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A device useful for the in vitro study of cell migration across a monolayer of epithelial cells while simultaneously measuring the transepithelial electrical resistance of the epithelial monolayer is provided and is characterized as including at least three components, a cylinder, a chamber and a punch. The cylinder is designed for tissue culture, the chamber provides a controlled space and/or environment for the cylinder and the punch is provided to cut the epithelial monolayer and its substrate.
1. An apparatus useful for the study of biological cell migration and for the measurement of electrical resistance across a monolayer of cells which comprises, an internal chamber, an external chamber, said internal chamber being a cylinder open at both ends, a substrate provided across and attached to one end of said internal chamber on which are growing a confluent layer of cells, said internal chamber being positioned within said external chamber such that said layer of cells on said substrate is contacted by a first fluid or solution at its basal aspect and by a second fluid or solution at its apical aspect within said chambers so that an electrical current may flow through said substrate and layer of cells, a pair of bridges consisting of one current passing bridge and one voltage recording bridge positioned above said layer within said internal chamber and an identical pair of said bridges located below said layer and substrate within said external chamber, said pair of bridges connected to circuit means for measuring voltage across said layer.
2. An apparatus of claim 1 wherein the cells are eptithelial cells.
3. An apparatus of claim 1 wherein the cells are endothelial cells.
4. An apparatus of claim 1 wherein the internal chamber is composed of a plastic.
5. An apparatus of claim 4 wherien the internal chamber is composed of Lexan.
6. An apparatus of claim 4 wherein the internal chamber is composed of Lucite.
7. An apparatus of claim 1 wherein the internal chamber is composed of glass.
8. An apparatus of claim 1 wherein the external chamber is composed of a plastic.
9. An apparatus of claim 9 wherein the external chamber is composed of Lexan.
10. An apparatus of claim 9 wherein the external chamber is composed of Lucite.
11. An apparatus of claim 1 wherein the external chamber is composed of glass.
12. An apparatus of claim 1 wherein said substrate provides a surface composed substantially of cellulose acetate or cellulose nitrate.
13. An apparatus of claim 1 wherein said substrate provides a surfce composed substantially of polycarbonate.
14. An apparatus of claim 1 wherein said substrate provides a surface composed substantially of connective tissue.
15. An apparatus of claim 1 wherein said internal chamber is positioned within the external chamber such that said first fluid or solution and one pair of bridges, consisting of one current passing bridge and one voltage recording bridge are isolated from said second fluid or solution and an identical pair of said bridges located within said external chamber.
16. An apparatus for the study of biologcial cell migration and for the measurement of electrical resistance across a monolayer of cells which comprises an internal chamber, an external chamber, said internal chamber being composed of a translucent or transparent material in the form of a hollow cylinder, a lip around the top outer edge of said cylinder, a groove in the lower outer edge of said internal cylinder, said groove being capable of holding an O-ring, and O-ring provided in said groove, a substrate stretched across the lower end of said internal cylinder held in place by said O-ring and on which are cells, said external chamber consisting of a well, a set-up station, a base and a lid, said well being in the shape of a hollow cylinder which tapers conically at one end, the material making up said hollow cylinder being transparent, said well of said cylinder provided with apertures for the egress of fluid at its conical tip, a current passing bridge and a voltage recording bridge provided within the tapered conical portion at said one end of said hollow cylinder, said well of said external chamber provided with means for holding a cylinder suspended inside the well, said lid being held in place by pins and screw-down nuts, said lid providing apertures for the introduction of fluids to said well and for the introduction and removal of fluid from the internal chamber and in which are located a pair of bridges consisting of one current passing bridge and one voltage recording bridge for said internal chamber, a set-up station adjacent to said well such that when said lid is placed on said threaded pins provided on either side of said chamber, accurate and reproducible setting of one voltage recording bridge for said internal chamber positioned within said external chamberis achievable so that as said layer of cells is contacted by a first fluid or solution at its basal aspect, and by a second fluid or solution at its apical aspect an electrical current flows therethrough and voltage measurement is recorded across said cell layer by said pair of bridgges consisting of one current passing bridge and one voltage recording bridge is positioned above said cell layer within said internal chamber and an identical pair of said bridge is located below said cell layer within said external chamber.
17. An apparatus of claim 16 wherein the cells are eplithelial cells.
18. An apparatus of claim 16 wherein the cells are endothelial cells.
19. An apparatus of claim 16 wherein the internal chamber is composed of a plastic.
20. An apparatus of claim 16 wherein the internal chamber is composed of Lexan.
21. An apparatus of claim 16 wherein the internal chamber is composed of Lucite.
22. An apparatus of claim 16 wherein the internal chamber is composed of glass.
23. An apparatus of claim 16 wherein the external chamber is composed of a plastic.
24. An apparatus of claim 16 wherein the external chamber is composed of Lexan.
25. An apparatus of claim 16 wherein the external chamber is composed of Lucite.
26. An apparatus of claim 16 wherein the external chamber is composed of glass.
27. An apparatus of claim 16 wherein said substrate provides a surface composed substantially of cellulose acetate or cellulose nitrate.
28. An apparatus of claim 16 wherein said substrate provides a surface composed substantially of polycarbonate.
29. An apparatus of claim 16 wherein said substrate provides a surface composed substantially of connective tissue.
30. A chamber useful for the study of biological cells, said chamber comprising as well, a set-up station, a base, a lid, said well being made of transparent material and being open at it upper end for the introduction of fluid thereinto and provided with an apertureat its lower end for the removal of flud, said well also being provided with apertures inw hich are located of a pair of electrical bridges consisting of one current passing bridge and one voltage recording bridge into the bottom of the well of said chamber, means for holding said lid in place, said lid being provided with apertures for the introduction and removal of fluids and in which are located of a pair of bridges into said well consisting of one current passing bridge and one voltage recording bridge and means associated with said lid to provide for a predtermined orientation of said voltage recording bridge in which lid is placed on said set-up station which is attached to said base.
This invention is directed to a device useful for the growing and/or testing of biological materials. More particularly, this invention is directed to a device for the in vitro study in cell migration across a monolayer of epithelial cells while simultaneously measuring the transepithelial electrical resistance of the epithelial monolayer.
In connection with studies of chronic inflammation, particularly granulomatous diseases, such a tuberculosis, sarcoidosis and the like, it is particularly useful to have a device and techniques of employing the same which are useful for a better understanding of the process of granuloma formation as well as the effects of drug therapy thereon.
Accordingly, it is an object of this invention to provide a device and techniques employing the same for the in vitro study of growing and/or testing biological materials, particulary for the in vitro study of cell migration across a monolayer of epithelial cells.
It is another object of this invention to provide a device for the in vitro study of cell migration across a monolayer of epithelial cells while simultaneously measuring the transepithelial electrical resistance of the epithelial monolayer.
It is another object of this invention to provide a device which has in connection with the above objects the capability of the operator employing the device being able to vary the type of the migrating cell and/or the type of epithelium as well as the type of substratum on which the epithelium is grown.
How these and other objects of this invention might be achieved are set forth in the accompanying disclosure made with reference to the accompanying drawings wherein:
FIG. 1 shows one embodiment of the invention in which two chambers are mounted on a base. A-recording chamber; B-set-up station; C-lid; D-screw-down nut; E-threaded pins; F and F'-upper and lower compartment voltage bridge aperture, respectively; G-lower compartment fluid outlet, H and H' upper and lower compartments current bridge aperture, respectively; M-plexiglas base; N-aperture for fluid addition and removal from upper compartment; P-aperture for fluid addition to lower compartment.
FIG. 2 is a top view of the embodiment shown in FIG. 1. B-set-up station; C-lid; D-screw-down nuts; E-threaded pins; F-upper compartment voltage bridge aperture: H-upper compartment current bridge aperture; H-plexiglas base; N-aperture for fluid addition and removal from upper compartment; P-aperture for fluid addition to lower compartment.
FIG. 3 is a top view of one chamber showing the recording chamber (A), E-threaded pins, the fluid outlet (G), voltage (F') and current (H') bridge apertures for lower compartment and the set-up station (B).
FIG. 4 is a side view of the chamber shown in FIG. 3. A-recording chamber; B-set-up station; E-delrin pin; F'-lower compartment voltage bridge aperture; G-lower compartment fluid outlet; H'-lower compartment current bridge aperture.
FIG. 5 is a side view of the chamber shown in FIG. 3 with the cylinder (J), lid (C), delrin pins (E) and screw-down nuts (D) in place. Voltage and current bridges are in place at F and F' and H and H', respectively. Viton O-ring (K) is in place around cylinder holding substrate (L) onto cylinder bottom. G-lower compartment fluid outlet. The right side of the figure shows set-up chamber (B) with voltage (F) and current (H) bridges in place.
FIG. 6 is a top view of one embodiment of the cylinder (J) with a large inside diameter.
FIG. 7 is a side view of the cylinder (J) shown in FIG. 6.
FIG. 8 is a top view of one embodiment of the cylinder (J) with a small inside diameter.
FIG. 9 is a cut away view of one embodiment of the punch, wherein the punch has been placed inside the cylinder (J) and is set to remove a plug of substrate (L) from the base of the cylinder. K-Viton O-ring.
FIG. 10 is a cut away view of a punch with a narrow outside diameter.
FIG. 11 is a view of one embodiment of the apparatus in which voltage and current bridges, F and F' and H and H', resepctively, are connected to electrodes and electrical equipment.
