Presence Of Polymorphonuclear Leukocytes Research Articles

Activation-Induced Cytidine Deaminase Expression in Patients with Chronic Myeloid Leukemia

Objective: The findings on chronic myeloid leukemia (CML) patients suggest that clonal tumoral cells tend to have additional mutations besides the formation of the BCR-ABL fusion gene. Previous studies have demonstrated abnormal activation-induced cytidine deaminase (AID) expression in various types of cancer, showing AID transcript levels to be elevated in the CML blast phase. The study aimed to investigate the AID gene expression levels in CML and to investigate the etiopathogenic role of AID, which is not yet fully understood. Materials and Methods: This study analyzed the AID transcript levels of 80 CML patients and 50 controls using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Results: The study found the AID transcript levels to be significantly elevated (p < 0.001) in the CML patients compared to the control group, with no significance in AID transcript levels with regard to the patients’ age, gender, clinical characteristics, or laboratory findings. No correlation was found between the AID and BCR-ABL transcript levels, while a positive correlation was present between AID transcript levels and presence of polymorphonuclear leukocytes (PMNL; r = 0.320, p = 0.021). No significant relationship occurred in AID transcription levels with the tyrosine-kinase-inhibitor (TKI) resistant mutation profile or cytogenetic response during TKI therapy. Conclusion: This study found AID expression levels to be significantly elevated in CML patients and AID to be able to contribute to the etiopathogenesis of CML

Bypassing Phase Variation of Lipooligosaccharide (LOS): Using Heptose 1 Glycan Mutants To Establish Widespread Efficacy of Gonococcal Anti-LOS Monoclonal Antibody 2C7.

The sialylatable lacto-N-neotetraose (LNnT; Gal-GlcNAc-Gal-Glc) moiety from heptose I (HepI) of the lipooligosaccharide (LOS) of Neisseria gonorrhoeae undergoes positive selection during human infection. Lactose (Gal-Glc) from HepII, although phase variable, is commonly expressed in humans; loss of HepII lactose compromises gonococcal fitness in mice. Anti-LOS monoclonal antibody (MAb) 2C7, a promising antigonococcal immunotherapeutic that elicits complement-dependent bactericidal activity and attenuates gonococcal colonization in mice, recognizes an epitope comprised of lactoses expressed simultaneously from HepI and HepII. Glycan extensions beyond lactose on HepI modulate binding and function of MAb 2C7 in vitro Here, four gonococcal LOS mutants, each with lactose from HepII but fixed (unable to phase-vary) LOS HepI glycans extended beyond the lactose substitution of HepI (lactose alone, Gal-lactose, LNnT, or GalNAc-LNnT), were used to define how HepI glycan extensions affect (i) mouse vaginal colonization and (ii) efficacy in vitro and in vivo of a human IgG1 chimeric derivative of MAb 2C7 (2C7-Ximab) with a complement-enhancing E-to-G Fc mutation at position 430 (2C7-Ximab-E430G). About 10-fold lower 2C7-Ximab-E430G concentrations achieved similar complement-dependent killing of three gonococcal mutants with glycan extensions beyond lactose-substituted HepI (lactose alone, LNnT, or GalNAc-LNnT) as 2C7-Ximab (unmodified Fc). The fourth mutant (Gal-lactose) resisted direct complement-dependent killing but was killed approximately 70% by 2C7-Ximab-E430G in the presence of polymorphonuclear leukocytes and complement. Only mutants with (sialylatable) LNnT from HepI colonized mice for >3 days, reiterating the importance of LNnT sialylation for infection. 2C7-Ximab-E430G significantly attenuated colonization caused by the virulent mutants.

Evaluation of BACTEC™ blood culture system for culture of normally sterile body fluids.

