Comprehensive validation of early diagnostic algorithms for myocardial infarction in the emergency department.

OBJECTIVE: To comprehensively evaluate diagnostic algorithms for myocardial infarction using a high-sensitivity cardiac troponin I (hs-cTnI) assay. PATIENTS AND METHODS: We prospectively enrolled patients with suspected myocardial infarction without ST-segment elevation from nine emergency departments in Japan. The diagnostic algorithms evaluated: (i) based on hs-cTnI alone, such as the European Society of Cardiology (ESC) 0/1-h or 0/2-h and High-STEACS pathways; or (ii) used medical history and physical findings, such as the ADAPT, EDACS, HEART, and GRACE pathways. We evaluated the negative predictive value (NPV), sensitivity as safety measures, and proportion of patients classified as low or high-risk as an efficiency measure for a primary outcome of type 1 myocardial infarction or cardiac death within 30 days. RESULTS: We included 437 patients, and the hs-cTnI was collected at 0 and 1 hours in 407 patients and at 0 and 2 hours in 394. The primary outcome occurred in 8.1% (33/407) and 6.9% (27/394) of patients, respectively. All the algorithms classified low-risk patients without missing those with the primary outcome, except for the GRACE pathway. The hs-cTnI-based algorithms classified more patients as low-risk: the ESC 0/1-h 45.7%; the ESC 0/2-h 50.5%; the High-STEACS pathway 68.5%, than those using history and physical findings (15-30%). The High-STEACS pathway ruled out more patients (20.5%) by hs-cTnI measurement at 0 hours than the ESC 0/1-h and 0/2-h algorithms (7.4%). CONCLUSIONS: The hs-cTnI algorithms, especially the High-STEACS pathway, had excellent safety performance for the early diagnosis of myocardial infarction and offered the greatest improvement in efficiency.

Effects of preemptive analgesia with intravenous acetaminophen on postoperative pain relief in patients undergoing third molar surgery: a prospective, single-blind, randomized controlled trial

BACKGROUND: The efficacy of preemptive analgesia in managing postoperative pain remains controversial. The aim of this study was to compare the efficacy of intravenous (IV) acetaminophen administered before or immediately after the surgical extraction of an impacted mandibular third molar. MATERIAL AND METHODS: This prospective randomized clinical trial included 120 patients. The patients were assigned to one of three groups: the preoperative-treatment group (pre-group), which received 1000 mg of IV acetaminophen 20 min before surgery; the postoperative-treatment group (post-group), which received 1000 mg of IV acetaminophen after surgery; the no-treatment group (control-group), which did not receive any analgesic. Rescue analgesic (60 mg loxoprofen) was issued to each patient, with instructions on self-administration if needed. For the rescue medication usage, the time of first loxoprofen usage and the total amount of loxoprofen consumption were obtained for a 17-hour period after surgery. We measured pain using the visual analogue scale at 1 hour and at 2, 3, 4, 5, and 15 hours after surgery. RESULTS: There was no significant difference in pain level among the three groups at any time interval. However, the pre-group demonstrated significantly lower rescue analgesic consumption and longer time until initial administration. CONCLUSIONS: Administration of IV acetaminophen before third molar surgery provides more effective pain control than postoperative administration and no treatment

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Effects of preemptive analgesia with intravenous acetaminophen on postoperative pain relief in patients undergoing third molar surgery: a prospective, single-blind, randomized controlled trial.

BACKGROUND: The efficacy of preemptive analgesia in managing postoperative pain remains controversial. The aim of this study was to compare the efficacy of intravenous (IV) acetaminophen administered before or immediately after the surgical extraction of an impacted mandibular third molar. MATERIAL AND METHODS: This prospective randomized clinical trial included 120 patients. The patients were assigned to one of three groups: the preoperative-treatment group (pre-group), which received 1000 mg of IV acetaminophen 20 min before surgery; the postoperative-treatment group (post-group), which received 1000 mg of IV acetaminophen after surgery; the no-treatment group (control-group), which did not receive any analgesic. Rescue analgesic (60 mg loxoprofen) was issued to each patient, with instructions on self-administration if needed. For the rescue medication usage, the time of first loxoprofen usage and the total amount of loxoprofen consumption were obtained for a 17-hour period after surgery. We measured pain using the visual analogue scale at 1 hour and at 2, 3, 4, 5, and 15 hours after surgery. RESULTS: There was no significant difference in pain level among the three groups at any time interval. However, the pre-group demonstrated significantly lower rescue analgesic consumption and longer time until initial administration. CONCLUSIONS: Administration of IV acetaminophen before third molar surgery provides more effective pain control than postoperative administration and no treatment.

