Photoprotective Effect of Carpomitra costata Extract against Ultraviolet B-Induced Oxidative Damage in Human Keratinocytes.

Natural marine products show various biological properties such as antiphotoaging, antioxidant, anticancer, and anti-inflammation. This study evaluated the protective effects of the brown alga Carpomitra costata (Stackhouse) Batters (Sporochnaceae) against ultraviolet B (UVB)-provoked damage in human HaCaT keratinocytes. C. costata extract (CCE) effectively reduced superoxide anion, hydroxyl radical, and UVB-stimulated intracellular reactive oxygen species (ROS) levels. CCE also restored the expression and activity of UVB-suppressed antioxidant enzymes. Furthermore, CCE decreased UVB-triggered oxidative damage to cellular components including DNA, protein, and lipid and defended the cells against mitochondrial membrane depolarization-medicated apoptosis. The results of this study indicate that CCE can safeguard human keratinocytes against UVB-induced cellular damage via a potent antioxidant mechanism. CCE may find utility as part of a therapeutic arsenal against the damaging effects of UVB radiation on the skin.

Gracilaria bursa-pastoris (Gmelin) Silva extract attenuates ultraviolet B radiation-induced oxidative stress in human keratinocytes.

The purpose of this study was to assess the protective effects of an ethanol extract derived from the red alga Gracilaria bursa-pastoris (Gmelin) Silva (GBE) on ultraviolet B (UVB)-irradiated human HaCaT keratinocytes. GBE exhibited scavenging activity against intracellular reactive oxygen species that were induced by either hydrogen peroxide or UVB radiation. In addition, both the superoxide anion and the hydroxyl radical were scavenged by GBE in cell-free systems. GBE absorbed light in the UVB range (280-320 nm) of the electromagnetic spectrum and lessened the extent of UVB-induced oxidative damage to cellular lipids, proteins, and DNA. Finally, GBE-treated keratinocytes showed a reduction in UVB-induced apoptosis, as exemplified by fewer apoptotic bodies. These results suggest that GBE exerts cytoprotective actions against UVB-stimulated oxidative stress by scavenging ROS and absorbing UVB rays, thereby attenuating injury to cellular constituents and preventing cell death.

Optimal dose of vancomycin for treating methicillin-resistant Staphylococcus aureus pneumonia in critically ill patients.

A prospective cohort study was performed to determine the optimal dose of vancomycin to maintain a serum trough concentration of at least 15 to 20 mg/l and to assess the efficacy of this target vancomycin concentration in the treatment of methicillin-resistant Staphylococcus aureus pneumonia. Vancomycin pharmacokinetic parameters were estimated using a CAPSIL software program from serum concentrations of 141 patients with pneumonia treated with vancomycin, regardless of methicillin-resistant Staphylococcus aureus status, at a 28-bed medical intensive care unit. Vancomycin trough concentrations and other pharmacokinetic parameters were compared between five groups of patients differing in their renal function: (1) creatinine clearance > or =60 ml/minute, (2) creatinine clearance 30 to 60 ml/minute, (3) creatinine clearance <30 ml/minute, (4) on intermittent haemodialysis, and (5) on continuous renal replacement therapy. More than 70% of patients failed to reach the recommended therapeutic serum trough concentrations: a higher dose of vancomycin is necessary to maintain serum trough concentration at 15 to 20 mg/l, particularly in critically ill patients with creatinine clearance above 60 ml/minute and in those on intermittent haemodialysis. Among patients with methicillin-resistant Staphylococcus aureus pneumonia, no significant differences were observed in the treatment success rate, length of intensive care unit stay, and intensive care unit mortality rate between patients with vancomycin trough concentrations of >20 mg/l, 15 to 20 mg/l and <15 mg/l.

Vascular endothelial growth factor expression of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin through nuclear factor-kappa B activation in endothelial cells.