Sir, Infectious diseases are now the world's biggest killer of children and young adults, it is also the basis of debilitating illness all through the year keeping children away from school and preventing adults from working.[1] Conventional methods of culture are now being replaced by automated culture systems, owing to reduced time for culture and ease of laboratory work using machines and higher isolation rate against conventional methods.[2] Lot of information is available for culturing blood samples using BACTEC™ culture, but there is a paucity of data regarding sterile body fluids culture. Hence, this study was done to compare isolation rate and the time needed for the detection of microorganisms between conventional methods of culture and BACTEC™ culture. This prospective study was conducted for a period of 2 years from July 2011 to July 2013 in the Department of Microbiology of a tertiary care hospital after obtaining the Institutional Ethical Committee approval. A total of 127 body fluid samples were inoculated into BACTEC™ peds Plus™/F vials, incubated in BACTEC™ machine. Direct plating of sample onto sheep blood agar, chocolate agar, MacConkey agar and incubated aerobically at 37°C for 48 h. Organisms isolated were identified using standard microbiological procedures and tested for antimicrobial susceptibility according to Clinical and Laboratory Standards Institute guidelines.[1,2] All data were analyzed using SPSS (IBM) Descriptive statistics were derived using frequency, percentage, and mean. In this study, 21 samples out of 127, showed a positive result as compared with 4 out of 127 by conventional method. The average time taken for growth in a conventional method was 26.5 h, while that in BACTEC™ is 14.25 h. This significant increase in isolation rate with reduced time to positivity gives BACTEC™ culture edge over conventional culture methods. This is quite a significant time difference considering that presumptive identification of the pathogen was informed to the clinician 12.25 h earlier than a conventional method. This would significantly affect the change of empirical antibiotic therapy given to the patient, eventually aiding in therapy and the final clinical outcome. Furthermore, the initial Gram's stain report of positive cultures and the association of it with the bacteria isolated were compared. Though few samples showed no bacteria, but polymorphonuclear leukocytes (PMNLs) and mononuclear cells on initial Gram's stain were seen. Alberts et al.,[3] states that presence of PMNLs and lymphocytes suggests infection, and it is responsible for phagocytosis and killing of invading bacteria. Consequently even low bacterial load in the sample, which is undetectable by conventional method, can be detected by BACTEC™.[4,5] Staphylococcus aureus, Streptococcus pneumoniae, Klebsiella pneumoniae and Pseudomonas aeruginosa were the frequently isolated bacteria in both methods. The fastidious bacteria like Group B and G Streptococcus, Burkholderia pseudomallei, Listeria monocytogenes, S. pneumoniae, Escherichia coli, Acinetobacter were isolated only from BACTEC™. Table 1 shows the distribution of bacteria isolated in various samples. L. monocytogenes, S. pneumoniae and Group B Streptococcus were also isolated from blood culture. Table 1 Distribution of organisms isolated from sterile body fluid samples In conclusion, BACTEC™ automated culture has higher isolation rate of pathogenic bacteria and lesser time for identification even with less bacterial load or count in the sterile body fluid sample. It also helps in guiding antimicrobial therapy especially in critically ill patients and eventually affecting patient's outcome.

P2.024 Evaluation of the Genital Microbiota in Men and Women Using an Automated System For Analysis of Microscopy Images of Wet and Stained Smears

BackgroundIn Russia, microscopy of the genital microbiota in both women and men are performed at laboratory, not at physician’s office. Microscopy is a simple and cheap method, which, together with...

Vulnerability of continence structures to injury by simulated childbirth

The goal of this study was to examine acute morphological changes, edema, muscle damage, inflammation, and hypoxia in urethral and vaginal tissues with increasing duration of vaginal distension (VD) in a rat model. Twenty-nine virgin Sprague-Dawley rats underwent VD under anesthesia with the use of a modified Foley catheter inserted into the vagina and filled with saline for 0, 1, 4, or 6 h. Control animals were anesthetized for 4 h without catheter placement. Urogenital organs were harvested after intracardiac perfusion of fixative. Tissues were embedded, sectioned, and stained with Masson's trichrome or hematoxylin and eosin stains. Regions of hypoxia were measured by hypoxyprobe-1 immunohistochemistry. Within 1 h of VD, the urethra became vertically elongated and displaced anteriorly. Edema was most prominent in the external urethral sphincter (EUS) and urethral/vaginal septum within 4 h of VD, while muscle disruption and fragmentation of the EUS occurred after 6 h. Inflammatory damage was characterized by the presence of polymorphonuclear leukocytes in vessels and tissues after 4 h of VD, with the greatest degree of infiltration occurring in the EUS. Hypoxia localized mostly to the vaginal lamina propria, urethral smooth muscle, and EUS within 4 h of VD. Increasing duration of VD caused progressively greater tissue edema, muscle damage, and morphological changes in the urethra and vagina. The EUS underwent the greatest insult, demonstrating its vulnerability to childbirth injury.

