Lysophosphatidylcholine as a prognostic marker in community-acquired pneumonia requiring hospitalization: a pilot study.

Clinical prediction indicators such as the pneumonia severity index (PSI) and CURB-65 score are useful, but they are complex and often not followed. Therefore, biomarkers that improve hospital outcome predictions are emerging. This study evaluated the prognostic value of a new sepsis biomarker, serum lysophosphatidylcholine (LPC) concentrations, in community-acquired pneumonia (CAP) patients. We prospectively collected blood samples from emergency department CAP patients on days 1 and 7 (post-admission) and analyzed their plasma LPC concentrations. We retrospectively reviewed patient medical records and analyzed correlations between plasma LPC concentrations and clinical parameters and hospital outcomes. A total of 56 CAP patients were included in this study; 24 (42.9 %) required intubation and 15 (26.8 %) died. The mean LPC concentrations on days 1 (p = 0.015) and 7 (p = 0.002) of hospitalization were significantly lower in the non-survivors. Day 1 LPC concentrations were inversely correlated with the PSI (ρ = -269) and CURB-65 scores (ρ = -386). For predicting hospital mortality, the day 1 LPC concentration was comparable with the CURB-65 or PSI scores. Day 1 LPC cut-off levels <29.6 µmol/L were associated with hospital CAP outcomes, including the need for mechanical ventilation, vasopressors, intensive care unit admission, and hospital mortality. Additionally, day 7 LPC concentrations were correlated with in-hospital mortality. Initial serum LPC concentrations predicted hospital outcomes in CAP patients requiring hospitalization. These values were correlated with prognostic markers, such as the PSI and CURB-65 scores. Additionally, follow-up LPC measurements predicted the clinical course of CAP patients.

Epidemiological analysis of critically ill adult patients with pandemic influenza A(H1N1) in South Korea.

A total of 245 patients with confirmed 2009 H1N1 influenza were admitted to the intensive-care units of 28 hospitals (South Korea). Their mean age was 55.3 years with 68.6% aged >50 years, and 54.7% male. Nine were obese and three were pregnant. One or more comorbidities were present in 83.7%, and nosocomial acquisition occurred in 14.3%. In total, 107 (43.7%) patients received corticosteroids and 66.1% required mechanical ventilation. Eighty (32.7%) patients died within 30 days after onset of symptoms and 99 (40.4%) within 90 days. Multivariate logistic regression analysis showed that the clinician's decision to prescribe corticosteroids, older age, Sequential Organ Failure Assessment score and nosocomial bacterial pneumonia were independent risk factors for 90-day mortality. In contrast with Western countries, critical illness in Korea in relation to 2009 H1N1 was most common in older patients with chronic comorbidities; nosocomial acquisition occurred occasionally but disease in obese or pregnant patients was uncommon.

Patient with positional wheezing due to endobronchial lipomatous hamartoma.

Endobronchial hamartoma is a rare form of pulmonary hamartoma, and endobronchial lipomatous hamartomas are even rarer. We describe the case of a 39-year-old man who presented with a two-year history of dyspnea on exertion and wheezing over the left chest only while lying on his left side. The patient was diagnosed with endobronchial lipomatous hamartoma occluding the left main bronchus. He underwent a superior segmentectomy of the left lower lobe, which promptly relieved the dyspnea and positional wheezing. To our knowledge, fewer than 10 such cases are cited in the English literature, none of which presented with positional wheezing. This patient represents the first case of focal and positional wheezing resulting from endobronchial lipomatous hamartoma.

Hyperthermal neutral beam sources for material processing.

Hyperthermal neutral beams have a great potential for material processes, especially for etching and thin film deposition for semiconductor and display fabrication as well as deposition for various thin film applications. Plasma-induced damage during plasma etching is a serious problem for manufacturing deep submicron semiconductor devices and is expected to be a problem for future nanoscale devices. Thermal and plasma-induced damage is also problematic for thin film depositions such as transparent conductive oxide films on organic light emitting diodes or flexible displays due to high temperature processes in plasma environments. These problems can be overcome by damage-free and low-temperature processes with hyperthermal neutral beams. We will present the status of the hyperthermal neutral beam development and the applications, especially, in semiconductor and display fabrication and introduce potential applications of thin film growing for optoelectronic devices such as light emitting diodes.