Vascular endothelial growth factor (VEGF) induces adhesion molecules on endothelial cells during inflammation. Here we examined the mechanisms underlying VEGF-stimulated expression of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin in human umbilical vein endothelial cells. VEGF (20 ng/ml) increased expression of ICAM-1, VCAM-1, and E-selectin mRNAs in a time-dependent manner. These effects were significantly suppressed by Flk-1/kinase-insert domain containing receptor (KDR) antagonist and by inhibitors of phospholipase C, nuclear factor (NF)-kappaB, sphingosine kinase, and protein kinase C, but they were not affected by inhibitors of mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) 1/2 or nitric-oxide synthase. Unexpectedly, the phosphatidylinositol (PI) 3'-kinase inhibitor wortmannin enhanced both basal and VEGF-stimulated adhesion molecule expression, whereas insulin, a PI 3'-kinase activator, suppressed both basal and VEGF-stimulated expression. Gel shift analysis revealed that VEGF stimulated NF-kappaB activity. This effect was inhibited by phospholipase C, NF-kappaB, or protein kinase C inhibitor. VEGF increased VCAM-1 and ICAM-1 protein levels and increased leukocyte adhesiveness in a NF-kappaB-dependent manner. These results suggest that VEGF-stimulated expression of ICAM-1, VCAM-1, and E-selectin mRNAs was mainly through NF-kappaB activation with PI 3'-kinase-mediated suppression, but was independent of nitric oxide and MEK. Thus, VEGF simultaneously activates two signal transduction pathways that have opposite functions in the induction of adhesion molecule expression. The existence of parallel inverse signaling implies that the induction of adhesion molecule expression by VEGF is very finely regulated.

Biochemical characterization of a thrombin-like enzyme and a fibrinolytic serine protease from snake (Agkistrodon saxatilis) venom.

A thrombin-like enzyme and a fibrinolytic serine protease were purified to homogeneity from the venom of a Korean snake Agkistrodon saxatilis emelianov. Both the purified enzymes migrated as a single protein band corresponding to 39 kDa in SDS-PAGE. However, the molecular mass was reduced to 28 kDa by enzymatic removal of the N-linked carbohydrates in those two different enzyme species. Although the thrombin-like enzyme and the fibrinolytic protease show homologous features in their molecular sizes and N-terminal amino acid sequences, yet they can be clearly distinguished from each other in terms of substrate specificity, susceptibility to inhibitors and fibrinogen degradation. It is postulated that these two enzymes are capable of functioning in a cooperative manner to effectively remove fibrinogen and consequently to reduce the blood viscosity.

Characterization and cDNA cloning of a platelet aggregation inhibitor.

A novel platelet aggregation inhibitor, sal-C, was purified to homogeneity from the venom of Korean snake (Agkistrodon halys brevicaudus). Several lines of experimental evidence clearly indicated that sal-C inhibits not only the collagen-induced platelet aggregation, but also the aggregation mediated by the cell surface glycoprotein IIb-IIIa (GP IIb-IIIa). We have isolated the cDNA encoding sal-C from the cDNA library of the snake venom gland and analyzed its complete nucleotide sequence. Sal-C is a single-chain polypeptide composed of 212 amino acids including 24 cysteines. The deduced polypeptide sequence of sal-C demonstrated considerable homology to previously described protein species of the collagen-induced platelet aggregation inhibitor family. Sal-C does not have the Arg-Gly-Asp (RGD) motif, but contains the Ser-Glu-Cys-Asp sequence. Interestingly, sal-C was found to inhibit GP IIb-IIIa binding to immobilized fibrinogen which is antagonized by the typical RGD motif of disintegrins.

Expression of chemokine genes in murine macrophages infected with Orientia tsutsugamushi.

