Total synthesis of (+)-albicanol and (+)-albicanyl acetate.

The drimane sesquiterpenes, (+)-albicanol (2) and (+)-albicanyl acetate (3), were synthesized from an optically active bicyclic diol [(+)-1] that had been obtained via the recently developed optical resolution of a general synthetic intermediate for drimane sesquiterpenes. The crucial step in the previous syntheses was markedly improved by the modified Wittig methylenation of a silyloxy ketone (7). The high overall yield (77% in 4 or 5 steps from (+)-1) by this total synthesis makes it possible to synthesize the other biologically active drimane sesquiterpenes.

Cloning of a full-length cDNA encoding ent-kaurene synthase from Gibberella fujikuroi: functional analysis of a bifunctional diterpene cyclase.

We report here the nucleotide sequence of a full-length cDNA encoding ent-kaurene synthase that was isolated by a reverse-transcription polymerase chain reaction from Gibberella fujikuroi (Gcps/ks). This cDNA encodes 952 amino acid residues with a relative molecular mass of 107 kDa. The sequence similarity between Gcps/ks and ent-kaurene synthase of the gibberellin A1-producing fungus, Phaeosphaeria sp. L487, is very high, suggesting that Gcps/ks is also a bifunctional diterpene cyclase. Its recombinant protein expressed in Escherichia coli converted geranylgeranyl diphosphate to copalyl diphosphate and ent-kaurene.

Induction of apoptosis by Coprinus disseminatus mycelial culture broth extract in human cervical carcinoma cells.

Extract of Coprinus disseminatus (pers. Fr.) (C. disseminatus) culture broth (EDCB) inhibits proliferation and induces apoptosis in the human cervical carcinoma cells at 5 microg/ml. To determine whether the cell death induced by the EDCB recruits caspases or not, one of the exclusive pathways in cell death, we examined caspase-3 activity in this cell death process. The activity of caspase-3 was remarkably increased when the cell was treated with EDCB, and this activity was nullified by Z-VAD-FMK, a well known caspase-3 inhibitor. From these results, we would expect the EDCB to contain substances with the ability to induce apoptosis in the human cervical carcinoma cells. The extent of the EDCB induced apoptosis is cell line-dependent.

Thermodynamics of Formation of and Adsorption at Interfaces with a Floating Lens.

The thermodynamic equations of the formation of and adsorption at interfaces were derived for a three-phase system having a floating lens with curved interfaces. The surface excess quantities were defined both in the H convention based on the two dividing surfaces and in the P convention based on the four dividing surfaces. Their physical significance and utilities were discussed. The equations were simplified to be applicable to the three-component systems with the plane A/W interface and a relatively large size of lens, such as the air/water/long-chain alcohol systems. The formulation and geometrical consideration of the two conventions with respect to this simple cases revealed that the H convention introduces the distance between the two dividing surfaces and the P convention introduces three kinds of distances between the four dividing surfaces including a kind of thickness of the reference oil lens phase in the A/W interfacial region. Furthermore, the general equations were simplified to be applicable to four-component systems such as the air/oil solution of a surface-active substance/water or the air/oil/aqueous solution of a surfactant with a floating oil lens. Copyright 1999 Academic Press.

Role of p53 in the ability of 1,2-diaminocyclohexane-diacetato-dichloro-Pt(IV) to circumvent cisplatin resistance.

Several reports indicate that the mechanism of resistance to cisplatin is multifactorial. However, DNA damage tolerance appears to be the more significant mechanism. It is clear that resistance in general is a major clinical concern, and a number of approaches have been taken to circumvent this clinical impediment. One approach is through analog development, and we have identified 1,2-diaminocyclohexane-diacetatodichloro-platinum(IV) as an analog with activity in cisplatin resistance. The activity is greatest against ovarian tumor cell lines where the latent, non-inducible wild-type p53 function can be reactivated by the analog. This functional activation of p53 also corresponds to a reduced threshold for tolerance to DNA damage induced by the analog. Interestingly, cell lines with mutant or null p53 are cross-resistant to the analog. The data indicate that cisplatin resistance due to an increase in DNA damage tolerance can arise through a loss of p53 function, and that functional activation of latent wild-type p53 by the analog facilitates cell death and circumvents this resistance mechanism.

