Lovastatin decreases the synthesis of inflammatory mediators during epileptogenesis in the hippocampus of rats submitted to pilocarpine-induced epilepsy.

Statins may act on inflammatory responses, decreasing oxidative stress and also reducing brain inflammation in several brain disorders. Epileptogenesis is a process in which a healthy brain becomes abnormal and predisposed to generating spontaneous seizures. We previously reported that lovastatin could prevent neuroinflammation in pilocarpine-induced status epilepticus (SE). In this context, this study investigated the long-lasting effects of lovastatin on mRNA expression of proinflammatory cytokines (interleukin-1ß, tumor necrosis factor α, interleukin-6) and the antiinflammatory cytokine IL-10 in the hippocampus during epileptogenesis by immunohistochemistry and real time polymerase chain reaction (RT-PCR) during the latent and chronic phases in the epilepsy model induced by pilocarpine in rats. For these purposes, four groups of rats were employed: saline (CONTROL), lovastatin (LOVA), pilocarpine (PILO), and pilocarpine plus lovastatin (PILO+LOVA). After pilocarpine injection (350mg/kg, i.p.), the rats were treated with 20mg/kg of lovastatin via an esophagic probe 2h after SE onset. All surviving rats were continuously treated during 15days, twice/day. The pilocarpine plus lovastatin group showed a significant decrease in the levels of IL-1ß, TNF-α, and IL-6 during the latent phase and a decreased expression of IL-1ß and TNF-α in the chronic phase when compared with the PILO group. Moreover, lovastatin treatment also induced an increased expression of the antiinflammatory cytokine, IL-10, in the PILO+LOVA group when compared with the PILO group in the chronic phase. Thus, our data suggest that lovastin may reduce excitotoxicity during epileptogenesis induced by pilocarpine by increasing the synthesis of IL-10 and decreasing proinflammatory cytokines in the hippocampus.

Oral administration of an HSP90 inhibitor, 17-DMAG, intervenes tumor-cell infiltration into multiple organs and improves survival period for ATL model mice.

In the peripheral blood leukocytes (PBLs) from the carriers of the human T-lymphotropic virus type-1 (HTLV-1) or the patients with adult T-cell leukemia (ATL), nuclear factor kappaB (NF-κB)-mediated antiapoptotic signals are constitutively activated primarily by the HTLV-1-encoded oncoprotein Tax. Tax interacts with the I κB kinase regulatory subunit NEMO (NF-κB essential modulator) to activate NF-κB, and this interaction is maintained in part by a molecular chaperone, heat-shock protein 90 (HSP90), and its co-chaperone cell division cycle 37 (CDC37). The antibiotic geldanamycin (GA) inhibits HSP90's ATP binding for its proper interaction with client proteins. Administration of a novel water-soluble and less toxic GA derivative, 17-dimethylaminoethylamino-17-demethoxygeldanamycin hydrochloride (17-DMAG), to Tax-expressing ATL-transformed cell lines, C8166 and MT4, induced significant degradation of Tax. 17-DMAG also facilitated growth arrest and cellular apoptosis to C8166 and MT4 and other ATL cell lines, although this treatment has no apparent effects on normal PBLs. 17-DMAG also downregulated Tax-mediated intracellular signals including the activation of NF-κB, activator protein 1 or HTLV-1 long terminal repeat in Tax-transfected HEK293 cells. Oral administration of 17-DMAG to ATL model mice xenografted with lymphomatous transgenic Lck-Tax (Lck proximal promoter-driven Tax transgene) cells or HTLV-1-producing tumor cells dramatically attenuated aggressive infiltration into multiple organs, inhibited de novo viral production and improved survival period. These observations identified 17-DMAG as a promising candidate for the prevention of ATL progression.

Activation of p53 by Nutlin-3a, an antagonist of MDM2, induces apoptosis and cellular senescence in adult T-cell leukemia cells.

It has been reported that the induction of cellular senescence through p53 activation is an effective strategy in tumor regression. Unfortunately, however, tumors including adult T-cell leukemia/lymphoma (ATL) have disadvantages such as p53 mutations and a lack of p16(INK4a) and/or p14(ARF). In this study we characterized Nutlin-3a-induced cell death in 16 leukemia/lymphoma cell lines. Eight cell lines, including six ATL-related cell lines, had wild-type p53 and Nutlin-3a-activated p53, and the cell lines underwent apoptosis or cell-cycle arrest, whereas eight cell lines with mutated p53 were resistant. Interestingly, senescence-associated-beta-galactosidase (SA-beta-gal) staining revealed that only ATL-related cell lines with wild-type p53 showed cellular senescence, although they lack both p16(INK4a) and p14(ARF). These results indicate that cellular senescence is an important event in p53-dependent cell death in ATL cells and is inducible without p16(INK4a) and p14(ARF). Furthermore, knockdown of Tp53-induced glycolysis and apoptosis regulator (TIGAR), a novel target gene of p53, by small interfering RNA(siRNA) indicated its important role in the induction of cellular senescence. As many patients with ATL carry wild-type p53, our study suggests that p53 activation by Nutlin-3a is a promising strategy in ATL. We also found synergism with a combination of Nutlin-3a and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), suggesting the application of Nutlin-3a-based therapy to be broader than expected.