The invention involves a device which permits the in vitro study of cell migration across a monolayer of epithelial cells while simultaneously measuring the transepithelial electrical resistance of the epithelial monolayer. One employing the device of this invention has the option of varying the type of migrating cell and/or the type of epithelium as well as the type of substratum in which the epithelium is grown. More particularly, the device of this invention comprises in combination three elements or parts, a cylinder, a chamber and a punch. The cylinder is designed for tissue culture. The chamber is made up of a well, setup station, base. The punch, such as a stainless steel punch, is designed to precisely cut off uniform discs without damaging or separating the epithelial monolayer from the substrate and to cut out the epithelial monolayer and its substrate when the cylinder is placed on a suitable support and the punch inserted and tapped with a hammer.
The device consists of three parts; the cylinder, the chamber and the punch. The cylinder is designed for tissue culture and is usefully composed of Lexan, a registered trademark for polycarbonate resin, which can be autoclaved and reused. Lexan is a polymeric material compartible with epithelial cell growth. The cells form a confluent monolayer and grow up the sides of the cylinder, forming a natural seal. Lexan is also translucent and allows visulatization of the fluid in the cylinder. Further, the cylinder is machined from one piece of Lexan which prevents separation of bonded sections.
The cylinder fits into various shaped and sized commercial tissue culture dishes. The lids of the tissue culture dishes can be placed on the culture dishes when the cylider either stands in a flat bottom tissue culture dish or multiwell that is at least 14 mm deep and larger than 20 mm in I.D., or the cylinder can hang by its rim in a smaller multiwell with I.D. of 16 mm. A slot in the rim allows easy fluid changes when the cylinders are hung in 16 mm multiwells. When the cylinder is suspended in the multiwell, fluid can reach the cells, growing on the substrate from both the apical and basal surface.
The cylinder is designed to use a variety of substrates. The outside diameter of the cylinder is 13 mm which is a standard commercial size for most filters. Millipore (cellulose acetate and nitrate) or nucleopore (polycarbonate) micropore filters can be glued to the bottom of the cylinder using a silicone adhesive sealant which is not toxic to the cells. The cylinder filter assembly can be autoclaved to provide sterility. The cylinder also allows filters of various pore sizes (0.1 μm-12.0 μm) and shapes (maze or straight) to be used. The pore size of the filters is important in determining the size cell which can migrate into the filter. A pore size of 0.45 μm excludes most cells but does allow them to accumulate on the apical surface of the filter. Pore sizes of 3.0 and 5.0 μm are used for chemotactic studies of neutrophils and monocytes, respectively.
The cylinder is designed so that amnion connective tissue can be pulled taut and securely attached to the bottom of the cylinder with a Viton O-ring (13 O.D., 10 I.D.). Viton is a registered trademark for a synthetic rubber material. The O-ring fits into a groove near the bottom of the cylinder. Viton is non-toxic to the cells and can be sterilized by autoclaving. Further, the bottom edges of the cylinder have been smoothed and rounded to prevent damage and tearing of the connective tissue. The cylinder also has a slot on one side in the groove to allow the O-ring to be removed easily. The design of the cylinder also allows other substrates to be used such as nylon mesh coated with a cross linked collagen gel, isolated epithelial sheets such as urinary bladder, mesentery and frog skin, walls of blood vessels, such as aorta, or dialysis membrane.
The cylinders can be designed in various sizes. The minimum I.D. is the spaced needed for two bridges and a needle for fluid exchange. At present, two sizes are useful (I.D. 6 or 9 mm) which hold 0.3 or 0.5 ml of fluid and exposes 0.32 or 0.71 cm2 of filter surface, respectively. The Lexan material of the cylinder is resistant to aldehyde fixatives. Therefore the cells can be fixed while still attached to the cylinder. This is especially convenient for ultrastructural tracer studies where it is desirable to fix the tissue while the two compartments are still separated. The cylinder can be reused after going through a cleaning procedure which involves quenching the free aldehydes in 1M glycine. The cylinder is designed to be able to grow epithelial cells from the respiratory, digestive, urinary and reproductive tracts as well as the endothelium lining blood vessels.
The chamber which is made of Lucite, a registered trademark for a polyacrylate material, comprises a well, set-up station, base and lid. (FIG. 1) The well forms the lower of two compartments when the cylinder with attached monolayer of epithelial cells is suspended in place. (FIG. 5,A) The set-up station is used to exactly set the length of the upper compartment voltage recording brdige (F) to 1mm above the epithelial monolayer. (FIG. 5,B) The base provides stability to the assembly. The lid has four holes. The current (H) and voltage (F) bridges going to the upper compartment occupy two holes. (FIG. 5) The other two holes (N and P) are used to remove or add fluid to the upper and lower compartments, respectively, without disturbing the monolayer.
The chamber can be used for simultaneous chemotactic and transepithelial electrical resistance studies. One can vary the type of migrating cell, epithelial monolayer, substrate or the fluid in both compartments. One can study either random, chemokinetic or directed (chemotactic) migration. The chamber design enables the transepithelial electrical resistance of the cell monolayer to be determined by measuring the voltage change across the monolayer. The chamber has two holes on the side of the well to hold one voltage and one current-passing bridge for the lower compartment. (FIG. 7,H) The holes are tapered slightly to hold the bridges securely and prevent leakage. They are placed on the side so they are near the electrodes and do not interfere with viewing the lower compartment.
This construction enables the voltage bridges to be easily and precisely set at 1 mm from either surface of the epithelium. The voltage bridge for the lower compartment is set by placing the bridge in the upper hole on the side of the well and lining up the end of the bridge with the center draining hole. The top voltage bridge is positioned by placing the lid on the pegs over the setup station by placing the bridge in the center hole and pushing it down until it is stopped at the flat surface of the set-up station. The bridge length is such that it will be precisely 1 mm above the apical surface of the epithelium when the lid with the bridge is moved to the pegs on either side of the well.
Before setting-up for recording, the lid with the electrodes can be rested on the set of pegs on either side of the set-up station while cylinder samples are changed. The tapered ends of the lid enable one to change the notched round rimmed cylinder without being restricted by the resting lid. The asymmetric design of the lid easily allows one to orient the bridge and fluid changing apertures in the correct direction. When the chamber is assembled, the design of the lid and chamber are such that fluid changes can be made without disturbing the monolayer. The lid is held in place by two delrin pins and nuts (FIG. 5, E and D). The well of the chamber (FIG. 1,A) is conical shaped with a drainage hole at the bottom (FIG. 1,G) which exits through the front of the chamber. Teflon, a regisered trademark for a polyfluororesin, tubing with an attached syringe is inserted into the exit pathway to prevent leakage and to remove fluid.
As previously mentioned, the chamber has holes in the lid which enable one to add and remove fluid from the upper compartment and add fluid to the lower compartment. When the slot in the rim of the cylinder is positioned beneath the fluid hole (FIG. 1, P), a needle inserted into this hole can reach the lower compartment. The chamber has space between the well and the cylinder walls to enable bubbles trapped beneath the cylinder to escape when the chamber is tipped. It is designed so that the top surface of the well has a cylindrical depression which matches the cylinder rim diameter and thickness. Its purpose is to center the insert within the well and prevent it from mvoing from side to side.
Chamber geometry (I.D.=15.7 mm) is such that when a large or small cylinder is filled with 0.5 or 0.3 ml, respectively, the fluid level in the upper and the lower compartments will be even when approximately 1.5 ml of fluid is added to the lower compartmtne. Equalizing the fluid levels prevents hydrostatic pressure fluctuations. As shown in the FIGS. 1, 2 and 11, the apparatus can be constructed with the chambers placed as mirrored images. The two chambers are mounted on a block of plexiglas (M) for support. This design enables two set-ups to run simultaneously.
The punch is made of stainless steel in two O.D.'s which fit the I.D. of the two different sized cylinders. It is designed to precisely cut uniform disks of tissue on substrate without damaging or separating the epithelial monolayer from the substrate. The punch is designed to cut the eipthelial monolayer and its substrate when the cylinder is placed on plexiglas and the inserted punch (FIG. 9) is trapped with a hammer. The knurled head of the punch of the punch allows the punch to be securely held by hand when hitting the head with a hammer. The cutting edge of the punch is slanted to prevent damaging the epithelial monolayer, and has a cutting edge which can be resharpened.
During the process of inflammation, leukocytes such as neutrophils and monocytes adhere to and migrate across the endothelium, a type of epithelium which lines blood vessels, and frequently a second epithelium which lines an infected organ. The epithelial cells are held together by tigh junctions which form an occluding belt around the circumferene of the cells. The transepithelial electrical resistance is a measure of the permeability of these junctions. Little is known about how leukocytes are able to traverse these barriers, the effect of epithelial permeability on the migration of these cells, the effect of leukocyte migration on epithelial permeability, or the role serum and connective factors play in this process. This device would enable one to grow in vitro either endothelium or the epithelia of organs of the respiratory, digestive, urinary or reproductive tract and study the interaction of neutrophils or monocytes with any of these tissues.
Studies23 have been carried out on the effects of biochemical compounds known as leukotrienes on endothelium and on the transendothelial migration of neutrophils. In one part of these studies human amnion tissue was attached across the bottom of a plastic cylinder. The tissue was prepared such that all connective tissue cells and epithelial cells were removed and only connective tissues remained, providing a support upon which a confident layer of human umbilical endothelial cells were grown. Transendothelial electrical resistance was measured across the confluent layer of endothelial cells. Further, neutrophil migration across the endothelial layer was studies in response to leukotriene stimulation. The connective tissue support layer and attached endothelial tissue was prepared for viewing in cross-section under the electron microscope, and neutrophils were observed in migration through the endothelial cell layer.
These studies can be performed in the absence of serum and connective tissue factors so that one can examine the direct effect of leukocytes on the integrity of endothelium or epithelium. One then has the option of adding various serum and connective tissue components in the absence and presence of leukocytes to investigate the specific as well as the cumulative effect of these factors.
