Disseminated mucocutaneous herpes simplex virus infection in an immunocompetent woman.

Disseminated mucocutaneous herpes simplex virus (HSV) infection in an immunocompetent person is quite rare. A 19-year-old healthy Japanese woman presented with painful, umbilicated vesicles and pustules on her genital region, both nipples and on the forearm 10 days after the last sexual contact with her partner who had cold sore at that time. Tzanck test and biopsy confirmed the diagnosis of disseminated mucocutaneous HSV infection. She did not have any visceral HSV disease. Skin lesions improved after treatment with acyclovir and erythromycin for seven days. We propose that like herpes gladiatorum, HSV dissemination in this case was acquired by close body contact.

Etretinate enhances the susceptibility of human skin squamous cell carcinoma cells to 5-aminolaevulic acid-based photodynamic therapy.

BACKGROUND: Photodynamic therapy (PDT) with 5-aminolaevulinic acid (5-ALA) is a noninvasive and effective treatment for superficial skin cancers. Etretinate, a derivate of vitamin A, with the chemical formula ethyl(2E,4E,6E,8E)-9-(4-methoxy-2,3,6-trimethylphenyl)-3,7-dimethyl-2,4,6,8-nona-tetraenoate, has been reported to have antitumour effects and to regulate the proliferation and differentiation of skin cancers. OBJECTIVE: In order to develop more efficient PDT, we investigated whether etretinate enhanced the cytotoxic action of ALA-based PDT against human squamous cell carcinoma cell line, HSC-5. METHOD: The in vitro cytotoxicity was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptotic cells were detected by double-staining with fluorescent annexin V and propidium iodide. Intracellular protoporphyrin IX (PpIX) converted from exogenous ALA was measured by a fluorescence meter. RESULTS: HSC-5 cells pretreated with a nontoxic concentration of etretinate became more susceptible to the cytotoxic action of ALA-based PDT. Etretinate-pretreated cells underwent apoptosis in response to ALA-based PDT. Etretinate pretreatment resulted in enhanced accumulation of ALA-dependent intracellular PpIX. CONCLUSIONS: The results suggest that etretinate enhances the susceptibility of HSC-5 cells to ALA-based PDT via the intracellular increase of ALA-dependent PpIX. Etretinate might be useful for improvement of ALA-based PDT.

Tea catechins decrease micellar solubility and intestinal absorption of cholesterol in rats.

A(-)-epicatechin (EC) and (-)-epigallocatechin (EGC) mixture and a mixture of their gallates (ECG and EGCG, respectively) markedly lowered lymphatic cholesterol absorption in rats with a cannulated thoracic duct. A mixture of ECG and EGCG was more effective in reducing cholesterol absorption than the EC and EGC mixture. These catechins also tended to decrease lymphatic absorption of triacylglycerols, although not so pronounced as in cholesterol absorption. An in vitro study on micellar solubility of cholesterol showed that these catechin mixtures precipitated cholesterol solubilized in mixed bile salt micelles in a dose-dependent manner. A mixture of ECG and EGCG more effectively precipitated micellar cholesterol than a mixture of EC and EGC. When purified EC, EGC, ECG and EGCG were used, EGCG was more effective in precipitating micellar cholesterol than ECG. The effect of EC and EGC was comparable and weaker than their gallate esters. The bile acid concentration in the micelles was not affected by these catechins. A positive correlation was observed between the amount of coprecipitated EGCG and cholesterol. These results clearly show that tea catechins, in particular their gallate esters, effectively reduce cholesterol absorption from the intestine by reducing solubility of cholesterol in mixed micelles. The observation accounts for the hypocholesterolemic effect of tea catechins.

GLP-I(7-36) amide augments Ba2+ current through L-type Ca2+ channel of rat pancreatic beta-cell in a cAMP-dependent manner.

