Assessment of flares in lupus patients enrolled in a phase II/III study of rituximab (EXPLORER).

The EXPLORER study was designed to assess the response to rituximab versus placebo in patients with moderate to severe extrarenal systemic lupus erythematosus (SLE) receiving background immunosuppression. The definition of response required reduced clinical activity without subsequent flares over 52 weeks, and the study did not meet its efficacy endpoint. The current exploratory analysis assessed flare rates in patients who achieved initial low disease activity response (British Isles Lupus Assessment Group [BILAG] C or better in all organs) during the study. Exploratory reanalysis of data from the EXPLORER trial was conducted, considering alternative definitions for flare. No difference was found between rituximab and placebo in preventing or delaying moderate to severe flares. However, when severe (BILAG A) flares alone were examined, rituximab reduced the risk of a subsequent first A flare (hazard ratio = 0.61; p = 0.052) and lowered mean ± SD annualized A flare rates (0.86 ± 1.47 vs. 1.41 ± 2.14; p = 0.038). Eighty-four (49.7%) rituximab-treated patients achieved low disease activity without subsequent A flares versus 31 (35.2%) placebo-treated patients (p = 0.027). Prednisone rescue for A flares was similar in rituximab- (24%) and placebo-treated (14%) patients (p = 0.204). This post hoc analysis evaluates the hypothesis that assessment of BILAG A flares may distinguish potential treatment effects with greater sensitivity than assessment of BILAG B flares.

Baseline autoantibody profiles predict normalization of complement and anti-dsDNA autoantibody levels following rituximab treatment in systemic lupus erythematosus.

B cells are thought to play a major role in the pathogenesis of systemic lupus erythematosus (SLE). Rituximab (RTX), a chimeric anti-CD20 mAb, effectively depletes CD20( +) peripheral B cells. Recent results from EXPLORER, a placebo-controlled trial of RTX in addition to aggressive prednisone and immunosuppressive therapy, showed similar levels of clinical benefit in patients with active extra-renal SLE despite effective B cell depletion. We performed further data analyses to determine whether significant changes in disease activity biomarkers occurred in the absence of clinical benefit. We found that RTX-treated patients with baseline autoantibodies (autoAbs) had decreased anti-dsDNA and anti-cardiolipin autoAbs and increased complement levels. Patients with anti-dsDNA autoAb who lacked baseline RNA binding protein (RBP) autoAbs showed increased complement and decreased anti-dsDNA autoAb in response to RTX. Other biomarkers, such as baseline BAFF levels or IFN signature status did not predict enhanced effects of RTX therapy on complement or anti-dsDNA autoAb levels. Finally, platelet levels normalized in RTX-treated patients who entered the study with low baseline counts. Together, these findings demonstrate clear biologic activity of RTX in subsets of SLE patients, despite an overall lack of incremental clinical benefit with RTX in the EXPLORER trial.

Pharmacist intervention in activated protein C therapy for severe sepsis: influence on health and economic outcomes.