Functional correlation between the nuclear localization of Fht1p and its flocculation and heat tolerance activities in budding yeast Saccharomyces cerevisiae.

Fht1p is involved in the flocculation and heat tolerance machinery of budding yeast Saccharomyces cerevisiae. Despite knowledge of its involvement in those phenotypes, a precise mechanism has yet to be discovered. To this end, we monitored the relationship between subcellular localization of Fht1p and its flocculation or heat tolerance function using newly developed expression vectors with a recombinant green fluorescent protein (GFP; S65T/S147P) of Aequorea victoria added at both the N- and C-terminus of Fht1p. The main fluorescent signal of the GFP tagged with either a wild-type Fht1p or mutants which preserve their flocculation function was detected in the nucleus, whereas signals of functionless mutants were dispersed to the cytoplasm.

Genome-wide expression changes induced by HTLV-1 Tax: evidence for MLK-3 mixed lineage kinase involvement in Tax-mediated NF-kappaB activation.

The Tax protein of human T-lymphotropic virus type 1 (HTLV-1), an oncoprotein that transactivates viral and cellular genes, plays a key role in HTLV-1 replication and pathogenesis. We used cDNA microarrays to examine Tax-mediated transcriptional changes in the human Jurkat T-cell lines JPX-9 and JPX-M which express Tax and Tax-mutant protein, respectively, under the control of an inducible promoter. Approximately 300 of the over 2000 genes examined were differentially expressed in the presence of Tax. These genes were grouped according to their function and are discussed in the context of existing findings in the literature. There was strong agreement between our results and genes previously reported as being Tax-responsive. Genes that were differentially expressed in the presence of Tax included those related to apoptosis, the cell cycle and DNA repair, signaling factors, immune modulators, cytokines and growth factors, and adhesion molecules. Functionally, we provide evidence that one of these genes, the mixed-lineage kinase MLK-3, is involved in Tax-mediated NF-kappa-B signaling. Our current results provide additional insights into Tax-mediated signaling.

Pleiotropic effects of HTLV type 1 Tax protein on cellular metabolism: mitotic checkpoint abrogation and NF-kappaB activation.

Tax protein expressed by human T cell leukemia virus type 1 (HTLV-1) is a strong trans-activator of its own LTR promoter; it also affects the function of multiple cellular genes involved in cell cycle control and transcription. One way in which Tax exerts its pleiotropic effects is through protein-protein interaction with cellular cofactors. By using yeast two-hybrid technology, we have isolated several cellular proteins that bind to Tax. Two of these are MAD1, a mitotic checkpoint control protein, and TXBP151, a suppressor of tumor necrosis factor alpha-induced apoptosis. Here we discuss findings describing the role of MAD1 in exit of cells from mitosis and TXBP151 in NF-kappaB activation.

Phosphorylation of the GTS1 gene product of the yeast Saccharomyces cerevisiae and its effect on heat tolerance and flocculation.

The GTS1 gene from the yeast Saccharomyces cerevisiae showed pleiotropic effects on yeast phenotypes, including an increase of heat tolerance in stationary-phase cells and an induction of flocculation. Here, we found that the GTS1 product, Gts1p, was partially phosphorylated at some serine residue(s) in cells grown on glucose. Studies using mutants of protein kinase A (PKA) and CDC25, the Ras-GTP exchange activator, showed that PKA positively regulated the phosphorylation level of Gts1p. Overexpression of Gts1p in a mutant with attenuated PKA activity did not show any increase of heat tolerance and partially decreased flocculation inducibility, suggesting that phosphorylation of Gts1p is required for induction of these phenomena.

A comparison between low-flow leak test and oxygen flush leak test.

PURPOSE: The clinical efficacy of two methods of preanesthetic leak test,namely the oxygen flush leak test (OFLT) and the low-flow leak test (LFLT), was compared regarding their ability to detect leakage in the anesthesia circuit and their accuracy. METHODS: Examinees comprised 16 staff anesthesiologists and 7 physicians undergoing anesthesia training at our institution. They performed the two leak tests on anesthesia machines with some intentional leaks (0.1-1.0 l.min(-1)). The leakage detection rates (LDR) were analyzed by the chi(2)-test. The ability to detect leaks was measured by recording how many leaks were detected by 50% (LDR(50)) and 95% (LDR(95)) of the examinees. RESULTS: The LDRs in the two tests were significantly different (chi(2)- analysis, P < 0.0001). Both LDR(50) and LDR(95) for the LFLT (0.23 and 0.41 l.min(-1), respectively) were smaller than the values for the OFLT (0.37 and 0.82 l.min(-1), respectively). The sensitivity and specificity of the LFLT (0.97 and 0.84, respectively) were higher than those of the OFLT (0.78 and 0.80, respectively). CONCLUSION: The LFLT was found to be superior to the OFLT regarding leak detection and reliability. We therefore recommend the LFLT for preanesthetic leak testing.