TTF-1/NKX2-1 binds to DDB1 and confers replication stress resistance to lung adenocarcinomas.

TTF-1, also known as NKX2-1, is a transcription factor that has indispensable roles in both lung development and physiology. We and others have reported that TTF-1 frequently exhibits high expression with increased copy number in lung adenocarcinomas, and also has a role as a lineage-survival oncogene through transcriptional activation of crucial target genes including ROR1 and LMO3. In the present study, we employed a global proteomic search for proteins that interact with TTF-1 in order to provide a more comprehensive picture of this still enigmatic lineage-survival oncogene. Our results unexpectedly revealed a function independent of its transcriptional activity, as TTF-1 was found to interact with DDB1 and block its binding to CHK1, which in turn attenuated ubiquitylation and subsequent degradation of CHK1. Furthermore, TTF-1 overexpression conferred resistance to cellular conditions under DNA replication stress (RS) and prevented an increase in consequential DNA double-strand breaks, as reflected by attenuated induction of pCHK2 and γH2AX. Our findings suggest that the novel non-transcriptional function of TTF-1 identified in this study may contribute to lung adenocarcinoma development by conferring tolerance to DNA RS, which is known to be inherently elicited by activation of various oncogenes.

Proposal of a new mechanism for the directional propagation of the action potential using a mimicking system.

A nerve conduction model is constructed by using some liquid-membrane cells that mimic the function of the K+ and Na+ channels. By imitating two types of Na+ channels (ligand-gated Na+ channels and voltage-gated Na+ channels), a new mechanism for the directional propagation of the action potential along the axon toward the axon terminal is proposed. When the nerve cell is excited by an external (outer) stimulus, it can be presumed that the ligand-gated channels work as power sources at the synapse to propagate the change in the membrane potential, and then the voltage-gated channels locally assist the propagation at each site of the axon (nodes of Ranvier).

Regulation of the lysophosphatidylserine and sphingosine 1-phosphate levels in autologous whole blood by the pre-storage leukocyte reduction.

BACKGROUND AND OBJECTIVES: The effect of leukoreduction and storage periods on the accumulation of bioactive lysophospholipids and substances in human autologous blood (AB units) has not been fully investigated. MATERIALS AND METHODS: The accumulation of bioactive lysophospholipids such as sphingosine 1-phosphate (S1P) and lysophosphatidylserine (LysoPS) in AB units during the storage was investigated. The time-dependent changes and the effect of the filtration in pre-storage leuckoreduction (LR) and unmodified samples derived from 46 AB units were analysed. Additionally, the changes of lysophospholipids and platelet releasate, namely ß-thromboglobulin (ß-TG), induced by exposure of whole blood (WB) or platelet-rich plasma (PRP) to the filter material were analysed. RESULTS: LysoPS, but not S1P levels, time-dependently and significantly increased in both unmodified and LR samples. LysoPS significantly decreased in LR compared with unmodified samples, whereas S1P increased in LR compared with unmodified samples. In addition, exposure of WB and/or PRP to the filter material in vitro resulted in increased levels of S1P, LysoPS and ß-TG. CONCLUSIONS: LR effectively reduced the accumulation of LysoPS in AB units. On the other hand, it increased concentrations of S1P due to platelet activation by exposure to the filter material. These suggest that increases of S1P levels in LR and LysoPS in the unmodified samples were mainly caused by the leukocytes and/or platelets and that LR was effective in inhibiting the accumulation of LysoPS.

Lung cancer associated with an azygos lobe successfully treated with video-assisted thoracoscopic surgery.