OBJECTIVE: To assess the health and cost outcomes of pharmacist intervention versus non-intervention in activated protein C (drotrecogin alpha) therapy for patients with severe sepsis. METHOD: This is a retrospective study. We reviewed the medical records of patients aged 18 years and older who were admitted to our hospital for severe sepsis from January 1, 2003 to December 31, 2007. Only patients who are prescribed activated protein C for the treatment of severe sepsis according to the reimbursement criteria can be reimbursed by the Taiwan Bureau of National Health Insurance (BNHI). Our hospital stipulated that the criteria check list must be evaluated by a clinical pharmacist and the prescribing physician as to whether the patient is eligible to receive activated protein C. To assess the influence of pharmacist intervention on outcomes, we divided eligible patients into two groups, pharmacist-intervention group (Group A; n = 19) and non-pharmacist intervention group (Group B; n = 19). Both groups received a 96-h intravenous infusion of activated protein C at 24 microg/kg/h. We defined evident severe sepsis as concurrent antibiotic plus ventilator and/or vasopressor use. We compared group characteristics, 28-day in-hospital mortality, length of stay and direct medical costs between the two groups. One-way ANOVA was used for analysis. RESULTS: 50% of patients in each group met the reimbursement criteria of the BNHI. Activated protein C therapy was initiated within 1.37 +/- 0.4 days and 7.21 +/- 7.8 days of admission to the ICU in Group A and Group B, respectively (p < 0.01). All of the patients in Group A (19/19) and 42.1% of the patients in Group B (8/19) received activated protein C within 12 - 48 h of admission to the Intensive care unit (ICU) (p < 0.01). 28-day mortality was lower for Group A than for Group B (26.7% and 43.8%, respectively). The length of stay in the ICU for patients in Group A was shorter than that in Group B (14.1 +/- 7.7 vs. 19.7 +/- 11.1, respectively; p < 0.079). Total direct medical costs for survivors in Group A were less than those in Group B (US$ 20,632.3 vs. US$ 24,785.8, respectively; p < 0.05). CONCLUSIONS: Pharmacist intervention in prescribing activated protein C for patients with severe sepsis might reduce direct medical costs and promote earlier initiation of therapy. The potential impact of pharmacist intervention on the timing of activated protein C therapy and the direct medical costs of treatment warrant further study.

Fatal amiodarone-induced hepatotoxicity: a case report and literature review.

OBJECTIVE: To report a fatal case of amiodarone-induced acute hepatotoxicity after intravenous amiodarone administration and similar fatal cases review. CASE SUMMARY: A 72-year-old woman with a history of hypertension, prior cardiovascular disease, atrial fibrillation and diabetes mellitus was admitted to the hospital with acute pyelonephritis and transferred to the intensive care unit due to cerebral infarction. An antidiabetic drug, a low dose of aspirin and intravenous amiodarone therapy was started. After receiving a second dose of amiodarone (1,200 mg; injection rate 1 mg/min), the woman developed ascites, jaundice, high levels of serum transaminases, decreased prothrombin time, and finally became unconscious. Immediately after treatment was discontinued, her extremely high hepatic parameters returned to normal. According to the Naranjo probability scale, this adverse reaction was highly probable. DISCUSSION: The occurrence of acute liver damage after intravenous amiodarone is rare but harmful. It can be induced by polysorbate 80, a solubilizer, by immunomediated centrilobular necrosis, or by the presence of a functional PPAR-I+/- gene. CONCLUSION: Amiodarone is an effective antiarrhythmic agent for preventing and treating atrial and ventricular arrhythmias. The molecular mechanism causing acute hepatic damage after amiodarone treatment is not clear. Therefore, amiodarone must be administered with care, and liver function should be monitored closely in patients treated with this drug.

Human monocyte colony-stimulating factor stimulates the gene expression of monocyte chemotactic protein-1 and increases the adhesion of monocytes to endothelial monolayers.