The device is designed so that one can simultaneously examine the interaction of two different cell types from a physiological, biochemical and morphological perspective. While the rapid migration of leukocytes to sites of inflammation is a very important defense mechanism in the body, sometimes the accumulation of leukocytes can be destructive. This is observed in diseases such as rheumatoid arthritis, glomerulonephritis and emphysema. This device can be used in the development of antiinflammatory drugs which either prevent the accumulation of leukocytes or interfere with their destructive properties. In addition, the device can be used to develop drugs which either cause or prevent increased vascular or epithelial permeability.
One need not always use the system to simultaneously study chemotaxis and epithelial permeability. One can use the device to grow endothelium and determine its transepithelial electrical resistance. This information will provde the research with a measure of the confluency and permeability of the monolayer without disturbing the cells. The tissue can then be used to study the process of metastasis. Relatively little is known about how or why certain cancer cells leave a primary tumor and migrate to another organ. In many ways this process is analogous to the migration of leukocytes and like leukocytes the cancer cells must also cross the endothelium.
Tumor cells leaving the primary tumor migrate into the blood by crossing the endothelium from the basal to the apical surface. They circulate in the blood stream and eventually leave the vasculature by crossing the endothelium, apically to basally, and develop in a new location.
The system is designed so that one can examine both aspects of the migratory process and perhaps determine ways to prevent it. In addition one can grow tumors in the connective tissue on which the endothelium is grown and examine how the metastasizing cells leave the primary site. One can also grow normal tissue from organs commonly involved in metastasis in the connective tissue beneath the monolayer of endothelial cells. This will provide a way to determine if there are attractants or growth factors in these organs which stimulate the accumulation and growth of tumor cells.
The migration of monocytes across endothelium can be observed during the process of atherosclerosis by growing an artificial vessel wall and studying the migration of monocytes into the vessel wall and their subsequent development into foam cells.
The system can also be modified to study chronic inflammation. Granulomatous diseases such as tuberculosis, sarcoidosis, schistosomiasis, leprosy and silicosis are major causes of morbidity and mortality worldwide. Foreign body granuloma can be formed by the addition of human peripheral monouclear cells to the apparatus with attached amnion connective tissue containing an absorbable synthetic suture. The device of this invention provides a way to study the process of granuloma formation, organization and repair, as well as the effects of drug therapy.
The process of inflammation is quite complex. It involves hemodynamic changes, increases in vascular permeability, and leukocyte exudation. Relatively little is known about the factors or mediators that control this phenomenon. Both slow-reacting substance (SRS), which consists of cysteine-containing 5(S)-Hydroxy-6(R)-S-gluthathionyl-7,9-trans-11,14-cis-eicosatetraenoic acid (LTC4), 5(S)-Hydroxy-6(R)-cysteinylglycyl-7,9-trans-11,14-cis-eicosatetraenoic acid (LTD4), and 5(S)-Hydroxy-6(R)-cysteinyl-7,9-trans-11,14-cis-eicosatetraenoic acid (LTE4), and 5(S)12(R)-Dihydroxy-6,24-cis-8,10-trans-eicosatetraenoic acid (LTB4) have been implicated in this process, LTC4 and LTD4 are vasoactive compounds that have been shown to produce transient constriction of arterioles and exudation from postcapillary venules in the hamster cheek pouch1 as well as rapid, transient pulmonary and systemic hypertension followed by a prolonged fall in blood pressure and decreased cardiac output in monkeys2. The closely related LTB4 stimulates white cell adhesion and extravasation in vivo3-5 and neutrophil chemotaxis in vitro.5-7 In addition, concentrations of LTB4 higher than those required for chemotaxis stimulate edema formation and enzyme release from leukocytes.6,8 How these compounds mediate their effects, however, is relatively unknown.
To determine possible mechanisms for the vasoactive actions of SRS, the effects of LTC4, LD4, and LTE4 on arachidonic acid (20:4) metabolism in endothelium were examined. Human endothelial cells were isolated from umbilical veins by collagenase treatment according to the method of Jaffe et al.9 Cells became confluent 3 to 4 days after plating. Experiments were performed on cells of either primary, first, or second passage. All cells examined contained factor VIII antigen as determined by indirect immunofluorescence.10 As shown by Wagner et al.,11 factor VIII antigen was apparent in the perinuclear region and in small rodlike structures throughout the cytoplasm.
To determine if SRS stimulated 20:4 metabolism in endothelium, each 35 mm dish of endothelial cells was washed in phosphate-buffered saline and was labeled with 1.0 uCi[3 H]20:4(91.2 Ci/nmol; New England Nuclear, Boston, Mass.) in 1 ml medium 199 containing 10% heat-inactivated human serum. After 16 hours, the cells were washed three times with phosphate-buffered saline solution, placed in medium 199 with the appropriate stimulus, and incubated at 37� C. Duplicate 50 μl aliquots of medium were removed at 10, 20, 30, and 60 minutes for radioactivity measurements as determined by liquid scintillation counting in Hydrofluor. Endothelial cells remained viable after a 60-minute exposure to LTC.sub. 4 as assessed by trypan blue exclusion. At this time the culture medium was removed and cells were washed with phosphate-buffered saline solution and scraped into 1 ml 0.5% Triton X-100. Duplicate aliquots were assayed for protein12 and tritium.
As previously reported,13 LTC4 at a concentration of 10-9 M caused little or no release of radiolabel above control levels. Maximal release occurred with 10-8 M to 10-6 M LTC4. At 30 minutes, 10-7 M LTC4 caused approximately 6% of the incorporated radiolabel to be released. This represented a release of radiolabel that was 400% of that observed in control cultures. Although the amount of radiolabel released varied with LTC4 concentration, the time course of tritium release was quite similar. LTC4 caused an increase in radiolabel release that reached maximum at 30 minutes.
At 60 minutes, the medium was extracted by the method of Unger et al.14 and total 20:4 metabolites were separated by reverse-phase High-pressure liquid chromatography (HPLC). As observed previously13 cyclooxygenase products eluted between 4 and 16 minutes and constituted about half (52.8%�9.3%, n=6) of the recovered radiolabel. Unreacted 20:4 (elution time 111 to 150 minutes) represented the second major peak, with approximately one third (32.7% �11.1%, n=6) of the released radiolabel, while lipoxygenase products (elution time 17 to 110 minutes) represented only approximately one seventh (14.8%�4.6%) of the released radiolabel. Control cultures released less radiolabel but produced a similar 20:4 metabolite profile.
The pooled cyclooxygenase products were rechromatographed on a Waters fatty acid analysis column to separate individual metabolites.15 The major cyclooxygenase product was 6-keto-PGF1 α, the breakdown product of protacyclin. In five separate experimetns, 6-keto-PGF1 α represented approximately 62.2%�7.6% of radiolabeled cyclooxygenase products, while a smaller peak of PGF2 α averaged 11.6%�4.6%. A peak of PGE2 (0% to 4%) was occasionally observed. Therefore, of the total released radiolabel, 63.3% was metabolized and 33% was converted to prostacyclin.
To determine if the three leukotrienes that comprise SRS equally stimulated 20:4 metabolism in endothelial cells, endothelial cultures were agin prelabeled with [3 H]30:4. Cells were exposed to 10-7 M to 10-9 M concentrations of LTC4, LTD4, and LTE4 and percent of released radiolabel was calculated. Maximal release of radiolabel was obtained with 10-7 M LTC4 and LTD4. Percent released in response to 10-7 M LTC4 was consistently higher than that released in response to 10-7 M LTD4. In fact, LTC4 promoted a higher level of release than did LTD4 at all concentrations (10-9 M to 4�10-7 M) tested. In contrast, LT4 at concentrations up to 10-6 M failed to promote an increase in tritium release above control values. Likewise, 10-6 M to 10-7 M LTB4 was ineffective in stimulating radiolabel release by endothelial cells.
As observed for LTC4, the major 20:4 metabolite synthesized byendothelial cells in response to LTD4 was prostacyclin. Thus the relative effectiveness of the leukotrienes (LTC4 >LTD4 >>>LTE4 =LTB4) indicated that specific structural features of the leukotriene molecule are necessary to promote prostacylcin synthesis. Removal of the glutamyl residue from LTC4 (conversion of LTC4 to LTD4) caused an approximate 50% loss in prostacyclin-stimulating activity and removal of the glycyl moiety (conversion of LTD4 to LTE4) rendered the molecule inactive. The need for the peptide backbone was further indicated by the fact that LTB4 was also ineffective in stimulating prostacyclin synthesis.
It was clear from the above studies that LTC4 and LTD4 increased 20:4 metabolism in endothelium and that LTC4 did not. While LTB4 does not appear to have an effect on 20:4 metabolism in endothelium, LTB4 is a potent chemoattractant for human neutrophils. To test the chemotactic activity of our synthetic LTB4, human neutrophils were isolated from citrated peripheral blood of normal volunteers by Ficoll/Hypaque and dextran sedimentation techniques.16 The resulting cell fraction contained approximately 98% neutrophils, with 98% to 99% viability as determined by trypan blue exclusion. Before use, the neutrophils were suspended in Gey's balanced salt solution with 2% bovine serum albumin, penicillin, 63 U/ml, and streptomycin, 138 g/ml. The chemotaxis assay was performed in a 48-well microchemotaxis chamber (Neuroprobe; Bethesda, Md.) according to the method of Harvath et al.17. Neutrophils were stimulated to traverse the 10 μm thick polyvinylpyrrolidine-free polycarbonate filter (3 μm pore) by the addition of 10-6 M to 10-9 M LTB4 or 10-7 M f-Met-Leu-Phe (Peninsula Laboratories, San Carlos, Calif.) to the lower compartment. After a 16-minute incubation at 37� C. in a 95% air/5% CO2 incubator, nonmigrating neutrophils were removed from the apical surface of the filter and neutrophils that had migrated to the lower filter surface were stained with hematoxylin. Four oil immersion fields (97X) were counted per well and the mean �SEM of each experimental group was determined from seven wells. The number of neutrophils responding to various concentrations of LTB4 were compared to those responding to 10-7 M f-Met-Leu-Phe and Gey's balanced salt solution. LTB4 at a concentration of 10-9 M elicited a neutrophil chemotactic response significantly greater (p<0.0005) than random migration. Maximal neutrophil migration occurred at 10-7 M LTB4. This response was significantly greater (p<0.0005) than that evoked by 10-7 M f-Met-Leu-Phe. The chemotactic response to 10-6 M LTB4 was significantly lower (p<0.0005) than that cuased by 10-7 M LTB4 but was similar in magnitude to that evoked by 10-8 M LTB4.