Preparation, characterization, and antitumor activity of new cisplatin analogs with homopiperazines: crystal structure of [PtII(1-methylhomopiperazine)(methylmalonato)].2H2O.

A series of new platinum(II) and (IV) complexes with homopiperazine have been synthesized and characterized by elemental analysis, infrared, and 195Pt nuclear magnetic resonance spectroscopic techniques. The complexes are of two types: [PtIILX] (where L = homopiperazine (hpip), 1-methylhomopiperazine (mhpip), or 1,4-dimethylhomopiperazine (dmhpip), and X = 1,1-cyclobutanedicarboxylato (CBDCA), or methylmalonato ligand) and [PtIV(L-)trans-(Y)2Cl2] (where Y = hydroxo, acetato, or chloro ligand). Among the complexes synthesized, the crystal structure of [PtII(mhpip)(methylmalonato)].2H2O was determined by the single crystal X-ray diffraction method. The crystallographic parameters were orthorhombic, P2(1)2(1)2(1) (no. 19), a = 7.2014(14), b = 7.3348(15), c = 26.971(5) A, and Z = 4. The structure refinements converged to R1 = 0.0641 and wR2 = 0.1847. In this complex, platinum has a slightly distorted square planar geometry with the two adjacent corners being occupied by two nitrogens of the mhpip ligand, whereas the remaining cis positions are coordinated with two oxygen atoms of the methylmalonato group. The mhpip ligand is in a boat conformation and forms five and six membered chelating rings with platinum. The intricate network of intermolecular hydrogen bonds holds the crystal lattice together. Some of these synthesized cisplatin analogs have good in vitro cytotoxic activity against the cisplatin-sensitive human ovarian A2780 (IC50 = 0.083-17.8 microM) and the isogenic cisplatin-resistant 2780CP (IC50 = 20.1-118.1 microM) cell lines.

Tumor stage in the proximal colon under conditions of a proximal shift of colorectal cancer with age.

BACKGROUND/AIMS: Although the incidence of proximal colon cancer is known to increase with age, the prognosis of proximal colon cancer remains unclear. METHODOLOGY: One thousand two hundred and five patients with colorectal cancer were analyzed with regard to tumor location, age and Duke's stage. RESULTS: When Duke's stage was compared with tumor location, an advanced Duke's stage was found more frequently in the proximal colon than in the distal colorectum (p<0.01). In Duke's A colorectal cancer, the incidence of tumors limited to within the submucosal layer decreased from 73.1% in patients under 65 years of age to 60.9% in patients over 65 years of age. CONCLUSIONS: Since advanced stage cancer was more frequently found in the proximal colon than in the distal colorectum, careful examination for the early detection of proximal colon cancer, particularly in the elderly, is called for.

Natural history of hepatectomy regarding liver function: a study of both normal livers and livers with chronic hepatitis and cirrhosis.

BACKGROUND/AIMS: The cirrhotic liver has been shown to regenerate after hepatectomy based on the findings of various imaging techniques. However, little has been reported so far regarding the restorative process using the liver function test in a long series after hepatectomy of both normal and injured livers. METHODOLOGY: A retrospective review was performed of 48 patients who underwent hepatic resection due to either primary or secondary liver cancers. They were histologically divided into three groups of 6 normal, 23 chronic hepatitis and 19 cirrhotic liver patients. In addition, the hepatitis group was divided into two groups according to the resected volume, which were 16 of a small resected volume and 7 of a large volume. For these patients, both a complete blood count and normal liver function test were performed before operation, and 7 and 14 days, 1, 3, and 6 months, and 1, 2, and 3 years, postoperatively. RESULTS: In the normal patient group, almost all test results recovered to their preoperative levels by 14 days to 1 month postoperatively. In the cirrhotic group, however, the red blood cell count and hematocrit levels did not recover to preoperative levels until 3 months postoperatively, while the GPT levels, the total bilirubin and albumin all recovered by one month postoperatively. However, the total cholesterol level needed 6 months to recover. In the large resection hepatitis group, the GPT level needed 7 days to recover, while the albumin and total cholesterol level needed three months to do so. In addition, the platelet level decreased significantly at 3 months postoperatively and thereafter it seemed to remain at a lower level. CONCLUSIONS: We demonstrated that a hepatic resection in patients with chronic liver disease requires a longer period of time to recover to preoperative levels based on the type of underlying liver parenchymal disease and the extent of resection.