Appropriate method of administration of propofol, fentanyl, and ketamine for patient-controlled sedation and analgesia during extracorporeal shock-wave lithotripsy.

PURPOSE: The aim of this study was to identify the appropriate method for administering propofol, fentanyl, and ketamine (PFK) for patient-controlled sedation and analgesia (PCSA) during extracorporeal shock-wave lithotripsy (ESWL). METHODS: Twenty-one unpremedicated patients were randomly assigned to three groups that received different drug administration regimens. (group 1: low loading dose and high demand bolus, group 2: high loading dose and demand bolus, group 3: high loading dose and low demand bolus). RESULTS: The patients in all groups were hemodynamically stable during ESWL. Oxygen desaturation was recognized in all groups, but was avoided by 2 l.min(-1) of oxygen supply via a nasal prong. The total administration dose of the drugs was significantly higher ( P < 0.05) in group 2 than in groups 1 and 3. The median level of sedation was the same, but the episodes of oversedation were not recognized in group 3 ( P < 0.05). A significant difference in the frequency of episodes of oversedation was found between groups 2 and 3 ( P < 0.05). The results were good or excellent for almost all patients, and were assessed as fair by only one patient in group 2. CONCLUSION: We concluded that the method used for group 3 is the most appropriate for administering PFK for PCSA during ESWL.

[Caesarean section in a patient with a history of HELLP syndrome].

A 36-year-old woman underwent emergency caesarean section following the diagnosis of HELLP syndrome. Four years earlier, after having undergone the same procedure for HELLP syndrome, she had experienced hypovolemic shock, renal failure, and disseminated intravascular coagulopathy during the postoperative period. This time, the patient showed bleeding, elevation of liver enzymes (ALT, AST, LDH) and a reduction of antithrombin III activity in the 36th week of pregnancy. Anesthesia was induced by thiamylal 4 mg.kg-1 and suxamethonium 1 mg.kg-1 and after delivery maintained by oxygen-nitrous oxide-isoflurane, and all procedures were performed without any incident. No major complications such as intraperitoneal bleeding, renal failure, or disseminated intravascular coagulopathy occurred during the postoperative period. It is suggested that caesarean section should be carried out as soon as possible after the diagnosis of HELLP syndrome is confirmed.

[Leak test for the internal circuit of anesthesia machines].

In spite of detailed periodic inspection performed by specialized engineers, a great number of anesthesia machines fail to meet the standard of low flow leak test because of leak in the internal circuit. To find out the background and the solution to the problem, we sent questionnaire to 11 major manufacturers and/or dealers each responsible for periodic inspection of anesthetic machines. According to the responses to the questionnaire, the manufacturers and/or the dealers had various methods of internal circuit leak test without a unified standard in detail. The mismatch of the test methods with those anesthesia machines equipped with check valve mechanism has also led to poor evaluation of internal circuit leak. Leak test must be standardized for its appropriate application to work effectively for the problem of leak in the internal circuit of anesthesia machines.

[The current state of leak in anesthetic machines detected by low flow leak tests].

To assess the current state of leak in anesthetic machines, we selected 66 units of anesthetic machines for inspection and repair from various medical institutions. Based on a newly designed inspection flow chart a low flow leak test for internal circuits of the anesthetic machines was performed. The conventional low flow leak test was also performed for smooth detection of leak for rational evaluation. Only 39% of the anesthetic machines met the standard of the low flow leak tests, and leak was detected in the remaining 61%. The average residual leak mounted to 0.97 l.min-1, with the maximum of 5.3 l.min-1. Canisters, corrugated tubes, and vaporizers were considered the primary causes of leak. After the inspection and repair, leak in 77.5% of the anesthetic machines either disappeared or decreased and the average residual leak dropped to 0.34 l.min-1. However, 47% of the anesthetic machines still failed to meet the standard of the low flow leak tests. To further improve the situation, more detailed inspection and repair are necessary especially for precise detection of the cause of leak in the internal circuit of anesthetic machines which often remains undetected.

[Patient-controlled sedation in a patient with respiratory dysfunction for reconstructive surgery of submandibular fracture under local anesthesia].