Scrub typhus, caused by Orientia tsutsugamushi infection, is characterized by local as well as systemic inflammatory manifestations. Inflammation is initiated by O. tsutsugamushi-infected macrophages and endothelial cells in the dermis. We investigated the regulation of chemokine induction in macrophage cell line J774A.1 in response to O. tsutsugamushi infection. The mRNAs for macrophage inflammatory proteins 1alpha/beta (MIP-1alpha/beta), MIP-2, and macrophage chemoattractant protein 1 were induced within 30 min, and their levels showed a transitory peak for 3 to 12 h. However, the lymphotactin, eotaxin, gamma interferon-inducible protein 10, and T-cell activation gene 3 mRNAs were not detected by RNase protection assays. Heat-killed O. tsutsugamushi induced a similar extent of chemokine responses. Induction of the chemokine genes was not blocked by the eukaryotic protein synthesis inhibitor cycloheximide, suggesting that de novo synthesis of host cell protein is not required for these transcriptional responses. The induction of chemokine mRNAs by O. tsutsugamushi was blocked by the inhibitors of NF-kappaB activation. Furthermore, O. tsutsugamushi induced the nuclear translocation and activation of NF-kappaB. These results demonstrate that heat-stable molecules of O. tsutsugamushi induce a subset of chemokine genes and that induction involves activation of the transcription factor NF-kappaB.

Linkage of the nuclear hormone receptor genes NR1D2, THRB, and RARB: evidence for an ancient, large-scale duplication.

The THRA gene encoding thyroid hormone receptor alpha shares an unusual partial overlap with the NR1D1 gene encoding the orphan receptor Rev-ErbAalpha. Though THRA and NR1D1 have close relatives in THRB and NR1D2, which encode TRbeta and Rev-ErbAbeta, these beta isoforms do not share an analogous overlap. Here we report that the human THRB and NR1D2 genes are separated by approximately 1 Mb on chromosome 3 and that these two genes are also linked to the RARB gene, which encodes retinoic acid receptor beta. Since previous results indicate that the THRA/NR1D1 locus is also linked to the RARA gene, these results suggest that the two receptor gene clusters were generated by a single large-scale duplication.

The reversed SoxS-binding site upstream of the ribA promoter in Escherichia coli.

The ribA gene, encoding GTP cyclohydrolase II in Escherichia coli, is a member of the soxRS regulon, which is induced by superoxide-generating agents. By evaluating lacZ expression driven by the ribA promoter carrying different lengths of upstream region in a monolysogen, we found that the superoxide-responsive element resides between 56 and 94 nucleotides upstream of the transcriptional start site. Purified SoxS protein bound to this region and protected nucleotides between positions -80 and -58 from degradation by DNase I. This region contains a putative SoxS-binding sequence (soxbox) in reverse orientation. The SoxS protein interacted specifically with four guanine residues within the soxbox sequence, as demonstrated by methylation interference analysis. These results clearly indicate that SoxS binds to the reversed soxbox sequence in the ribA gene, while in other known genes of the soxRS regulon it binds to the normally oriented soxbox. Possible modes of interaction between SoxS and RNA polymerase are discussed.

[Coronary artery bypass grafting in an octogenarian with chronic myelomonocytic leukemia].

Chronic myelomonocytic leukemia is a disease of the elderly. It tends to have a variable clinical course, as the patient's state is immunologically dysunctional. There has been reluctance to perform open cardiac procedures because of concern about early postoperative sepsis leading to death. A 84-year-old man was admitted for the management of effort angina. PTCA was performed twice. He had left nephrectomy for Grawitz tumor nine years ago and additionally, he had been diagnosed as having chronic myelomonocytic leukemia since the next year. Preoperative laboratory assessment revealed that the total white blood cell counts were 2500 with 25 per-cent of granulocytes, a hematocrit of 31.1%, and platelet counts were 10.0 x 10(4). At the night of the treatment of his granulocytopenia with injection of granulocyte stimulating factor, he complained of continuous anterior chest pain with ST depression on ECG. Emergency single CABG was performed using a saphenous vein graft under the diagnosis of impending myocardial infarction. Postoperative course was uneventful. This is the first case report of CABG in octogenarian with chronic myelomonocytic leukemia in the world.