The stimulation of the human umbilical vein endothelial cell (HUVEC) with recombinant human monocyte-derived colony-stimulating factor (MCSF) increased the gene expression of monocyte chemotactic protein (MCP-1). Northern blot analysis indicated that 50 U/ml of MCSF is the optimal concentration for this effect. The elevation of MCP-1 mRNA started as early as 1 h after stimulation and was maintained for at least 8 h. An increased MCP-1 level in MCSF-treated HUVEC was also demonstrated at the protein level by immunocytochemical staining using a polyclonal MCP-1-specific antibody. HUVEC activated by 50 U/ml of MCSF for 5 h showed a stronger immunofluorescence staining than control cells. Micropipette separation of THP-1 monocytes from HUVEC showed that the activation of both THP-1 and endothelium by MCSF led to an increase in the separation force by more than three times (36.2 +/- 6.7 x 10(-4) vs. 9.6 +/- 3.6 x 10(-4) dyn). An increased adhesiveness was also observed after MCSF activation of peripheral blood monocytes and HUVEC (16.7 +/- 2.7 x 10(-4) vs. 5.2 +/- 0.9 x 10(-4) dyn). The increased adhesive force in both systems was blocked by the use of anti-MCP-1 (5.5 +/- 0.8 x 10(-4) and 6.8 +/- 1.1 x 10(-4) dyn). Similar results were obtained in experiments in which only HUVEC, but not monocytes, were activated by MCSF. This increased adhesion of untreated monocytes to MCSF-activated HUVEC was also blocked by the addition of anti-MCP-1. In contrast, experiments in which only THP-1 or peripheral blood monocytes, but not HUVEC, were treated with MCSF did not show a significant increase of adhesion between these cells. These results indicate that MCSF augments monocyte-endothelium interaction primarily by its action on the endothelial cell and that this function is probably mediated through an increased expression of MCP-1. The MCSF/MCP-1-dependent adhesive mechanism might be operative in the arterial wall in vivo to lead to the trapping of the infiltrated monocyte-macrophage in the subendothelial space during atherogenesis.

Inhibition of DNA2 nuclease as a therapeutic strategy targeting replication stress in cancer cells.

Replication stress is a characteristic feature of cancer cells, which is resulted from sustained proliferative signaling induced by activation of oncogenes or loss of tumor suppressors. In cancer cells, oncogene-induced replication stress manifests as replication-associated lesions, predominantly double-strand DNA breaks (DSBs). An essential mechanism utilized by cells to repair replication-associated DSBs is homologous recombination (HR). In order to overcome replication stress and survive, cancer cells often require enhanced HR repair capacity. Therefore, the key link between HR repair and cellular tolerance to replication-associated DSBs provides us with a mechanistic rationale for exploiting synthetic lethality between HR repair inhibition and replication stress. DNA2 nuclease is an evolutionarily conserved essential enzyme in replication and HR repair. Here we demonstrate that DNA2 is overexpressed in pancreatic cancers, one of the deadliest and more aggressive forms of human cancers, where mutations in the KRAS are present in 90-95% of cases. In addition, depletion of DNA2 significantly reduces pancreatic cancer cell survival and xenograft tumor growth, suggesting the therapeutic potential of DNA2 inhibition. Finally, we develop a robust high-throughput biochemistry assay to screen for inhibitors of the DNA2 nuclease activity. The top inhibitors were shown to be efficacious against both yeast Dna2 and human DNA2. Treatment of cancer cells with DNA2 inhibitors recapitulates phenotypes observed upon DNA2 depletion, including decreased DNA double strand break end resection and attenuation of HR repair. Similar to genetic ablation of DNA2, chemical inhibition of DNA2 selectively attenuates the growth of various cancer cells with oncogene-induced replication stress. Taken together, our findings open a new avenue to develop a new class of anticancer drugs by targeting druggable nuclease DNA2. We propose DNA2 inhibition as new strategy in cancer therapy by targeting replication stress, a molecular property of cancer cells that is acquired as a result of oncogene activation instead of targeting currently undruggable oncoprotein itself such as KRAS.

First Report of Formosan Michelia Seedling Root Rot Caused by Pythium splendens in Taiwan.