A 70-year-old male with respiratory dysfunction, chronic obstructive pulmonary disease associated with asthmatic component, received patient-controlled sedation (PCS) using propofol for reconstructive surgery of submandibular fracture under local anesthesia. PCS produced sedation levels between eyelid closure with prompt response to verbal commands and drowsiness. Respiratory disturbance and hemodynamic change were not recognized. The patient was very cooperative, and felt comfortable during the operation. Propofol PCS is an appropriate method for a patient with respiratory dysfunction under local anesthesia.

[Differential epidural block for definitive diagnosis of snapping hip joint].

Snapping hip joint syndrome is defined as snapping phenomenon accompanied by motion of the hip joint. It occurs from various causes, and its diagnosis and treatment are often difficult. We gave anesthesia for surgery against snapping hip joint under patient's active motion for definitive regional diagnosis. Differential epidural block, which precludes pain while maintaining motor function, was performed by 0.25% bupivacaine, 0.5% lidocaine, and additional administration of fentanyl by 25 micrograms. Although the mechanism of differential nerve block remains to be unknown, the case suggests that effective pain block preserving motor function may be achieved by differential epidural block in combination with low concentration of local anesthetics and small dose of opioid.

[Is patient-controlled sedation good for elderly patients?].

The purpose of this study was to evaluate the safety and advantage of intra-operative patient-controlled sedation (PCS) in elderly patients. Propofol PCS was compared with anesthesiologist-controlled sedation (ACS) during knee arthroplasty under epidural anesthesia. Eleven elderly patients scheduled for unilateral knee total or partial arthroplasty were divided randomly into PCS group (n = 6) and ACS group (n = 5). Epidural anesthesia was performed to produce an appropriate level of sensory block (T 10 through S). Firstly a mixture of pentazocine 0.2 mg.kg-1 and 2% mepivacaine 6-9 ml was injected to the epidural space, and anaesthesia was maintained using 2% mepivacaine afterward. Patients in both groups received propofol 0.3 mg.kg-1 i.v. as a loading dose and 0.6 mg.kg-1.h-1 continuous infusion. Furthermore patients in PCS group received propofol PCS (bolus: 0.2 mg.kg-1, lockout time: 3 min). Patients in ACS group were administered propofol continuously and infusion rates were regulated to maintain a sedation score 3 (Wilson et al) by anesthesiologist. Respiratory rate, blood pressure, heart rate, SpO2, arterial blood gas analysis and plasma levels of propofol were measured 4 times during and after the surgery. Satisfaction of patients and surgeons was questioned. Patients in PCS group received a mean propofol dose of 1.9 +/- 0.1 mg.kg-1 during procedures with a mean duration of 147 min. On the other hand patients in ACS group received propofol 2.9 +/- 0.3 mg.kg-1 with 142 min of procedures. Satisfaction of patients and surgeons, the incidence of complication were similar between the groups. For elderly patients who undergo epidural anesthesia, PCS is a safe and effective technique providing similar good sedation as with ACS.

[Where is a leak point detected by "the low flow leak test" of anesthetic machines?].

"The low flow leak test" is recommended for pre-anesthetic inspection of anesthetic machines. We carried out anesthesia compression tests as a standard. Even in that case, often the low flow leak test does not meet the standard. We investigated the point where there is a leak in the anesthetic machine. Observing the leak that fluctuates each time there is detachment or attachment of the canister, the primary cause of the leak is thought to be related to the canister. It is important to carry out an inspection of the canister if the low flow leak test does not meet the standard.

Identification and characterization of a novel trans-membrane protein gene, pdh1, from Schizosaccharomyces pombe.

We have cloned a new gene, pdh1, from genomic DNA of fission yeast Schizosaccharomyces pombe. pdh1 is actively transcribed as 1400-nucleotide mRNA in vegetatively growing cells and can code for a 226 amino acid polypeptide (pdh1p). Computational structural prediction has revealed that the pdh1p is a highly hydrophobic protein with seven transmembrane domains. The prediction has also detected a possible C-kinase phosphorylation site within the longest hydrophilic loop.

Biliary hamartomas simulating multiple hepatic metastasis on imaging findings.

We encountered a 62-year-old female with advanced gastric cancer, in whom multiple nodular lesions in the liver, which were depicted as small low density areas, less than 5 mm in diameter, on computed tomography and demonstrated as a heterogenic pattern on ultrasonograms. This patient was suspected to have multiple liver metastases of gastric cancer. She deteriorated gradually and died of respiratory failure due to lung metastasis. At autopsy these hepatic nodular lesions were biliary hamartomas. Biliary hamartoma is a lesion usually identified microscopically, and there have been only 8 cases including ours, which were depicted on images. Along with a remarkable advance in imagings, it should become more important to differentiate biliary hamartoma from malignant hepatic neoplasms on images.