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PubMed2004

Planar electrochromatography.

Recent developments in planar electrochromatography (PEC) in both the normal-phase and the reversed-phase modes, and at both atmospheric and elevated pressure, are reviewed. Other forced-flow techniques in planar chromatography are also briefly covered. Mobile phase migration in PEC is primarily due to electroosmotic flow, which is controlled by the applied electric field. Capillary mediated flow is an important secondary contributor to migration, and occurs because the layer is unsaturated as a consequence of liquid evaporating from the layer due to Joule heating. The magnitude of the electric field and the concentration of ions in solution are important variables that control both electroosmotic flow and Joule heating. Separations are faster and more efficient than those obtained by conventional planar chromatography, provided appropriate experimental conditions are selected. With inappropriate conditions, either mobile phase accumulates on the surface of the sorbent layer, or Joule heating causes excessive evaporation. The former results in poor spot shape, and the latter can cause the layer to dry. Good separations are obtained when there is a balance between these two effects. The problems associated with mobile phase accumulating on the surface of the sorbent layer, and with excessive evaporation of mobile phase, do not occur with pressurized planar electrochromatography. This technique is performed at high pressure, under conditions that allow heat to be removed form the sorbent layer. This allows the use of a substantially higher electric field than in PEC, and results in a high mobile phase flow rate.

Chromatography, Micellar Electrokinetic Capillary↗
PubMed2001

Altered expression of L-selectin (CD62L) on polymorphonuclear neutrophils of children vertically infected with human immunodeficiency virus type 1.

The expression of the leukocyte adhesion molecule L-selectin (CD62L) on polymorphonuclear neutrophils (PMN) and circulating levels of the soluble form of the receptor (sCD62L) were determined for a group of HIV-1-infected children, categorized as having mild or severe disease, and a group of uninfected control children. The fluorescence intensity of CD62L on PMN was significantly reduced in the HIV-1-infected children with mild disease compared to the uninfected controls. The proportion of lymphocytes expressing CD62L, as well as their corresponding fluorescence intensities, was significantly reduced in both the mild and the severe disease groups compared to the uninfected children, while peripheral levels of sCD62L were significantly elevated in the HIV-1-infected children with mild and severe disease compared to the controls. Altered cell migration resulting from reduced expression of CD62L may be an important contributor to the increased susceptibility to secondary microbial infections seen in HIV-1-infected children.

AIDS-Related Opportunistic Infections↗
PubMed2003

p19Arf-p53 tumor suppressor pathway regulates cell motility by suppression of phosphoinositide 3-kinase and Rac1 GTPase activities.

The p19(Arf)-p53 tumor suppressor pathway plays a critical role in cell-cycle checkpoint control and apoptosis, whereas Rho family small GTPases are key regulators of actin structure and cell motility. By using primary mouse embryonic fibroblasts that lack Arf, p53, or both, we studied the involvement of the p19(Arf)-p53 pathway in the regulation of cell motility and its relationship with Rho GTPases. Deletion of Arf and/or p53 led to actin cytoskeleton reorganization and a significant increase in cell motility. The endogenous phosphoinositide (PI) 3- kinase and Rac1 activities were elevated in Arf(-/-) and p53(-/-) cells, and these activities are required for p19(Arf)- and p53-regulated migration. Reintroduction of the wild type Arf or p53 genes into Arf(-/-) or p53(-/-) cells reversed the PI 3-kinase and Rho GTPase activities as well as the migration phenotype. These results suggest a functional relationship between an established tumor suppressor pathway and a signaling module that controls actin structure and cell motility and show that p19(Arf) and p53 negatively regulate cell migration by suppression of PI 3-kinase and Rac1 activities.

Animals↗
PubMedDate not supplied

Migration of intravitreal silicone oil through a Baerveldt tube into the subconjunctival space.

A 28-year-old patient developed proliferative diabetic retinopathy with florid rubeosis iridis and ultimately required the placement of a Baerveldt tube to control his secondary glaucoma. Eighteen months later, he underwent a pars plana vitrectomy, scleral buckle, lensectomy, and membrane peeling to reattach a severe diabetic retinal detachment. Ultimately, some of the 5000 cs silicone oil migrated through the tube to the episcleral region under the plate of the Baerveldt device. The oil intermittently blocked the shunt, causing elevated intraocular pressure. Despite ultimate surgical removal of the oil from around the tube and plate, a substantial amount remained encapsulated in the subconjunctival space. Prevention of this complication includes placement of a short tube well anterior to the iris in the inferior portion of the anterior chamber.

