Activation of Wnt signaling has been implicated in tumorigenesis, and epigenetic silencing of Wnt antagonist genes has been detected in various cancers. In the present study, we examined the expression and methylation of DICKKOPF (DKK) family genes in gastrointestinal cancer cell lines. We found that all known DKK genes were frequently silenced in colorectal cancer (CRC) cells (DKK1, 3/9, 33%; DKK2, 8/9, 89%; DKK3, 5/9, 56% and DKK4, 5/9, 56%), but not in normal colon mucosa. DKK1, -2 and -3 have 5' CpG islands, and show an inverse relation between expression and methylation. DKK methylation also was frequently observed in gastric cancer (GC) cell lines (DKK1, 6/16, 38%; DKK2, 15/16, 94% and DKK3, 10/16, 63%), but was seen less frequently in hepatocellular carcinoma and pancreatic cancer cell lines. DKKs also were frequently methylated in primary CRCs (DKK1, 7/58, 12%; DKK2, 45/58, 78% and DKK3, 12/58, 21%) and GCs (DKK1, 15/31, 48%; DKK2, 26/31, 84% and DKK3, 12/31, 39%). Against a background of CTNNB1 or APC mutations, Dickkopfs (Dkks) were less effective inhibitors of Wnt signaling than secreted frizzled-related proteins, though over-expression of Dkks suppressed colony formation of CRC cells with such mutations. Our results demonstrate that DKKs are frequent targets of epigenetic silencing in gastrointestinal tumors, and that loss of DKKs may facilitate tumorigenesis through beta-catenin/T-cell factor-independent mechanisms.

The study demonstrated for the first time that total spasm is a risk factor, independent of other factors, for cardiovascular events in patients with vasospastic angina. Treatment with benidipine showed a better prognosis than that with diltiazem.

SummaryWe examined the influence of the concentration of sevoflurane and the degree of muscle block at the time of reversal on the activity of neostigmine. Ninety ASA 1-2 patients were anaesthetised with 0.2, 0.7 or 1.2 MAC of sevoflurane (30 patients each) in 66% nitrous oxide in oxygen. The electromyographic (EMG) response of the adductor digiti minimi was monitored at 20-s intervals after train-of-four stimulation of the ulnar nerve. The initial neuromuscular block was produced by vecuronium 100 mg.kg ¹1 . When the amplitude of the first response (T1) values had recovered to 10%, 25% or 40% of the control, neostigmine 40 mg.kg ¹1 was administered. The train-of-four ratio values were recorded at 1-min intervals during the subsequent 15-min period. Higher endtidal concentrations (p < 0:0001) and more pronounced block at the time of reversal (p < 0:0001) were associated with a delayed recovery in the train-of-four ratio. In addition, the train-of-four ratio 15 min after neostigmine administration was more dependent on the sevoflurane concentration than on the degree of block present (p < 0:0001 Previous studies have shown that volatile anaesthetics can impair neostigmine reversal of neuromuscular block [1][2][3][4]. Since the termination of anaesthesia can reduce this impairment to a great degree, it has been suggested that the administered concentration of volatile anaesthetics during reversal is an important determinant of the activity of neostigmine [3,4]. However, in these studies, reversal was attempted at only one degree of neuromuscular block [1][2][3][4]. The depth of neuromuscular block at the time of reversal is also a major factor in the activity of neostigmine [5][6][7]. Reversal at different depths of neuromuscular block should exhibit a different degree of the impairment produced by the volatile anaesthetics. An examination of how these two major factors affect the extent and speed of reversal with neostigmine was thus undertaken. Sevoflurane was selected as the volatile anaesthetic agent because we have previously shown that it impairs the neostigmine reversal of vecuronium block more profoundly than isoflurane [4]. Materials and methodsAfter obtaining approval from the local ethics committee and written informed consent, we studied 90 women of physical status ASA 1 or 2, aged 18-65 years, height 144-171 cm and weighing 41-71 kg. They required anaesthesia for minor elective surgery and patients scheduled for upper abdominal surgery were not studied. None of the patients had neuromuscular, hepatic or renal diseases and none was receiving any drugs known to affect neuromuscular function. The patients received no premedication. On arrival in the operating theatre, ECG and noninvasive blood pressure monitors were attached. The left forearm and hand were immobilised and prepared for stimulation of the ulnar nerve at the wrist using surface electrodes. The evoked electromyogram (EMG) was recorded from the adductor digiti minimi using a Datex NMT-100 Relaxograph. We used the amplitude of the first respon...

One MAC sevoflurane and isoflurane anaesthesia impair edrophonium reversal of vecuronium block to a similar degree.

A case of skin injuries due to stings by crown-of-thorns starfish, Acanthaster planci, in a 53-year-old Okinawan woman is reported. She went to a beach to gather shellfish on 8 April 2001 and fell to the ground with her left palm on a crown-of-thorns starfish that happened to be close to her. She hurried to the emergency section of our hospital. An emergency doctor sterilized the wound and administered an antibiotic, an analgesic agent and an injection of a tetanus antitoxin. He tried to remove the remaining spines from the palm with great difficulty. Because swelling and subcutaneous indurations of the left palm had persisted thereafter, oral and topical administration of corticosteroid started on 13 April. Physical examination at the dermatology section revealed approximately 10 stab wounds of the left palm with pus, subcutaneous bleeding and many abrasions around them. X-rays of the left hand showed foreign bodies, 2-10 mm in size, located on the lesions. The patient was treated with a topical injection of 2 mg triamcinolone acetonide (Kenacort-A), diluted fivefold with 1% Xylocaine, once a week. Some of the foreign body granulomatous lesions improved but pain and subcutaneous indurations persisted in most of the lesions. Because the X-ray photographs showed many remaining spines, surgical excision to remove them was performed under local anesthesia 3 months after the injury. All the symptoms improved after the operation. Scanning electron microscopic examination of the spines revealed that their tips had fragile lattice-like structures.