Although LTB4 has been reported to stimulate adherence and migration of neutrophils across postcapillary venules and larger venules,3 it was not clear if it was directly responsible for this response. In attempting to answer this quenstion, there was used an in vitro model based on a system developed for the transepithelial migration of leukocytes,18 also studies that used amnion connective tissue19, and another model system20. The system employed involved growing human umbilical vein endothelial cells on connective tissue derived from human amnion. The amnion was separated from the chorion and attached to the bototm of plastic cylinders (inner diameter 9.5 mm) with the amnion epithelium facing outward. Cylinders were submerged for 2 hours in 0.25M NH4 OH to destroy any possible connective tissue cells (although none have been observed) and the amnion epithelium. The amnion epithelium was scraped off with cotton swabs and cell-free connective tissue was extensively washed in phosphate-buffered saline solution and preincubated in medium 199 with 20% heat-inactivated human serum for 1 hour. Primary or first-passaged human umbilical endothelial cells (5�105 cells/ml) were plated onto the connective tissue surface. Cells were fed every other day with medium 199 with 205 heat-inactivated human serum. Six to seven days after plating, the cells formed a thin confluent monolayer with transendothelial electrical resistance (a measure of endothelial permeability) as high as 50 ohms cm2. Endothelial cultures were washed three times with medium 199 before each experiment. Neutrophils, suspended in medium 199 at 3.6�106 cells/ml, were added to the upper compartment and were stimulated to traverse endothelium and underlying connective tissue for 30 minutes in response to 10-7 M synthetic LTB4. At 5, 10, and 30 minutes after the addition of the neutrophils, cylinders were removed from the 37� C. incubator, washed three times in medium 199 to remove nonadherent neutrophils, and fixed in 2.5% glutaraldehyde in 0.1M phosphate buffer (pH 7.3). Tissue was postfixed ion 1% osmium tetroxide in 0.1M phosphate buffer (pH 7.3), dehydrated, and embedded in Epon 812. One-micron cross sections were made and the number of neutrophils that adhered to or migrated across each millimeter of endothelium was determined. At 5 minutes, a few neutrophils were adherent to the endothelial surface (2.6�0.4 neutrophils/mm endothelium) and a few neutrophils had started to migrate across the endothelium (1.3�0.4 neutrophils/mm endothelium). After 10 minutes, there was a significant increase in both neutrophil adherence to (33.0�6.1 neutrophils/mm endothelium) and migration across (45.0�5.6 neutrophils/mm endothelium) the endothelium. Examination of the tissue by transmission electron microscopy revealed neutrophils adherent to the apical endothelial surface, in the process of traversing the endothelium, and caught between the basal surface of the endothelium and the endothelial basal lamina (FIG. 3). After 15 minutes of incubation, neutrophils had penetrated the basal lamina and by 20 minutes they had migrated deeper into the connective tissue. At 30 minutes there was a decline in the number of neutrophils adherent to the nedothelial surface (19.2�1.0 neutrophils/mm endothelium), whereas the number that had accumulated in the connective tissue (240.0�18.1 neutrophils/mm endothelium) was now approximately 500% that observed at 10 minutes. Thus, in the absence of serum and connective tissue factors, LTB4 was quite capable of stimulating neutrophil adherence to and migration across endothelium.
The ability of 10-7 M f-Met-Leu-Phe and LTB4 to stimulate transendothelial migration of neutrophils was observed. As had been seen inthe micropore filter chemotactic assay, LTB4 was again able to stimulate significantly (p<0.0005) more neutrophils to undergo chemotaxis and to traverse endothelium than were stimulated by f-Met-Leu-Phe.
Tests are carried out to study the effects of leukotrienes on human endothelium and on the transendothelial migration of human neutrophils. Results thus far indicate the LTC4 and LTD4 (10-6 M to 10-8 M) promote endothelial cells to metabolize 20:4 and that the major metabolite formed was prostacyclin. Unlike thrombin21 and histamine,22 which produce maximal levels of prostacyclin release within 10 minutes, LTC4 and LTD4 induced prostacyclin synthesis over an approximate 30-minute period. Direct comparisons of LTC4, LTD4, LTE4, and LTB4 on a molar basis indicate that LTC4 was approximately 50% more effective than LTD4 in stimulating prostacyclin synthesis and that LTE4 and LTB4 were ineffective. And finally, it was possible through the use of an in vitro model system to demonstrate that LTB4 was able to stimulate neutrophil adherence to and migration across an endothelial monolayer in the absence of serum and connective tissue factors, thus suggesting that LTB4 is able to directly mediate these events.
1. Dahlen S-E, Bjork J, Hendquist P, Arfors K-E, Hammarstrom S, Lindgren J-A, Samuelsson B: Leukotrienes promote plasma leakage and leukocyte adhesion in postcapillary venules. In vivo effects with relevance to the acute inflammatory response. Proc Natl Acad Sci USA 78:3887, 1981
2. Smedegaard G, Hedquist P, Dahlen S-E, Revenas B, Hammerstrom S, Samuelsson B: Leukotriene C4 affects pulmonary and cardiovascular dynamics in monkey. Nature 295:327, 1982.
3. Lindbom L, Hedqvist P, Dahlen S-E, Lindgren J-A, Arfors K-E: Leukotriene B.sub. 4 induces extravasation and migration of polymorphonuclear leukocytes in vivo. Acta Physiol Scand 116:105, 1982.
4. Bray MA, Ford-Hutchinson AW, Smith MJH: Leukotriene B4 : an inflammatory mediator in vivo. Prostaglandins 22:213, 1981.
5. Lewis RA, Goetzl EJ, Drazen JM, Soter NA, Austen KF, Corey EJ: Functional characterization of synthetic leukotriene B and its sterochemical isomers. J Exp Med 154:1243, 1981.
6. Palmblad J, Malmsten C, Uden A-M, Radmark O, Samuelsson B: Leukotriene B4 is a potent and sterospecific stimulator of neutrophil chemotaxis and adherence. Blood 58:658, 1981.
7. Goetzl EJ, Pickett WC: Novel structural determinants of the human neutrophil chemotactic activity of leukotriene B. J Exp Med 153:482, 1981.
8. Bjork JH, Hedqvist P, Arfors K-E: Increase in vascular permeability induced by leukotriene B4 and the role of polymorphonuclear leukicytes. Inflammation 6:189, 1982.
9. Jaffe EA, Nachman RL, Becheer CG, Minick CR: Culture of human endothelial cells derived from umbilical veins. J Clin Invest 52:2745, 1973.
10. Jaffe EA, Hoyer LW, Nachman RL: Synthesis of antihemophilic factor antigen by cultured human endothelial cells. J Clin Invest 52:2757, 1973.
11. Wagner DD, Olmsted JB, Marder VJ: Immunolocalization of von Willebrand protein in Weibel-Palade bodies of human endothelial cells. J Cell Biol 95:355, 1982.
12. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with Folin phenol reagent. J Biol Chem 193:265, 1951.
13. Cramer EB, Pologe L, Pawlowski NA, Cohn ZA, Scott WA: Leukotriene C promotes prostacyclin synthesis by human endothelial cells. Proc Natl Acad Sci USA 80:4109, 1983.
14. Unger WG, Stanford IF, Bennett A: Extraction of protaglandins from human blood. Nature 233:336, 1971.
15. Alan I, Ohuchi K, Levine L: Determination of cyclooxygenase products and prostaglandin metabolites using high pressure liquid chromatography and radioimmunassay. Anal Biochem 93:339, 1979.
16. Boyum A: Isolation of mononuclear cells and granulocytes fromhuman blood: isolation of mononuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedi mentationat 1G. Scand J Clin Lab Invest [Supp]21:77, 1968.
17. Harvath L, Falk W, Leonard E: Rapid quantitation of neutrophil chemotaxis: use of a polyvinylpyrrolidone-free polycarbonate membrane in a multiwell assembly. J Immunol Methods 37:39, 1980.
18. Cramer EB, Milks L, Ojakian g: Trnasepithelial migration of human neutrophils: An in vitro model system. Proc Natl Acad Sci USA 77:4069, 1980.
19. Folz CM, Russo FG, Siegal GP, Terranova VP, Liotta LA: Interactions of tumor cells with whole basement membrane in the presence or absence of endothelium. Prog Clin Biol Res 89:353, 1982.
20. Furie MB, Cramer EB, Naprstek BL, Siverstein SC: Cultured endothelial cell monolayers that restrict the transendothelial passage of macromolecules and electrical current. J Cell Biol 98:103, 1984.
21. Weksler BB, Ley CW, Jaffe EA: Stimulation of endothelial call prostacyclin production by thrombin, trypsin and ionophore A23187. J Clin Invest 62:923, 1978.
22. Baenziger NL, Force LE, Becherer PR: Histamine stimulates prostacyclin synthesis in cultured human umbilical vein endothelial cells. Biochem Biophys Res Commun 92:1435, 1980.