Adult↗
PubMedDate not supplied

Directed migration of Ascogregarina taiwanensis (Apicomplexa: Lecudinidae) in its natural host Aedes albopictus (Diptera: Culicidae).

Directed migration of trophozoites from the midgut toward the Malpighian tubules is essential for Ascogregarina taiwanensis (Apicomplexa: Lecudinidae) to complete its developmental cycle within the natural host Aedes albopictus. We have obtained a 275-bp actin cDNA fragment amplified from extracted mRNAs of migrating trophozoites, suggesting the involvement of actin in trophozoite motility. Down-regulation on the migration of the trophozoite was seen in mosquito larvae fed with cytochalasin D, ML-7, and BDM, indicating that myosin, in the form of an actomyosin system, may also be involved in driving motility of the trophozoite. The "protruding apparatus" (PA) formed at the anterior end of trophozoites during the migrating stage had significant deposits of actin by immunofluorescent microscopy. Moreover, PA formation was enhanced in response to elevated levels of 20-hydroxyecdysone (20-HE) in cultures of alimentary canals in which the trophozite was contained. Thus, 20-HE may also promote expression of actin and perhaps myosin simultaneously.

Actins↗
PubMed1994

Impaired early bone formation in periodontal fenestration defects in dogs following application of insulin-like growth factor (II). Basic fibroblast growth factor and transforming growth factor beta 1.

Effects of a topically applied growth factor combination on fibroblast migration, collagen fiber formation and bone regeneration were studied in standardized periodontal defects in 4 beagle dogs. Following elevation of facial mucoperiosteal flaps, fenestration defects, 3 mm in diameter, were made through the cortical bone and into the dentin of maxillary and mandibular teeth. Collagen sponges, impregnated with 200 ng insulin-like growth factor II, 20 ng basic fibroblast growth factor and 6 ng transforming growth factor beta 1 were fitted to defects randomly in right or left quadrants and the flaps repositioned and sutured. Contralateral control defects received the collagen with vehicle only. Experimental procedures were staggered to allow observations of healing 3, 7, 10, and 14 days after surgery. Histometric analysis showed no differences in fibroblast and collagen density between control and growth factor defects. Bone regeneration was significantly greater in control than in growth factor defects 10 and 14 days after surgery. The rate of healing generally appeared more affected by intra-dog variations or procedural variations than by the growth factor combination.

Alveolar Process↗
PubMed1992

Characterization of the heat shock response in Brucella abortus and isolation of the genes encoding the GroE heat shock proteins.

In an effort to define the heat shock response in the bovine intracellular pathogen Brucella abortus, a rough variant lacking extensive lipopolysaccharide was pulse-labeled with [35S]methionine following exposure to elevated temperatures. The major heat shock proteins observed following sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography migrate at 70, 62, 18, and 10 kDa. The maximum response was observed between 42 and 46 degrees C and within 2 to 3 h of the shif in temperature and varied slightly for the different proteins. Accumulation of the 62-kDa heat shock protein (62-kDa Hsp) was observed to continue for up to 5 h following the shift in temperature. In an effort to better define the heat shock response and its potential relationship with protective immunity, genes encoding the major heat shock proteins were isolated from recombinant libraries constructed from B. abortus S19 and S2308 and sequenced. The 62-kDa Hsp shares more than 60% amino acid homology with members of the GroEL family and is immunoprecipitated with polyclonal antibodies to Escherichia coli GroEL and monoclonal antibodies to mycobacterial Hsp 65. Western blot (immunoblot) analysis with pooled sera from vaccinated and infected cattle revealed that the 62-kDa Hsp is a predominantly recognized antigen. The roles of these gene products during environmental stress and in protective immunity against brucellosis are under investigation.

Amino Acid Sequence↗
PubMed2006

Analysis of inflammation.