The whole-cell patch-clamp method was used to examine the effect of glucagon-like peptide I (GLP-I)(7-36) amide on the activation process of L-type Ca2+ channels of rat pancreatic beta-cells. After depolarization, GLP-I (1-100 nmol/l) caused action potentials in cells exposed to a glucose-free solution for 10 min. The percentage of cells producing action potential depended on the concentration of GLP-I. In some cells, GLP-I caused action potentials without the prior depolarization of the membrane. In cells exposed to the glucose-free solution for longer than 30 min, or in cells that were deprived of ATP by a means of the conventional whole-cell configuration, GLP-I (20 nmol/l) did not cause the electrical excitation. Application of GLP-I augmented the maximum Ba2+ current (IBa) through L-type Ca2+ channels and shifted the current voltage curve to the left. Values of changes in the maximum IBa depended on GLP-I concentration. Application of dibutyryl cAMP (dbcAMP, 1 mmol/l) also augmented IBa. In cells pretreated with Rp-cAMP, dbcAMP did not change the magnitude of IBa. Also in cells pretreated with Rp-cAMP, GLP-I failed to augment IBa. These results suggest that in pancreatic beta-cells, GLP-I, by a cAMP-dependent mechanism, increases opening of L-type Ca2+ channels. cAMP-dependent augmentation of Ca2+ entry as well as cAMP production itself by GLP-I plays a crucial role in controlling insulin secretion.

Protein kinase C-dependent and -independent inhibition of Ca(2+) influx by phorbol ester in rat pancreatic beta-cells.

Phorbol esters were used to investigate the action of protein kinase C (PKC) on insulin secretion from pancreatic beta-cells. Application of 80 nM phorbol 12-myristate 13-acetate (PMA), a PKC-activating phorbol ester, had little effect on glucose (15 mM)-induced insulin secretion from intact rat islets. In islets treated with bisindolylmaleimide (BIM), a PKC inhibitor, PMA significantly reduced the glucose-induced insulin secretion. PMA decreased the level of intracellular Ca(2+) concentration ([Ca(2+)](i)) elevated by the glucose stimulation when tested in isolated rat beta-cells. This inhibitory effect of PMA was not prevented by BIM. PMA inhibited glucose-induced action potentials, and this effect was not prevented by BIM. Further, 4alpha-phorbol 12,13-didecanoate (4alpha-PDD), a non-PKC-activating phorbol ester, produced an effect similar to PMA. In the presence of nifedipine, the glucose stimulation produced only depolarization, and PMA applied on top of glucose repolarized the cell. When applied at the resting state, PMA hyperpolarized beta-cells with an increase in the membrane conductance. Recorded under the voltage-clamp condition, PMA reduced the magnitude of Ca(2+) currents through L-type Ca(2+) channels. BIM prevented the PMA inhibition of the Ca(2+) currents. These results suggest that activation of PKC maintains glucose-stimulated insulin secretion in pancreatic beta-cells, defeating its own inhibition of the Ca(2+) influx through L-type Ca(2+) channels. PKC-independent inhibition of electrical excitability by phorbol esters was also demonstrated.

New xenografts of human megakaryoblastic cell line (MEG-01) for evaluating anti-tumor agents.

A human megakaryoblastic cell line (MEG-01) was successfully transplanted into athymic nude mice. MEG-01 cells (5 x 10(7)) were inoculated subcutaneously in KSN-nu/nu mice, none of which were pretreated with irradiation or chemotherapeutic agents. All mice developed solid tumors at the site of injection after incubation for 10-14 days reaching a size of 200-400 mm2 (product of cross-sectional diameters) after 30 days. These tumors, designated as MEG-01/nu, were transplanted into other nude mice. The transplanted tumors infiltrated the liver and spleen, and leukemic megakaryoblastic cells appeared in the blood of some transplanted mice. Cells resuspended from MEG-01/nu tumors exhibited almost the same megakaryocytic characteristics as the original MEG-01 cells, and underwent in vitro differentiation to a mature form of megakaryocyte upon addition of phorbol diesters. MEG-01/nu was evaluated for sensitivity to cytosine arabinoside, vincristine, and daunorubicin in vitro and in vivo. Daunorubicin exhibited significant anti-tumor activity against MEG-01/nu in vivo, while cytosine arabinoside did so in vitro. Vincristine showed no activity against these cells. This cell line may provide a useful model for testing the in vivo efficacy of anti-tumor agents and immunotoxins, and for studying the pathophysiological mechanisms of human megakaryoblastic leukemia.