Appraisal of a manual blunt dissection for an intraoperative diagnosis of extrahepatic cancer invasion.

BACKGROUND/AIMS: Liver cancer extension to the adjacent organs does not necessarily indicate tumor invasion. It is very hard to diagnose extrahepatic cancer invasion to the adjacent organs using preoperative imaging. This study was undertaken, therefore, to determine the real cancer invasion using a manual dissection. METHODOLOGY: Of the 51 consecutive patients with liver cancer, 6 cases with extrahepatically growing tumors were suspected to have cancer invasion both preoperatively and intraoperatively. Thus, we diagnosed whether or not any real extrahepatic cancer invasion was present by either removing the tumour by hand or by performing a blunt gauze dissection. RESULTS: Three hepatocellular carcinomas and one metastatic cancer were stripped off from the adjacent organ using a manual blunt dissection. However, the other two tumors could not be removed in the same manner and therefore a combined resection of the invaded portion had to be performed along with the hepatectomy. A postoperative detailed pathological examination revealed no cancer cells on the surface of the adjacent organ in the former 4 tumors. Cancer invasion was recognized, however, on the outside of the liver in the latter 2 combined resected tumors. CONCLUSIONS: An accurate intraoperative diagnosis of extrahepatic cancer invasion can be made using manual blunt dissection.

Phytochrome regulates gibberellin biosynthesis during germination of photoblastic lettuce seeds.

Germination of lettuce (Lactuca sativa L.) seed is regulated by phytochrome. The requirement for red light is circumvented by the application of gibberellin (GA). We have previously shown that the endogenous content of GA1, the main bioactive GA in lettuce seeds, increases after red-light treatment. To clarify which step of GA1 synthesis is regulated by phytochrome, cDNAs encoding GA 20-oxidases (Ls20ox1 and Ls20ox2, for L. sativa GA 20-oxidase) and 3beta-hydroxylases (Ls3h1 and Ls3h2 for L. sativa GA 3beta-hydroxylase) were isolated from lettuce seeds by reverse-transcription polymerase chain reaction. Functional analysis of recombinant proteins expressed in Escherichia coli confirmed that the Ls20ox and Ls3h encode GA 20-oxidases and 3beta-hydroxylases, respectively. Northern-blot analysis showed that Ls3h1 expression was dramatically induced by red-light treatment within 2 h, and that this effect was canceled by a subsequent far-red-light treatment. Ls3h2 mRNA was not detected in seeds that had been allowed to imbibe under any light conditions. Expression of the two Ls20ox genes was induced by initial imbibition alone in the dark. The level of Ls20ox2 mRNA decreased after the red-light treatment, whereas that of Ls20ox1 was unaffected by light. These results suggest that red light promotes GA1 synthesis in lettuce seeds by inducing Ls3h1 expression via phytochrome action.

A new technique for repairing a blunt injured duodenum.

Repairing a blunt injured duodenum remains a problem for surgeons because of the difficulty in early diagnosis, associated organ injuries, and frequent anastomotic site leakage. We have devised a simpler and safer procedure for transecting the ligament of Treitz and performing a debridement of the duodenum The duodenum is freed and exposed by transecting the ligament of Treitz. The injured site is then resected with appropriate debridement. The remnant stumps of the duodenum and the jejunum are anastomosed. The advantages of our method are less invasion, a more physiologic state, and no retention of food in the repaired duodenum. This technique reduces tension in the repaired wound and also reduces mortality and morbidity.

Thermodynamic Study on the Surface Formation of the Mixture of Water and Ethanol

The formation of the water + ethanol mixture/air surface was studied by measuring the surface tension as a function of temperature and the mole fraction of ethanol in the mixture. The surface composition, the entropy Deltas , and energy Deltau of surface formation were evaluated by adopting the thermodynamic equations. It was shown that the surface was enriched in ethanol molecules compared with the bulk solution. The values of Deltas and Deltau were examined closely by estimating the contribution from the nonideal mixing in the bulk solution and that at the surface to Deltas and Deltau . It was concluded that the surface formation is driven by entropy at lower x 2 and by energy at higher x 2 , respectively, and that the surface formation becomes more energetically favorable with rising temperature.

Does a long pre-operative hospital stay before hepatectomy improve liver dysfunction in HCC patients with chronic liver disease?