Primary lung cancer associated with an azygos lobe is extremely rare. Herein, we report the case of a 64-year-old woman with lung adenocarcinoma arising in the right upper lobe with an azygos lobe. The patient underwent a right upper lobectomy and lymph node dissection with video-assisted thoracoscopic surgery. Video-assisted thoracoscopic surgery lobectomy for lung cancer of this variant has yet to be reported. We demonstrate the intraoperative findings and the resected pulmonary lobe. This is the first case of primary lung cancer associated with the azygos lobe treated by lobectomy with video-assisted thoracoscopic surgery.

A novel technology for on-site cupric oxide recovery from cupric chloride etchant waste.

For on-site copper recovery in print circuit board factories, we propose a novel technology to obtain cupric oxide with a low content ratio of chloride from high chloride concentration waste, such as cupric chloride etchant waste. Our technology is designed to avoid formation of double salt and accumulation of cupric hydroxide. In the proposed method, etchant waste mixed with hydrogen peroxide solution is added to sodium hydroxide solution by stepwise addition. We performed lab-scale experiments on the influence of reaction pH conditions on the content ratio of chloride in recovering cupric oxide. The results show that recycled cupric oxide tends to contain a lower content ratio of chloride under higher starting temperatures and higher final pH conditions of the reaction. We also confirmed the optimized conditions; the starting temperature of the sodium hydroxide solution is higher than 70 degrees C, and the final pH of the reaction is 11.5 to 12. Based on the optimized temperature and pH conditions, we also performed a pilot trial to recover cupric oxide from real etchant waste. Then, we successfully obtained cupric oxide with a content ratio of chloride in 80 mg-Cl/kg-CuO.

Vascular endothelial growth factor expression by hyalocytes and its regulation by glucocorticoid.

AIM: Tumour necrosis factor-alpha (TNF-alpha) is one of the major inflammatory cytokines involved in the pathogenesis of various vitreoretinal diseases. The authors investigated the effect of hypoxia, TNF-alpha and dexamethasone on vascular endothelial growth factor (VEGF) expression by cultured hyalocytes. METHODS: Hyalocytes were isolated from bovine vitreous. Hypoxic and TNF-alpha-dependent effects on cultured hyalocytes were investigated using several assays to determine VEGF protein expression, hypoxia-inducible factor (HIF)-1alpha protein levels, HIF-1alpha-DNA-binding ability and VEGF mRNA stability. The effects of dexamethasone on VEGF expression and its intracellular signalling under hypoxic or TNF-alpha stimulated conditions were also examined. RESULTS: Hypoxic conditions and TNF-alpha stimulation induce VEGF expression in hyalocytes. These stimuli also stabilise HIF-1alpha protein and increase its DNA-binding ability. Dexamethasone significantly inhibits both HIF-1alpha protein levels and HIF-1alpha-DNA-binding activity, and also decreases the hypoxic- and TNF-alpha -dependent induction of VEGF expression in hyalocyte. However, dexamethasone has no significant effect on the stability of VEGF mRNA. CONCLUSIONS: Hyalocytes may be involved in various vitreoretinal diseases by increasing HIF-1alpha protein stability and HIF-1alpha-DNA binding, and thus increasing VEGF production under pathological conditions. Dexamethasone seems to be capable of inhibiting hypoxic and TNF-alpha dependent VEGF production, presumably via its inhibitory effects on HIF-1alpha protein levels and its DNA-binding activity.

Stearoyl-CoA desaturase mRNA expression during bovine adipocyte differentiation in primary culture derived from Japanese Black and Holstein cattle.

Stearoyl-CoA desaturase (SCD) catalyzes the synthesis of monounsaturated fatty acids (MUFAs). In cattle, the MUFAs are related to softness and flavor of meat. In order to investigate gene expression profile during bovine preadipocyte differentiation, we isolated stromal-vascular cells from perirenal adipose tissues of Japanese Black and Holstein steers. Gene expression level of adipocyte type fatty acid binding protein (FABP4), SCD, sterol regulatory element binding protein 1 (SREBP1) and CCAAT/enhancer binding protein alpha (C/EBP-alpha) were elucidated by real-time PCR assay. The levels of SCD mRNA expression were significantly increased to 10.8 and 6.3-fold in Japanese Black and Holstein, respectively, on day 1 of the culture. The difference in SCD expression between the two breeds may reflect differences in the fat development characteristics of the cattle breeds. Although transcription factors SREBP1 and C/EBP-alpha are supposed to regulate SCD expression, expression levels of the two factors were not completely consistent with that of SCD.