In the past, inflammation has been associated with infections and with the immune system. But more recent evidence suggests that a much broader range of diseases have telltale markers for inflammation. Inflammation is the basic mechanism available for repair of tissue after an injury and consists of a cascade of cellular and microvascular reactions that serve to remove damaged and generate new tissue. The cascade includes elevated permeability in microvessels, attachment of circulating cells to the vessels in the vicinity of the injury site, migration of several cell types, cell apoptosis, and growth of new tissue and blood vessels. This review provides a summary of the major microvascular, cellular, and molecular mechanisms that regulate elements of the inflammatory cascade. The analysis is largely focused on the identification of the major participants, notably signaling and adhesion molecules, and their mode of action in the inflammatory cascade. We present a new hypothesis for the generation of inflammatory mediators in plasma that are derived from the digestive pancreatic enzymes responsible for digestion. The inflammatory cascade offers a large number of opportunities for development of quantitative models that describe various aspects of human diseases.

Animals↗
PubMed1999

Nitric oxide and C-type atrial natriuretic peptide stimulate primary aortic smooth muscle cell migration via a cGMP-dependent mechanism: relationship to microfilament dissociation and altered cell morphology.

Migration of aortic smooth muscle cells is thought to be of essential importance in vascular restenosis, remodeling, and angiogenesis. Recent studies have shown that NO donors inhibit the migration of subcultured aortic smooth muscle cells. However, there is evidence that NO elicits opposite effects on cell proliferation in primary versus subcultured cells, indicating fundamental differences among different models of aortic smooth muscle cell cultures. The purpose of the current study was to investigate the effect of NO donors on migration of primary cultures of rat aortic smooth muscle cells and to compare and contrast their response with those in subcultured cells. A second purpose was to investigate some of the underlying mechanisms associated with NO-induced effects on cell migration. We report that 2 NO donors, S-nitroso-N-acetylpenicillamine (SNAP) and 2, 2-(hydroxynitrosohydrazino)bis-ethanamine, stimulated the migration of primary cells in a wounded-culture model as well as in a transwell migration model. The effect of NO donors was mimicked by 2 cGMP analogues and C-type natriuretic peptide and blocked by a specific inhibitor of guanyl cyclase, 1H-(1,2,4)oxadiazolo[4,3, -a]quinoxalin-1-one, indicating the involvement of cGMP as second messenger. Moreover, neither NO donors nor cGMP analogues altered migration of primary cultures stimulated by either FBS or angiotensin II. In contrast to its effect in primary cultures, SNAP did not alter basal or stimulated migration of subcultured cells, except at a relatively high concentration of 1 mmol/L, at which migration was inhibited. The migration-stimulatory effect of NO donors and cGMP was associated with altered cell morphology and dissociation of actin filaments, consistent with recent studies indicating that cell morphology and cytoskeletal organization influence cell migration. The results suggest the possible involvement of NO-induced cell migration in vascular injury or remodeling, representing conditions in which vascular NO levels would be expected to be elevated.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed2001

Tumor necrosis factor-alpha induces fibronectin synthesis in coronary artery smooth muscle cells by a nitric oxide-dependent posttranscriptional mechanism.

Postcardiac transplant coronary arteriopathy is associated with tumor necrosis factor-alpha (TNF-alpha) induction of fibronectin-dependent smooth muscle cell (SMC) migration into the subendothelium, resulting in occlusive neointimal formation. Because expression of inducible nitric oxide synthase (iNOS) is elevated in neointimal formation after transplantation and upregulated in vascular SMCs by TNF-alpha, we investigated whether TNF-alpha induction of fibronectin synthesis in coronary artery (CA) SMCs is mediated by nitric oxide (NO). TNF-alpha caused a dose-dependent increase in reactive oxygen and nitrogen intermediates in CA SMCs (P<0.05). This correlated with increased NO production (P<0.05) and fibronectin synthesis (P<0.05). TNF-alpha induction of fibronectin synthesis was abrogated by the NOS inhibitor N(G)-monomethyl-L-arginine (L-NMMA) (P<0.05) or the flavonoid-containing enzyme inhibitor diphenyleneiodonium (DPI) (P<0.05) and reproduced with the NO donor S-nitroso-N-acetyl-penicillamine (SNAP) (P<0.05). Northern blotting showed no effect of TNF-alpha on steady-state fibronectin mRNA levels. TNF-alpha increased expression of light chain 3 (LC-3), a protein shown previously to facilitate fibronectin mRNA translation through its interaction with an adenosine-uracil rich element (ARE) in the 3'-untranslated region of fibronectin mRNA. RNA gel mobility shift and UV cross-linking assays using CA SMC lysates revealed protein binding complexes with radiolabeled oligonucleotide containing the ARE, similar to those generated with recombinant LC-3. One of these complexes increased after TNF-alpha treatment, an effect inhibited with L-NMMA or DPI. These data demonstrate a novel paradigm whereby cytokines regulate mRNA translation of extracellular matrix proteins through NO-dependent modulation of RNA binding protein interaction with mRNA.

