Atopic dermatitis phenotype with early onset and high serum IL-13 is linked to the new development of bronchial hyperresponsiveness in school children.

BACKGROUND: Atopic dermatitis (AD) is characterized by a heterogeneous clinical spectrum, and some forms of AD are associated with the initial steps of allergic march. The aims of this study were to determine AD phenotypes in school-age children and investigate their associations with the allergic march in each cluster. METHODS: We included 242 children (6-8 years) with current AD from the Children's HEalth and Environmental Research study, a 4-year prospective follow-up study with 2-year survey intervals. Latent class analysis was used. Serum IL-13 and thymic stromal lymphopoietin (TSLP) levels at the time of enrollment were measured using ELISA. RESULTS: We identified four current AD phenotypes in children, characterized as 'early onset with low atopy' (26.4% of the sample; group 1), 'early onset with high atopy and high eosinophil percentages' (48.3%; group 2), 'late onset with low atopy' (9.9%; group 3), and 'late onset with high atopy and normal eosinophils' (15.3%; group 4). Although groups 2 and 4 demonstrated high atopic burden, children in group 2 showed the persistence of AD and eosinophilia associated with a high prevalence of new cases of bronchial hyper-responsiveness and asthma symptoms during follow-up. The serum IL-13 level was significantly increased in the early-onset AD groups, but there was no significant difference in the serum TSLP levels across all four groups. CONCLUSION: An allergic march-associated AD phenotype exists that is characterized by early onset of AD with its persistence, increased serum IL-13 levels, high atopy, and a persistently increased blood eosinophil percentage.

Lateral spread response monitoring during microvascular decompression for hemifacial spasm. Comparison of two targets of partial neuromuscular blockade.

AIM: The aim of the present study was to determine (1) whether successful intraoperative electromyography monitoring for lateral spread response (LSR) is possible with partial neuromuscular blockade (NMB) in subjects undergoing microvascular decompression (MVD) for hemifacial spasm and (2) the adequate level of NMB to achieve that goal. MATERIAL AND METHODS: A total of 61 patients in whom LSR was monitored during MVD were enrolled in the study. Patients were randomly allocated to two groups: group TOF in which the NMB target was maintenance of two train-of-four (TOF) counts and group T1 in which the NMB target was maintenance of a T1/Tc ratio of 50 % (T1: first twitch height of TOF and Tc: control twitch height). The adductor pollicis brevis muscle was used to monitor TOF responses. The frequency of successful LSR monitoring, defined as successful baseline establishment and maintenance of LSR until surgical decompression, was compared between the two groups. RESULTS: Of the 61 patients 2 were excluded from the study so that 30 patients in group TOF and 29 patients in group T1 were analyzed. The success rate of LSR monitoring was clinically acceptable and significantly higher in group T1 than in group TOF, i.e. n = 15 (50.0 %) in group TOF versus n = 24 (82.8 %) in group T1 (P = 0.008), corresponding to a 32.8 % higher success rate in group T1 than group TOF (95 % CI: 13.9-51.7 %). Mean vecuronium infusion dose was smaller and mean TOF count was higher in group T1 than group TOF with a TOF count = 2 (1) in group TOF versus 3 (1) in group T1 (P = 0.003). Mean sevoflurane and remifentanil infusion doses were not different between groups. There was no incidence of spontaneous movement during microscopy in either group. CONCLUSION: Maintenance of partial NMB with a target T1/Tc ratio of 50 % resulted in a clinically acceptable success rate of LSR monitoring and surgical condition during MVD. Maintenance of partial NMB with a target T1/Tc ratio of 50 % rather than TOF count of two during LSR monitoring for MVD can therefore be recommended.

Control of protein phosphatase 2A by simian virus 40 small-t antigen.