Analysis of the grade of esophagitis, chromendoscopical and histological findings of esophagus in patients with gastroesophageal reflux disease before and after the therapy

The symptoms of gastroesophageal reflux disease (GERD) are among the most common complaints for which patients are indicated for visiting gastroenterologist. It occurs as a result of the effect made by gastric reflux contents that moves into the esophagus. The prevalence of all forms of GERD is 40%. The aim of this study was to analyze the grade of esophagitis, chromendoscopical and histological findings of esophagus in patients with GERD before and after the therapy. A prospective study included 90 patients with symptoms of GERD, divided into 2 groups depending on whether they had endoscopic signs of gastroezophageal reflux (group ERD), or not (group NERD). All the patients had esophagogastroduodenoscopy, chromoendoscopy staining, test for Helicobacter pylori and histological findings of the esophagus. In the patients with Helicobacter pylori infection eradication therapy was done. Esophagitis-B level was present in most of the patients. Among the groups, roughly the same number responded to positive findings on chromoendoscopy. After the therapy, chromoendoscopy was significantly negative in both groups of the patients comparing to chromoendoscopy before the therapy (p = 0.00001). Multiplication and elongation of papilla, basal cell hyperplasia, vascular dilatation, increasing of mitotic activity and the presence of polymorphonuclear leukocyte cells were statistically more frequent histological findings in the group ERB compared to the group NERB. After the therapy, the patients in both groups had statistically less histological findings of appropriate esophageal parameters. Chromoendoscopy combined with the standard endoscopy increases the sensitivity and specificity for reflux disease. Histology in the reflux disease is associated with endoscopic and clinical findings so that the localization of taking biopsies and histological criteria of pathohistological changes must be clearly defined. Multiplication and elongation of papilla, basal cell hyperplasia and the presence of polymorphonuclear leukocytes are the most relevant criteria in the diagnosis NERD.

Mechanisms related to endothelial nitric oxide synthase (eNOS) uncoupling in myocardial ischemia/reperfusion (MI/R)

MI/R results in endothelial and cardiac contractile dysfunction in the presence of polymorphonuclear leukocytes (PMNs). The role of eNOS in reperfusion injury was studied in a rat PMN‐induced MI/R injury model. Isolated hearts subjected to ischemia (20 min) and reperfused (45 min) with activated PMNs were studied. Cell permeable protein kinase C (PKC) epsilon activator (ε+) or inhibitor (ε‐) peptides were used to increase or decrease eNOS activity respectively. Tetrahydrobiopterin (BH4) or dihydrobiopterin (BH2) are cofactors of eNOS that regulate nitric oxide or superoxide (SO) release respectively. The combination of PKC ε+ (10µM)/BH4 (5µM) (n=7) given during reperfusion significantly restored post‐reperfusion cardiac function compared to I/R PMN hearts (n=11, P <0.01) or PKC ε+ (n=6) or BH4 (n=6) by itself. By contrast, the combination of PKC ε+/BH2 (100µM) (n=6) compromised post‐reperfusion cardiac function suggesting that increased eNOS activity compromises cardiac function when eNOS becomes uncoupled by BH2 or can be cardioprotective when eNOS is coupled to BH4. Moreover, the combination of PKC ε‐ (5µM)/BH2 (n=6) is cardioprotective compared to I/R PMN hearts (P <0.01) or BH2 (n=7) by itself suggesting that decreased eNOS activity during reperfusion attenuates SO release from eNOS. This study was supported by NHLBI Grant 2R15HL‐076235‐02A1 and the Center for the Chronic Disorders of Aging at PCOM.

