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S Sela

Publications and source records attributed to S Sela.

At least 19 recordsLinked to original sources
PubMed2000

Effect of lipoteichoic acid on the uptake of Streptococcus pyogenes by HEp-2 cells.

Lipoteichoic acid (LTA) is thought to play a role in the interactions between Streptococcus pyogenes and host cells. We have examined the effect of exogenous LTA on the adherence and entry of S. pyogenes JRS4 strain into HEp-2 epithelial cells. LTA markedly inhibited bacterial entry in a concentration-dependent manner, up to 250 microg ml(-1). In contrast, LTA had only a slight inhibitory effect on adherence. LTA also inhibited the entry but not adherence of Salmonella typhimurium strain into HEp-2 cells. Binding experiments showed a dose-dependent binding of LTA to cells up to 10 microg ml(-1). Confocal laser microscopy imaging and analysis revealed that LTA was internalized by the epithelial cells and colocalized with F-actin. These results might imply that, following binding, exogenous LTA enters HEp-2 cells and exerts a cytotoxic effect that interferes with bacterial internalization. A possible target for LTA activity might be the actin cytoskeleton, which is known to be essential for bacterial uptake.

Bacterial Adhesion↗
PubMed2000

Mutation in csrR global regulator reduces Streptococcus pyogenes internalization.

Transposon (Tn 916) mutagenesis was employed to identify genes in group A streptococcus (GAS) that are involved in bacterial internalization by epithelial cells. One mutant displayed significantly reduced internalization efficiency and was therefore selected for further characterization. The mutant harbored a single Tn 916 insertion in csr, a genetic locus encoding a two-component regulatory system. Mutations in csr were found to derepress hyaluronic acid (HA) capsule synthesis. Since capsule expression has been previously reported to interfere with internalization of GAS, it was possible that the transposon exerted its inhibitory effect either by derepression of capsule synthesis, or by another mechanism. To study the effect of the csr mutation on bacterial internalization, isogenic mutants deficient in either csrR, hasA or both were generated. The hasA mutant adhered to and internalized into HEp-2 cells significantly better than the parent and the csrR mutant strains. The internalization efficiency of the double mutant (csrR(-)/hasA(-)) was reduced by seven-fold compared to that of the hasA mutant. These findings suggest that csrR affects streptococcal entry by modulating capsule expression as well as by another, yet unknown, mechanism.

Bacterial Adhesion↗
PubMed2000

Capsule impedes adhesion to and invasion of epithelial cells by Klebsiella pneumoniae.

The adhesion of K21a, K26, K36, and K50 capsulated Klebsiella strains to ileocecal (HCT-8) and bladder (T24) epithelial cell lines was significantly lower than that of their corresponding spontaneous noncapsulated variants K21a/3, K26/1, K36/3, and K50/3, respectively. Internalization of the bacteria by both epithelial cell lines was also significantly reduced. Similarly, a capsule-switched derivative, K2(K36), that exhibited a morphologically larger K36 capsule and formed more capsular material invaded the ileocecal epithelial cell line poorly compared to the corresponding K2 parent strain. None of the capsulated strains exhibited significant mannose-sensitive type 1 fimbriae, whereas two of the noncapsulated variants K21a/3 and K50/3 exhibited potent mannose-sensitive hemagglutinating activity. Although hemagglutinating activity that could be attributed to mannose-resistant Klebsiella type 3 fimbriae was weak in all strains, in several cases the encapsulated parent strains exhibited lower titers than their corresponding noncapsulated variants. Although the level of adhesion to the ileocecal cells is not different from adhesion to bladder cells, bacterial internalization by bladder cells was significantly lower than internalization by ileocecal cells, suggesting that bladder cells lack components required for the internalization of Klebsiella.

Bacterial Adhesion↗
PubMed1999

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Journal Article↗
PubMed1999

Interaction between erythropoietin and peripheral polymorphonuclear leukocytes in hemodialysis patients.