Animals↗
PubMed1995

HIV/AIDS in the rural United States: epidemiology and health services delivery.

This integrated research review addresses the epidemiology of rural human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) and the organization, financing, and delivery of health services for rural persons living with HIV or AIDS (PLWHIVs, PLWAs). Several abstracting services, indexing services, and bibliographies were searched. An annotation form served as the guideline for data extraction. Several conclusions emerged from this review. Epidemiological evidence indicates that there has been a dramatic increase in the relative proportion of rural HIV/AIDS incident cases over the past 5 years. Explanations for the rural increase focus on injection drug use, heterosexual behavior, and sexually transmitted disease levels. Dramatically elevated rates of infection in rural Black women are indicated. Rural areas experience important levels of in-migration of HIV/AIDS-infected individuals. The health services literature suggests that rural providers and institutions have limited resources and little experience with PLWHIVs or PLWAs.

Acquired Immunodeficiency Syndrome↗
PubMed2002

[Bacterium culture and flora analysis of gunshot wound on limbs of dogs in hot and humid environment].

OBJECTIVE: To investigate the time of bacterial invasion into the blood and conduct flora analysis of the blood and secretions in the gunshot wound in the limbs of dogs in hot and humid environment. METHODS: Gun-shot wound was induced in 8 dogs who were subsequently assigned at random into hot and humid environment (HHE) group and normal environment (NE) group for observation. At the time points of 0, 4, 6, 8, 12, 24 h after injury, body temperature was measured and bacterial culture and flora analysis performed. RESULTS: Body temperature elevation occurred earlier and lasted for longer time in HEE group than in NE group. Bacterial invasion into the blood occurred at 12 h after the injury in NE group, but at 8 h in HHE group. In the wound tracts of the dogs, surface bacteria were found in both groups. Surface bacteria were also found in the blood of dogs in both groups, but in HHE group, even intestinal flora were identified. CONCLUSION: Body temperature elevation and bacterial invasion into the blood occurred at an earlier time in hot and humid environment where migration of intestinal bacteria into the blood easily takes place, indicating the early application of broad-spectrum antibiotics under such environmental conditions.

Animals↗
PubMed1992

A postsynaptic role for single-stranded DNA-binding protein in recA protein-promoted DNA strand exchange.

The single-stranded DNA-binding protein (SSB protein) is required for efficient genetic recombination in vivo. One function for SSB protein in DNA strand exchange in vitro is to remove secondary structure from single-stranded DNA (ssDNA) and thereby aid in the formation of recA protein-saturated presynaptic complexes. In the preceding paper (Lavery, P. E., and Kowalczykowski, S. C. (1992) J. Biol. Chem. 267, 9307-9314) we demonstrated that DNA strand exchange can occur in the presence of volume-occupying agents at low magnesium ion concentration, where secondary structures are reduced. Our results suggest that SSB protein is not acting during presynapsis under these conditions, yet the DNA strand exchange reaction is stimulated by the addition of SSB protein. In this study we present biochemical evidence which suggests that SSB protein stimulates DNA strand exchange by binding to the ssDNA displaced from joint molecules, thereby stabilizing them and allowing branch migration to extend the region of heteroduplex DNA. Therefore, our results indicate dual roles for SSB protein at elevated magnesium ion concentration; it functions during presynapsis, removing secondary structure from ssDNA, as indicated previously, and it also functions postsynaptically, binding to the ssDNA displaced from joint molecules.