Premier épisode de rejet aigu après transplantation rénale : étude des caractéristiques histopathologiques en fonction du risque immunologique du patient

Renal allograft biopsies (n=34) of two different populations of patients according to the immunological risk (high versus low-risk) have been compared retrospectively. The presence of polymorphonuclear leukocytes in peritubular capillaries was more frequent in the high-risk group. The C4d staining was positive in 10% of the low-risk patients and in 50% of the high-risk patients (P=0.03). There were more early graft loss, renal infarctions, interstitial hemorrhage, severe glomerulitis, neutrophilic glomerulitis and Banff III grade rejection in the positive C4d group. In conclusion, half of the immunized patients had a humoral rejection, patients with a C4d positive rejection had more early graft loss and more severe histological lesions.

The combination of protein kinase C epsilon activator (PKC ε +) and tetrahydrobiopterin (BH4) exerts cardioprotective effects in ischemia/reperfusion injury (I/R) when given during reperfusion

In the presence of polymorphonuclear leukocytes (PMNs) during reperfusion, following myocardial ischemia, cardiac contractile dysfunction is observed. Isolated ischemic (20 min) and reperfused (45 min) rat hearts in the presence of activated PMNs were studied. A cell permeable peptide (PKC ε+) that enhances interaction of PKC ε with endothelial nitric oxide synthase (eNOS) to increase nitric oxide (NO) release, and BH4, an essential cofactor of eNOS, were given during the first five min of reperfusion. In hearts given PKC ε+ (10 μM) and BH4 (5 μM) (n=7) with PMNs, left ventricular developed pressure (LVDP) significantly recovered to 92% ± 11% of baseline values compared to I/R + PMN hearts (n=5) (48% ± 7%) (p<0.05) and the maximal rate of LVDP (+dP/dtmax) significantly recovered to 83% ± 7% compared to I/R + PMN hearts (34% ± 4%) (p<0.01) at 45 min reperfusion. The cardioprotective effects of PKC ε+ and BH4 were associated with an approximate 50% decrease in PMN infiltration in post‐reperfused hearts as confirmed by myeloperoxidase assay. These results suggest that the combination of PKC ε+ and BH4 attenuates PMN‐induced cardiac contractile dysfunction after I/R, in part by promotion of eNOS to increase NO release and inhibition of PMN infiltration in post‐reperfused hearts. This study was supported by NHLBI Grant 1R15HL‐76235‐01 and the Center for the Chronic Disorders of Aging at PCOM.

Influence of different types of surgical suture material on the intensity of tissue reaction in oral cavity

Throughout the history the most diverse suture material have been used for closing and suturing surgical wounds. The four basic features of suture material are described: knot safety, stretch capacity, tissue reactivity and wound safety. Tissue reaction, even the minimum one, which develops during the first to seven days after applying the suture in the tissue. The aim of this study was to investigate influence of a monofilament suture material (nylon) on the intensity of local tissue reaction in experimental conditions, and to compare it with the multifilament suture used in the routine practice of oral surgery (silk). This investigation is a prospective experimental study carried out on Wistar rats. The experiment included 30 animals, in which Black Silk (thickness 4-1) and Nylon (thickness 4-0) were applied in the upper and lower jaw, respectively. To monitor tissue reaction on different suture materials the following parameters were used: coagulum formation, presence of polymorphonuclear leukocytes, presence of macrophages and granuloma, formation of epithelial bridge and connective tissue, collagen synthesis, granulomatous tissue formation and presence of fibrous tissue. After comparing parameters for the intensity of tissue reaction to the investigate suture materials by suturing the oral mucosa, certain advantage could be given to the monofilament suture materials.