Formosan michelia (Michelia compressa (Maxim.) Sargent) is a valuable evergreen tree in Taiwan that is distributed from low to medium (200 to 1,800 m) altitudes. In many nurseries in Taiwan, Formosan michelia seedlings grow poorly or wilt. The etiology of the disease observed in April 2004 in a nursery in Jinshan was investigated. Diseased seedlings with chlorotic leaves and decayed feeder roots lost leaves, died back, and then wilted. The putative pathogen, Pythium splendens Braun, was isolated and identified on a morphological basis (1). P. splendens was isolated from the roots of diseased seedlings on 2% water agar with 100 ppm of ampicillin. Isolates increased daily on potato dextrose agar at 24°C by 27 to 30 mm and on malt extract agar (MEA) by 23 to 25 mm. No zoosporangia and zoospores were produced. The main hyphae were as much as 9 µm wide on MEA. Hyphal swellings were abundant, globose, smooth, terminal, and 33 to 42 µm in diameter, often with dark, densely granulated contents. Attempted matings of four P. splendens isolates in V8 medium failed. To prove pathogenicity, the four isolates were cultured in 300-ml flasks containing 150 ml of 2% malt extract medium at room temperature for 14 days. The mycelia were homogenized in sterile water at 4,500 rpm for 5 min. The suspension was adjusted to 5 × 106 hyphal swellings per ml. Roots of the 2-month-old seedlings were immersed in the suspension for 2 h, whereas the control seedlings were immersed in sterilized water. Five seedlings of each of three replicates were inoculated with one of the four isolates for a total of 60 seedlings. Controls were replicated in the same way. The inoculated plants were transplanted into plastic flowerpots containing sterilized peat and moss and kept in the greenhouse at 20 to 24°C. After 14 days, inoculated seedlings developed symptoms like those of the original plants. The putative pathogen was reisolated from the roots of inoculated plants. Cultures are maintained at the Forest Pathology Lab of the National Taiwan University. To our knowledge, this is the first report of proof of pathogenicity of P. splendens on Formosan michelia seedlings. Reference: (1) A. J. Van der Plaats-Niterink. Stud. Mycol. 21:151, 1981.

Indoor environmental risk factors and childhood asthma: a case-control study in a subtropical area.

This study examined the relation between indoor environmental factors and childhood asthma in a subtropical area. A hospital-based case-control study was performed in Kaohsiung, Taiwan, between July of 1995 and June of 1996. Eighty-six children seen in the out-patient clinic of our university hospital and who had a first-time diagnosis of asthma made by a pediatrician were the test group; 86 control subjects were selected from children attending the Childhood Orthopaedic Clinic in the same hospital and who had no previous diagnosis of asthma or asthma symptoms and no history of physician confirmed atopic diseases. The control subjects were matched with test case children on the basis of gender and age. Information was obtained from parents using a structured questionnaire. Of the many indoor environmental factors included in this study, only home dampness showed an association with asthma (adjusted odds ratio=1.77; 95% confidence intervals, 1.24-2.53). We conclude that dampness in the home is a new public health risk factor related to asthma in subtropical areas.

First Report of Sclerotium rolfsii on Island Ash Seedlings in Taiwan.

Island ash (Fraxinus formosana Hay.) is a large, semideciduous tree in Taiwan. It is used for forestation, a shade tree, and producing wood for furniture. During the summer of 2001, a sudden wilt of 1-year-old plants was observed in a nursery in northern Taiwan. Initial symptoms included stem necrosis at the soil line and yellowing and tan discoloration of the leaves. As stem necrosis progressed, infected plants wilted and died. Necrotic tissues were covered with white mycelium that differentiated into reddish brown, spherical (1 to 2 mm in diameter) sclerotia. Sclerotium rolfsii was consistently recovered from the surface of symptomatic stem sections that were disinfected for 1 min in 0.5% NaOCl and then plated on potato dextrose agar (PDA) amended with 100 ppm of ampicillin. Pathogenicity of two S. rolfsii isolates was confirmed on 1-year-old island ash seedlings grown in 12.7 cm- (5-in) plastic pots in a sterilized mixture of peat moss and vermiculite (3:1). Seedlings were inoculated with mycelia and sclerotia of the pathogen grown on PDA. Three plants each were inoculated with four disks (5 mm) of agar with mycelium and three were inoculated with 10 sclerotia that were placed on the soil surface around the base of each plant. Noninoculated plants served as controls. All plants were kept in a growth chamber at 25 to 35°C and >95% relative humidity. The test was repeated once. All inoculated plants developed symptoms within 14 days, while control plants remained symptomless. Sclerotia developed on infected tissues, and S. rolfsii was reisolated from symptomatic tissues. This disease has been observed on many species of plants (1), but to our knowledge, this is the first report of Southern blight of Island ash seedlings caused by S. rolfsii in Taiwan. Reference: (1) Y. P. Tsai ed. List of Plant Diseases in Taiwan. The Plant Protection Society of the Republic of China and The Phytopathological Society of the Republic of China, 1991.