Thimerosal modulates the agonist-specific cytosolic Ca2+ oscillatory patterns in single pancreatic acinar cells of mouse.

Modulation of the agonist-specific cytosolic Ca2+ oscillatory pattern by thimerosal has been investigated in single pancreatic acinar cells using patch-clamp perforated whole-cell recording to measure the calcium-dependent chloride current (I(C1)(Ca2+)). 1 microM thimerosal, which fails to evoke Ca2+ oscillation alone, clearly changed the pattern of Ca2+ oscillation from pulsatile spikes (evoked by low concentrations of activators) to sinusoidal or transient oscillations. The mimetic action of thimerosal was independent of extracellular Ca2+, was blocked by extracellular application of dithiothreitol or 10 mM caffeine, as well as by internal perfusion with heparin; but was unaffected by ruthenium red. We conclude that thimerosal modulates the agonist-specific cytosolic Ca2+ oscillatory patterns mediated by sensitizing the InsP3-induced Ca2+ release.

Influence of 1 alpha,25-dihydroxyvitamin D-3 on growth regulation through epidermal growth factor receptor in human breast cancer cell lines.

We investigated the relationship between epidermal growth factor (EGF) dependent cell growth and antiproliferative effects of 1 alpha,25-dihydroxyvitamin D-3 (1,25(OH)(2)D-3) in hormone responsive breast cancer cell lines in vitro. MCF-7 breast cancer cells and GMC-M, which is a serum-independent, hormone receptor-positive subtype derived from MCF-7, were used in this study. EGF stimulated the growth Of both cell lines, and 1,25(OH)(2)D-3 inhibited the EGF-stimulated cell growth in a dose dependent fashion. But treatment with 1,25(OH)(2)D-3 did not change the EGF receptor (EGFR) level significantly in either cell line. GMC-M had a higher level of EGFR and was more sensitive to EGF than MCF-7. These results suggest that other mechanisms of action, which are different from EGFR modulation, concern with the growth inhibitory effect of 1,25(OH)(2)D-3, and that 1,25(OH)(2)D-3 will be a new effective treatment for breast cancer irrespective of EGFR.

Diametrically opposite effects of estrogen on the excitability of female rat medial and lateral preoptic neurons with axons to the midbrain locomotor region.

Electrical stimulation of the midbrain locomotor region (MLR) in 76 ovariectomized, urethan-anesthetized female rats elicited antidromic action potentials in 252 preoptic neurons. Thresholds and refractory periods for the activation ranged from 60 to 1550 microA and 1.3 to 5.0 ms, respectively. The probability distribution for the peak-to-peak amplitude (2-14 mV) or the overall duration (0.7-4.4 ms) was bell-shaped, whereas that for the latency (1.8-33.5 ms) was distinctively bimodal with a division at 12.0 ms. Two groups of preoptic neurons of a similar soma size therefore project to the MLR presumably via different routes. In 121 neurons with latencies < or = 12.0 ms, estrogen lowered the antidromic activation thresholds (nested analysis of variance, P < 0.02), but 131 neurons with latencies > 12.0 ms had their thresholds increased (P < 0.005) and refractory periods prolonged (P < 0.02) by estrogen. Even though both overlapped in part, many potentials with the shorter latencies were recorded from the medial part of the lateral preoptic area (mLPO), lateral to the recording sites of the longer-latency potentials in the medial preoptic area (MPO). The observed antagonistic effects of estrogen on the two groups of preoptic neurons with axons to the MLR may contribute to increased locomotor activity in female rats in estrus.

Intensive individualized induction therapy with behenoyl cytarabine, daunorubicin and 6-mercaptopurine followed by intensive consolidation including intermediate-dose continuous cytarabine, mitoxantron, etoposide and vinca alkaloids in acute myeloid leukemia in adults.

