Leukemia cell to endothelial cell communication via exosomal miRNAs.

Recent findings indicate that specific microRNAs (miRNAs), such as those of the miR-17-92 cluster, may be responsible for regulating endothelial gene expression during tumor angiogenesis. Secreted miRNAs enclosed in exosomes also have an important role in cell-cell communication. To elucidate whether miRNAs secreted from neoplastic cells transfer into endothelial cells and are functionally active in the recipient cells, we investigated the effect of exosomal miRNAs derived from leukemia cells (K562) on human umbilical vein endothelial cells (HUVECs). As K562 cells released the miR-17-92 cluster, especially miR-92a, into the extracellular environment, K562 cells, transfected with Cy3-labeled pre-miR-92a, were co-cultured with HUVECs. Cy3-miR-92a derived from K562 cells was detected in the cytoplasm of HUVECs, and the Cy3-miR-92a co-localized with the signals of an exosomal marker, CD63. The expression of integrin α5, a target gene for miR-92a, was significantly reduced in HUVECs by exosomal miR-92a, indicating that exogenous miRNA via exosomal transport can function like endogenous miRNA in HUVECs. The most salient feature of this study is the exosome, derived from K562 cells with enforced miR-92a expression, did not affect the growth of HUVECs but did enhance endothelial cell migration and tube formation. Our results support the idea that exosomal miRNAs have an important role in neoplasia-to-endothelial cell communication.

High expression of N-acetylglucosaminyltransferase IVa promotes invasion of choriocarcinoma.

BACKGROUND: Gestational trophoblastic diseases (GTDs) are related to trophoblasts, and human chorionic gonadotropin (hCG) is secreted by GTDs as well as normal placentas. However, the asparagine-linked sugar chains on hCG contain abnormal biantennary structures in invasive mole and choriocarcinoma, but not normal pregnancy or hydatidiform mole. N-acetylglucosaminyltransferase-IV (GnT-IV) catalyses ß1,4-N-acetylglucosamine branching on asparagine-linked oligosaccharides, which are consistent with the abnormal sugar chain structures on hCG. METHODS: We investigated GnT-IVa expression in GTDs and placentas by immunohistochemistry, western blot, and RT-PCR. We assessed the effects of GnT-IVa knockdown in choriocarcinoma cells in vitro and in vivo. RESULTS: The GnT-IVa was highly expressed in trophoblasts of invasive mole and choriocarcinoma, and moderately in extravillous trophoblasts during the first trimester, but not in hydatidiform mole or other normal trophoblasts. The GnT-IVa knockdown in choriocarcinoma cells significantly reduced migration and invasive capacities, and suppressed cellular adhesion to extracellular matrix proteins. The extent of ß1,4-N-acetylglucosamine branching on ß1 integrin was greatly reduced by GnT-IVa knockdown, although the expression of ß1 integrin was not changed. In vivo studies further demonstrated that GnT-IVa knockdown suppressed tumour engraftment and growth. CONCLUSION: These findings suggest that GnT-IVa is involved in regulating invasion of choriocarcinoma through modifications of the oligosaccharide chains of ß1 integrin.

Downregulated plasma miR-92a levels have clinical impact on multiple myeloma and related disorders.

Recent studies have demonstrated that one-third of known microRNAs (miRNAs) are stably detectable in plasma. Therefore, we assessed plasma miRNAs to investigate the dynamics of oncomir 17-92a, which is highly expressed in multiple myeloma (MM) patients. The plasma miR-92a level in symptomatic MM patients was significantly downregulated compared with normal subjects (P<0.0001), regardless of immunoglobulin subtypes or disease stage at diagnosis. In contrast, miR-92a levels in peripheral blood CD8(+) or CD4(+) cells from MM patients were lower than those of normal subjects, and the miR-92a levels of the cells tended to correlate with plasma miR-92a levels. The plasma miR-92a level in the complete remission group became normalized, whereas the partial response (PR) and very good PR groups did not reach the normal range. In smoldering MM, the plasma miR-92a level did not show a significant difference compared with normal subjects. Our findings suggest that measurement of the plasma miR-92a level in MM patients could be useful for initiation of chemotherapy and monitoring disease status, and the level may represent, in part, the T-cell immunity status of these patients.

Long-term survival of young women receiving fertility-sparing surgery for ovarian cancer in comparison with those undergoing radical surgery.