First Report of Sclerotium rolfsii on Star-cluster (Pentas lanceolata) in Taiwan.

Star-cluster (Pentas lanceolata (Forssk.) Deflers) has recently become popular as a bedding plant in Taiwan. During the summer of 2000, a sudden wilt of 60-day-old plants was observed in a nursery in Tainan City (southern Taiwan). Initial symptoms included stem necrosis at the soil line and yellowing and tan discoloration of leaves. As stem necrosis progressed, infected plants wilted and died. Necrotic tissues were covered with white mycelium that differentiated into reddish brown, spherical (1 to 2 mm in diameter) sclerotia. Sclerotium rolfsii was consistently recovered from the surface of symptomatic stem sections that were disinfected for 1 min in 0.5% NaOCl and plated on potato dextrose agar amended with 100 ppm streptomycin sulfate. Pathogenicity of three isolates of S. rolfsii was confirmed by inoculating 90-day-old plants of P. lanceolata that were grown in pots. Three plants each were inoculated with a 5-mm plug of agar with mycelium or two sclerotia of the pathogen. Inoculum was placed on the soil surface against the stem of each plant. Three noninoculated plants served as controls. All plants were kept in a growth chamber at 20 to 30°C with relative humidity >85%. The pathogenicity test was repeated. Inoculated plants developed symptoms within 7 days, while control plants remained symptomless. Sclerotia developed on infected tissues and S. rolfsii was reisolated from symptomatic tissues. Although this disease has been observed on many species of plants (1), to our knowledge, this is the first report of southern blight of P. lanceolata caused by S. rolfsii in Taiwan. Reference: (1) Tsai, Y. P., ed. List of Plant Diseases in Taiwan. The Plant Protection Society of the Republic of China and The Phytopathological Society of the Republic of China. 1991.

Cyclic strain induces redox changes in endothelial cells.

Our previous studies have shown that cyclic strain to endothelial cells (ECs) increases reactive oxygen species (ROS) that act as second messengers. The potential impact of these enhanced ROS levels on ECs was examined by studying the antioxidant activities and heme oxygenase-1 (HO-1) expression in strained ECs. Cyclic strain to ECs increased lipid peroxidation and augmented oxidation of low-density lipoproteins. ECs subjected to strain increased their superoxide dismutase activities. Concomitantly, glutathione peroxidase activities increased in 3 to 6 hr and returned to basal level 24 hr after continuous cyclic strain treatment. A decrease of glutathione (GSH) was accompanied with an increase of oxidized glutathione (GSSH) level in ECs 3 to 6 hr after strain treatment. This was followed with a return of both GSH and GSSH to basal levels in 24 hr. Consistently, H2O2 treatment of ECs decreased the GSH/GSSG ratio. ECs pretreated with catalase abolished the strain-induced change in GSH/GSSG. Strain treatment, similar to H2O2 exposure, induced HO-1 expression in a time-dependent manner. This induction was inhibited after treating ECs with catalase or free radical scavenger. ECs treated with N-acetyl-cysteine abolished HO-1 gene induction. Our results suggest that cyclic strain-induced ROS cause a transient increase of glutathione peroxidase activity that results in a decrease of GSH level in ECs and that this decrease is crucial to HO-1 induction.