23. Cramer EB, Migliorisi, g, Pologe, G, Abrahams, E, Pawlowski, E, Cohn, Z, Scott, WA: Effect of Leukotrienes on Endothelium and the Transendothelial Migration of Neutrophils. J Allerg & Clinc Immun 74:386, No. 3, Part 2, 1984.
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People who are depressed may be unable to enjoy positive experiences, according to research that counters previous theories that the disorder is caused primarily by negative emotions. "Since depression is characterized by negative thinking, it is easy to assume that depressed people learn the negative lessons of life better than nondepressed people -- but that's not true," said study co-author Laren Conklin, an Ohio State University graduate student in psychology.
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Two weeks ago the Bronson Town Council reached a stalemate when it came to the appointment of someone to fill the seat left vacant by the resignation of Berlon Weeks.
The Levy County Sheriff's Office reports the following arrests:
• Dec. 14, Iverson Tyrone Aiken, 23, Cross City, battery, resisting office. Bond $35,000.
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Ratings based on 37 Verified Reviews
They paid and stayed. We double-checked.
4.4 Room cleanliness
4.1 Service & staff
4.3 Room comfort
4.3 Hotel condition
They paid and stayed.
We double-checked.
We double-checked.
for Everyoneby A verified travelerfrom Madrid, SPain
Exellent luxury and healthy hotelPosted Aug 14, 2014
Pros: The best are gym, SPA, the location and the rooms
Cons: breafast was not included in our rate, and is really expensive and they don't have a wide variety
Location: location is premium, in front of the ocean
the location is exceptional, in front of the sea; the hotel is really nice, wonderful suite-rooms, the SPA and gym incredible. THe food is really good, organic, healthy, although is appropriate for adults, but not to travel with young boys..as they don't appreciate it.
for Everyoneby Jasminefrom New York, NY
Canyon Ranch SpaPosted Jun 23, 2014
This place was totally serene, staff is friendly, super clean facility, and classes were amazing. I can't wait to go back, hopefully this year.
for Everyoneby A verified traveler
Loved itPosted Jun 21, 2014
The service, food, spa and gym/classes were awesome! I would love to come back!!
Recommendedby Luis
Posted Jun 15, 2014 on Hotels.com
for Everyoneby carlosfrom houston tx
Fist class servicePosted May 18, 2014
The food was great and the service unreal I loved the ocean view the spa is great to have in the same hotel he gym view was great I loved this place
for Business Travelersby A verified traveler
Posted May 4, 2014 on Hotels.com
for Business Travelersby A verified traveler
Posted May 1, 2014 on Hotels.com
for Everyoneby A verified traveler
Excellent hotel close to beach with awesome spa!Posted Apr 1, 2014
Pros: The experience as a whole was very relaxing...lot to do, but also many opportunities to relax and be pampered if you want. The room was so lovely!
Cons: The TV didn't work well...but, otherwise, it was a great experience.
Location: You need a car or travel by cab if you want to go to South Beach. If you want to simply relax, then there's no real reason to leave the hotel.
Really loved the Aquavena experience and most of the classes I took. The beach was lovely as were the pools. The staff was outstanding! Really helpful and very nice. And, the room was awesome!
for Personal Eventby MBfrom New York, NY
Expensive, good location, not worth itPosted Mar 29, 2014 on Hotels.com
Stayed at plenty of 5star hotels. While very nice, this was not a wow. And the rates are very high. We went to relax, and the hotel is quiet and peaceful. However, I think there are other nicer hotels for the money, and the main draw here is the spa (which when I used the jacuzzi, gave me a rash). I think they have great wellness programs, but if you aren't going specifically for that, other hotels in Miami offer more for less.
for Business Travelersby Claufrom Argentina
Un placer la estancia !!!!Posted Mar 4, 2014 on Hotels.com
Todo divino!!! Las piletas no muy grandes pero con el mar q hay para q las piletas. El spa lo más impresionante relaja te y divino de todo. Un placer !!!!! Volveré


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Vancouver Stealth defender Matt Beers assessed one-game suspension after review of Week 18 incidents. (Photo: Dan Brodie)
The National Lacrosse League has announced a summary of discipline stemming from incidents occurring during the Week 18 finale of the 2014 NLL regular season for games played on Friday, April 25 and Saturday, April 26.
Vancouver Stealth defenseman Matt Beers has been assessed a one-game suspension stemming from a game misconduct for multiple major penalties in the same game against the Calgary Roughnecks on April 26. Beers was assessed a major penalty for fighting at the 15:00 mark of the second quarter and a major penalty for spearing at 13:28 of the fourth quarter. With the Two Majors in One Game game misconduct in the team's finale (Rule 38.2), Beers has been assessed a one-game suspension for violation of Game Misconduct Assessed In Teams Final Game (Rule 40.4).
Toronto Rock defenseman Ethan O'Connor has been assessed a game misconduct penalty stemming from a boarding incident on Minnesota Swarm forward Shayne Jackson at approximately the 10:08 mark of the fourth quarter of their game on April 26. A major penalty was called on the field, and upon league review of the incident, O'Connor has been assessed a game misconduct penalty for boarding (Rule 74.3)
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The Westin San Diego
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TravelPod Member ReviewsThe Westin San Diego
I love this hotel!!! When you go on vacation, you expect to make experiences on the places you visit...and the hotel is just somewhere you stay. We weren't expecting much for our stay here, but each of our encounter with the staff added a great deal of our experience during our vacation.
We weren't very impress with the room. It felt small and our extra payment for a bay-view upgrade was not worth it. There was a huge parking lot in our view as well as two huge buildings blocking the bay.
Our hotel experience began at the Coast Restaurant for breakfast. The staff welcomed us and were constantly offering us something to drink. Our waiter, Cindy, made sure we were well taken care of, all our needs were met, and were satisfied with the food. We exchanged several travel experiences and she also recommended a few place for us to visit in San Diego. Not only that, but later that day, we came back to our room to see that she had left 2 complimentary voucher for us to visit Coast Restaurant again. We came back the next day and received more hospitality service from Juan. I loved that he remember our name and greeted us by name when we entered the restaurant.
This review is the subjective opinion of a TravelPod member and not of TravelPod.com.
Historical Traveler Reviews The Westin San Diego
Very poor customer communication
Having to leave the hotel earlier than expected due to health problems at home, find that a 'penalty' of one night is charged which had NOT been stated when I booked out. Only found out when checking card statement. No CD player, no robe in room. Also Reception should be clear as to who is booked in a room - confusion ensured when my wife phones and there is a suggetion that there is more than one person booked into the same room at the same time! Do divorce claims against the hotel hold?? On a better note, the concierge was very helpful in finding an urgent flight back home.
Great location and service
My wife and I spent 3 great nights at the Wyndham Emerald Plaza. The location could have not been better since it was close to Gaslamp, Seaport Village and Downtown. Our room was a King Junior Suite which was extremely large and comfortable. We had windows on three sides of a half octagon shaped room which gave us a great view of the bay and an aircraft carrier parked at North Island Naval Air Station. At night when we were walking around, the lighting of the hotel was like a beacon so we were able to easily find our way back. We ate at the hotel twice and the food was very good. One downside was that the atrium area was being renovated so we had to steer clear of that, but the results when finished should be awesome. We are members of the By Request program so we were greeted with several ammenities upon checkin such as a snack and beverage of our choice waiting in our room plus extra towels and pillows without having to ask for it,We will definitely be coming back here again especially once the new Petco Park is opened in Spring 2004.
Poor Customer Service!
We recently stayed at the Wyndham Emerald Plaza July 18-20th. We were extremely disappointed with the customer service. We had reserved a room for 4 people (2 beds), non-smoking months prior to our arrival yet when we checked in we were told all they had available in the entire hotel was one smoking room with one King size bed. Not sure why we even made a reservation if it was not going to be honored! We were actually taken to this room which was the size of a large storage closet and smelled like an ashtray. Not quite cozy for four people. Only after speaking to the manager were we given a room that still had only one king bed. We were provided with a lovely roll-away bed for my 6'1" husband! We were told there were CD players in each of the rooms which were not there, we were also told that there were only phone books available at the front desk which we later discovered was misinformation as we had one in our room. My husband requested the front desk hail a cab for us and he was told that she would have to wait over 20 minutes on the phone to get one!!!!! The carpet in our room was stained, the music jukebox service on the TV never worked, and an old eaten room-service tray from our neighbors sat outside our doorway through the night into the next morning until we moved it! All in all, an embarassment for the hotel chain!
Not up to par
My husband and I stayed here a few weeks ago and were very disappointed. The hotel nickeled and dimed us every step of the way.
The bathroom in the first room we were given was dirty so we were moved to a different room where the toilet constantly ran, keeping us up all night. We decided to leave the next day instead of carrying through our three night reservation. The hotel had the audacity to charge us for leaving early. When we got back home we wrote a letter and the manager did credit us for the fee however, we would never recommend this hotel to any of our family or friends.
Great Hotel
My family stayed at the Wyndham San Diego North Hotel over the week of July 4th. We loved it. The rooms were nice and clean. The pool was great. I would have liked a balcony though.
A good experience
We stayed at the wyndham last weekend. We were very pleased. We secured a very inexpensive($80 a night)rate at the last minute on Hotwire. We were treated very well. Had a room on the 20th floor overlooking the bay,were not charged for the roll away bed and had a good experience. The parking however is totally inconvenient. Valet only and no place else close to park. All in all a good experience.
Great place to stay!!
Me and my 3 friends stayed there for 6 nights at the Emerald Plaza. We got a really good deal and it was faboulous. It is in the heart of downtown. We didn't have a car so the hotel is right next to the trolley stop. The concierge was very helpful when we asked him how to get to Coronado beach. The room was great, and the bathroom was spacious and beautiful. The only complaint was the pool wasn't really that deep, but the whirlpool was nice. Also there was a fee for using the gym. All in all though it was very nice and I would deffinately stay there again if I could get a great deal again.