Dual regulation of the paraquat-inducible gene pqi-5 by SoxS and RpoS in Escherichia coli.

The pqi-5 gene, producing a probable membrane protein of unknown function, has been reported to be a member of the soxRS regulon. The SoxRS-dependent induction of pqi-5 by paraquat occurs only during the exponential phase. The expression of pqi-5 increased in the absence of paraquat during the stationary phase or under conditions of carbon or phosphate starvation. This increase was regulated at the transcriptional level by RpoS (sigma S), which recognized the second promoter (P2) approx. 5 nucleotides upstream from the promoter (P1) used at the exponential phase. Studies with a series of 5' deletions revealed that the paraquat-responsive element resides between -52 and -42 nucleotides upstream from the P1 start site, whose nucleotide sequence matches closely to other SoxS-binding sequences. The stationary-phase induction required sequences up to position -42, which correspond to the 5' border of the putative -35 hexamer for the P2 promoter. The binding of the purified SoxS protein to the pqi-5 promoter upstream sequences was demonstrated by gel mobility-shift and DNase I protection assays. The transcription from P1 promoter by E sigma D was activated by purified SoxS in vitro, as was observed in vivo. The dual regulation of pqi-5 by SoxS at the exponential phase and RpoS at the stationary phase is the first to be reported among the members of the soxRS regulon, suggesting that this gene might indeed play some role under stressful conditions.

Regulation of the ribA gene encoding GTP cyclohydrolase II by the soxRS locus in Escherichia coli.

We isolated a promoter that is inducible by paraquat, a superoxide-generating agent, from Escherichia coli using the promoter-probe plasmid pRS415. Sequence analysis revealed that the promoter derives from the ribA gene encoding GTP cyclohydrolase II, which is the first enzyme in the biosynthetic pathway of riboflavin. We fused the lacZ gene with the ribA promoter to monitor the expression of the gene in the single-copy state. LacZ expression from the ribA promoter was induced about eight-fold by 200 microM paraquat. Other known superoxide generators, menadione and plumbagin, also induced the expression of beta-galactosidase in the fusion strain. On the other hand, no significant induction was observed following treatment with hydrogen peroxide, ethanol or heat shock. Induction of beta-galactosidase was significantly reduced by the introduction of a delta sox-8::cat or soxS3::Tn10 mutation into the fusion strain, indicating that the ribA gene is a member of the soxRS regulon. The transcriptional start site was determined by primer extension analysis and putative binding sites for SoxS in both orientations were identified. GTP cyclohydrolase II activity in soluble extracts of E. coli increased more than three-fold on treatment with paraquat. This increase was dependent on the soxRS locus, and reflects the increase in transcript levels. However, flavin pools did not change significantly. A possible role for ribA induction during superoxide stress is discussed.

Isolation of a novel paraquat-inducible (pqi) gene regulated by the soxRS locus in Escherichia coli.

We have isolated promoters inducible by paraquat, a superoxide radical-generating agent, from Escherichia coli, using promoter-probing plasmid pJAC4 (Y.S. Koh and J.H. Roe, Korean J. Microbiol. 31:267-273, 1993). One promoter clone pqi-5 (pqi denotes paraquat-inducible gene) was mapped at 21.8 min on the E. coli chromosome by using the Kohara phage library. We constructed an operon fusion of the lacZ gene with the pqi-5 promoter to monitor the expression of the gene in the single-copy state. LacZ expression was induced about 7- to 13-fold by 77 to 780 microM paraquat. Other known superoxide generators such as menadione, phenazine methosulfate, and plumbagin also induced the expression of beta-galactosidase in this fusion strain. On the other hand, no significant induction was observed with treatment with hydrogen peroxide, ethanol, and heat shock. Induction of beta-galactosidase was significantly reduced by introducing a delta sox-8::cat or soxS3::Tn10 mutation into the fusion strain, indicating that pqi-5 is a member of the soxRS regulon. A DNA fragment containing the pqi-5 promoter was cloned and sequenced from the Kohara phage E2E5. We identified two pqi-5 open reading frames (ORFs); ORF-A encodes a predicted protein of 342 amino acids, and ORF-B is truncated at the cloning site. The transcription start site from the pqi-5 promoter was determined by primer extension and S1 nuclease protection analyses. Northern (RNA) and S1 analyses indicated that there are two kinds of pqi-5 transcript; one covers ORF-A only and the other covers ORF-A and possibly also ORF-B.