Cloning and heterologous expression of a novel ligninolytic peroxidase gene from poroid brown-rot fungus Antrodia cinnamomea.

A novel ligninolytic peroxidase gene (ACLnP) was cloned and characterized from a poroid brown-rot fungus, Antrodia cinnamomea. The genomic DNA of the fungus harboured two copies of ACLnP, with a length of 2111 bp, interlaced with 12 introns, while the full-length cDNA was 1183 bp, with a 66 bp signal peptide and an ORF of 990 bp. The three-dimensional molecular structure model was comparable to that of the versatile peroxidase of Pleurotus eryngii. ACLnP was cloned into vector pQE31, successfully expressed in Escherichia coli strain M15 under the control of the T5 promoter and produced a non-glycosylated protein of about 38 kDa, pI 5.42. The native and recombinant ACLnP was capable of oxidizing the redox mediator veratryl alcohol, and also decolorized bromophenol blue and 2,6-dimethoxyphenol dyes, implicating a functional extracellular peroxidase activity. The significance of discovering a functional ACLnP gene in A. cinnamomea in terms of wood degradation and colonization capacity in its unique niche is discussed.

Pulsatile and steady flow induces c-fos expression in human endothelial cells.

The effects of pulsatile and steady fluid flow on the mRNA levels of proto-oncogenes c-fos, c-jun, and c-myc in cultured human umbilical vein endothelial cells (HUVEC) were investigated. c-fos mRNA levels in stationary cultures were very low. A 1 Hz pulsatile flow with an average shear stress of 16 dynes/cm2 induced a dramatic increase of c-fos mRNA levels in HUVEC 0.5 h after the onset of flow, which declined rapidly to basal levels within 1 h. Steady flow with a similar shear stress also induced a transient increase of c-fos mRNA levels, but to a lesser extent. In addition, increased c-fos mRNA levels were observed when low shear (2-6 dynes/cm2) was replaced by high shear (16-33 dynes/cm2). Pulsatile and steady flow caused a slight increase of c-jun and c-myc mRNA levels. The role of pulsatility was also investigated in platelet-derived growth factor (PDGF) expression. Pulsatile flow induced a transient increase of PDGF A- and B-chain mRNA levels with peaks at 1.5-2 h. Pulsatile flow, which was more stimulatory in mediating c-fos expression, however, was less stimulatory than steady flow in mediating PDGF expression. By using various inhibitors, protein kinase C was found to be an important mediator in flow-induced c-fos expression, with the involvement of G proteins, phospholipase C, and intracellular calcium. Protein kinase C was previously shown as a possible major mediator in flow-induced PDGF expression which, at least partly, appeared to follow the induction mechanism of c-fos, suggesting a possible connection between c-fos and PDGF induction. However, the c-fos antisense treatment, which significantly inhibited c-fos transcription, failed to block the flow-induced PDGF expression, suggesting that flow-induced c-fos expression may not play an important role in the mechanism of flow-induced PDGF expression. The difference in the induction of c-fos and PDGF expression under pulsatile as compared to steady flow indicates that a complex, flow-mediated regulatory mechanism of gene expression exists in HUVEC. The increased expression of these proto-oncogenes mediated by flow may be important in regulating long-term cellular responses.

Shear stress increases endothelial platelet-derived growth factor mRNA levels.

We have investigated the effect of shear stress on platelet-derived growth factor (PDGF) A and B chain mRNA levels in cultured human umbilical vein endothelial cells (hUVEC). The levels of both PDGF A and B mRNA in hUVEC were increased by a physiological shear stress (16 dyn/cm2), reaching a maximum approximately 1.5-2 h after the onset of shear stress and returning almost to control values at 4 h. The peak levels showed a more than 10-fold enhancement for PDGF A mRNA and a 2- to 3-fold increase for PDGF B mRNA (P less than 0.05). PDGF A mRNA also showed a shear-dependent increase from 0 to 6 dyn/cm2 (P less than 0.05) and then plateaued from 6 to 51 dyn/cm2. PDGF B mRNA levels were elevated as shear stress increased from 0 to 6 dyn/cm2 then declined gradually to a minimum at 31 dyn/cm2 (P less than 0.05) and increased again when shear stress rose to 51 dyn/cm2 (P less than 0.05). PDGF, a potent smooth muscle cell mitogen and vasoconstrictor, released from the endothelium may regulate the blood flow in vivo. The shear stress-dependent elevation of PDGF A and B mRNA in endothelial cells may be involved in the adaptation of blood vessels to flow mediated by the endothelium.