Not worthy in my opinion
My wife and I just spent three nights in San Diego and stayed at the Wyndham Emerald Plaza. We shopped online and found a good airfare/hotel package deal there. While checking in, I requested to be put in as high a floor as possible with a water view. I got no response. They gave me a room on the fifth floor of a 26 story hotel. I would've had a water view if I was 100 feet higher up. I could understand if there was no more rooms available but they didn't make that known. The front desk staff was short and rude. Some of the bellmen were very nice; one man gave us a ride to the zoo since he didn't have anyone to take to the airport. After a day of sightseeing, our room was not clean at 5:30 pm. I got a complimentary breakfast for that. I don't think I would stay here again and I really can't say it deserves a four star rating, especially after walking into the Grand Hyatt down the street where the people there greeted us with open arms even though we didn't have a room there.
It's decent
Stayed here in November 2002. After seeing the great photos of the hotel, we were really excited to stay here.
It's a decent place, but can't say it was anything special, considering it is a four-star hotel. It seems a little 'cold' -- it does not have the warm, fuzzy feeling of some other hotels.
The front desk staff was adeduately friendly, but nothing 'above and beyond.' The bellman really hustled, and didn't like the fact that we wanted to bring our bag in ourselves. He's right, it was his job to do so, but we were the guests, and we have the right to say no.
Our room was pleasantly spacious, and we had a wonderful view of the city and ocean. Visited the health spa, and there was a fee to use the gym -- c'mon. It was only open for another half hour at that point.
Parking is about $20/day, but there are in and out privileges.
Location is pretty good -- a few blocks from the heart of the gas lamp district.
To summarize, it's clean, it's spacious, it's well located -- but our experience was nothing out of the ordinary.
Gem of a Place
The Emerald Plaza is in the heart of downtown and is fabulous! We spent three nights. Room service helped a friend put together a welcome basket for our arrival. The Front Desk was quick and friendly. Everyone in the hotel smiles! Wonderful.
TripAdvisor Reviews The Westin San Diego
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2 days trip to San Diego
... br>
As I was the only one above 25 it was up to me to drive (it's much cheaper then..). I rent a "normal" car but the car rental gave me a Dodge Grand Caravan with almost 300 PS. Funny...by the way the first time ever that I've driven an automatic one. Crazy, they got plenty of lanes, sometimes up to 8-9 in one direction. Other cars overtake you from both sides, though the speed limit does not exceed 65 mph ..:) most often I drove 70 mph and was ...
San Diego with a dash of Mexico
... was an exaggeration however it still was pretty fun and cool. Yes almost everything there is made of lego and there are some huge lego recreations of characters, people and cities which were amazing. We didn't get to go on most rides as the lines were huge but we did go on a couple. We also got to see how lego was made .
Next day we drove into the city and parked next to the Midway Museum. The USS Midway is a massive ...
Stay Classy San Diego!
... around 6 hours today so we are spending most of the day on the coach. We will be taking a scenic drive through the Cleveland National Forest, the Anza Borrego Desert and the Sonoran Desert too. We will stop a few times for snacks and lunch so will keep you updated with all the cool things we are doing! I'm feeling tired and Paul has a sore throat but other than that we are doing really well. ...


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Matthew Immanuel Walker - December 18, 2013To William "Bill" and Veronica Walker of Jakarta, Indonesia, on Friday, Nov. 22, 2013, a son, Matthew Immanuel, weighing 7 pounds, 4 ounces, and was 19¾ inches in length. He was welcomed home by brothers, Delon and Farell.
Grandparents are Terry and Orpha Walker of Gothenburg and Mariwati Rusli of Jakarta.
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Everton midfielder Marouane Fellaini faces a race against time to be fit for the Merseyside derby at the end of October after it was confirmed the Belgium international had suffered a knee injury and will be unavailable for as much as three weeks, ESPN has reported.
The Belgium Football Association claimed Fellaini suffered the injury during Everton's 2-2 Premier League draw with Wigan Athletic on Saturday, and, after undergoing a scan on Tuesday, was ruled out of his country's World Cup qualification match against Serbia on Friday and the meeting with Scotland early next week.
After the international break Everton travel to Loftus Road to meet Queens Park Rangers and then welcome Liverpool to Anfield a week later, for the first derby meeting of the season.
This season Fellaini has made eight appearances for Everton, scoring three goals, including the winner in the opening round of the season against Manchester United. The 24-year-old moved to Goodison Park from Standard Liege in 2008.
See what the expert tipsters are tipping on OLBG
Stoke vs Chelsea - Premier League preview and team news
Chelsea outcast Salah targeted by Spartak Moscow
Lallana: Liverpool are turning the corner
Everton closing in on Marseille's Ayew
Southampton vs Everton - Premier League preview and team news
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The Real Housewives of Orange County's Vicki Gunvalson has announced she is officially dating Brooks Ayers again and that he is currently living with her as he battles the return of his non-Hodgkin lymphoma cancer. Vicki also reveals that at the time of the interview her daughter Briana did not know about Brooks moving in and that her RHOC co-star Tamra Barney has yet to reach out to her. Keep reading to watch the clip!
Current Category: Real Housewives of Orange County
On again off again frenemies Gretchen Rossi and Tamra Barney have a long history together as co-stars on The Real Housewives of Orange County, but where do they stand today?
The Real Housewives of Orange County star Vicki Gunvalson's on-again, off-again boyfriend Brooks Ayers is currently enmeshed in a lawsuit, but the 47-year-old revealed today that his legal battles aren't his biggest concern right now as he has been diagnosed with Stage 3 cancer.
As we reported earlier, The Real Housewives of Orange County star Vicki Gunvalson's controversial affirmation-loving boyfriend Brooks Ayers was caught on tape admitting that he cheated on Vicki with lots of women, including prostitutes. The tape, recorded by a wary business associate of Brooks in a Las Vegas hotel room, was obtained by The Enquirer and a portion of it has now made its way online via Radar Online.
It appears that The Real Housewives of Orange County OG Vicki Gunvalson's affirmation-loving boyfriend Brooks Ayers has once again been caught with his foot in his mouth thanks to the magic of recording equipment. The new recording allegedly features Brooksy admitting to hiring hookers and filling the love tanks of more women than he can count while dating Vicki. Keep reading for all the details, including a clip from RHOC Season 8 in which Lauri says Brooks was dating a "stripper slash hooker."
Part two of The Real Housewives of Orange County Reunion brought along some serious name calling between cast members. Tamra's BFF Vicki called her a "liar" and Lizzie said she is "nasty." Read on for details plus a video of both confrontations!
In the latest round of the Tamra Judge vs. Lizzie Rovsek post-Birthdaygate boxing match, Lizzie throws a couple high-road quote punches before a left hook insinuating Tamra's sick daughter excuse might not have been legit. Plus, get the latest casting reports that have Lizzie out and Tamra back in!
Nooooooooooooo!!!! It is being reported that Bravolebrity stalwart Tamra Judge has been fired from The Real Housewives of Orange County after seven seasons in the wake of a series of Facebook posts, in one of which Tamra railed against Bravo producers, and in another she posted cast mate Lizzie Rovsek's cell phone number. Keep reading for Tamra's full post (her Facebook page has been shut down), as well as her and Lizzie's reaction to the fall out.
On last night's episode of The Real Housewives of Orange County it was revealed that Tamra Judge's son Ryan Vieth suffers from the genetic blood disorder hemochromatosis, which requires him to make weekly visits to the doctor's office to have blood drained. Keep reading to find out more about his disorder, read Tamra's comments on whether or not it is linked to his HGH use, and see lots of photos and commentary from Ryan from his visits to the doctor over the last few months.
The ladies of The Real Housewives of Orange County began filming the Season 9 Reunion yesterday and we're getting lots of behind the scenes photos that have me thinking Bravo will be titling this episode "Boobs 'n' Bangs!" Keep reading to see Lizzie and Vicki's love tanks on display and check out Heather's dramatic new "banged up" look!


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L'Oreal • True Match Super-Blendable Makeup SPF 17 • Liquid
(2830 reviews)
|Average Rating.
|Would buy this product again.
feralstrumpet on 9/4/2010 2:23:00 PM more reviews by feralstrumpet
Age: 36-43 Skin: Combination, Fair Hair: Red Eyes: Green
This is my HG foundation. I wear the N2 or classic ivory. It evens out my skin tone very well and gives a nice matte finish that's not too dense. It's very easy to blend and looks like my own skin, only better.
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csaetern07 on 9/4/2010 1:08:00 PM more reviews by csaetern07
Age: 19-24 Skin: Normal, Olive Hair: Brown Eyes: Brown
I'm in a shade W6- sun beige (NC35) and the shades are definitely TRUE to MATCH. This foundation is my face in a bottle, its absolutely crazy! My problem areas are my cheeks because I have acne scarring from my teenage years which has faded over the time. I don't need much, a little bit goes a LONG way. This foundation can cake up on you when using the wrong tools. I first used a universal face brush and it was streaking and took forever to blend away but using a stippling brush, the finish is PERFECT. This foundation I would say is light to medium coverage but it is very buildable.
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cheylestar on 9/4/2010 10:52:00 AM more reviews by cheylestar
Age: 19-24 Skin: Combination, Tan Hair: Brown Eyes: Brown
PROS: I like this product because it has various colors to blend with the many different skin tones out there. Even as the seasons change, I always find the perfect matching tone for my skin all year around. Super easy to blend, very smooth & natural looking finish. Coverage is amazing, even when I got burned it covered the darkest spot on my face which was pretty dark. I like to use the powder in this same product to finish the natural & clean look. The price is very affordable, & the package is ok but yes it would be less messier & more convienant with a pump instead. Does not break your skin out unless you don't wash your face at least once daily. If you have dry to normal skin, I would definately recommend this product to you.