Universal Definition of Myocardial Infarction

Myocardial infarction is a major cause of death and disability worldwide. Coronary atherosclerosis is a chronic disease with stable and unstable periods. During unstable periods with activated inflammation in the vascular wall, patients may develop a myocardial infarction. Myocardial infarction may be a minor event in a lifelong chronic disease, it may even go undetected, but it may also be a major catastrophic event leading to sudden death or severe hemodynamic deterioration. A myocardial infarction may be the first manifestation of coronary artery disease, or it may occur, repeatedly, in patients with established disease. Information on myocardial infarction attack rates can provide useful data regarding the burden of coronary artery disease within and across populations, especially if standardized data are collected in a manner that demonstrates the distinction between incident and recurrent events. From the epidemiological point of view, the incidence of myocardial infarction in a population can be used as a proxy for the prevalence of coronary artery disease in that population. Furthermore, the term myocardial infarction has major psychological and legal implications for the individual and society. It is an indicator of one of the leading health problems in the world, and it is an outcome measure in clinical trials and observational studies. With these perspectives, myocardial infarction may be defined from a number of different clinical, electrocardiographic, biochemical, imaging, and pathological characteristics. In the past, a general consensus existed for the clinical syndrome designated as myocardial infarction. In studies of disease prevalence, the World Health Organization (WHO) defined myocardial infarction from symptoms, ECG abnormalities, and enzymes. However, the development of more sensitive and specific serological biomarkers and precise imaging techniques allows detection of ever smaller amounts of myocardial necrosis. Accordingly, current clinical practice, health care delivery systems, as well as epidemiology and clinical trials all require a …

Treatment of men with urethritis negative for Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum and Ureaplasma urealyticum

Some patients with symptomatic non-gonococcal urethritis (NGU) are negative for Chlamydia trachomatis, mycoplasmas and ureaplasmas. The optimal antimicrobial chemotherapy for such NGU has not fully been elucidated, though many studies of antimicrobial chemotherapies for C. trachomatis-positive NGU have been performed. We assessed the efficacy of antimicrobial agents that are active against C. trachomatis on non-mycoplasmal, non-ureaplasmal and non-chlamydial NGU (NMNUNCNGU). One hundred men whose first-pass urine samples were negative for C. trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum, and Ureaplasma urealyticum were treated with levofloxacin, gatifloxacin, minocycline, or clarithromycin for 7 days. Urethritis symptoms and the presence of polymorphonuclear leukocytes (PMNL) in urethral smears were assessed before and after treatment. Eighty-eight (88.0%) of 100 men with NMNUNCNGU showed no signs of urethral inflammation after treatment, but two men complained of some symptoms of urethritis. Twelve (12.0%) of 100 men had significant numbers of PMNL in urethral smears, but five of these 12 men had no symptoms of urethritis. The efficacy for normalization of urethral smears was 90.7% for clarithromycin, 89.7% for levofloxacin, 87.5% for gatifloxacin, and 75.0% for minocycline. The 12 men who showed signs of urethral inflammation were retreated with levofloxacin, gatifloxacin, minocycline or clarithromycin for an additional 7 days. The 10 men who returned after the second treatment had negative urethral smears. Our present findings suggest that antimicrobial agents active against C. trachomatis are effective against NMNUNCNGU and that a 7-day treatment regimen with an appropriate antimicrobial agent may be sufficient to manage patients with NMNUNCNGU.

Tetrahydrobiopterin (BH4) exerts cardioprotective effects in ischemia/reperfusion injury when given at reperfusion

Myocardial ischemia followed by reperfusion (I/R) in the presence of polymorphonuclear leukocytes (PMNs) results in a marked cardiac contractile dysfunction. The effects of BH4 (cofactor of endothelial nitric oxide (NO) synthase) or dihydrobiopterin (BH2, i.e., oxidized BH4) were examined in isolated ischemic (20 min) and reperfused (45 min) rat hearts reperfused with PMNs. I/R+PMN hearts perfused with BH4 (10μM) during the first 5 min of reperfusion (n=6), left ventricular developed pressure (LVDP) significantly recovered to 85±8% of baseline compared to control I/R+PMN hearts (53±7%, n=9) at 45 min post-reperfusion (P<0.01). Moreover, the cardioprotection of BH4 was blocked by a NO synthase inhibitor NG–nitro-L-arginine methyl ester (50 μM). By contrast, I/R+PMN hearts perfused with BH2 (100 μM, n=7), LVDP only recovered to 64±9% of baseline, which was similar to control I/R + PMN hearts. The effects of BH4 and BH2 on I/R+PMN cardiac contractile function may be correlated with increased endothelial NO release by BH4 and decreased NO release by BH2, respectively, from non ischemic rat aortic segments. In summary, our preliminary data show that BH4, not BH2, significantly attenuates PMN-induced decreased LVDP following I/R, suggesting that hearts receiving BH4 during the first 5 min of reperfusion may promote eNOS to produce NO to maintain endothelial function to limit reperfusion injury. This study was supported by NHLBI Grant 1R15HL-76235-01 and Center For Chronic Disorders Of Aging. Philadelphia College of Osteopathic Medicine.