Shear-induced platelet-derived growth factor gene expression in human endothelial cells is mediated by protein kinase C.

Our previous studies have shown that steady shear stress causes a transient increase of platelet-derived growth factor (PDGF) A and B chain mRNA levels in human umbilical vein endothelial cells (HUVEC). In the present study, we elucidated the signaling pathway of shear stress in HUVEC by examining the roles of protein kineses, intracellular calcium, cyclooxygenase, and guanine nucleotide-binding proteins (G proteins) in the PDGF gene induction by shear. The protein kinase C inhibitors, H7 and staurosporine, strongly inhibited the shear-induced PDGF gene expression in HUVEC. In contrast, HA1004, a cAMP- and cGMP-dependent protein kinases inhibitor, was only slightly inhibitory. BAPTA/AM, an intracellular calcium chelator, partially (50%) inhibited the shear-induced PDGF gene expression. The cyclooxygenase inhibitors, ibuprofen and indomethacin, were slightly inhibitory. A 35-50% inhibition of shear-induced PDGF gene expression was found with GDP-beta-S, an inhibitor of G proteins. These results suggest that shear-induced PDGF gene expression in HUVEC is mainly mediated by protein kinase C activation and requires intracellular calcium. Furthermore, G proteins seem to be involved in this process, whereas prostaglandin synthesis via cyclooxygenase pathway is not. We propose a mechanism of shear-induced PDGF gene expression in HUVEC: Shear stress, either directly or indirectly (G protein-mediated), enhances the membrane phosphoinositide turnover via phospholipase C, producing diacylglycerol, an activator of protein kinase C. The activated protein kinase C then triggers the subsequent PDGF gene expression.

Fluid shear stress induces a biphasic response of human monocyte chemotactic protein 1 gene expression in vascular endothelium.

The focal distribution of atherosclerotic lesions in the arterial tree is related to the local shear stress generated by blood flow, but the molecular basis of the atherogenic response of endothelial cells in these lesion-prone areas is still unclear. We report that shear stress mediates a biphasic response of monocyte chemotactic protein 1 (MCP-1) gene expression in vascular endothelial cells (EC). Northern blot analysis indicated that the level of MCP-1 mRNA in human umbilical vein EC (HUVEC) subjected to a shear stress of 16 dynes/cm2 (1 dyne = 10 microN) for 1.5 hr increased by 2- to 3-fold when compared with static cells. The MCP-1 gene expression decreased to the basal level at 4 hr and then declined further to become completely quiescent at 5 hr after the onset of shear. Once the gene expression was fully suppressed, it remained quiescent even after static incubation for 1.5 hr and would not respond to reshearing after this static incubation. However, if the postshearing incubation extended from 1.5 to 24 hr, the MCP-1 mRNA returned to the basal level and was then able to increase after the reapplication of shear stress. Nuclear run-on experiments showed that the shear-induced increased MCP-1 mRNA in HUVEC was regulated at the transcriptional level. By using cycloheximide, it was shown that de novo protein synthesis was not necessary for the induction of MCP-1 by shear stress. The biphasic response of MCP-1 gene expression was found in experiments in which the applied shear stress was 6, 16, or 32 dynes/cm2, and it was observed not only in HUVEC but also in HeLa cells, glioma cell lines, and skin fibroblasts. This in vitro study demonstrates that the response of MCP-1 gene to shear stress represents an immediate early gene activation and suggests that this gene is probably suppressed in EC that have been exposed to a constant shear stress.