No effect of fluvastatin on the bone mineral density of children with minimal change glomerulonephritis and some focal mesangial cell proliferation, other than an ameliorating effect on their proteinuria.

AIM: There are conflicting data regarding the clinical benefit of the effect of HMG-CoA reductase inhibitors (statins) in osteoporosis. We have reported that fluvastatin (a statin) is effective in improving proteinuria and renal function in childhood IgA nephropathy with mild histological findings and moderate proteinuria. The aim of the present study was to clarify the effect of fluvastatin on the bone mineral density, bone metabolic markers, proteinuria, and renal function of children with minimal change glomerulonephritis with some focal mesangial cell proliferation whose glomeruli did not stain positive for IgA and on moderate proteinuria. PATIENTS AND METHODS: We conducted a prospective controlled study of 36 children who had recently been diagnosed with normocholesterolemic minimal change glomerulonephritis with some focal mesangial cell proliferation and moderate proteinuria, and in whom strenuous exercise was restricted. The 36 patients were randomly assigned to receive 20 mg of fluvastatin (group 1) or 5 mg/kg of dipyridamole (group 2) for two years. RESULTS: By the end of the trial, there was no difference in BMD between the groups, and there were no changes in the four bone metabolic parameters. However, the urinary protein, hematuria and BUN levels had significantly decreased in group 1 compared to baseline, and the serum total protein and albumin levels and creatinine clearance had significantly increased in group 1 compared to baseline and group 2. CONCLUSIONS: The results of this study suggest that fluvastatin therapy has an antiproteinuric effect and improves renal function in moderately proteinuric patients with mild histological glomerulonephritis, but does not increase BMD.

Estimation of estrogenic and antiestrogenic activities of selected pesticides by MCF-7 cell proliferation assay.

Estrogenic activities of 20 selected pesticides-which are used for agricultural production as insecticides, fungicides and herbicides-were examined by estrogen receptor (ER)-dependent MCF-7 cell proliferation assay. Among them, chlordecone, dicofol, methoxychlor, gamma-HCH, fenarimol, EPN, triadimefon, and triadimenol had estrogenic activities, all of which were suppressed by the addition of pure antiestrogen ICI 182,780. The first 5 compounds exhibited binding capacities to ERalpha. The antiestrogenic activity of a compound was examined by estimating its suppressive effect on cell proliferation induced by 30 pM 17beta-estradiol. Strongly suspected antiestrogens were captan and myclobutanil, both of which were found to have the capacity to bind to ERalpha and which might exert their activities by competing at the level of ERalpha. Antiestrogenic activities of nitrofen, fenitrothion, fenarimol and triadimefon were also suggested. Affinities of the compounds for ERalpha and/or androgen receptor (AR) were lower than those of synthetic estrogen (diethylstilbestrol) and testosterone (mibolerone), respectively. Fenitrothion had the highest affinity to AR. Chlordecone, dicofol, methoxychlor, nitrofen, fenarimol, myclobutanil and pyridate had capacities to bind both ERalpha and AR. Chlordecone and pyridate were much more effective as competitors of estrogen binding to ERalpha than androgen binding to AR and, conversely, nitrofen was a more effective competitor of androgen binding to AR.

Vitamin D deficiency rickets due to soybean milk.

We describe vitamin D deficiency rickets in an infant nursed with soybean milk (not specifically designed for infants) instead of cow's milk-based infant formula. Symptoms included irritability and carpopedal spasm when crying. The infant's condition improved with nutrition by vitamin-enriched cow's milk-based infant formula, oral administration 1 alpha-hydroxy vitamin D3 and exposure to sunlight. Content analysis of the milk showed very low calcium, phosphate, magnesium and vitamin D levels compared to cow's milk-based infant milk formulas. This case highlights the unsuitability of soybean milk as the sole provider of infant nutrition and demonstrates the false perception that soybean milk is a healthy food for infants. It is necessary to be cautious about not only health claims for soybean milk, but also today's health and natural food booms. Social enlightenment and correction of such misperceptions are necessary.