CONS: For me, I have super oily skin & I've had lots of trouble finding something that controls just that. This is ok for the first maybe 2hours max. Even when I finish it with the powder, not many hours later I look in the mirror & I have an oily shine that I just hate touching up every 2 hours. It does get everywhere if you aren't careful. It always stains my boyfriend's collar when we hug , even hours after it has already dried. If you have oily skin, I do not recommend this product to you.
I will continue to buy this product until I find something that works just as well minus the oily effect.
1 out of 1 people found this review helpful. Was this review helpful to you? Yes No
Pasha8888 on 8/30/2010 11:22:00 AM more reviews by Pasha8888
Age: 19-24 Skin: Other Hair: Other Eyes: Other
So far, this is the first foundation that doesn't make my pores look huge on my upper cheeks. I have large pores, and most foundations accentuate them, but not this one! I love it. I like the thin consistency because it still looks like skin as opposed to a mask of makeup!
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*TOP REVIEWER* natalielouiseee on 8/29/2010 6:52:00 AM more reviews by natalielouiseee
Age: 18 & Under Skin: Normal, Fair, Cool Hair: Brunette, Curly, Coarse Eyes: Blue
It's a good foundation, but not for me. I'm an NC10 in winter, NW15 in summer and I can't find any shades that match my skintone, without adding moisturiser to them, in which case it still makes my skin look orange!
The palest shade they have is a little too pale for me, so I use it with ALOT of moisturiser and use it as an under eye concealer and it's fabulous.
The only reason it get's three lippies is because it works well for me as an under eyeconcealer, it's affordable and it does give good coverage as a foundation, but there are no shades that suit my skintone.
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*TOP REVIEWER* Aishteru85 on 8/29/2010 3:23:00 AM more reviews by Aishteru85
Age: 25-29 Skin: Very Oily, Tan, Warm Hair: Brown, Straight, Coarse Eyes: Brown
Skin type: Oily/Pimple-Prone
Well, i was looking around to replace my revlon colorstay as, they have stopped carrying my shade, at least where i'm at, so i decided to try this. Basically I was looking for a foundation, much like revlon, gives medium to full coverage, controls oil, long-wearing, rub-proof, and doesnt melt when i sweat. But this really let me down.
-Its got a wide range of shade
-contains suncreen
- 30 ml can last awhile.
-provides medium to full coverage
-by 11am, the foundation on my chin and forehead are gone.
-my face is super duper oily. like i can fry an egg on it by mid-day.
-fades off pretty quickly.
-and when i sweat, my sweat turn orange...eww
-and its not rub proof.
bottomline: this is probably suitable for those with normal or dry skin...oily skin should stay away
4 out of 5 people found this review helpful. Was this review helpful to you? Yes No
*TOP REVIEWER* ashleydutta on 8/28/2010 4:40:00 AM more reviews by ashleydutta
Age: 25-29 Skin: Combination, Fair, Warm Hair: Blond, Wavy, Fine Eyes: Green
I just bumped my rating for this down a notch. I do like it, and can see myself picking it up again when I need an affordable foundation. but it's not perfect...guess for $11 how could I expect it to be?
Here's what's good about it:
of course, color selection is great - it's really easy to find your match.
it blends into skin really quickly and easily - they are right when they call it super blendable. It looks very natural on skin too - it doens't even look like you're wearing foundation!
Here's what's not so good:
it doesn't wear too well - after about 2 or 3 hours it starts kind of sliding off, looking as if I don't have any on anymore...and settles in pores.
I can't put it near my eyes b/c it settles in the fine lines and makes my under eyes look really aged.
of course, application is messy since there's no pump.
well that's about it. it's great to have on hand though, for when you need to put something on quickly to run to the grocery store/post office etc. no need in wasting my $50 foundations on days like that, so this is good to have!
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*TOP REVIEWER* azraelle on 8/27/2010 4:51:00 AM more reviews by azraelle
Age: 19-24 Skin: Very Oily, Fair, Cool Hair: Brunette, Wavy, Fine Eyes: Hazel
I remeber this very well, I used it when I was 16, my first foundation, which my mom bought me. It had a pump back then but let me tell you it was a pain to blend this, as even the lightest shade was too dark, so I had give this up, though I really liked the results and I needed some touch-ups after 6 hours or so but I guess my skin was much oilier back then and it would preform better now, but the shade-issue didn´t change, so I wont repurchase this. But all in all this is a really good drugstore-product and if you can find a shade that fits you - go for it!
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Aquablue66 on 8/27/2010 3:51:00 AM more reviews by Aquablue66
Age: 44-55 Skin: Combination, Fair Hair: Blond Eyes: Brown
Well, i have to say i can't live without this product!
Was out with my neighbors today with the kids playing and one of the gals said my skin looked radiant - this is my skin care secret,lol.
2 out of 2 people found this review helpful. Was this review helpful to you? Yes No
Rkhan715 on 8/27/2010 3:38:00 AM more reviews by Rkhan715
Age: 25-29 Skin: Acne-prone, Fair-Medium Hair: Black Eyes: Brown
I bought this in W3 nude beige. I am asian/middle eastern with prominent yellow undertones. For reference, i am a NC25 in mac studio fix liquid foundation and a NC30 in mac studio fix powder foundation. I was shocked that it matched my skin perfectly! I am very hard to match and it is next to impossible to find foundation (liquid or powder) that actually matches my skin and not be too pink, light or dark. i also have post acne marks and it covered them up okay, but not enough...i would use a concealer. I dont know why everyone has a issue with blending...it blends beautifully! Maybe the bottle you got was defective. I just quickly put it on using a sponge to see if it matched okay, but my normal routine would be use a stippling brush like mac 187 or a flat top brush like elf's studio powder brush (my HG). So far so good...havent worn it enough to see if it will oxidize or its staying powder.
2 out of 2 people found this review helpful. Was this review helpful to you? Yes No
Top Rated Liquid
Kanebo Sensai Fluid Finish foundation56 reviews
Cargo Liquid Foundation57 reviews


text:
Simple Service Manager 0.001
A simple service manager
Developer comments
This is my first real project with PyGTK so it is a little rough. While this tool should not break your system, I MAKE NO GUARANTEES.
It works with both Upstart and System V but can only disable/enable services. It cannot edit run-levels.
- last updated on:
- November 9th, 2009, 8:06 GMT
- price:
- homepage:
- gtk-apps.org
- license type:
- GPL (GNU General Public License)
- developed by:
- Stebalien
- category:
- ROOT \ Desktop Environment \ Tools
Add your review!


text:
Sunshine Villas
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Day 125, Update on Lab
... consists of the digestion of the samples with concentrated sulfuric acid at 400C Celsius in the presence of a catalyze of copper, followed by distillation. To calculate the amount of brute protein we measure the amount of total N2 of the sample and multiplying it by a known empirical factor. This assumes that the protein contain about 16% of N2.
The method can be divided in 3 phases:
1) Digestion of the sample with concentrate of ...
123, Angel Shark vs Human
... that it swam away. We decided to follow it. I took out my gopro and started filming. The shark was swimming out into the open ocean, I was on its left side, diving to the bottom and staying close to it to film. Antonio was on its right side. He swam a little faster, and then dove down. At this point he was right above the sharks head more or less. What might have happened is that the shark felt trapped by us humans (me on the left, and Antonio on the right and above) that ...
Day 118, Lab Update
Folch Method to quantify lipids
Prepare a mixture of chloroform-metano (2:1 v/v) cotaining 0.01% of BHT. Once the solvent has evaporate with a current of nitrogen, the lipids are weighed and the stored in an atmosphere of nitrogen and dissolved in chloroform to prent oxidation.
Day 95, Solar Oven
A dear friend of mine came to visit from the cloudy and rainy UK. We decided to spend the whole weekend at the beach, so a camping agenda was set. We went to Montana de Arena, and had 2 very sunny days and lots of fresh cold water. The highlight of the trip was the use of a solar oven I bought on the internet. With the pictures it ...
Day 91, Cleaning Tanks
... and bleach and a broom to scrub off all the alea growth. 4 air tubes and 1 large tube be cleaned. The other 5 tanks had to be siphones, emptied but still enough water for the fish, and srubbed slowly pushing all debree to the center, that is where the filter was. Then these tank were refilled. The filters, a large tube (with more empty space) in the middle with a mesh screen. We have two types of ...


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Seven privacy settings you should change immediately in iOS 8
Let's face it. Software has holes. And hackers love to exploit them. New vulnerabilities appear almost daily. If you have software - we all do - you need to keep tabs on the latest vulnerabilities.
Featured Stories
Articles about Security
Bonus Windows updates fix other Windows updates
In addition to the new Patch Tuesday updates, Microsoft reissued two older updates to fix problems in them and tightened up SSL security in Internet Explorer.
Microsoft patches Windows, IE, Office and Exchange
Seven updates include one to Exchange Server which was scheduled for last month and withheld.
Apple releases iOS 8.1.2
Apple has begun pushing iOS 8.1.2 update to iPhone and iPad users.
Adobe fixes Flash zero day, plus bugs in Acrobat, Reader and ColdFusion
The Acrobat updates are regularly-scheduled but the Flash and ColdFusion updates are a surprise. One Flash vulnerability is being exploited in the wild.
Box launches 'Trust' security program with Symantec, Splunk, others
Now the soon-to-be public business is taking initiative by launching its own security coalition, dubbed Box Trust.
IT Staff Systems/Data Access Policy
IT professionals such as system/network administrators, help desk staff and management personnel are routinely provided full access to company servers, network devices and data so they can effectively...
from TechProResearch »
POODLE not fixed? Some TLS systems vulnerable
The original POODLE bug which dogged SSLv3 was built into the protocol, but some TLS implementations make a mistake which causes the same effect.