Forty-one consecutive adult patients with acute myeloid leukemia (AML) were treated with an intensive individualized induction therapy of behenoyl cytarabine, daunorubicin, and 6-mercaptopurine, 29 patients (71%) achieved complete remission (CR). Patients then received three courses of intensive consolidation therapy, including intermediate-dose continuous cytarabine (400 mg/m2, for 5 days) and non-cross resistant drugs such as mitoxantron, etoposide and vincristine. During the course of the consolidation therapy, three patients died of infections and one died of myocardial infarction. Four patients underwent allogeneic bone marrow transplantation. The patients then received six courses of moderately intensive maintenance therapy for 1 year. The predicted 5-year continuing CR and disease-free survival rates of the CR patients were 62% (95% confidence limit, 41% to 83%) and 53% (33% to 73%), respectively. Although the number of patients in this study is small, the present study indicated that it may be possible to cure a fairly large proportion of AML patients by chemotherapy alone, if intensive induction therapy is followed by intensive consolidation therapy.

Axon-sparing lesion of the preoptic area enhances receptivity and diminishes proceptivity among components of female rat sexual behavior.

Stereotaxic infusion of ibotenic acid deleted neurons in the medial preoptic area (POA) in the ovariectomized female rats. A well-circumscribed lesion was infiltrated by astrocytes; local axons of passage were spared. Following estrogen priming and progesterone supplement, the females with the lesion had higher lordosis quotients than the vehicle-infused controls, when males successfully mounted them. On the other hand, the treatment did not induce solicitation in females with the lesion nor reduced their rejection of male partners. Meanwhile, gradual and persistent suppression of the lordosis reflex followed electrical stimulation through electrodes placed in the POA lesion. Except that the females with the POA lesion needed less estrogen to obtain comparable prestimulation quotients with the controls, the lesioned and control animals responded similarly to the stimulation. Because an adjunct neural transection dorsal to the POA lesion abolished the stimulus-bound suppression of lordosis, the effect was due to the activation of axons of passage that presumably descend from the septum. It is concluded that the POA is the major target for estrogen in eliciting proceptive behavior; local POA neurons as well as septal efferents appear to inhibit the lordosis reflex, the principal receptive component in female rat sexual behavior.

Acute myeloblastic leukemia (M2) with translocation (7;11) followed by marked eosinophilia and additional abnormalities of chromosome 5.

We present an 18-year-old woman who was diagnosed with acute myeloblastic leukemia (AML M2), and in whom chromosome analysis of bone marrow cells revealed t(7;11), an abnormality rarely found in leukemias with a differentiation potency. She relapsed 1 year after complete remission was achieved by chemotherapy. Bone marrow examination then revealed a t(7;11) abnormality in 48 of 50 metaphases examined, even when there were less than 7.5% leukemic blasts in the marrow, indicating that the morphologically normal cells were derived from leukemic blasts. The number of leukemia clones with the additional abnormalities in chromosome 5 increased, with concurrent development of eosinophilia, fever, asthma-like symptoms, erythema, itching, and hepatosplenomegaly. Elevation of interleukin 5 (IL-5) in serum and an enhanced expression of IL-5 mRNA were also detected. The increase in IL-5 may have been produced by an abnormality on chromosome 5.

Effect of toremifene on the growth, hormone receptors and insulin-like growth factor-1 of hormone-dependent MCF-7 tumors in athymic mice.

Toremifene given in different sizes of silastic capsules was used to treat MCF-7 tumors in athymic mice. Toremifene inhibited the estradiol-stimulated growth of MCF-7 tumors in athymic mice. Average serum concentrations of toremifene obtained using a sustained-release preparation of the drug (in 0.5-, 1.0-, and 2.0-cm silastic capsules) increased gradually in a capsule-size-dependent fashion. Much higher levels of toremifene or N-demethyl-toremifene were detected in tumors (target tissues of estrogen) as compared with muscles (non-target tissues of estrogen). The concentration of toremifene in serum (i.e., 10-30 ng ml-1) was sufficient to inhibit the estrogen-stimulated growth of MCF-7 tumors at physiological (i.e., 200-400 pg ml-1) serum estradiol concentrations in premenopausal women. No significant difference in estrogen receptor (ER) levels was found between the estradiol-alone group and the toremifene-treated groups. However, the ER levels in the toremifene-alone group and the no-treatment group (no toremifene or estradiol) tended to increase as compared with the estradiol-alone group. Toremifene blocked the estradiol-induced increase in progesterone receptor levels in a dose-dependent fashion. Insulin-like growth factor-1 (IGF-1) levels in the MCF-7 tumors significantly decreased in the toremifene-alone group as compared with the estradiol-alone group. These results show the antiestrogenic action of toremifene on hormone-dependent MCF-7 tumors in athymic mice.