The effect of erythropoietin (EPO) on the oxidative stress and inflammation caused by polymorphonuclear leukocytes (PMNLs) in chronic hemodialysis (HD) patients was investigated in vivo and in vitro. The studies were performed on isolated PMNLs from peripheral blood of healthy controls and HD patients before and following 6 weeks of EPO treatment. The oxidative stress was expressed by the rate of superoxide release from phorbol 12-myristate 13-acetate stimulated PMNLs, and the inflammatory state was evaluated by in vitro PMNL survival, in addition to white blood cell and PMNL counts of the enrolled subjects. Following 6 weeks of EPO treatment, in HD patients, the rate of superoxide release from PMNLs as well as WBC and PMNL counts fell significantly when compared with the pretreatment values. PMNLs from HD patients and healthy controls incubated in vitro with increasing amounts of EPO displayed a significant reduction in their rates of superoxide release and a significant improvement in survival. We have concluded that EPO interacts with PMNLs, attenuating their primed state in HD patients, thus reducing oxidative stress and the extent of inflammation. To the best of our knowledge, this attenuation of the primed state of PMNLs by EPO is a new finding.

Cell Survival↗
PubMed1999

Closure of the ductus venosus in premature infants: findings on real-time gray-scale, color-flow Doppler, and duplex Doppler sonography.

OBJECTIVE: Our objective was to use gray-scale, color-flow, and duplex Doppler sonography to study the anatomy, flow pattern, and time of closure of the ductus venosus in healthy premature infants. SUBJECTS AND METHODS: We prospectively examined the ductus venosus in 130 premature infants whom we divided into two groups: Group I comprised 27 neonates of gestational age 28-32 weeks, and group II comprised 103 neonates of gestational age 33-36 weeks. Neonates who had undergone umbilical vessel manipulation were excluded from the study. All examinations included gray-scale, color-flow, and duplex Doppler sonography. Patency, length, color flow, and Doppler characteristics of the ductus venosus were recorded. Neonates were examined 1-2 days after birth, 6-7 days after birth, and subsequently every 3-4 days until ductus closure was observed. The time of closure of the ductus for the two groups was compared using the chi-square test. RESULTS: The ductus venosus was patent during the initial examination in 128 of the 130 neonates. Doppler waveform was venous with little variation in velocity. Ductus length slightly exceeded 1 cm in both groups. We found a statistically significant difference in the percentage of infants having a patent ductus venosus after the initial examination: At 1 week after birth, ductus patency was shown in 85% of the infants in group I and in 56% of the infants in group II; at 2 weeks, the respective percentages were 42% and 14%; and at 3 weeks, 27% and 0%. CONCLUSION: The ductus venosus is patent 1-2 days after birth in virtually all premature infants. From 6 days after birth and onward, a significantly greater percentage of smaller premature infants (i.e., 28-32 weeks' gestational age) have a patent ductus venosus than do larger premature infants (i.e., 33-36 weeks' gestational age).

Blood Flow Velocity↗
PubMed1999

Why do we fail with penicillin in the treatment of group A streptococcus infections?

Acute pharyngotonsillitis caused by beta-haemolytic group A streptococcus (GAS) is a common childhood disease. Phenoxymethyl penicillin remains the drug of choice, as no resistance has been reported so far. Nevertheless, the failure of penicillin to eradicate streptococci from the throat occurs in up to 35% of patients with pharyngotonsillitis, and might present clinical concern. Various explanations have been proposed over the years to account for this perplexing phenomenon. Among these are coexistence of oropharyngeal beta-lactamase-producing bacteria that degrade penicillin, growth interference by aerobic and anaerobic commensals, penicillin tolerance, reinfection, and poor antibiotic compliance. Although GAS has been considered an extracellular pathogen, recent studies have demonstrated that strains of this bacterium can internalize epithelial cells both in vitro and in vivo. The intracellular niche may protect the bacterium from penicillin that does not gain high intracellular concentration. In support of this hypothesis, GAS strains were shown to survive 4-7 days inside cultured epithelial cells. In addition, it was found that GAS strains isolated from patients with eradication failure harbour the internalization-associated gene, prtF1/sfbI, in higher prevalence than do strains recovered from patients with successful eradication. Thus, internalization and intracellular survival represent a novel explanation for penicillin eradication failure.

Humans↗
PubMed1998

Participation of peripheral polymorphonuclear leukocytes in the oxidative stress and inflammation in patients with essential hypertension.