Soluble, monomeric simian virus 40 (SV40) small-t antigen (small-t) was purified from bacteria and assayed for its ability to form complexes with protein phosphatase 2A (PP2A) and to modify its catalytic activity. Different forms of purified PP2A, composed of combinations of regulatory subunits (A and B) with a common catalytic subunit (C), were used. The forms used included free A and C subunits and AC and ABC complexes. Small-t associated with both the free A subunit and the AC form of PP2A, resulting in a shift in mobility during nondenaturing polyacrylamide gel electrophoresis. Small-t did not interact with the free C subunit or the ABC form. These data demonstrate that the primary interaction is between small-t and the A subunit and that the B subunit of PP2A blocks interaction of small-t with the AC form. The effect of small-t on phosphatase activity was determined by using several exogenous substrates, including myosin light chains phosphorylated by myosin light-chain kinase, myelin basic protein phosphorylated by microtubule-associated protein 2 kinase/ERK1, and histone H1 phosphorylated by protein kinase C. With the exception of histone H1, small-t inhibited the dephosphorylation of these substrates by the AC complex. With histone H1, a small stimulation of dephosphorylation by AC was observed. Small-t had no effect on the activities of free C or the ABC complex. A maximum of 50 to 75% inhibition was obtained, with half-maximal inhibition occurring at 10 to 20 nM small-t. The specific activity of the small-t/AC complex was similar to that of the ABC form of PP2A with myosin light chains or histone H1 as the substrate. These results suggested that small-t and the B subunit have similar qualitative and quantitative effects on PP2A enzyme activity. These data show that SV40 small-antigen binds to purified PP2A in vitro, through interaction with the A subunit, and that this interaction inhibits enzyme activity.

AH/PH domain-mediated interaction between Akt molecules and its potential role in Akt regulation.

The cytoplasmic serine-threonine protein kinase coded for by the c-akt proto-oncogene features a protein kinase C-like catalytic domain and a unique NH2-terminal domain (AH domain). The AH domain is a member of a domain superfamily whose prototype was observed in pleckstrin (pleckstrin homology, or PH, domain). In this communication, we present evidence that the AH/PH domain is a domain of protein-protein interaction which mediates the formation of Akt protein complexes. The interaction between c-akt AH/PH domains is highly specific, as determined by the failure of this domain to bind AKT2. The AH/PH domain-mediated interactions depend on the integrity of the entire domain. Akt molecules with deletions of the NH2-terminal portion (amino acids 11 to 60) and AH/PH constructs with deletions of the C-terminal portion of this domain (amino acids 107 to 147) fail to interact with c-akt. To determine the significance of these findings, we carried out in vitro kinase assays using Akt immunoprecipitates from serum-starved and serum-starved, platelet-derived growth factor-stimulated NIH 3T3 cells. Addition of maltose-binding protein-AH/PH fusion recombinant protein, which is expected to bind Akt, to the immunoprecipitates from serum-starved cells induced the activation of the Akt kinase.

Calcitonin gene-related peptide protects rats from cerebral ischemia/reperfusion injury via a mechanism of action in the MAPK pathway.

The aim of the present study was to investigate the protective function and underlying mechanism of calcitonin gene-related peptide (CGRP) on cerebral ischemia/reperfusion damage in rats. Adult male Wistar rats were selected for the establishment of an ischemia/reperfusion injury model through the application of a middle cerebral artery occlusion. Animals were randomly divided into 6 groups of 24 animals. Drugs were administered according to the design of each group; animals were administered CGRP, CGRP8-37, PD98059 and SB20358. The neurobehavioral scores of the rat cerebral ischemia model in each group were calculated. The infarction range of the rat brain tissues was observed by 2,3,5-triphenyltetrazolium chloride staining. The expression levels of three proteins, phosphorylated c-Jun N-terminal kinase (JNK)/JNK, phosphorylated extracellular signal-regulated protein kinase (ERK)/ERK and p-p38/p38, in the mitogen-activated protein kinase (MAPK) pathway in the brain tissues was detected by western blotting. The results showed that CGRP could improve the neurobehavioral function of the ischemic rats and reduce the infarction range. Western blotting results confirmed that the function of the CGRP was mediated mainly through the reduction of the JNK and p38 phosphorylation and the promotion of ERK phosphorylation. Therefore, the present study confirmed that an increase in the exogenous CRGP could effectively improve ischemia/reperfusion injury of the brain tissue. The mechanisms of action were achieved through a reduction in JNK and p38 phosphorylation and an increase in ERL phosphorylation in the MAPK pathway. These mechanisms were interdependent.

Experience with continuous infusion of recombinant activated factor VII in the Asia-Pacific region.

There is increasing interest in continuous infusion of recombinant activated factor VII (rFVIIa) as a convenient and safe alternative to intermittent bolus therapy. In the Australian patients reported in this paper, cost savings of up to 25% in the first 12 h of treatment with continuous infusion of rFVIIa have been achieved safely, suggesting that substantial overall savings are possible. However, in the Thai patient reported, a dose reduction of 35% in the first 12 h was associated with poor haemostatic control, suggesting that a dose reduction of >25% may be inadvisable. The indications for treatment in the five Australian patients were: retroperitoneal haemorrhage (n = 3); right forearm compartment syndrome (n = 1); wrist haemarthrosis and median nerve compression (n = 2); sublingual haematoma (n = 1); and cerebral (mid-brain) haemorrhage (n = 1). Treatment was effective in four out of five patients (six bleeding episodes) and there was one treatment failure where treatment had been substantially delayed. The Thai patient was treated as part of a prospective, uncontrolled, observational study of 34 bleeding episodes in 22 patients in the Asia-Pacific region. Treatment was judged ineffective after 24 h, but full haemostatic control was subsequently achieved with intermittent rFVIIa therapy.