Adenosine Triphosphatases↗
PubMed1979

Lipoprotein abnormalities in cholestasis. I. Electrophoretic and ultracentrifugal analyses.

The alterations of lipid composition in sera of patients with liver diseases, particularly intrahepatic cholestasis and biliary obstruction, were studied by ultracentrifugation and polyacrylamide-gel disc-electrophoresis of lipoproteins and apoproteins. The elevation of serum cholesterol in intrahepatic cholestasis was greater than in biliary obstruction. The appearance of lipoprotein X in obstructive disease accounted for most of the increased cholesterol. The level of non-lipoprotein X cholesterol in intrahepatic cholestasis was significantly elevated, this being in part ascribed to the appearance of a new class of cholestatic lipoprotein, Slow-migrating HDL. The electrophoretic pattern of lipoprotein in cholestasis was generally characterized by a decrease in alpha band intensity and, in some types of cholestasis, by the appearance of Slow-migrating HDL. In addition, other abnormal lipoproteins exhibiting the characteristics of triglyceride-rich LDL (LP-Y), LP-X-like HDL and LDL-like HDL were found in some cases of intrahepatic cholestasis and biliary obstruction.

Apolipoproteins↗
PubMed1993

[Acute pancreatitis associated with biliary lithiasis].

The physical characteristics of gallstones and anatomical predisposing factors strongly suggest that acute pancreatitis associated with gallstones is due to a transient blockage of the ampulla of Vater by migrating stones. The diagnosis of acute pancreatitis is based on clinical probabilities and is supported by a threefold elevation in serum amylase. In cases of acute biliary pancreatitis the initial serum amylase value is higher and its decline more rapid than in pancreatitis due to other causes. The diagnostic sensitivity and specificity of serum amylase are higher in cases of acute biliary pancreatitis. The early detection of gallstones in acute pancreatitis by standard imaging techniques may be misleading; ultrasonography fails to identify the gallbladder in almost one-third of patients. The reported accuracy of systems for the clinico-biochemical detection of gallstones in acute pancreatitis is 75 percent. Their lack of specificity prohibits their use as the sole means of detecting gallstones, but they may allow the selection of patients who require further investigations. Ranson's multiple laboratory criteria are generally recognized as providing the best early assessment of severity in acute pancreatitis; a modified scoring system is required for patients known to have pancreatitis due to gallstones. The timing of surgery in gallstone pancreatitis is no longer controversial. In patients with benign pancreatitis the time of surgery appears to have little effect on the outcome, whereas in patients with severe pancreatitis early surgery results in a significant increase in morbidity and mortality rates. Surgery should be performed in the first days following hospital admission, soon after the pancreatitis has subsided. Further investigations are warranted before advocating a widespread use of early endoscopic sphincterotomy in the treatment of severe gallstone acute pancreatitis.

Acute Disease↗
PubMed1998

[Significance of antibodies to herpesviridae viruses detectable in rheumatic diseases].

AIM: To assay antibodies to cytomegalovirus (CMV), herpes simplex virus type 1 and 2 (HSV-1, HSV-2) and Epstein-Barr virus (EBV) in rheumatic patients and to clarify clinical correlations. MATERIALS AND METHODS: A total of 66 patients were examined: 7, 19, 6, 3, 5, 2 and 24 with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), reactive arthritis (ReA), scleroderma systematica (SS), erythema nodosum (EN), hemorrhagic vasculitis (HV), active or chronic viral infection (A/CVI), respectively. Clinical, laboratory tests, tests for specific IgM- and IgG-antibodies to CMV, HSV-1, HSV-2, EBV, x-ray examinations were performed. RESULTS: IgG-antibodies to CMV were detected in 79%, VCA-IgG-antibodies to EBV in 70.3%, EA-IgG-antibodies to EBV in 56.6%, IgG-antibodies to HSV-1 in 42.1% of patients. Active CMV infection was diagnosed in 27.8%, active EBV infection in 56.6%, combination of CMV and EBV infection in 46.9% of cases. High titers of antibodies to CMV and EBV correlated with such symptoms as fever, arthritis, myalgia, carditis, hepatomegalia, migrating erythematous eruption. Acute-phase indices were related to high titers of antibodies to CMV and EBV. Elevated titers of antibodies to CMV and EBV were registered both in untreated patients and in patients treated with corticosteroids, nonsteroid antiinflammatory drugs and aminoquinoline drugs. CONCLUSION: In differential diagnosis of rheumatic diseases it is necessary to consider possibility of CMV and EBV infections. If these are detected, antiviral measures should be taken.