Effect of fluvastatin and dipyridamole on proteinuria and renal function in childhood IgA nephropathy with mild histological findings and moderate proteinuria.

AIM: In recent reports, some kinds of HMG-CoA reductase inhibitors were able to decrease proteinuria and to improve renal function. Here we aimed to clarify the effect of fluvastatin (an HMG-CoA reductase inhibitor) on proteinuria and renal function in children with mild IgA nephropathy. PATIENTS AND METHODS: We conducted a prospective controlled study of 30 children who had been recently diagnosed with normocholesterolemic IgA nephropathy following the detection of a minor lesion or of focal mesangial proliferation and moderate proteinuria. The 30 patients were randomly assigned to receive both of 20 mg of fluvastatin and 5 mg/kg of dipyridamole (group 1), or 5 mg/kg of dipyridamole only (group 2) for 1 year. RESULTS: By the end of the trial, urinary protein, hematuria, BUN and serum creatinine levels had significantly decreased in the patients of group 1 as compared to baseline. Serum total cholesterol, triglyceride and LDL cholesterol levels had significantly decreased, while serum total protein and albumin, and creatinine clearance had significantly increased in group 1 as compared to baseline and group 2. The urinary protein level had significantly decreased in the group 2 patients as compared to baseline, but only slightly. CONCLUSIONS: The results of this study suggest that fluvastatin and dipyridamole treatment yields an antiproteinuric effect and leads to the amelioration of renal function in moderately proteinuric patients with mild histological IgA nephropathy.

Cell death induced by the phenolic antioxidant tert-butylhydroquinone and its metabolite tert-butylquinone in human monocytic leukemia U937 cells.

2-tert-Butyl-4-hydroquinone (TBHQ), a phenolic antioxidant used as a food additive, and its metabolite 2-tert-butyl-1,4-benzoquinone (TBQ) were both cytotoxic in human monocytic leukemia U937 cells, TBQ being the more strongly cytotoxic. Both compounds induced caspase activity towards DEVD-MCA as a substrate and the cleavage of poly(ADP-ribose) polymerase in cells. Enzyme activities of caspase-3,-7,-6 and -9 seemed to be induced, and procaspases-3 and-7 were processed to active forms in cells treated with TBHQ and TBQ. They induced nuclear condensation and fragmentation in some cells. Electron microscopic examination revealed severe disruption of mitochondrial structure and the formation of intracellular vacuoles. Morphological changes were more marked in the cells treated with TBHQ than TBQ. Mitochondrial transmembrane potential was disrupted. Cytochrome c was released from mitochondria to cytosol and ATP level was moderately decreased by the treatment of cells with these chemicals. Cellular glutathione (GSH) appeared to contribute to defense against cell death induced by TBQ, but its contribution was not marked in the case of TBHQ. TBHQ and TBQ exhibited the apoptotic features in various assays, but the mode of cell death may not be defined as a typical apoptosis or necrosis.

Development of efficient strategies for the production of genetically modified pigs.

Although pronuclear DNA micro-injection has long been the most reliable method to produce transgenic pigs, the efficiency of production of transgenic offspring is generally plagued by 1% of the DNA-injected embryos. Therefore, a problem with this method is the need for large numbers of pronuclear stage embryos. One great advancement would be the use of in vitro-matured (IVM) oocytes for the purpose of transgenic pig production. High developmental competence of IVM oocytes was proven by transfer of parthenogenetic IVM oocytes. A combined method of sperm vectors with the IVM of oocytes would make the production of transgenic pigs remarkably feasible. Rate of blastocyst formation following intracytoplasmic sperm injection (ICSI) by frozen sperm was over 20%, and transgene was expressed in approximately 50% of blastocysts generated. Somatic cell nuclear transfer would enable more efficient and sophisticated genetic modification of the pig. Simultaneous comparison between two nuclear transfer methods by electro-fusion and intracytoplasmic injection revealed clear differences in the pattern of nuclear remodeling and development of the reconstructed embryos. To specify the donor cell type that allows efficient genetic modification and easy reprogramming or to establish such cell lines is a critical issue in pig cloning. We tested pre-adipocytes from the subcutaneous adipose tissue of adult pigs for nuclear transfer. Cell cycle synchronization by differentiation induction is unique to the pre-adipocytes. Frequency of apoptosis was low in the cells synchronized by differentiation induction compared with other synchronization methods, including serum starvation, confluency, and chemical treatment. It would be of great worth if cryopreserved clone embryos were available. We have demonstrated that cryopreservation of in vitro-produced porcine embryos as well as clone blastocysts is possible by our unique method.