[A case report of carotenemia].

Carotenemia is a common disease in children, and it is mainly due to excessive intake of food enriched with carotene. A fourteen-year-old girl, whose skin had been discolored yellow for several months, was brought to Kaohsiung Medical College Hospital for examination. Physical examination revealed she had a marked yellow color skin, especially on the palms and soles; conjunctiva was not anemic; sclera was not icteric; breath and heart sounds were normal; liver and spleen were not enlarged. Laboratory examination revealed normal results for the liver function test, however, serum carotene was elevated to 460 micrograms/dl in contrast to 80 micrograms/dl from a same-aged healthy child as a control (normal range of the serum carotene: 50-250 micrograms/dl). Tracing back her past history, she had taken a lot of oranges daily for more than one year. After consulting a nutritionist and changing her eating habits to a balanced diet for eight weeks, the serum carotene level decreased to 270 micrograms/dl (same-aged healthy child: 30 micrograms/dl). The skin color was also improved.

Immunotherapy for recurrent spontaneous abortions in a Chinese population.

The efficacy of immunotherapy for the treatment of recurrent spontaneous abortions was tested in patients selected from the same ethnically homogeneous population of Chinese in Taiwan in whom the immunogenetics of gestational trophoblastic tumors and of recurrent spontaneous abortion had been studied. The patients, who included both primary and secondary aborters, were randomly assigned to three groups: those who were immunized with their own lymphocytes (controls) (49); those who were immunized with their husbands' lymphocytes (39); and those who were immunized with third party lymphocytes (11). The data were analyzed individually for the primary and secondary aborters and collectively for both groups combined. The number of babies born, the number of current pregnancies, and the number of recurrent abortions were not statistically significantly different between the control and the immunized groups, and a similar small number of congenital abnormalities (4-9%) occurred in both the control and immunized groups. The increase in the blocking effect for the mixed lymphocyte reaction was not related to the success of the postimmunization pregnancies. Thus, this study does not show any significant improvement in the rate of livebirths in women immunized with their husbands' lymphocytes or with third party lymphocytes compared to that in a placebo-controlled group of women.

Increased incidence of asthma and pulmonary dysfunction after severe lower respiratory tract infection in infancy.

In the present study we tried to define the effect of lower respiratory tract infections upon pulmonary function and/or asthma in childhood. Thirty-five children with history of pneumonia in infancy were followed five to ten years later; all were asked to respond questionnaire, received physical examination and were diagnosed for pulmonary function. The results follow: 13 children (37%) had developed asthma, a significantly higher percentage than normal prevalence among students in this area. Simple pulmonary function test, pulmonary function test after distilled water mist and after hypertonic saline (4.5%) mist all showed abnormal values in VC (17%, 14%, 29% respectively), in IVC (46%, 51%, 53%), in FVC (20%, 23%, 24%), in FEVl (17%, 23%, 29%), in FEF25-75% (37%, 49%, 47%), in FEF75% (26%, 23%, 29%) and in FEVl/VC (20%, 14%, 29%). Methacholine challenge test (PC20) showed a marked decrease of PC20 in asthmatic children; each was less than 5 mg/ml (mean value; 0.99 mg/ml). Family-allergy in at least one parent and wheeze were the two significant risk factors. Nevertheless, in 22 non-family-allergy children, the occurrence of asthma was also higher than general prevalence (18.2% vs 5.6%). Wheezing was evident in viral infections in infancy, but bacterial culture from sputum or throat swabs failed to find pathogenic bacteria. These results indicate that while the genetic factor may be important, viral infections may be more important because, even in non-family-allergy children, the occurrence of asthma was higher for infants infected in early infancy than the general prevalence for age-matched students.