursor foot depends on whether both the short and the long chains are in the wetting regime. Diluting the concentration of the strongly wetting component in a mixture with a weakly wetting component decreases the rate of diffusion of the wetting material from the bulk to the surface and limits the spreading rate of the precursor foot, but the bulk spreading rate actually increases when both components are present. This may be due to the strongly wetting material pushing out the weakly wetting material as it moves toward the precursor foot.

Journal Article↗

A unique monoclonal antibody mNI-11 rapidly enhances spread formation in human umbilical vein endothelial cells.

We previously reported a novel monoclonal antibody (MAb), designated mNI-11, recognizing an adhesion-associated antigen distinct from any previously reported ones. In this article, this adhesion-associated antigen with a molecular weight of about 97 kDa was found to be strongly expressed on human umbilical vein endothelial cells (HUVECs) by fluorescence-activated cell sorter (FACS) analysis. Expression of this antigen on HUVECs was slightly increased in response to the exposure to tumor necrosis factor-alpha (TNF-alpha) or phorbol myristate acetate (PMA). As a biological function exerted by this antigen, it was of great interest that immobilized mNI-11 directly and rapidly enhanced the spread formation of HUVECs, whereas MAbs binding other adhesion-associated antigens such as mNI-58A (anti-CD11a), L130 (anti-CD18), L133.1 (anti-CD31), L178 (anti-CD44), L25.3 (anti-CD49d), or LB-2 (anti-CD54) did not carry such activity under the same conditions. The HUVECs spread formation enhanced by mNI-11 was completely blocked in the presence of a microfilament formation inhibitor, cytochalasin D (CyD), a Ca2+ calmodulin inhibitor, W-7, EDTA, and was partially blocked by a microtubule formation inhibitor, nocodazole, a protein kinase C (PKC) inhibitor, H-7, and a protein synthesis inhibitor, cycloheximide (CHX). However, a protein tyrosine kinase (PTK) inhibitor, genistein, did not affect the spread formation under the same conditions. Taken together, it was suggested that the spread formation of HUVECs enhanced by mNI-11 was mainly associated with the influx of Ca2+ and microfilament reorganization. In addition, the novel property associated with mNI-11 to enhance the spread formation of HUVECs was possibly mediated through its reaction against a unique epitope on HUVECs.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

Evaluation of electrosurgical interference to low-power spread-spectrum local area net transceivers.

OBJECTIVE: To study whether an electrosurgery device interferes with the operation of a low-power spread-spectrum wireless network adapter. METHODS: Nonrandomized, unblinded trials with controls, conducted in the corridor of our institution's operating suite using two portable computers equipped with RoamAbout omnidirectional 250 mW spread-spectrum 928 MHz wireless network adapters. To simulate high power electrosurgery interference, a 100-watt continuous electrocoagulation arc was maintained five feet from the receiving adapter, while device reported signal to noise values were measured at 150 feet and 400 feet distance between the wireless-networked computers. At 150 feet range, and with continuous 100-watt electrocoagulation arc five feet from one computer, error-corrected local area net throughput was measured by sending and receiving a large file multiple times. RESULTS: The reported signal to noise (N = 50) decreased with electrocoagulation from 36.42+/-3.47 (control) to 31.85+/-3.64 (electrocoagulation) (p < 0.001) at 400 feet inter-adapter distance, and from 64.53+/-1.43 (control) to 60.12+/-3.77 (electrocoagulation) (p < 0.001) at 150 feet inter-adapter distance. There was no statistically significant change in network throughput (average 93 kbyte/second) at 150 feet inter-adapter distance, either transmitting or receiving during continuous 100 Watt electrocoagulation arc. CONCLUSIONS: The manufacturer indicates "acceptable" performance will be obtained with signal to noise values as low as 20. In view of this, while electrocoagulation affects this spread spectrum network adapter, the effects are small even at 400 feet. At a distance of 150 feet, no discernible effect on network communications was found, suggesting that if other obstructions are minimal, within a wide range on one floor of an operating suite, network communications may be maintained using the technology of this wireless spread spectrum network adapter. The impact of such adapters on cardiac pacemakers should be studied. Wireless spread spectrum network adapters are an attractive technology for mobile computer communications in the operating room.

Electricity↗

Regional differences in intestinal spreading and pH recovery and the impact on salmon calcitonin absorption in dogs.