BACKGROUND/AIMS: We retrospectively evaluated whether or not hepatic dysfunction improved in patients with chronic liver disease who had been waiting to undergo a hepatectomy after admission. MATERIALS AND METHODS: Fifty-two hepatocellular carcinoma patients had been admitted for more than 2 weeks prior to undergoing a hepatectomy. They had a liver function test twice, at admission and just before surgery, during the hospitalization period. Twenty-six of them were histologically diagnosed as having chronic hepatitis while the remainder had liver cirrhosis. In the liver function test, the serum levels of albumin, total bilirubin, glutamic pyrubic transaminase (alanine transaminase), total cholesterol and the Child grade were examined. RESULTS: First, including the pre-operative treatment cases for small tumors under angiography, the total bilirubin and transaminase levels improved in the chronic hepatitis patients with a statistically significant difference, but no difference was observed in the Child grade. In the examined cirrhotic patients, no significant difference was shown in the tests. Second, after excluding the pre-operative treatment cases, we performed the same investigation as that for chronic liver disease cases and only the transaminase level significantly improved. CONCLUSIONS: A long pre-operative hospital stay might only by justified in patients with a high level of transaminase corresponding to chronic active hepatitis.

Increased incidence of proximal colon cancer in the elderly.

To define the characteristics of colorectal cancer with regard to site distribution, age, and sex, due to the significantly increased incidence of colorectal cancer in Japan, we investigated 1,205 Japanese patients with colorectal cancer between 1975 and 1994. When we compared site distribution for age and sex in the entire 20-year period, tumors in the proximal colon were more frequent in elderly patients than in young ones (p < 0.05); this tendency was true only for females (p < 0.01), but not for males. The increased incidence of proximal colon cancer in elderly patients was also found in the second 10-year period (p < 0.05). Because elderly patients are characterized by an increasingly high incidence of proximal colon cancer in recent years, more concerted efforts for the early detection of proximal colon cancer, particularly in the elderly, are called for.

cDNA cloning and characterization of gibberellin-responsive genes in photoblastic lettuce seeds.

Two cDNA clones, cLRG5 and cLRG11, that respond to gibberellin (GA) were isolated from seeds of photoblastic lettuce (Lectuca sativa L. cv. Grand Rapids) by differential screening. Northern blot analysis indicated that the levels of LRG5 and LRG11 mRNAs were raised to slightly higher levels 10 h after the start of GA treatment and the levels were maintained at least for further 8 h, while those in the control seeds gradually decreased. Red light irradiation had effects similar to GA treatment. The cLRG5 insert encodes a putative polypeptide of 380 amino acids that is highly homologous to alcohol dehydrogenases from several higher plants. With regard to the cLRG11 insert, no homologous gene has been reported.

[A study on bacteria within stones in urolithiasis].

The bacteria in 37 stones obtained from 37 patients with urinary stone diseases, that is, 11 renal stones (containing 2 staghorn calculi), 21 ureteral stones, 4 bladder stones and 1 urethral stone, were studied, according to the Nemoy & Stamey's method. The stones were collected by partial nephrectomy (1 case), nephrolithotomy (1 case), pyelolithotomy (1 case), percutaneous nephrolithotripsy (PNL) (6 cases), 12 ureterolithotomies, transurethral ureterolithotripsy (2 cases), cystolithotripsies (4 cases) and spontaneous deliveries (10 cases). According to a stone analysis by infrared spectrophotometer revealed 30 were noninfection stones (81.1%) and 7 infection stones (18.9%). Of these 37 stones, 5 stones (13.5% of 11 stones) had bacteria within them. These stones consisted of 4 infection stones (57.1% of all infection stones) and 1 noninfection stone (3.3% of all noninfection stones). Out of 4 patients having bacteria within their stones, urine culture of whom were carried out before stones were collected, only one patient (25%) had the same species of bacterium (E. coli) both within the stone and in urine. The E. coli within the stone and that in urine, however, showed quite different reactions to some antibiotics. The fact that all organisms within stones cannot be detected by urine culture before collecting stones of the patients in our present study, suggests that patients undergoing endourological surgery, such as extracorporeal shock wave lithotripsy and PNL, may have a risk of complications, such as severe urinary tract infection and urosepsis, if the possibility of the presence of organisms within stones is not taken into consideration.