Oxidative stress and inflammation have recently been linked to endothelial damage in essential hypertension (EH). Activated peripheral polymorphonuclear leukocytes (PMN) damage surrounding tissue by releasing reactive oxygen species (ROS) and proteolytic enzymes before self-necrosis. PMN necrosis further exacerbates inflammation and promotes chemotaxis and PMN recruitment. The number and properties of PMN from untreated EH patients is the focus of the present study. Oxidative stress was assessed by measuring the rate of superoxide anion release from separated, phorbol ester-stimulated PMN and the redox state of plasma glutathione. Inflammation was estimated indirectly by determining PMN number and their in vitro survival. PMN from EH patients (n = 37) released superoxide anion faster (P < .0001) than those of normotensives (NC, n = 37), 17.7 +/- 1.14 v 9.54 +/- 0.51 nmol/10 min/10(6) cells. The redox state of glutathione was twofold higher in EH plasma (P < .02) indicating systemic oxidative stress. PMN survival in vitro correlates linearly with the rate of superoxide release (r2 = 0.60, P < .02) and PMN count of EH patients, although in the normal range, were significantly higher (P < .0001), indicating necrosis and recruitment. Hypertensive plasma significantly reduced NC PMN viability, whereas normal plasma significantly increased EH PMN viability. What our studies show is that EH is accompanied by a primed state PMN that does not correlate with the levels of blood pressure. PMN priming in EH patients reflects an in vivo exposure to a constant stimulus ending in oxidative stress, increased self-necrosis, and cell recruitment. Oxidative stress and inflammation will result in endothelial damage and atherosclerosis in the long run.

Adult↗
PubMed1998

Protein F1 is required for efficient entry of Streptococcus pyogenes into epithelial cells.

It was recently reported that strains of Streptococcus pyogenes are capable of inducing entry of the bacterium into epithelial cells; however, nothing is known regarding the gene(s) and the underlying mechanism(s) involved. Using isogenic mutants of S. pyogenes JRS4 strain that are defective in the expression of each of the surface proteins F1 and M6, it was demonstrated that both are required for efficient internalization. Expression of F1 on the surface of a poorly invading S. pyogenes strain significantly enhances its internalization efficiency. Protein F1-mediated internalization is inhibited by UR, the nonrepetitive fibronectin-binding domain of this protein, and to a lesser extent, by the repetitive fibronectin-binding domain, RD2. Polyclonal anti-human fibronectin antibodies completely abolish F1-mediated internalization; increasing fibronectin concentrations result in a significant enhancement of bacterial uptake. The findings shown here suggest that protein F1 mediates streptococcal internalization and that the M6 protein is required for more efficient entry of the bacterium.

Adhesins, Bacterial↗
PubMed1998

Renal ischemia-reperfusion injury: contribution of nitric oxide and renal blood flow.

The contributions of nitric oxide (NO) and renal blood flow (RBF) were examined in ischemia-reperfusion injury in the rat kidney. The function of both kidneys was assessed by glomerular filtration rate (GFR), and fractional excretion of sodium (FENa), calculated before, during unilateral renal artery clamping (45 min), and following reperfusion (90 min). RBF was measured in the same model by ultrasonic flowmetry. Intrarenal NO levels were modulated by administration of S-nitroso-N-acetylpenicillamine (SNAP), L-arginine, acetylcholine, and the NO synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME). SNAP increased GFR from 0.20 +/- 0.04 ml/min in control ischemic kidney to 0.38 +/- 0.06 ml/min and reduced FENa from 19.3 +/- 3.4 to 9.5 +/- 1.8%. Similar results were observed when L-arginine was administered. Acetylcholine had no effect on GFR or FENa. RBF was fully restored within 60 min following reperfusion, with no change in the rate of recovery by L-arginine. L-NAME aggravated the ischemia-reperfusion injury, preventing full restoration of RBF, further reducing GFR and worsening FENa. In conclusion, ischemia-reperfusion injury ends in low intrarenal levels of NO. We propose that this low NO level results from damage to the endothelial receptor signal transduction process and is not due to impaired NO synthase activity or to changes in RBF.

Acetylcholine↗
PubMed1997

Insertional inactivation of streptolysin S expression is associated with altered riboflavin metabolism in Streptococcus pyogenes.

Transposon Tn916 mutagenesis was used to create a mutant of Streptococcus pyogenes M type 3, designated ISS417, in which the ability to produce streptolysin S (SLS) and several other exoproteins was impaired. Concomitantly, the mutant became dependent upon riboflavin for growth and was able to grow in Todd Hewitt broth (THB) when supplemented with riboflavin or riboflavinrich yeast extract. The parent strain was apparently able to utilize THB-derived components as a substitute for riboflavin, while the mutant was not. Although the parent strain grew well in synthetic medium, it was unable to produce SLS, except when it was supplemented with a small amount of THB. Thus, a component of THB was able to "trigger" SLS formation in the parent strain. The mutant grew well in this medium, but was unable to produce SLS even when it was supplemented with THB. Southern hybridization analysis revealed that the ISS417 mutant harbours a single transposon insertion in its chromosome. Phage transduction experiments showed that the riboflavin dependency and the inability to make SLS phenotypes are co-transducible. The pleotrophic properties of the ISS417 mutant differ from those reported for insertional inactivation of the mga locus which regulates production of a number of surface proteins in S. pyogenes and the sar locus which regulates production of a number of exoproteins in Staphylococcus aureus. In view of the possibility that there exist a genetic linkage between the riboflavin biosynthetic pathway and expression of the oxygen-stable SLS, we hypothesize that SLS has a role in the growth economy of S. pyogenes.