Molecular cloning and cultivar specific expression of MAP kinases from Capsicum annuum.

Two MAP kinases, MK1 and MK2, were cloned from Capsicum annuum (pepper) cv. Subicho using a parsley MAP kinase gene as a heterologous probe. MK1 and MK2 encode stress-inducible protein kinases that can contribute to the response to wounding, UV-C, and cold. MK1 has a 92% amino acid identity with WIPK of tobacco. It was transcriptionally induced in response to wounding. In contrast, no detectable MK1 transcript was found in unwounded leaves of pepper. MK2 has a high level of amino acid homology to MAP kinases, such as NTF4 and SIPK and was constitutively expressed in all tissues. Both MK transcripts were downregulated by UV-C treatment. Each MK protein activation was independently wound-inducible in a cultivar dependent manner. MK1 is phosphorylated in cv. Pungchon but not cv. Subicho; whereas, the MK2 protein activation by wounding is restricted to cv. Subicho. In addition, de novo synthesis of the MK1 protein and tyrosine phosphorylation was rapidly and transiently induced in cv. Pungchon by wounding. In contrast, it is highly unlikely that the MK1 protein is produced in cv. Subicho, even though there is an abundant expression of MK1 mRNA after wounding in this cultivar. In Escherichia coli, which overexpresses MK1, autophosphorylation is observed at conserved threonine and tyrosine phosphorylation sites.

Bronchial hyperreactivity in patients with endobronchial tuberculosis.

Some patients with endobronchial tuberculosis (EBTB) have wheeze on physical examination and normal chest PA, which mimic bronchial asthma. Non-specific bronchial challenge tests have been used to confirm the presence of bronchial hyperreactivity, which is a hallmark of bronchial asthma. To evaluate the effect of endobronchial tuberculous inflammation on bronchial responsiveness to histamine, the provocation concentrations of histamine required to reduce FEV1 by 20% of the pre-challenge baseline (PC20) were compared between patients with EBTB, patients with symptomatic bronchial asthma and normal, healthy controls. PC20 in EBTB patients (17.2 +/- 2.3 mg ml-1) and normal controls (19.5 +/- 1.4 mg ml-1) were significantly higher than in bronchial asthma patients (0.99 +/- 0.15 mg ml-1). PC20 in EBTB patients was not affected by disease location in the bronchial tree was not correlated with FVC or FEV1. In conclusion, one should consider the possibility of EBTB for differential diagnosis from bronchial asthma, if airway responsiveness appears normal in wheezy patients.

In vitro efficacy of recombinant diphtheria toxin-murine interleukin-4 immunoconjugate on mouse glioblastoma and neuroblastoma cell lines and the additive effect of radiation.

OBJECT: The prognosis for patients with primary malignant brain tumors is poor despite aggressive treatment, and tumor recurrence is common regardless of the chosen therapy. Although multimodal treatment does not provide a cure, it is necessary to determine which treatment modalities have the greatest cytotoxic effect and can potentially prolong survival. Immunotoxin therapy is a novel approach for the treatment of tumors, and it has been successfully used in the central nervous system. Because the interleukin (IL)-4 receptor is commonly expressed on brain tumor cells, the purpose of this study was to evaluate the cytotoxic effect of using a modified diphtheria toxin-murine IL-4 (DT390- mIL4) immunoconjugate for the treatment of murine brain tumor cell lines and to determine whether the addition of radiation therapy could potentiate the effect of this agent. METHODS: Spontaneous murine glioblastoma (SMA-560) and two neuroblastoma (Neuro-2a and NB41A3) cell lines were treated using DT390-mIL4 at different concentrations, and the anti-mouse IL-4 monoclonal antibody (11B11) was used for blocking its cytotoxicity. Other SMA-560 and Neuro-2a cell lines were treated using 500 cGy of radiation 3 hours before DT390-mIL4 treatment. Cytotoxity was evaluated using a trypan blue viability assay. The immunoconjugate exhibited a dose-dependent cytotoxic effect with 50% inhibitory concentration values of 0.56 x 10(-9) M in SMA-560, 1.28 x 10(-9) M in Neuro-2a, and 0.95 x (-10) M in NB41A3 cell lines. The cytotoxicity of DT390-mIL4 was specifically blocked by an excess of 11B11. Cytotoxicity was additive when the DT390-mIL4 at 10(-9) M immunoconjugate administration was followed by radiation therapy. CONCLUSIONS: These results indicate that the IL-4 receptor can be a target for diphtheria toxin fusion proteins and that radiation can potentiate the effects of DT390-mIL4. The development of multimodal approaches to treat malignant brain tumors with agents that have different mechanisms of action may be beneficial.

