Structure-activity relationships of antifungal phenylpropanoid derivatives and their synergy with n-dodecanol and fluconazole.

trans-Anethole (anethole) is a phenylpropanoid; with other drugs, it exhibits synergistic activity against several fungi and is expected to be used in new therapies that cause fewer patient side effects. However, the detailed substructure(s) of the molecule responsible for this synergy has not been fully elucidated. We investigated the structure-activity relationships of phenylpropanoids and related derivatives, with particular attention on the methoxy group and the double bond of the propenyl group in anethole, as well as the length of the p-alkyl chain in p-alkylanisoles. Antifungal potency was largely related to p-alkyl chain length and the methoxy group of anethole, but not to the double bond of its propenyl group. Production of reactive oxygen species also played a role in these fungicidal activities. Inhibition of drug efflux was associated with the length of the p-alkyl chain and the double bond of the propenyl group in anethole, but not with the methoxy group. Although a desirable synergy was observed between n-dodecanol and anethole or p-alkylanisoles with a length of C2-C6 in alkyl chains, it cannot be explained away as being solely due to the inhibition of drug efflux. Similar results were obtained when phenylpropanoid derivatives were combined with fluconazole against Candida albicans.

Down-scattered neutron imaging detector for areal density measurement of inertial confinement fusion.

A custom developed (6)Li glass scintillator (APLF80+3Pr) for down-scattered neutron diagnostics in inertial confinement fusion experiments is presented. (6)Li provides an enhanced sensitivity for down-scattered neutrons in DD fusion and its experimentally observed 5-6 ns response time fulfills the requirement for down-scattered neutron detectors. A time-of-flight detector operating in the current mode using the APLF80+3Pr was designed and its feasibility observing down-scattered neutrons was demonstrated. Furthermore, a prototype design for a down-scattered neutron imaging detector was also demonstrated. This material promises viability as a future down-scattered neutron detector for the National Ignition Facility.

L-2,5-dihydrophenylalanine, an inducer of cathepsin-dependent apoptosis in human promyelocytic leukemia cells (HL-60).

L-2,5-Dihydrophenylalanine (DHPA), a phenylalanine analogue, induced apoptosis in human promyelocytic leukemia cells (HL-60). This apoptosis was demonstrated by morphological changes of the cells, such as fragmentation of nuclei and chromatin condensation, and by some evidence found in biochemical analysis, such as DNA ladder and activation of caspase 3. The DHPA-induced apoptosis was prevented by a pan-caspase inhibitor, Z-VAD-fmk, and a cysteine protease inhibitor, E-64d, which inhibits calpains and cathepsin B and L. A calpain inhibitor, Z-LL-H, did not affect this apoptosis. A cathepsin B specific inhibitor, CA074-Me, prevented only chromatin condensation. However, E-64d and a cathepsin L specific inhibitor, Z-FY(t-Bu)-dmk, protected the cells from both chromatin condensation and oligonucleosomal DNA fragmentation. As proceeding to the apoptotic process, the activities of both cathepsin B and L increased gradually. These results indicated that DHPA was an inducer of cathepsin-dependent apoptosis in HL-60 cells.

[Circadian changes in remnant-like particle-cholesterol (RLP-C), and its diagnostic value for postprandial increase in remnant lipoprotein in diabetes mellitus].

To investigate whether remnant-like particle-cholesterol (RLP-C) increases in the postprandial state, and if so, whether such response can be used for detecting postprandial increase in remnant lipoprotein, we measured RLP-C levels at two points (before breakfast, and after lunch), or seven points (before and after each meal, and midnight) in 36 diabetic patients. delta RLP-C was calculated by subtracting RLP-C level before breakfast from that at each point. delta RLP-C after lunch was defined as delta RLP-CL and maximum delta RLP-C as delta RLP-Cmax. Daily profiles of RLP-C (n = 22) showed that RLP-C increased during the daytime in 6 patients (27.3%; responders), but not in the others (nonresponders). In the histogram of delta RLP-Cmax, we could easily distinguish responders (delta RLP-Cmax = 6 approximately 12 mg/dl) from nonresponders (< 4 mg/dl). By using delta RLP-CL of 4 mg/dl as the cut-off value, we could also separate two groups without overlap. With this cut-off value, 11 out of 36 patients (30.6%) were diagnosed as responders. High performance liquid chromatography (HPLC) analysis with cholesterol-monitoring revealed that VLDL-sized RLP increased markedly after lunch in responders. In conclusion, RLP-C increases in some diabetic patients, and delta RLP-CL is useful for detecting postprandial increase in remnant lipoprotein.