Oral Clindamycin and Histologic Chorioamnionitis in Women With Abnormal Vaginal Flora

Oral clindamycin reduced late miscarriage and preterm birth in asymptomatic women with bacterial vaginosis or intermediate flora. We investigated whether clindamycin reduced the incidence of histologic chorioamnionitis as a mechanism for these beneficial effects. This was a subanalysis of 126 participants from a larger randomized controlled trial. We compared the incidence of histologic chorioamnionitis between the clindamycin and placebo groups. Histologic chorioamnionitis was diagnosed by the presence of polymorphonuclear leukocytes, separately in the amnion and chorion, decidua, fetal surface of the placenta, the walls of fetal chorionic vessels, umbilical cord, or in the subchorionic fibrin layer. Microbiologic cultures were done on swabs from the space between the chorion and amnion layers. Histopathologic results were available for 122 placentas, 62 (51%) and 60 (49%) in the clindamycin and placebo groups, respectively. There were no significant differences in inflammation between the groups in the decidua (41% compared with 43%), membranes (25% compared with 41%), fetal vessels (16% compared with 14%), or subchorionic fibrin (32% compared with 34%). Adjusting for gestational age, ethnic origin, or history of miscarriage did not alter the results. There were no significant differences in the outcomes of pregnancy between women with and without inflammation, either before or after adjustment for treatment group. Although oral clindamycin reduced late miscarriage and preterm birth in women with abnormal vaginal flora, this effect is unlikely to be mediated through a reduction in the incidence of histologic chorioamnionitis. The relatively small size of the groups, however, does not allow us to rule out a real effect, especially given the lower rate of membrane inflammation observed in the clindamycin group. I.

Protein Kinase C epsilon (PKC ε+) peptide activator exerts cardioprotective effects in ischemia/reperfusion injury when given prior to ischemia

Ischemia followed by reperfusion (I/R) in the presence of polymorphonuclear leukocytes (PMNs) results in cardiac contractile dysfunction. Enhancing endothelial nitric oxide (NO) release prior to ischemia attenuates cardiac contractile dysfunction following I/R. A cell-permeable PKC ε peptide activator enhances NO release when given to rat aortic segments. We studied isolated rat hearts following ischemia (20 min) and reperfusion (45 min) in the presence of activated PMNs. In hearts given PKC ε+ (5 μM, n = 6) prior to I/R and reperfused with PMNs, left ventricular developed pressure (LVDP) and the maximal rate of LVDP (+dP/dtmax) recovered to 90% ± 10% and 89% ± 9% of baseline values, respectively, at 45 min post-reperfusion, and these cardioprotective responses were blocked by NG-nitro-L-arginine methyl ester (L-NAME). I/R hearts receiving PMNs alone (n = 10) recovered to 58% ± 4% and 49% ± 4% of baseline values for LVDP and +dP/dt max respectively. The effect of PKC ε+ on LVDP and +dP/dtmax was significant (p<0.01) compared to I/R + PMN hearts at 45 min post-reperfusion. The PKC ε+ significantly increased endothelial NO release (p<0.01) from a basal value of 2.12 ± 0.39 (n = 11) pmol NO/mg tissue to 4.76 ± 0.36 pmol NO/mg tissue for 5 μM (n = 36), and this response was blocked by L-NAME. These results suggest that PKC ε+ attenuates PMN induced post-I/R cardiac contractile dysfunction, in part, by increasing endothelial NO release prior to ischemia. This study was supported by NHLBI Grant 1R15HL-76235-01.