Detection of minimal residual disease in Philadelphia chromosome positive acute lymphoblastic leukemia: rationale for bone marrow transplantation from the polymerase chain reaction point of view.

Bone marrow transplantation (BMT) is performed as curative therapy for acute lymphoblastic leukemia (ALL). In most patients, BMT is performed at the time of remission which implies that the number of leukemic cells is less than 5% of all hematopoietic cells, namely, 0 to 10(10) leukemia cells in the body. Thus, some patients may well undergo BMT despite the fact that no leukemic cells are left in the body. In this respect, more accurate diagnosis of complete remission status would be to the patients' benefit. To detect minimal residual disease (MRD) not found by light-microscopy, further strategies are required after achieving hematological remission. Cytogenetic methods, Southern blot analysis and conventional immunological techniques can all provide accurate diagnosis, however, the sensitivity of these techniques for the detection of MRD is just as low as that of the light microscopy. Recently, polymerase chain reaction (PCR) has become available for the detection of low levels of chimeric bcr-abl transcripts in Philadelphia chromosome positive (Ph1) ALL patients. With this assay, investigators have reported MRD in patients after chemotherapy or BMT. Most patients who achieve hematological remission after conventional chemotherapy still have bcr-abl transcript detectable by PCR, confirming the general concept that this particular leukemia needs BMT in order to cure the disease. Some patients who had MRD prior to BMT continued disease free survival > 1 year after BMT with a negative PCR result and in these patients, MRD seems to have been eradicated by the BMT procedure.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential regulation of estrogen-dependent sexual development of rat brain by growth factors.

Intraventricular infusion of antiserum to nerve growth factor (ANGF), but not that to insulin, epidermal growth factor nor normal rabbit serum, resisted estrogen-induced behavioral defeminization in the female rat neonates. A significant number of the ANGF-treated rats showed lordosis as adults despite neonatal estrogen, but positive feedback of estrogen on serum luteinizing hormone was absent. Sexual phenotype in behavioral and gonadotropic functions may be under differential development regulation.

Stria terminalis conveys a facilitatory estrogen effect on female rat lordosis reflex.

The effects of bilateral cut of the stria terminalis (ST) on the dosage of estradiol benzoate (EB) needed to induce the lordosis reflex in the ovariectomized female rat were examined. Seven different doses ranging 0.5-20 micrograms were given s.c. in oil, daily for 2 days in a counterbalanced order, at 3-week intervals. The log dose-response relationship was linear in both ST-cut and sham-operated control rats. A significantly larger dose of 9.3 +/- 1.4 (mean +/- SEM) micrograms/day of EB was needed in the ST-cut rats (n = 8) to obtain a 50% lordosis quotient than in the controls (n = 7), in which a 2.0 +/- 0.3 micrograms/day dose had a similar effect. The slope of the dose-effect curve was significantly steeper in the ST-cut rats than in the controls. The ST mediates facilitatory effects of estrogen on the lordosis reflex that originate presumably in the amygdala.

Suppression of the lordosis reflex of female rats by efferents of the medial preoptic area.