Bone growth oscillation: longitudinal metabolic process of bone growth in congenital adrenal hyperplasia and nonendocrine short stature.

To clarify the longitudinal metabolic process of bone growth in children, we observed the relationship between the level of serum osteocalcin (OC), a marker of bone metabolism, and growth velocity in 10 prepubertal patients with congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency and 9 prepubertal patients with nonendocrine short stature (NESS), but no major hormonal abnormalities influencing bone metabolism. Observations were made every 6 months over a 7-year period. In patients with CAH who exhibited a wide variation in growth velocity during the course of the investigation, the levels of OC fluctuated over a wide range, suggesting metabolically variable bone growth. In contrast, in patients with NESS who exhibited a relatively stable growth velocity, the OC level remained within a narrow range, suggesting metabolically stable bone growth. The meaning of such divergent metabolic processes of bone growth observed in CAH and NESS and its relationship to actual bone structure or bone intensity should be further investigated.

Production of active bovine cathepsin C (dipeptidyl aminopeptidase I) in the methylotrophic yeast Candida boidinii.

The heterologous production of active bovine cathepsin C (CTC; dipeptidyl aminopeptidase I) was investigated. Attempts to express CTC in Escherichia coli were hampered by formation of inclusion bodies that were partially degraded. To overcome this impediment, secretion of recombinant CTC was attempted in the methylotrophic yeast Candida boidinii. A DNA fragment encoding bovine procathepsin C was synthesized based on preferred codon usage in C. boidinii and placed downstream of the C. boidinii proteinase A signal sequence resulting in secretion of active CTC into the culture medium. The gene was expressed under the control of the methanol-inducible formate dehydrogenase gene promoter. Production levels were significantly improved by using a protease-deficient strain, changing medium composition, and by lowering the temperature of induction. When the recombinant C. boidinii was grown for 90 h in a jar-fermenter, active CTC was secreted with a yield of up to approximately 12 mg/l.

2-Amino-3-carboxy-1,4-naphthoquinone affects the end-product profile of bifidobacteria through the mediated oxidation of NAD(P)H.

Glucose metabolism of bifidobacteria in the presence of 2-amino-3-carboxy-1,4-naphthoquinone (ACNQ), a specific growth stimulator for bifidobacteria, and ferricyanide (Fe(CN)(6)(3-)) as an extracellular electron acceptor was examined using resting cells of Bifidobacterium longum and Bifidobacterium breve. NAD(P)H in the cells is oxidized by ACNQ with the aid of diaphorase activity, and reduced ACNQ donates the electron to Fe(CN)(6)(3-). Exogenous oxidation of NADH by the ACNQ/Fe(CN)(6)(3-) system suppresses the endogenous lactate dehydrogenase reaction competitively, which results in the remarkable generation of pyruvate and a decrease in lactate production. In addition, a decrease in acetate generation is also observed in the presence of ACNQ and Fe(CN)(6)(3-). This phenomenon could not be explained in terms of the fructose-6-phosphate phosphoketolase pathway, but suggests rather that glucose is partially metabolized via the hexose monophosphate pathway. This was verified by NADP(+)-induced reduction of Fe(CN)(6)(3-) in cell-free extracts in the presence of ACNQ. Effects of the ACNQ/Fe(CN)(6)(3-) system on anaerobically harvested cells were also examined. Stoichiometric analysis of the metabolites from the pyruvate-formate lyase pathway suggests that exogenous oxidation of NADH is an efficient method to produce ATP in this pathway.