Radiologic assessment in pulmonary lobar transplantation.

UNLABELLED: Pulmonary lobar transplantation provides a clue to the acute donor shortage. To examine the experimental and clinical applicability of lobar transplantation, the authors observed the extent of lung expansion and infiltrate in the allografted lobe through the sequential analysis of the early chest roentgenograms. MATERIALS AND METHODS: Twenty two mongrel dogs weighting 17 kg on average were used. Donor lung bloc was taken and flushed with Euro-Collins solution. The left lower lobar bloc was procured and implanted in the pneumonectomized recipient dog. The anastomosis was performed in the order of the pulmonary vein, artery, and bronchus. To assess the radiological pattern in the lobar allograft, a grading system was designed according to the extent of lung expansion and infiltrate. RESULTS: A) Expansion pattern: Good to excellent lung expansion was seen on postoperative day 0 in 6 out of 10 dogs; on day 1, 4/7; day 2, 3/12; day 3, 1/1; and day 4, 1/3, respectively. Radiographs on day 6, 7, and 12 also showed good expansion in one dog. B) Lung opacity pattern: Clear to minimal infiltrates were seen on day 0 in 8 out of 10 dogs; day 1, 7/17; day 2, 2/12; and day 4, 1/3. The same appearances were detected in a single dog on day 6, 7, and 12. C) Expansion-opacity correlation pattern: Radiographs on postoperative day 0 showed good expansion with mild infiltrates, and excellent expansion with minimal infiltrates were observed on day 1 in 3 out of 17 dogs, day 2, 1/12; and day 4, 1/3, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Nosocomial pneumonia in medico-surgical intensive care unit.

Cases of hospital acquired pneumonia occurring during the 1st 12 months of Medico-Surgical ICU (Intensive care unit, MSICU) in operation were evaluated retrospectively to determine its incidence, common causative pathogens, outcome and radiological patterns with the new hospital setting providing a unique relatively aseptic environment. Among the 920 admitted patients, 73 episodes of nosocomial pneumonia on 63 patients were identified and the incidence rate was 7%. The most common pathogens were Pseudomonas. Staphylococcus, Serratia, and Enterobacter in the order of frequency of occurrence, and the gram-negative pathogens comprised 70%. Nosocomial pneumonia was more common after use of antibiotics due to such pathogens as Enterobacter, Acinetobacter, and Candida which caused poor outcome. Enterobacter had the greatest tendency to be related with poor outcome and Serratia the least. Overall mortality was 25%. Bronchopneumonia was the most common type of pneumonia caused by any pathogen except Acinetobacter which caused a mixed type of nosocomial pneumonia.

[A case report of Konno procedure for Ebstein's anomaly with subaortic stenosis after Hardy procedure and repair of coarctation and VSD].

A rare case of Ebstein's anomaly with coarctation and VSD is reported. Severe subaortic stenosis and LV dysfunction were progressed after PA banding and coarctation repair at four months old and Hardy procedure and VSD closure at five month old. Konno procedure was successfully performed with SJM 21A at one year and nine months old. Since coarctation type VSD may cause subaortic stenosis after its closure due to posteriorly deviated infundibular septum, anterosuperior margin of VSD would occasionally better to be eliminated at the time of VSD closure. This is the youngest one of Konno procedure to our knowledge.