Business security leaders fear loss of cyber war: IBM
According to IBM's latest CISO study, 60 percent of security leaders feel as though their organizations are outgunned in the supposed cyber war.
Two stealthy Linux malware samples uncovered, following in Windows variants' tracks
Two new variants of malware targeting government and business targets have been unearthed.
Advertisers to lose $6.3 billion to fraud in 2015
According to new research, advertisers are being swindled out of billions by cybercriminals looking to cash in on Internet advertising.
Blackphone launches app store for personal security and privacy
Together with the launch of updated custom Android software PrivatOS, the handset maker has revealed a new store dedicated to security and privacy applications.
Online shopping drives fraud penetration up: APCA
The Australian Payments Clearing Association has blamed the increase in online shopping for the rise in the total rate of fraud on Australian cards and cheques.
Cyber-espionage expected to surge in 2015: McAfee Labs
Cyber-warfare has been batted around everywhere from IT circles to popular culture, almost reaching a fever pitch recently surrounding suspicion and reports regarding the breach at Sony Pictures.
Microsoft to host Office 365, Dynamics CRM in Australia
Microsoft has announced that it will locally host Office 365 and Dynamics CRM Online in Australia, and extend its Cloud Solutions Provider program.
Breach epidemic's lurid chapter: who pays?
The Target breach case has the potential to define who has to write the checks to pay for the cleanup following a major breach.
Sony PlayStation back online after attackers claim credit for outage
A group calling themselves Lizard Patrol have claimed responsibility for a recent Sony PlayStation Network outage.
Twelve Days of Congress: gifts for the government that has everything (2013 Gift Guide)
How much do you love your country? Enough to shop until you drop, choosing just the right gifts? If you're stumped on exactly what to give your favorite nation, we have the answers. Come on in!
14 enormously useful products for the DIY-ITer (2013 Gift Guide)
Welcome to the official 2013 edition of our ZDNet DIY-IT Gift Guide. This year, in honor of 2013, we present to you 14 interesting and useful products that the DIY-ITer in your life will find particularly useful, fun, or cool.
Ada's Technical Books in Seattle
Ada's Technical Books in Seattle recently relocated, adding a cafe and more technical science, security, sci-fi and engineering books than before into its breathtaking new location.
ZDNet's top products and tech trends for 2013
Which products, platforms, and big ideas made the most impact in 2013?
Top tech blunders, catastrophes and epic fails of 2013
Thanksgiving is over, and we're already deep into December. It's time for a look back at all the blunders, catastrophes, epic fails and major screw-ups of 2013.
The world's biggest data breaches and hacks of 2013
From Facebook to Adobe, 2013 has been a tough year for companies looking to defend against cybercrime.
2013 in Review: Tech Turkeys of the year
And now, for your reading pleasure, the very worst from the tech world in 2013.
NSA mass surveillance leaks: Timeline of events to date
The U.S. government mass surveillance scandal may be the biggest ongoing story of the year. In this updating timeline, you can explore the full scope of the Edward Snowden leaks, which have implicated the world's most powerful nations in the worldwide spying operation.
The top ten scams to watch out for this Christmas
On the fifth day of Christmas, my true love gave to me, an app which stole my identity.
Presidential gadgets: What technology does Obama use?
The most tech-savvy U.S. president to date, Barack Obama uses the best devices and technology for the job, in and outside of the White House. Here's a look at some of the devices, platforms and technology he uses to carry out his day-to-day presidential duties.
Halloween '13: What tech frightens us most
In their latest October 2013 survey released just in time for All Hallows' Eve, IT recruiting firm Modis surveyed 1000 Americans to find out what potential tech disasters are keeping them up at night.
7 enterprise security improvements in iOS 7
iOS 7 is a major step forward in enterprise mobile security. Apple has institutionalized security techniques for which, until now, enterprises had to go to an independent MDM/MAM vendor.
NSA, Linux and handcuffs: Success in Serbia for Balkan hacker conference BalCCon
Groundbreaking Balkan hacker conference BalCCon: First Contact brought hackers from all over the world to its successful launch and makes plans for next year's edgy event.
24 hours in Serbia at Balkan hacker conference BalCCon
This weekend the Balkan Computer Congress BalCCon: First Contact is putting Balkan hackers center stage with international hackers at the first-ever Balkan hacker conference.
Gallery: Hacker Playing Cards from Firehost
The limited edition security themed playing cards from Firehost handed out at Black Hat USA have hackers as jokers, and more hidden surprises in the artwork than you'd expect.
NAB developing PlayBook-specific apps
The National Australia Bank (NAB) has revealed that it is currently building apps specifically for Research In Motion (RIM)'s new BlackBerry PlayBook tablet, while voicing ongoing concerns over the security of Android-based banking.
Content owners don't back AFACT: iiNet
Content providers are stepping away from the Australian Federation Against Copyright Theft (AFACT)'s pursuit of ISPs over copyright infringement, and are open to iiNet's piracy mediator model, according to iiNet's chief regulatory officer Steve Dalby.
AusCERT 2011: the good, the iPad, the insecure
The AusCERT 2011 is over and the sun has set on the exhibition floor for another year. ZDNet Australia brings you the highlights of the conference.
Testing Qantas' next-generation systems
Qantas' push to make check-in faster and easier is certainly intriguing, but does it actually work? We put the premium airline's new RFID luggage tags to the test.
Smartphones: the perfect bugging device
Security experts at the AusCERT 2011 Conference in Queensland this week warned that serious attacks on mobile phones are expected before the end of this year, and that those attacks will involve tracking users, not just stealing their money.
Silent victims thwart cybercops: Qld Police
Police could pursue more online criminals if more victims reported the crimes committed against them, according to one of Australia's highest-profile investigators.
AusCERT 2011: firms ignore ID theft risk
Businesses don't take the risk of identity theft seriously enough, according to Welsh identity theft victim, activist and comedian Bennett Arron.
AusCERT kicks off 10th conference
The tenth annual Australian Computer Emergency Response Team (AusCERT) conference takes place in Queensland's Gold Coast this week.
Apple to fix iOS location tracking 'bug'
Apple has finally broken its silence on the iPhone tracking controversy, posting a lengthy FAQ on its website last night.
AFACT loses iiNet piracy appeal case
The Federal Court has dismissed the appeal for the landmark iiNet copyright case brought against it by the Australian Federation Against Copyright Theft (AFACT).
The Future Of... Credit Cards
They're a modern-day must. Anyone who's ever forgotten their credit card at home or worse, lost it, knows exactly how crucial that piece of plastic truly is. With rampant fraud and ID theft though, most cardholders are equally aware of the risks. But what if the account number on your card disappeared when not in use? ZDNet correspondent Sumi Das explores payment innovations from Dynamics, Square and Bling Nation.
PGP Personal for Windows 8.0
PGP 8.0 is an industrial-strength encryption program with all the features necessary to protect your files and online communications.
PGP Personal for Windows 8.0
PGP 8.0 is an industrial-strength encryption program with all the features necessary to protect your files and online communications.
McAfee VirusScan Home Edition 7.0
McAfee VirusScan 7.0 is a top-notch virus killer for first-time users, but unless you use Outlook Express or Eudora, VirusScan 6.0 users won't need to upgrade.
Panda Antivirus Platinum 7.0
This virus hunter's new interface is simple to use, but Panda Antivirus Platinum stumbles on support and detection skills.
Norton Internet Security 2003
Norton Internet Security 2003 is a solid package for the price, especially as an upgrade.
Norton AntiVirus 2003
For zapping viruses, worms and other malicious code, you can't go wrong with Norton AntiVirus 2003. However, current NAV users need not upgrade.
Detecting network intrusions: PureSecure
Detecting network intrusions: PureSecure
SpamKiller 2.90
An effective and easy-to-use tool for fighting spam, SpamKiller 2.90 is a must for anyone whose inbox is overrun by unwanted mail.
ZoneAlarm Pro 3.0
ZoneAlarm Pro 3.0 is as good as personal firewalls get. Although you'll still need an anti-virus product to completely protect your PC, we think that the extras in ZoneAlarm Pro are worth the cash -- free alternative or not.
McAfee Internet Security 4.0
Most people won't need the extra security tools in McAfee Internet Security 4.0. You're better off buying the standalone VirusScan 6.0, or Norton AntiVirus 2002, and downloading a free firewall such as ZoneAlarm.
Norton Internet Security 2002
We love the firewall and anti-virus combo, but if all you want is a way to keep hackers at bay, Norton Internet Security 2002 costs too much. Get it if you need anti-virus protection too; otherwise, download the free ZoneAlarm firewall.
VirusScan 6.0
VirusScan is not the best virus slayer on the street. Go for Norton instead and install a free firewall such as ZoneAlarm.
Norton AntiVirus 2002 8.0
There's no better virus killer than Norton AntiVirus -- but unless you're moving to Windows XP, don't bother with this minor upgrade.
PGP Personal Security 7.03
PGP Personal Security will only be effective if you understand what it's doing and make it part of an overall security scheme -- just installing and running it won't provide a miracle cure.
VirusScan Wireless
Buy a standard desktop anti-virus package instead.


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"Searching your library's specialized subject encyclopedia collection helps refine your search; save time, and get to the book or journal you need more quickly and accurately.
Reference Universe contains 35 million data elements to over 45,000 specialized reference works. The elements are matched to your library's electronic or print holdings, offering you a customized view."
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The cold hard truth is that anti-gunners DO reap the silent benefits of gun ownership in any area where firearms are in the hands and homes of responsible gun owning citizens.
That is because the criminal element has no way to differentiate between individuals and households "with & without" & "for and against" firearms.
Criminals are then much more cautious/hesitant than if the entire population is defenseless with regard to deadly force.