Bacterial Proteins↗
PubMed1997

Proteins M6 and F1 are required for efficient invasion of group A streptococci into cultured epithelial cells.

Group A streptococci were recently shown to be capable of invading human epithelial cell monolayers. Cell invasion might be an important virulence trait of streptococci that enable the pathogen to gain entry into deeper tissues after initial binding to host cells. Nothing is known concerning the nature of streptococcal components that mediate invasion. Using isogenic mutants of strain JRS4 that are defective in the expression of either M6 protein or protein F1, or both proteins, it was demonstrated that both adhesins are required for efficient invasion. Further more, expression of protein F1 on the surface of a non-invasive strain rendered the latter invasive, suggesting that protein F1 is directly involved in the invasion process.

Adhesins, Bacterial↗
PubMed1997

Molecular and structural differences between rat brain D-1 and renal DA-1 dopamine receptors.

Renal DA-1 dopamine receptors in proximal tubules (PTs) of the Wistar-Kyoto (WKY) rat display pharmacological binding properties which are different from central nervous system (CNS) striatal D-1 dopamine receptors. In general, the renal DA-1 receptors display affinity binding values of dopaminergic drugs which are 6-36-fold less than those seen for brain D-1 receptors. The renal and brain DA receptors also displayed differential sensitivity toward the alkylating agent, N-ethylmaleimide (NEM). Inactivation of 50% of DA-1 renal receptors was achieved at lower concentrations of NEM (5.2 microM), relative to brain D-1 receptors (140 microM). Western blot analyses of rat pituitary GH4C1 cells, transfected with human CNS D-1 receptor cDNA, with human anti-D-1 dopamine receptor antiserum, detected a single polypeptide with M(r) of 66 kDa. In PTs, a specific polypeptide of higher molecular weight (M(r) = 72 kDa) was seen. Surprisingly, in rat striatal membranes, the D-1 antiserum failed to detect any proteins within this molecular weight range. Photoaffinity labeling studies with a DA-1 selective photoligand, identified the identical protein by autoradiography and Western blots in kidney, but not in striate. Together, these data indicate that renal DA-1 dopamine receptors have distinct molecular properties relative to brain D-1 dopamine receptors.

Animals↗
PubMed1997

Dysfunctional D1A receptor-G-protein coupling in proximal tubules of spontaneously hypertensive rats is not due to abnormal G-proteins.

BACKGROUND: Dysfunctional dopamine neurotransmission and defective D1A receptor-G protein coupling exist in renal proximal tubules (RPT) of the spontaneously hypertensive rat (SHR). OBJECTIVE: To determine whether the G proteins in SHR are abnormal, preventing formation of agonist high affinity sites in SHR. METHODS: We examined the expression levels of the alpha-subunits of G proteins, as well as D1A receptor receptor coupling to exogenously added normal G proteins, in RPT of SHR and the normotensive Wister-Kyoto (WKY) rat. RESULTS: In the presence of 110 mmol/l NaCl, the D1A dopamine receptor-selective agonist SKF R-38393 binds both to high- and to low-affinity sites on solubilized and reconstituted D1A receptors extracted from renal proximal tubules of normotensive Wistar-Kyoto (WKY) rats. In the spontaneously hypertensive rat (SHR), SKF R-38393 bound to a single site on the reconstituted receptor with affinity values corresponding to the low-affinity state of the receptor. Western blot analyses indicated that the alpha-subunit of the guanine nucleotide binding protein (G-protein), Gs, was expressed at similar levels, whereas G(o)alpha was not expressed in proximal tubule membranes from WKY rats and SHR. Pretreatment of proximal tubule membranes with the alkylating agent N-ethylmaleimide in the presence of SKF R-38393 inactivated alpha-subunits of endogenous G-proteins, but not D1A receptors, resulting in loss of high-affinity binding sites in WKY rats. These N-ethylmaleimide-treated D1A receptors from WKY rats, when reconstituted with exogenous sources of G-proteins, were able to couple to these exogenous G-proteins, with complete restoration of high-affinity sites. Moreover, the affinity values and the proportion of these hybrid sites were similar to those of untreated receptors, and these affinity sites were regulated by guanine nucleotide analogs. Reconstitution of D1A receptors from SHR with the same exogenous G-proteins failed to similarly induce formation of the high-affinity binding sites in the hybrid reconstituted systems, and SKF R-38393 continued to bind in a single low-affinity state of the receptor. CONCLUSION: These results demonstrate that the absence of G-protein coupling in SHR is due to intrinsic defects within the receptor protein, rather than to any abnormalities of the endogenous G-proteins themselves.