[The effect of epidermal growth factor on keratocytes during healing of corneal penetrating incision].

We investigated the effect of epidermal growth factor (EGF) on keratocytes during corneal wound healing focusing on cell proliferation. A penetrating linear incision was made in the center of rabbit corneas. The corneas were then treated with eye drops of recombinant human EGF (10 micrograms/ml) or physiological saline (control) four times a day. After 1, 2, 3, and 7 days, the corneas were excised, labeled with 3H-thymidine (10 microCi/ml) at 37 degrees C for 4 hours and subjected to autoradiography. The results demonstrated increased number of keratocytes incorporating 3H-thymidine on the 1st, 2nd, 3rd and 7th days of healing and accelerated stromal wound healing in corneas treated with EGF compared with the controls. Thus, EGF stimulates of proliferation keratocyte and promotes corneal stromal wound healing.

Establishment and molecular characterization of a novel leukemic cell line with Philadelphia chromosome expressing p230 BCR/ABL fusion protein.

The cell line AR230 was established from the peripheral blood mononuclear cells of a patient with chronic myeloid leukemia and t(9;22) translocation bearing a variant type of BCR/ABL rearrangement. AR230 expresses a BCR/ABL fusion protein with a molecular mass of 230 kilodaltons (kDa) due to the insertion of 180 amino acids encoded by 3' exons of BCR (b4 to c3). An immune complex kinase assay showed that the 230-kDa BCR/ABL protein ahd autophosphorylation activity. Immunoprecipitation analysis revealed a stable complex of GRB2 and 230-kDa BCR/ABL proteins, indicating that the Ras activation pathway is involved in the process of transformation. AR230 expressed another short transcript consisting of a BCRc2/ABL junction, which is associated with a stop signal shortly after the junction. To our knowledge, this is the first cell line expressing a 230-kDa fusion product of BCR/ABL. AR230 will be useful for studying the biological function of divergent BCR/ABL proteins.

[HLA tissue types in patients with acute tubulointerstitial nephritis accompanying uveitis].

Four cases of idiopathic acute tubulointerstitial nephritis (TIN) associated with uveitis (so-called TINU syndrome) were experienced between 1986 and 1990. Patients' ages ranged from 14 to 42 years old and three were female and one was male. All cases showed general symptoms, such as general malaise, anorexia and weight loss. All patients had initially TIN and became ill uveitis four to eight months after the onset of TIN. All cases had mild proteinuria, mild anemia, the lower serum levels of potassium, hyper gamma-globulinemia and the reduced glomerular filtration rate with the increased beta 2-microglobulin in urine and serum. All renal biopsies specimens showed mild edema and diffuse infiltration of inflammatory mononuclear cells in the interstitium without any glomerular or vascular abnormalities. Furthermore, numerous CD4 positive cells, CD8 positive cells and CD11c positive cells were seen in the interstitium. Of four patients, three cases were treated with both oral administration and eye drop of prednisolone (PSL), another one case was therapied with eye drop PSL only. In all cases TIN had good prognosis, but two patients had recurrences of uveitis. All patients underwent tissue typing for HLA-A, B, C and DR antigens. Three patients had identical HLA-Cw3 and all four cases revealed identical HLA-A24(9). These results suggest that immunological mechanism, especially cell-mediated, and HLA system may play an important role in the occurrence of TINU syndrome.

Endothelin-renin-angiotensin-atrial natriuretic peptide system in ovaries: an intraovarian ERAANP system.