Protein Kinase C βII Peptide Inhibitor Exerts Cardioprotective Effects in Rat Cardiac Ischemia/Reperfusion Injury

Ischemia followed by reperfusion (I/R) in the presence of polymorphonuclear leukocytes (PMNs) results in a marked cardiac contractile dysfunction. A cell-permeable protein kinase C (PKC) betaII peptide inhibitor was used to test the hypothesis that PKC betaII inhibition could attenuate PMN-induced cardiac dysfunction by suppression of superoxide production from PMNs and increase NO release from vascular endothelium. The effects of the PKC betaII peptide inhibitor were examined in isolated ischemic (20 min) and reperfused (45 min) rat hearts with PMNs. The PKC betaII inhibitor (10 microM; n = 7) significantly attenuated PMN-induced cardiac dysfunction compared with I/R hearts (n = 9) receiving PMNs alone in left ventricular developed pressure (LVDP) and the maximal rate of LVDP (+dP/dt(max)) cardiac function indices (p < 0.01). The PKC betaII inhibitor at 10 microM significantly increased endothelial NO release from a basal value of 1.85 +/- 0.18 pmol NO/mg tissue to 3.49 +/- 0.62 pmol NO/mg tissue from rat aorta. It also significantly inhibited superoxide release (i.e., absorbance) from N-formyl-L-methionyl-L-leucyl-L-phenylalanine-stimulated rat PMNs from 0.13 +/- 0.01 to 0.02 +/- 0.004 (p < 0.01) at 10 microM. Histological analysis of the left ventricle of representative rat hearts from each group showed that the PKC betaII peptide inhibitor-treated hearts experienced a marked reduction in PMN vascular adherence and infiltration into the postreperfused cardiac tissue compared with I/R + PMN hearts (p < 0.01). These results suggest that the PKC betaII peptide inhibitor attenuates PMN-induced post-I/R cardiac contractile dysfunction by increasing endothelial NO release and by inhibiting superoxide release from PMNs.

Protein kinase C-ζ inhibition exerts cardioprotective effects in ischemia-reperfusion injury

Ischemia followed by reperfusion (I/R) in the presence of polymorphonuclear leukocytes (PMNs) results in marked cardiac contractile dysfunction. A cell-permeable PKC-zeta peptide inhibitor was used to test the hypothesis that PKC-zeta inhibition could attenuate PMN-induced cardiac contractile dysfunction by suppression of superoxide production from PMNs and increase nitric oxide (NO) release from vascular endothelium. The effects of the PKC-zeta peptide inhibitor were examined in isolated ischemic (20 min) and reperfused (45 min) rat hearts reperfused with PMNs. The PKC-zeta inhibitor (2.5 or 5 microM, n = 6) significantly attenuated PMN-induced cardiac dysfunction compared with I/R hearts (n = 6) receiving PMNs alone in left ventricular developed pressure (LVDP) and the maximal rate of LVDP (+dP/dt(max)) cardiac function indexes (P < 0.01), and these cardioprotective effects were blocked by the NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester (50 microM). Furthermore, the PKC-zeta inhibitor significantly increased endothelial NO release 47 +/- 2% (2.5 microM, P < 0.05) and 54 +/- 5% (5 microM, P < 0.01) over basal values from the rat aorta and significantly inhibited superoxide release from phorbol-12-myristate-13-acetate-stimulated rat PMNs by 33 +/- 12% (2.5 microM) and 40 +/- 8% (5 microM) (P < 0.01). The PKC-zeta inhibitor significantly attenuated PMN infiltration into the myocardium by 46-48 +/- 4% (P < 0.01) at 2.5 and 5 microM, respectively. In conclusion, these results suggest that the PKC-zeta peptide inhibitor attenuates PMN-induced post-I/R cardiac contractile dysfunction by increasing endothelial NO release and by inhibiting superoxide release from PMNs thereby attenuating PMN infiltration into I/R myocardium.