Synthesis and in vitro cytotoxicity of 4-alkyl- or 4-arylaminosubstituted cyclopenta[c]quinoline derivatives.

Twelve 4-substituted cyclopenta[c]quinoline derivatives were synthesized and evaluated in vitro cytotoxicity against four human cancer cell lines (HOP62, SK-OV-3, MD-MB-468 and T-47D). The compounds 6c and 6e bearing p-anisidine and pyrrolidine side chain were more active than the others.

Synthesis and in vitro cytotoxicity of 1-azaanthraquinone-3-carboxamides.

Five 1-azaanthraquinone-3-carboxamides were synthesized and evaluated in vitro cytotoxicity against four human cancer cell lines. The most active compound, 7b, exhibited cytotoxic activity comparable to doxorubicin.

Psammaplin A, a natural bromotyrosine derivative from a sponge, possesses the antibacterial activity against methicillin-resistant Staphylococcus aureus and the DNA gyrase-inhibitory activity.

Psammaplin A, a natural bromotyrosine derivative from an associated form of two sponges (Poecillastra sp. and Jaspis sp.) was found to possess the antimicrobial effect on the Gram-positive bacteria, especially on methicillin-resistant Staphylococcus aureus (MRSA). The minimal inhibitory concentration of psammaplin A against twenty one MRSAs ranged from 0.781 to 6.25 microg/ml, while that of ciprofloxacin was 0.391-3.125 microg/ml. Psammaplin A could not bind to penicillin binding protein, but inhibited the DNA synthesis and the DNA gyrase activity with the respective 50% (DNA synthesis) and 100% (DNA gyrase) inhibitory concentration 2.83 and 100 microg/ml. These results indicate that psammaplin A has a considerable antibacterial activity, although restricted to a somewhat narrow range of bacteria, probably by inhibiting DNA gyrase.

Synthesis and in vitro cytotoxicity of 2-alkylaminosubstituted quinoline derivatives.

Eight 2-alkylaminosubstituted 5,8-dimethoxy-4-methylquinolines and nine 2-alkylaminosubstituted or 2,6-disubstituted 4-methylquinoline-5,8-diones were synthesized and evaluated in vitro cytotoxicity against four human cancer cell lines (HOP62, SK-OV-3, HCT15 and SF295).

Regulatory mechanism of polarized membrane transport by glucocorticoid in renal proximal tubule cells: involvement of [Ca2+]i.

We examined the effect of glucocorticoids on brush border membrane transporters and, furthermore, the involvement of Ca2+ in its action in the primary cultured rabbit renal proximal tubule cells (PTCs). Dexamethasone (DEX, 10(-9) M) decreased Pi uptake by 17%; whereas DEX affected neither alpha-methyl-glucopyranoside (alpha-MG) uptake nor Na+ uptake. The DEX-induced inhibition of Pi uptake was due to a decrease of V(max). In contrast, other steroid hormones such as progesterone, testosterone, and 17beta-estradiol (10(-9) M) did not induce inhibition of Pi uptake. In order to examine the involvement of Ca2+ in DEX-induced inhibition of Pi uptake, PTCs were treated with A 23187 (10(-6) M, Ca2+ ionophore). A 23187 also inhibited Pi uptake, mimicking DEX action in Pi uptake. Treatments with W-7 (10(-4) M, calmodulin dependent kinase inhibitor), KN-62 (10(-6) M, Ca2+/calmodulin-dependent protein kinase II inhibitor), and BAPTA/AM (10(-6) M) or TMB-8 (10(-4) M) (intracellular Ca2+ mobilization blockers) blocked the DEX-induced inhibition of Pi uptake. However, nifedifine, methoxyverapamil (10(-6) M, L-type Ca2+ channel blockers), and EGTA (1 mM, extracellular Ca2+ chelator) did not block it. In conclusion, DEX inhibited Pi uptake via, in part, Ca2+/calmodulin pathway mediated by intracellular Ca2+ mobilization in the PTCs.

Effect of alanine-producing bacteria on the growth of chicks.