Alkylating Agents↗
PubMed1996

Heparin induces apoptosis in human peripheral blood neutrophils.

Programmed cell death, by apoptosis, has been shown to play an important role in the regulation of haemopoiesis. Using trypan blue exclusion for distinguishing intact membranes, flow cytometry for detection of sub G1 peak and in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL), this study shows that heparin induces apoptosis in vitro in human peripheral blood neutrophils. The known anti-proliferative effect of heparin in several in vitro cell systems has therefore to be interpreted in the light of apoptosis. In addition, apoptosis may help explain the anti-inflammatory effects resulting from the interaction between vessel wall heparan sulphate and chemoattracted peripheral blood neutrophils.

Apoptosis↗
PubMed1996

Sulfhydryl groups of renal D1A dopamine receptors: differential sensitivity of receptors to N-ethylmaleimide in normotensive and hypertensive rats.

OBJECTIVES: To determine whether sulfhydryl groups are present on D1A receptors of spontaneously hypertensive rats (SHR) and to test the hypothesis that failure of agonists to bind to such receptors is linked to microstructural changes involving sulfhydryl groups. METHOD: Alkylation of renal proximal tubule membranes by N-ethylmaleimide caused 70% loss of D1A dopamine receptor binding sites in the normotensive Wistar-Kyoto (WKY) rat and the SHR. RESULTS: The concentration of N-ethylmaleimide (IC50) required to produce half-maximal loss of receptor binding was 5.2 and 1200 mumol/l in WKY rats and SHR, respectively. Previous receptor occupancy of WKY rat D1A sites by the D1A agonist SKF R-38393 completely protected the binding sites from N-ethylmaleimide-mediated inactivation. Occupancy with the D1A antagonist SCH 23390 partially protected the binding sites and produced a 500-fold increase in the IC50 of N-ethylmaleimide. In SHR, receptor occupancy either by SKF R-38393 or by SCH 23390 failed to protect the D1A sites from N-ethylmaleimide or to alter the IC50 of N-ethylmaleimide-mediated inactivation. CONCLUSION: These results indicate that D1A dopamine receptors both of WKY rats and of SHR contain sulfhydryl groups at or near the ligand binding site, which display differential sensitivity to N-ethylmaleimide.

Animals↗
PubMed1996

Absence of DA1/DA2 dopamine receptor interactions in proximal tubules of spontaneously hypertensive rats.

The impact of defective DA1 dopamine receptors in proximal tubules (PT) of the spontaneously hypertensive rat (SHR) on DA1/DA2 receptor interactions was assessed with the DA1-selective photoaffinity ligand, (+/-)-7-[125I]iodo-8-hydroxy-3-methyl-1-(4-azidophenyl)- 2,3,4,5-tetrahydro-1H-3-benzazepine ([125I]MAB). In PT membranes from both normotensive (Wistar-Kyoto, WKY) and spontaneously hypertensive rats (SHR), [125I]MAB was specifically incorporated into a polypeptide with an M(r) of 74,000 Da, corresponding to the DA1 receptor. The labeling of this band by [125I]MAB in both SHR and WKY was not prevented by SKF-82526, a potent DA1-selective agonist. However, in the presence of the DA2 antagonist, (-)-sulpiride, but not DA2 agonist, LY-171555, SKF-82526 abolished photoincorporation of [125I]MAB into the 74,000-Da band in WKY. In SHR, (-)-sulpiride failed to enhance the ability of SKF-82526 to compete with [125I]MAB for binding to the 74,000-Da subunit. In competition binding studies with SKF-82526, (-)-sulpiride induced the formation of agonist high-affinity binding sites in WKY but not in SHR. These data suggest that in membranes of SHR, but not WKY, DA1/DA2 dopamine receptor interactions are lacking.

Affinity Labels↗