Pro-tumour necrosis factor cleavage enzyme in macrophage membrane/particulate.

Tumour necrosis factor (TNF) is synthesized initially as a membrane-bound precursor which is then cleaved to yield soluble, mature protein. The 26,000 MW TNF precursor isolated from the lysate of activated RAW 264.7 (mouse macrophage) cells by immunoprecipitation was used to identify pro-TNF cleavage enzyme in the same cells. A significant amount of mature protein was formed in samples containing Nonidet P-40 (NP-40)-lysed cells, whereas sonicated cells showed negligible activity. Most of the cleavage activity in macrophages was localized in the membrane/particulate fraction and remained largely insoluble after sonication or treatment with 2 mM EDTA/1 M NaCl, indicating that the enzyme is associated with the membrane/particulate fraction. The crude cleavage activity in membrane/particulate was partially inhibited by a spectrum of serine, cysteine and aspartate proteinase inhibitors, whereas secretion of TNF from activated macrophages was inhibited exclusively by serine and serine/cysteine proteinase inhibitors. This result suggested that, among heterogenous pro-TNF cleavage activities, the enzyme responsible for the processing of TNF is a serine proteinase. Pro-TNF cleavage activity was present in non-stimulated macrophages and decreased significantly 8 hr after lipopolysaccharide (LPS) stimulation, suggesting that it is negatively regulated after an initial burst of TNF synthesis.

Tosylphenylalanine chloromethyl ketone inhibits TNF-alpha mRNA synthesis in the presence of activated NF-kappa B in RAW 264.7 macrophages.

Serine proteinase inhibitors such as N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) and N alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK) were shown to inhibit production of tumour necrosis factor-alpha (TNF-alpha) in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. The proteinase inhibitors were also reported to inhibit activation of the transcription factor nuclear factor-kappa B (NF-kappa B) by blocking the signalling pathway for stimuli-induced phosphorylation of the inhibitory subunit (I kappa B alpha) and thus preventing its degradation. In RAW 264.7 cells TPCK and TLCK significantly suppressed LPS-induced increase in TNF-alpha mRNA, induction of nuclear kappa B-binding activity and degradation of I kappa B alpha. TPCK and TLCK effectively blocked TNF-alpha mRNA synthesis even when they were added after LPS stimulation. In these cells, however, the inhibitory modes of the two inhibitors were found to be different: while addition of TLCK suppressed I kappa B alpha degradation and reduced NF-kappa B activity, a comparable decrease in the nuclear kappa B-binding activity or in I kappa B alpha degradation was not observed in cells treated with TPCK. Our results show that TPCK inhibits LPS-induced TNF-alpha mRNA synthesis in the presence of activated NF-kappa B and suggests that mechanisms other than NF-kappa B activation are involved in the transcriptional regulation of the TNF-alpha gene.

Cloning and bacterial expression of sesquiterpene cyclase, a key branch point enzyme for the synthesis of sesquiterpenoid phytoalexin capsidiol in UV-challenged leaves of Capsicum annuum.

Sesquiterpene cyclase, a branch point enzyme in the general isoprenoid pathway for the synthesis of phytoalexin capsidiol, was induced in detached leaves of Capsicum annuum (pepper) by UV treatment. The inducibility of cyclase enzyme activities paralleled the absolute amount of cyclase protein(s) of pepper immunodetected by monoclonal antibodies raised against tobacco sesquiterpene cyclase. A cDNA library was constructed with poly(A)+ RNA isolated from 24 h UV-challenged leaves of pepper. A cDNA clone for sesquiterpene cyclase in pepper was isolated by using a tobacco 5-epi aristolochene synthase gene as a heterologous probe. The predicted protein encoded by this cDNA was comprised of 559 amino acids and had a relative molecular mass of 65,095. The primary structural information from the cDNA clone revealed that it shared 77%, 72% and 49% identity with 5-epi aristolochene, vetispiradiene, and cadinene synthase, respectively. The enzymatic product catalyzed by the cDNA clone in bacteria was identified as 5-epi aristolochene, as judged by argentation TLC. RNA blot hybridization demonstrated the induction of an mRNA consistent with the induction of cyclase enzyme activity in UV-treated pepper.