In a recent investigation of the ovary, high levels of endothelin-1 (ET-1), the renin-angiotensin system (RAS), and atrial natriuretic peptide (ANP) were identified. It was determined that ET, RAS, and ANP, alone or together, affected ovarian function. It is important that mutual relationships of these vasoactive and hormonal peptides, which coexist in the ovary, are examined to determine their in vivo functions. This study, using immature rats treated with pregnant mare's serum gonadotropin (PMS), examined the effects of ET-1, angiotensin II (Ang II) and ANP, individually or in combination, on steroidogenesis by granulosa cells (GCs) cultured for 72 h. ET-1 Ang II, and ANP, alone or combined, also had a mutual effect on steroidogenesis. Concomitantly with previous findings on the ovary, the authors propose that the intraovarian endothelin-renin-angiotensin-ANP system (ERAANPS) functions as a novel intraovarian regulator system.

A proposal of ovarian ERAANPS (endothelin-renin-angiotensin-atrial natriuretic peptide system) and effects of tokishakuyakusan, keishibukuryogan, shakuyakukanzoto and unkeito on the ERAANPS.

We have previously shown the presence in a high concentration of endothelin-1 (ET) in the corpus luteum and renin-angiotensin system (RAS) and binding sites for atrial natriuretic peptide (ANP) in the ovarian follicle. The present study was undertaken to identify the existence of ET, renin, angiotensin II and the binding site for ANP in the ovary at proestrus and examine in vivo the effects of herbal medicines [Tokishakuyakusan (TS), Keishibukuryogan (KB), Shakuyakukanzoto (SK) and Unkeito (UT)] on them. ET, all components of RAS and binding sites for ANP were found at high levels in the ovary. TS, KB, SK and UT decreased the ET levels in ovary, while components of RAS and binding sites for ANP have the propensity to increase. However, ET, renin, angiotensin II and ANP levels in plasma were not at all affected before and after treatment with TS, KB, SK or UT. Taken together with previous observations showing the existence of ET, RAS and the binding site for ANP in the ovary, we propose here the ERAANPS (endothelin-renin-angiotensin-ANP system) in the ovary as a functional regulator. Further, these results suggest that TS, KB, SK or UT may regulate the ovarian ERAANPS.

Effects of tokishakuyakusan, keishibukuryogan, shakuyakukanzoto and unkeito on ovarian endothelin, renin and angiotensin II in pregnant mare's serum gonadotropin-treated immature rats.

We have previously proposed the ovarian ERAANPS (endothelin-rein- angiotensin-atrial natriuretic peptide system). The present study was undertaken to examine in vivo the effects of herbal medicines [Tokishakuyakusan (TS), Keishibukuryogan (KB), Shakuyakukanzoto (SK) and Unkeito (UT)] on endothelin-1 (ET), renin and angiotensin II (A II) in the ovaries, of immature rats treated with 10 IU PMS for 48 h. ET and all components of renin-angiotensin system (RAS) were found at high levels in the ovary. Concomitant treatment with PMS plus TS, KB, SK or UT, especially TS and UT, tended to decrease the ET levels in ovary, while components of RAS tended to increase. However, ET, renin and A II levels in plasma were not at all affected after treatment with TS, KB, SK or UT. These results suggest that TS, KB, SK or UT may regulate the ovarian ERAANPS.

Successful treatment of a senile diabetic woman with cataract with goshajinkigan.

A 65-year-old woman with diabetes mellitus (DM) and suffering a cataract in the left eye, that was resistant to Western medical treatment, was treated with goshajinkigan (GJ), a herbal medicinal drug, in combination with eye drops. This treatment remarkably improved the visual disturbance due to the senile cataract. Ten days after commencement of the GJ treatment, her visual acuity did not deteriorate any further. There was also no further deterioration of DM condition. These results suggest that GJ may be valuable in the treatment of diabetic women with cataracts.

[A case of acute tubulo-interstitial nephritis and uveitis syndrome].