Antibodies, Viral↗
PubMed2006

Elevated serum level of YKL-40 is an independent prognostic factor for poor survival in patients with metastatic melanoma.

BACKGROUND: YKL-40 is a growth factor for connective tissue cells and stimulates migration of endothelial cells. Cancer cells, macrophages, and neutrophils secrete YKL-40. Its function in cancer is unknown. High serum YKL-40 levels have been associated with a poor prognosis in patients with several solid tumors. The prognostic impact of serum YKL-40 in metastatic melanoma was evaluated. METHODS: YKL-40 was measured in serial serum samples from 110 patients with metastatic melanoma obtained immediately before and during treatment and from 245 healthy subjects. RESULTS: Patients had higher serum YKL-40 values than healthy subjects (P < 0.001). Pretreatment serum YKL-40 was elevated in 45% of the patients and correlated to site of metastases (P = 0.03) and poor performance status (P = 0.002). Multivariate Cox analysis showed that serum YKL-40 (hazard ratio [HR] = 1.9; 95% confidence interval [CI], 1.2-2.8; P = 0.004) and serum lactate dehydrogenase (LDH) (HR = 1.9; 95% CI, 1.2-2.9; P = 0.004) were independent prognostic factors for survival. A combination variable of elevated serum YKL-40 and LDH quadrupled the risk of early death (HR = 4.4; 95% CI, 2.5-7.7; P < 0.001) compared with patients with normal levels. The combination of YKL-40 and LDH had a stronger prognostic impact than the American Joint Committee on Cancer (AJCC) Stage IV classification. Furthermore, serum samples were available from 12 patients during followup. In 9 of 11 patients a significant increase in serum YKL-40 was observed together with disease progression. In one patient with a lasting complete response, serum YKL-40 remained normal. CONCLUSIONS: An elevated serum YKL-40 was an independent prognostic factor for poor survival in patients with metastatic melanoma. When combining serum YKL-40 and LDH, patients could be separated into three prognostic groups based on the number of elevated biomarkers. The findings should be validated in an independent study.

Adipokines↗
PubMed2005

Increase in macrophage migration inhibitory factor levels in lacrimal fluid of patients with severe atopic dermatitis.

BACKGROUND AND AIMS OF THE STUDY: Atopic dermatitis is a chronic inflammatory skin disorder that often involves some ophthalmic features. Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine that is associated with the generation of cell-mediated immune responses. Although serum MIF levels may be elevated in severe atopic dermatitis, the quantity of MIF in regional ocular fluid remains unknown. We measured MIF levels in tears (lacrimal fluid) of patients with atopic dermatitis. PATIENTS AND METHODS: Tear samples were collected from 16 patients with atopic dermatitis, 10 patients with allergic conjunctivitis, and 15 healthy control subjects. The clinical severity of atopic dermatitis was evaluated according to the Scoring Atopic Dermatitis (SCORAD) index. The index was calculated by summing the following scores: extent criteria, intensity criteria, and subjective symptoms. Macrophage migration inhibitory factor levels were determined by a human MIF enzyme-linked immunosorbent assay. All comparisons were two-tailed, and P values <0.01 were considered as statistically significant. RESULTS: The mean MIF concentration in lacrimal fluid collected from healthy control subjects was 0.69+/-0.2 ng/ml. The mean tear MIF levels were 17.87+/-6.3 ng/ml in moderate-to-severe atopic dermatitis (SCORAD> or =15, P=0.002), 0.93+/-0.08 ng/ml in mild atopic dermatitis (SCORAD<15), and 2.76+/-0.86 ng/ml in allergic conjunctivitis (P=0.008). CONCLUSIONS: A proinflammatory cytokine MIF level was elevated in tears as well as serum in cases of severe atopic dermatitis. These results suggest that MIF may play an important role in the induction or enhancement of ophthalmic features related to severe atopic dermatitis.

Adolescent↗