Acute appendicitis in patients with acute leukemia.

The decision to operate for abdominal pain in patients with leukopenia can be exceedingly difficult. Surgical exploration may be the only effective way to differentiate acute appendicitis from other causes, but it involves considerable risk of infectious complications due to immunosuppression. Leukemic patients, who presented significant RLQ pain, had been indicated for operation, despite having advanced disease or having had received chemotherapy or steroids. Four adult leukemia patients, complicated by acute appendicitis, were reviewed. Two patients were in induction chemotherapy, one receiving salvage chemotherapy due to relapse and the other was in conservative treatment. Two patients were acute myelocytic leukemia (AML), one had acute lymphocytic leukemia (ALL), and the other had aleukemic leukemia. All patients underwent appendectomy and recovered without complication. Our experience supports the theory that the surgical management of appendicitis in acute leukemia is the most effective way, in spite of leukopenia.

Expression and amplification of cellular oncogenes in human developing placenta and neoplastic trophoblastic tissue.

To confirm the expression of cellular oncogenes during normal development, their differential RNA levels in developing human placenta have been studied using radioactive probes such as v-abl, v-erbA, v-fms, v-mos, v-myc, N-ras and v-src. The c-mos and N-ras genes are expressed and amplified at high levels especially in term placenta, while c-abl, and c-erbA are expressed constantly during development. These findings indicate that c-mos and N-ras genes may be closely linked to normal differentiation, although c-abl and c-erbA may participate in overall developmental processes. In contrast, transcripts of c-myc and c-src are enhanced at first trimester and decreased sequentially thereafter, showing that these genes may play a role in early proliferation. Expression patterns of c-fms gene are same as that of c-myc and c-src except reelevation at term. In addition, to characterize the effect of cellular oncogene expression has been also examined in hydatidiform mole and tumor cells such as BeWo and choriocarcinoma. All cellular oncogenes examined in this study were significantly overexpressed. Thus, our results suggest that cellular oncogene activation may be strongly associated with neoplastic change of trophoblast.

Levels of paraquat in fatal intoxications.

We describe here fatal levels of paraquat in plasma of victims due to the ingestion of the herbicide, paraquat, and the relationship between those levels and survival times after ingestion. We determined paraquat levels in plasma of 106 paraquat poisoning cases in Korea between June 1992 and December 1996 using a visible spectrophotometric method based on the alkali-dithionite reaction. Among 55 cases with known survival times, plasma paraquat levels of victims who died within one day of ingestion ranged from 2.3 to 636.6 microg/ml (average 127.6 microg/ml) while those of the victims who died between 1 approximately 4 days ranged from 0.9 to 25.1 microg/ml (average 7.0 microg/ml). Since preparations containing 24.5% paraquat dichloride are still used in Korea, our data for Korean victims who died within one day were 8.5 times higher than those in Baselt and Cravey's report for the same survival times. Our data also suggest that victims in whom a plasma paraquat concentration of more than 30 microg/ml was detected are likely to have died within 24 h after ingestion.

Forensic toxicological implication of acute fatal poisoning cases due to benfuracarb ingestion.

We describe here three cases involving acute fatalities due to benfuracarb ingestion and the forensic toxicological implications. Benfuracarb, a carbamate insecticide and its main metabolite carbofuran, were detected using thin layer chromatography (TLC) and gas chromatography/mass spectrophotometry (GC/MS) after extraction with ethyl acetate and then quantified using gas chromatography (GC) equipped with NPD. The blood levels of benfuracarb and carbofuran were in the range of 0.30-2.32 microg/ml and 1.45-1.47 microg/ml, respectively. Benfuracarb was not detected in urine, but carbofuran was detected in the range of 0.53-2.66 microg/ml.