Actinobacillus actinomycetemcomitans-induced periodontal disease in mice: patterns of cytokine, chemokine, and chemokine receptor expression and leukocyte migration

Although the pathogenesis of periodontal disease (PD) is not well known, cytokines, chemotactic factors and inflammatory cells are certainly involved in the disease outcome. Here, we characterized the evolution of the PD induced by Actinobacillus actinomycetemcomitans in mice, showing that oral inoculation of these bacteria leads to the migration of leukocytes to periodontal tissues and marked alveolar bone resorption. We found the expression of pro-inflammatory and Th1-type cytokines including TNF-α, IFN-γ and IL-12 in periodontal tissues after infection with A. actinomycetemcomitans, from the early stages after infection and throughout the course of the disease. Similar kinetics of expression were found for the chemokines CCL5, CCL4, CCL3 and CXCL10 and for the receptors CCR5 and CXCR3, all of them linked to the Th1-type pattern. The expression of the Th2-type mediators IL-10, CCL1 and their receptors CCR4 and CCR8 was detected only after 30 days of infection, determining a time-dependent mixed pattern of polarized immune response. The chemokine expression was correlated with the presence of polymorphonuclear leukocytes, macrophages, CD4 and CD8 lymphocytes, and B cells in the inflammatory infiltrate. Interestingly, during the predominance of the Th1-type response, a sharp increase in the number of inflammatory cells and intense bone loss was seen. By contrast, after the increased expression of Th2-type mediators, the number of inflammatory cells remained constant. Our data demonstrate that mice subjected to oral inoculation of A. actinomycetemcomitans represent a useful model for the study of PD. In addition, our results suggest that expression of cytokines and chemokines can drive the selective recruitment of leukocyte subsets to periodontal tissues, which could determine the stable or progressive nature of the lesion.

Gö 6983 exerts cardioprotective effects in myocardial ischemia/reperfusion.

Ischemia followed by reperfusion (I/R) in the presence of polymorphonuclear leukocytes (PMNs) results in cardiac contractile dysfunction. Inhibiting protein kinase C (PKC) inhibits the release of superoxide from PMNs. The compound Gö 6983 is an inhibitor of all five PKC isoforms present in PMNs. Therefore, we hypothesized that Gö 6983 could attenuate PMN-induced cardiac dysfunction by suppression of superoxide production from PMNs. We studied isolated rat hearts following ischemia (20 minutes) and reperfusion (45 minutes) infused with activated PMNs. In hearts reperfused with PMNs and Gö 6983 (100 nM, n = 7), left ventricular developed pressure (LVDP) and the rate of LVDP (+dP/dt max) recovered to 89 +/- 7% and 74 +/- 2% of baseline values, respectively, at 45 minutes postreperfusion compared with I/R hearts (n = 9) receiving PMNs alone, which only recovered to 55 +/- 3% and 45 +/- 5% of baseline values for LVDP and +dP/dtmax, respectively (P < 0.01). Gö 6983 (100 nM) significantly reduced PMN adherence to the endothelium and infiltration into the myocardium compared with I/R + PMN hearts (P < 0.01), and significantly inhibited superoxide release from PMNs by 90 +/- 2% (P < 0.01). In the presence of PMNs, Gö 6983 attenuated post-I/R cardiac contractile dysfunction, which may be related in part to decreased superoxide production.

Romboencefalitis: una forma de infección por Listeria monocytogenes en el sistema nervioso central

Listeria monocytogenes infections can involve the central nervous system in the form of a rhomboencephalitis. Three possible cases of rhomboencephalitis by Listeria monocytogenes are reported (2 females, aged 44 and 49 and a man of 36 years old). The three cases were preceded by an unspecific prodrome of headache, vertigo and fever in absence of a meningeal syndrome. The neurological stage was defined by the unilateral involvement of cranial nerves and the cerebellum and a clear inflammatory cerebrospinal fluid (CSF) with the presence of polymorphonuclear leukocytes, and normal glucose and protein levels. A magnetic nuclear resonance (MRI) showed the appearance of characteristic images, present in the bulboprotuberancial region. These images are one of the most constant features of this disease, reported in the literature. The early diagnosis of rhomboencephalitis was based on the clinical picture, the study of the CSF and the MRI, allowing the use of antimicrobials, prior to microbiological identification. Therefore, the risk of brain stem and cardiac complications of the disease is reduced.