We report herein the case of a 14-year-old female who has acute tubulo-interstitial nephritis (AIN) associated with bilateral diffuse uveitis. She was admitted for the evaluation of "proteinuria", following general fatigue and weight loss about 2 weeks ago. Her laboratory data showed mild anemia, hyper gamma-globulinemia, mild proteinuria, and the reduced glomerular filtration rate with the increased urinary excretion of beta 2-microglobulin. The histological examination obtained by renal biopsy showed mild edema and diffuse infiltration of mononuclear cells in interstitium without any glomerular or vascular abnormalities, which were compatible with AIN. As for the etiology of AIN, clinical investigations could not reveal any specific causes, such as bacterial and viral infections, drugs and systemic diseases. About 4 months after the onset of nephritis, she also became to suffer from bilateral diffuse uveitis. Therefore, the diagnosis of the acute tubulo-interstitial nephritis and uveitis syndrome (TINU syndrome) (Vanhaesebrouck et al., 1985) could be confirmed. In her clinical course, it was noteworthy that uveitis relapsed frequently in spite of systemic administration of prednisolone, and it took two years until uveitis cured, whereas the AIN subsided spontaneously prior to the specific treatment. In this case, characteristic findings of granulomatous uveitis was closely similar to those of sarcoidosis, which has been rarely reported in TINU syndrome. In this respect, the involvement of immune processes, especially cell-mediated, was suggested as the possible pathogenesis in this case.

Treatment with hachimijiogan, a non-ergot Chinese herbal medicine, in two hyperprolactinemic infertile women.

Two hyperprolactinemic infertile women, one with and one without a pituitary adenoma, who were resistant to bromocriptine treatment, were treated orally with Hachimijiogan, a Chinese herbal medicine. This treatment reduced the serum prolactin level, resulting in a normal ovulatory cycle and pregnancy, without side effects.

Hachimijiogan treatment is effective in the management of infertile women with hyperprolactinemia or bromocriptine-resistant hyperprolactinemia.

Twenty-seven infertile women with hyperprolactinemia were treated with Hachimijiogan (5.0 to 10.0 g/day), Chinese herbal medicine. In 15 out of 18 patients, who were treated with Hachimijiogan, the serum prolactin levels dropped to 19.4 +/- 10.5 ng/ml during Hachimijiogan treatment, and the levels in 8 patients remained essentially unchanged during 6 months after the cessation of Hachimijiogan treatment. Three patients with 100 to 300 ng/ml did not drop to the level below 30 ng/ml. Four of 6 patients with amenorrhea ovulated. Eleven patients conceived and delivered normally. The hormonal profiles during or after pregnancy were normal. These results indicate that Hachimijiogan is a new safe treatment for hyperprolactinemic women.

Follicle-stimulating hormone induces deoxyribonucleic acid polymerase alpha before deoxyribonucleic acid synthesis in immature rat ovarian follicles perfused in vitro.

To examine the effects of follicle-stimulating hormone (FSH) on cell proliferation and cell division in ovaries, deoxyribonucleic acid (DNA) polymerase activities and DNA concentrations were determined in ovarian follicles of immature, hypophysectomized, estradiol-treated rats perfused in vitro. Ovine FSH (2-20 micrograms/mL) significantly enhanced the DNA polymerase alpha activity in the follicles perfused for 180-300 minutes, whereas the DNA polymerase beta activity and DNA concentration showed no significant changes. It is concluded that FSH causes the induction of DNA polymerase alpha before DNA synthesis during follicle growth in immature rat ovaries.

Complement-dependent cytotoxic factor to megakaryocyte progenitors in sera from patients with idiopathic thrombocytopenic purpura.

The influence of sera from patients with idiopathic thrombocytopenic purpura (ITP) was examined on colony formation from megakaryocyte (M) progenitors. Though incubation of marrow cells in Iscove's modified Dulbecco's medium (IMDM) containing 50% sera from several ITP patients stimulated M-colony formation in 8 of 13 cases, incubation in the sera from the patients and in baby rabbit serum as a source of complement significantly suppressed the colony formation. Experiments showed that sera of immunoglobulin G from ITP patients had significant complement-dependent cytotoxicity to M-progenitors in normal marrow cells or in the marrow cells from corresponding patients, but not to CFU-e, BFU-e or CFU-gm. Cytospin preparations of individually collected M-colonies from marrow cells treated with ITP patients' sera and complement revealed a reduction of megakaryocyte colonies containing cells of multilineages. These results indicate that autoantibodies detected in ITP patients can bind not only to platelets and megakaryocytes, but may also bind to M-progenitors.