In freely moving, estrogen- and progesterone-treated ovariectomized female rats, electrical stimulation of the medial preoptic area, which had been isolated dorsally from the septum by a horizontal knife cut (roof cut), caused immediate interruption of the lordosis reflex, a principal component of female reproductive behavior in this species. Neither the knife cut nor the stimulation affected their proceptive interactions with the males. Lordosis was interrupted in a graded manner in response to increased stimulus intensity, with a threshold at 30 microA. The optimal frequency was at 50-100 Hz. Lordosis was reinstated promptly by the termination of current application. The rapid time course distinguished the stimulation effect in the rats with the roof cut from that in the intact control animals, which was slow in its onset and recovery. In the sham-operated animals, an adjunct bilateral cut of the stria terminalis was sufficient to restore the rapid response as in the roof-cut animals. The roof-cut animals were sensitive to lower doses of estrogen than those without the knife cut; therefore, the preoptic area appears to be a separate entity from the septum in the inhibitory control of lordosis. The elimination of facilitatory neural components for this reflex, which enter the preoptic area dorsally via the stria terminalis, might be responsible for the prompt and exaggerated stimulation effect in the roof-cut animals.

Interruption of the lordosis reflex of female rats by ventral midbrain stimulation.

The lordosis reflex, dorsiflexion of the vertebral column, is an estrogen-dependent, essential element of female sexual behavior in rodents. Unilateral electrical stimulation of the midbrain ventral tegmental area through a chronically implanted electrode in freely moving, estrogen-primed ovariectomized female rats caused a rapid and strong suppression of the lordosis reflex in response to either male mounts or manual cutaneous stimuli. The interruption occurred in a graded manner to increased stimulus intensity, with a threshold at 30 microA. The optimal frequency was at 75-125 Hz. After the termination of electrical stimulation, lordosis performance returned promptly to the pre-stimulation level. No aversive response accompanied the blockade of lordosis. Electrical stimulation specifically blocked lordosis, without disrupting the proceptive components of female sexual behavior. In 10 animals tested, concomitant injection of dopamine receptor blocker pimozide tended to offset the effects of electrical stimulation in 2 cases. Interruption of the lordosis reflex might be mediated by projections from the ventral tegmental area, which activate a descending pathway inhibitory to the lordosis reflex arc at or below the lower brain stem.

Relationship between grade of intraductal component of breast cancer within main tumor and extent of intraductal spread in surrounding tissue.

Ninety-three specimens obtained by quadrantectomy of invasive ductal carcinoma of the breast were examined to evaluate the correlation between the grade of the intraductal component within the tumor and the extent of intraductal spread in the surrounding tissue. We used immunohistochemistry of a-smooth muscle actin to visualize the contour of intraductal components. The grade of the intraductal component within the tumor was classified into 4 groups (td0 to td3) by counting number of intraductal components; the extent of intraductal spread in the surrounding tissue was also divided into 4 groups (sd0 to sd3) according distance from the tumor margin. Fifty-eight (89.2%) of 65 cases with low-grade td were low-grade sd, and 17 (60.7%) cases of high-grade td were high-grade of sd. The grade of the intraductal component correlated with the extent of intraductal spread in the surrounding tissue, significantly (p < 0.001, Spearman rank correlation test). From these results, we believe that investigation of intraductal components within the tumor is useful for estimating the extent of intraductal spread in the surrounding tissue, and excision specimens from diagnostic lumpectomy should be examined for the extent of the intraductal component, the results being important in estimating the risk of local recurrence.

Clinical significance of first line treatment in recurrent breast cancer.

From 1962 to 1992, 279 patients with recurrent breast cancer were treated. Of these, the cases with resected lesions, imperfectly evaluated cases and cases where information was lacking were excluded, and 185 recurrent breast cancers were evaluable at the first line or subsequent treatment. Sixty-seven (36.2%) out of 185 cases responded to first line treatment, while the remaining 118 cases did not respond. In the 67 responder cases at the first line treatments, 16 (23.9%) responded to second line or third line, however, in the 118 nonresponder cases, only 13 (11.0%) responded to next or subsequent treatment. The incidence of responder in the second line or third line treatment was significantly higher in the first line responder group than in the first line nonresponder group. Furthermore, overall survival of first line responders was significantly better than that of first line nonresponders. There was no significant difference in the survival after recurrence between responders and nonresponders in the first line chemotherapy, chemoendocrine treatment or radiotherapy, however, a significant difference was seen between the two groups in the first line endocrine treatment. These results suggest that first line treatment may select the treatment sensitive (especially, truly hormone-dependent) recurrent breast cancers which show a better prognosis.