The effects of alanine-producing bacteria administered orally on the growth of chicks was investigated. The chicks were given diets containing adequate amounts of only the essential amino acids as sources of nitrogen: a basal diet (BD); a BD plus 40 g of urea per kilogram of diet (UD); a UD with bacteria named AR-8-3; a UD with bacteria named AB-605; and a BD plus 59.4 g and plus 118.7 g of L-alanine per kg of diet, respectively, from Day 8 to Day 15. The growth of chicks fed diets containing urea or alanine was significantly faster than that of the BD controls; no significant difference in growth was detected among the groups given the UD and bacteria. Since viable bacteria could not be detected from the intestinal content or from excreta for both groups given the bacteria, the establishment and achievement of alanine-producing activity by the bacteria in the gastrointestinal tract were considered doubtful. The supplement with 59.4 g of alanine per kg of diet improved the growth rate, but the higher inclusion (118.7 g of alanine per kg of diet) resulted in no further improvement.

Responses of the pancreatic digestive enzyme secretion to amino acids, glucose and cholecystokinin in chicks.

1. Pancreatic secretion of digestive enzymes, amylase, trypsinogen and chymotrypsinogen, was studied in response to the wing vein injection of digestive end products, various single amino acids and glucose, with or without cholecystokinin (CCK) in chicks. 2. Among amino acids administered, only phenylalanine significantly (P less than 0.05) increased trypsinogen and chymotrypsinogen secretions, while other amino acids did not. 3. Simultaneous injection of single amino acids with CCK increased digestive enzyme secretion to various extents depending on the kind of amino acids whereas the injection of glucose with CCK did not affect when compared with that of CCK alone. 4. By varying doses, a synergetic action of CCK plus amino acid on the secretion of pancreatic digestive enzymes was observed at 0.5 mM for valine and 5 mM for arginine.

Molecular cloning and sequence analysis of the catalytic subunit of bovine type 2A protein phosphatase.

We have isolated a cDNA clone corresponding to the Mr 38,000 catalytic subunit of bovine type 2A protein phosphatase. The cDNA was isolated from a bovine adrenal gland cDNA library through the use of oligonucleotide probes whose sequences were based on partial amino acid sequence obtained from cyanogen bromide fragments of the purified cardiac enzyme. The entire 1724-base-pair cDNA has been sequenced and found to contain an open reading frame coding for a protein of 325 amino acids having a calculated molecular weight of 37,320. The deduced amino acid sequence contains the experimentally determined sequences of five different cyanogen bromide peptides. Transfection of COS-m6 cells with the cloned cDNA resulted in transient expression of a protein that could be detected by immunoblot analysis with a monoclonal antibody directed against the purified cardiac protein phosphatase. The expressed protein had the same apparent molecular weight as the purified enzyme when analyzed by NaDodSO4/polyacrylamide gel electrophoresis, suggesting that this clone contains the entire coding region of the phosphatase mRNA. The cloned cDNA hybridizes to a mRNA of 2.0 kilobases in bovine heart and adrenal gland. Under conditions of reduced stringency, the cDNA also hybridizes to a mRNA species of 1.2 kilobases in cardiac tissue.

Effect of short chain fatty acids on the performance and intestinal weight in germ-free and conventional chicks.

1. In experiment 1, the performance and tissue weights of germ-free (GF) and conventional (CV) chicks fed on diets containing 25.4 g acetic acid/kg diet (AD) or 25.4 g kaolin/kg diet (KD) were investigated. Body weight gain in GF chicks was significantly higher on the AD, but significantly lower on the KD compared with their CV counterparts. The values for food efficiency, protein retention and energy retention followed a similar pattern to that of the body weight gain. 2. The weights of all sections of the intestine except the colon were significantly greater in CV chicks. In CV but not in GF birds the jejunum and ileum were heavier from birds fed on the AD than from those on the KD diet. 3. In experiment 2, the influence of butyric acid administration on the weight of some organs in chicks was investigated. The weight of duodenum, jejunum and ileum was significantly increased by intraperitoneal administration of butyric acid (2 ml of 100 mM solution/d) for 4 d, but no significant effect was observed by oral administration. 4. It might be suggested that short chain fatty acids such as acetic and butyric acids formed by bacterial action in the crop and subsequently absorbed are at least partly responsible for the heavier gut weight in CV birds.

Enhanced release of cholecystokinin by soya-bean trypsin inhibitor in chickens.