PURPOSE: To investigate the regional influence of intestinal spreading and pH recovery on the performance of drug and excipient delivery systems and their impact on the oral absorption of a model peptide drug, salmon calcitonin (sCT), in conscious beagle dogs. METHODS: Male beagle dogs were surgically prepared with subdermal Intestinal Access Ports (IAP). The catheter from one port was placed in the duodenum and the other in the ileum. Fluoroscopy and Heidelberg pH capsule studies were performed to characterize intestinal spreading and pH recovery, respectively. Three treatments were performed: (1) a radiopaque dye and citric acid (CA) were infused into the intestinal segments, (2) a radiopaque powder capsule containing CA was given orally, and (3) capsules containing CA and sCT were given orally. Regular blood samples were collected and analyzed by radioimmunoassay (RIA) to determine the absorption characteristics of sCT. RESULTS: Since sCT is an excellent substrate for the pancreatic serine protease trypsin, the rate of degradation of sCT in the GI lumen is dependent upon the regional pH, activity of digestive enzymes and the concentration of sCT at the site of absorption. Fluoroscopy results clearly showed that when the radiopaque dye was infused into the duodenum and capsule disintegration occurred early, there was significant dilution and spreading of the excipients throughout a large section of the upper small intestine (USI). However, when the radiopaque dye was infused into the ileum and capsule disintegration occurred in the lower small intestine (LSI), the excipients moved along as a bolus (i.e., plug). The pH monitoring results were consistent with the fluoroscopy results. The pH dropped only momentarily and rose quickly in the USI consistent with well-stirred mixing kinetics. In the LSI, dilution and spreading were minimal and the drop in pH was greater and persisted for a longer period of time. Plasma levels of sCT were maximal when disintegration occurred in the LSI. CONCLUSIONS: Since significantly less dilution and spreading occurred in the LSI, the exposure of the intestine to pharmaceutical excipients and sCT was more concentrated resulting in a higher fraction of sCT absorbed. The results of this study demonstrate that intestinal mixing kinetics have a dramatic impact on the ability of pharmaceutical excipients to modulate the oral bioavailability of peptide drugs like sCT.

Administration, Oral↗

A healthy diet rich in carotenoids is effective in maintaining normal blood carotenoid levels during the daily use of plant sterol-enriched spreads.

Blood cholesterol levels are affected by diet and in particular by the type and amount of fat intake. In recent years, vegetable oil spreads containing plant sterols/stanols (as their fatty acid esters) have been developed. Numerous clinical trials on spreads with added plant sterols/stanols have shown that they have much greater cholesterol-lowering properties than conventional vegetable oil spreads. Plant sterols decrease both dietary and biliary cholesterol absorption in the small intestine, with a consequential increase in excretion of cholesterol. It is also recognized that plant sterol/stanol-enriched, cholesterol-lowering spreads, if consumed regularly, may induce a 10-20% decrease in plasma carotenoids, adjusted for changes in plasma lipids. A 10-20% decrease in plasma carotenoids falls well within the seasonal variation observed in individuals. Our current understanding of the physiological functions of carotenoids does not indicate any health risk associated with the slight decrease in their blood levels due to the intake of plant sterol/stanol. The questions that have been raised, though, are how plant sterols/stanols affect plasma carotenoid levels, and in addition, what quantity of fruits and vegetables (the richest dietary sources of carotenoids) would have to be consumed to improve plasma carotenoid levels? The current mini-review covers the cholesterol-lowering effect of plant sterols, their mechanisms of action and effect on blood carotenoids, and concludes with the potential heath benefits of daily intake of plant sterol-enriched spreads.

Anticholesteremic Agents↗

Evidence from gabbro of the Troodos ophiolite for lateral magma transport along a slow-spreading mid-ocean ridge.

The lateral flow of magma and ductile deformation of the lower crust along oceanic spreading axes has been thought to play a significant role in suppressing both mid-ocean ridge segmentation and variations in crustal thickness. Direct investigation of such flow patterns is hampered by the kilometres of water that cover the oceanic crust, but such studies can be made on ophiolites (fragments of oceanic crust accreted to a continent). In the Oman ophiolite, small-scale radial patterns of flow have been mapped along what is thought to be the relict of a fast-spreading mid-ocean ridge. Here we present evidence for broad-scale along-axis flow that has been frozen into the gabbro of the Troodos ophiolite in Cyprus (thought to be representative of a slow-spreading ridge axis). The gabbro suite of Troodos spans nearly 20 km of a segment of a fossil spreading axis, near a ridge-transform intersection. We mapped the pattern of magma flow by analysing the rocks' magnetic fabric at 20 sites widely distributed in the gabbro suite, and by examining the petrographic fabric at 9 sites. We infer an along-axis magma flow for much of the gabbro suite, which indicates that redistribution of melt occurred towards the segment edge in a large depth range of the oceanic crust. Our results support the magma plumbing structure that has been inferred indirectly from a seismic tomography experiment on the slow-spreading Mid-Atlantic Ridge.

Journal Article↗