Whether ingestion of soya-bean trypsin inhibitor (SBTI) alters the release of cholecystokinin (CCK) was investigated in chickens. A meal of an adequate diet supplemented with SBTI (0, 100, or 1000 mg/kg) was given through a stomach tube, followed by CCK determination with specific CCK-8 antibody. The plasma CCK level increased from a basal level (control diet) of 9.6 +/- 0.6 to 13.4 +/- 0.6 and 18.1 +/- 0.8 fmol/ml plasma at 90 min after feeding the diet supplemented with 100 and 1000 mg SBTI, respectively. Since the SBTI supplementation did not affect crop emptying rates significantly, it was concluded that SBTI by itself enhanced CCK release into circulation in a dose-dependent fashion.

Enhanced release of cholecystokinin by dietary amino acids in chicks (Gallus domesticus).

1. The effect of dietary amino acids and protein on cholecystokinin (CCK) release into plasma was investigated in chicks by feeding a meal through a stomach tube, followed by the CCK determination with specific CCK-8 antibody. 2. The results showed that both isolated soya protein and an amino acid mixture simulating the amino acid composition of the soya protein increased the release of CCK, though to a lesser extent with a delayed response in the former, when added to a protein-free diet. 3. Among amino acids added singly to the protein-free diet, phenylalanine was more efficient than arginine and valine, exerting a response almost identical to the complete amino acid mixture.

Responses of the pancreatic digestive enzyme secretion to various combinations of amino acids and cholecystokinin in chicks (Gallus domesticus).

1. Effect of amino acid administration on pancreatic secretion of digestive enzymes, amylase, trypsinogen and chymotrypsinogen was studied after wing vein injection of an amino acid (AAs) mixture (Thr, Lys, Phe, Leu, Ile, Glu, Val, His, and Met) or combinations of selected amino acids, i.e. Thr + Phe + Ile, Thr + Phe, Thr + Ile or Phe + Ile, in the presence of cholecystokinin (CCK) in chicks. 2. Time course changes of enzyme output were similar in all treatment groups having a peak within 10-30 min, except for Phe + Ile that resulted in delayed induction of the enzyme release as shown by significant increases in the last 20 min compared with those in the rest. 3. When increases in enzyme outputs for the first 30 min were compared, it was shown that the three enzyme responses brought about by the administration of the AAs mixture was almost entirely accounted for by the combined injection of Thr + Phe. 4. Neither Thr + Ile nor Phe + Ile was as effective as Thr + Phe in inducing the output of these pancreatic enzymes. 5. The present results suggest that Thr and Phe may have a specific regulatory role in the secretion of pancreatic digestive enzymes in chicks when administered simultaneously.

Effect of phenylalanine on pancreatic amylase secretion in chicks (Gallus domesticus).

1. Effect of phenylalanine (Phe) on pancreatic amylase secretion in growing chicks was investigated in four experiments. 2. In Experiment 1, birds were injected through a wing vein with 0.25 ml Phe at 0, 0.1, 0.5, 2.5 and 12.5 mM in physiological saline. No significant difference was observed in amylase secretion among treatments. 3. Effect of various concentrations of Phe with cholecystokinin (CCK, 0.31 Crick unit) on amylase secretion was investigated in Experiment 2. Amylase secretion increased with time, although no significant effect was detected in Phe treatment. 4. Efficacy of Phe and tyrosine (Tyr) injection with CCK on amylase secretion was compared. There was no significant difference between Phe and Tyr treatments. 5. Birds were injected intraperitoneally with dl-p-chlorophenylalanine (p-CP), which is an inhibitor of phenylalanine hydroxylase, or saline 1 day before the collection of pancreatic amylase in Experiment 4. Both chicks showed increased amylase secretion with CCK (0.31 Crick unit), whereas the response was at a drastically reduced rate in chicks with the p-CP treatment.

Influence of dietary sorbose on lipogenesis in gold thioglucose-injected obese mice.

1. The influence of dietary sorbose on food intake and fatty acid synthesis of the liver and epididymal white adipose tissue (EWAT) was investigated in gold thioglucose (GTG)-injected obese mice from 12 to 14 weeks of age. 2. Sorbose was supplemented to a semi-purified diet at a level of 200 g/kg diet at the expense of sucrose. 3. On the last day of the experiment, fatty acids synthesis in the liver and EWAT was measured using an i.p. injection [1-14C]sodium acetate. 4. The decreases in body weight and food intake by dietary sorbose in GTG-injected obese mice were greater than those in control mice. 5. Lipid content and fatty acid synthesis in the liver and EWAT of control mice were not influenced by dietary sorbose. 6. In GTG-injected obese mice, the reduction of food intake by dietary sorbose suppressed fatty acid synthesis and lipid deposition in both liver and EWAT.