OBJECTIVES: To compare the clinical outcome of patients with stage I epithelial ovarian cancer (EOC) who received with fertility-sparing surgery (FSS) with those who underwent radical surgery (RS). METHODS: After a central pathological review and search of the medical records from multiple institutions, a total of 572 patients were retrospectively evaluated. All patients were divided into three groups: group A {FSS (n=74); age, ≤ 40}; groups B and C [RS; age, 40 ≥{(B), n=52}; 40<{(C), n=446}]. RESULTS: Five-year overall survival (OS) and disease-free survival (DFS) rates of patients in the groups were as follows: group A, 90.8% (OS)/87.9% (DFS); group B, 88.3% (OS)/84.4% (DFS); group C, 90.6% (OS)/85.3% (DFS), respectively (OS, P=0.802; DFS, P=0.765). Additionally, there was no significant difference in OS and DFS among the three groups stratified to stage IA or IC (OS (IA), P=0.387; DFS (IA), P=0.314; OS (IC), P=0.993; DFS (IC), P=0.990, respectively). Furthermore, patients with a grade 1-2 or 3 tumours in the FSS group did not have a poorer prognosis than those in the RS group. CONCLUSIONS: Stage I EOC patients treated with FSS showed an acceptable prognosis compared with those who underwent RS.

Ambulation-promoting effect of peppermint oil and identification of its active constituents.

Various plant-derived essential oils (EOs) have traditionally been used in the treatment of mental disorders, despite a lack of scientific evidence. In a previous study, we demonstrated that certain EOs possess behavioral effects, a finding that supports our original hypotheses that EOs possess psychoactive actions. The present study was conducted in order to obtain further evidence to support our hypothesis. Peppermint oil, a type of EO, is believed to be effective for treating mental fatigue. When the oil was administered intraperitoneally to ICR mice, the ambulatory activity of mice increased dramatically. We identified alpha-pinene, beta-pinene, (R)-(+)-limonene, 1,8-cineol, isomenthone, menthone, menthol, (R)-(+)-pulegone, menthyl acetate and caryophyllene as constituent elements of peppermint oil by GC-MS analysis. We then examined the effect of each constituent element of peppermint oil on ambulatory activity in mice. Intraperitoneal administration of 1,8-cineol, menthone, isomenthone, menthol, (R)-(+)-pulegone, menthyl acetate and caryophyllene significantly increased ambulatory activity in mice, suggesting that these chemicals are the behaviorally active elements of peppermint oil. Intravenous administration of these substances to mice induced a significant increase in ambulatory activity at much lower doses. The present study provides further evidence demonstrating that EOs possess pharmacological actions on behavior. In addition, our finding revealed that the action of peppermint oil comes from its constituent elements.

Behavioral effects of plant-derived essential oils in the geller type conflict test in mice.

The present study was conducted to further explore plant-derived essential oils that possess an anticonflict effect using the Geller type conflict test in ICR mice. The benzodiazepine anxiolytic diazepam increased the response (lever pressing) rate during the alarm period (i.e., an anticonflict effect), but the 5-HT1A partial agonist buspirone did not. Oils of juniper, cypress, geranium and jasmine did not produce any effect in this test. Frankincense oil decreased the response rate during the safe period at 1600 mg/kg, but did not exhibit any effect on the response rate during the alarm period. In contrast, lavender oil increased the response rate during the alarm period in a dose-dependent manner in the same manner as diazepam. These results indicate that not only rose oil but also lavender oil possess an anticonflict effect in mice.

Anticonflict effects of plant-derived essential oils.

The present study examined the pharmacological actions of four different plant-derived essential oils (rose, ylang-ylang, camomile, orange) in two types of conflict tests using ICR mice. In the Vogel conflict test, in which any drinking behavior of the mice was punished by an electric shock, the benzodiazepine agonist, diazepam (DZ), increased the number of electric shocks the mice received. This number increased after administration of rose oil. In contrast, ylang-ylang, camomile, and orange oil did not produce such an effect in this test. In the Geller conflict test where lever-pressing of mice was reinforced by food pellets and then punished by electric shock, response (lever-pressing) rate during the alarm period was increased as well by the positive control drug, DZ. Similarly, the response rate during the alarm period increased after administration of rose oil. Here as well, ylang-ylang, camomile, and orange oils did not produce an anticonflict effect. In the Vogel conflict test, the anticonflict effect of DZ was reversed by the benzodiazepine antagonist, flumazenil (Ro15-1788) (FL). However, the effect of rose oil in this test was not antagonized by FL. The present study showed that rose oil possesses anticonflict effects, and that the effects are not mediated by the benzodiazepine binding site of the GABA(A) receptor complex. Such pharmacological actions may at least partially account for human behavioral effects attributed to essential oils.

Effects of psychoactive drugs in the Vogel conflict test in mice.

This study examined effects of various psychoactive drugs on the Vogel conflict test, where drinking behavior is punished by electric shocks, in ICR mice to clarify the pharmacological features of this method in mice. A benzodiazepine anxiolytic diazepam and a barbiturate pentobarbital produced significant anticonflict effects, which mean that these drugs increased the number of electric shocks mice received during 40-min test session. On the other hand, yohimbine (alpha2-receptor antagonist), caffeine (adenosine-receptor antagonist), scopolamine (muscarinic cholinergic antagonist), cyclazocine (sigma-receptor antagonist), cimetidine (H2-receptor antagonist), baclofen (GABA(B)-receptor agonist), MK-801 (NMDA-receptor antagonist), buspirone (5-HT1A-receptor agonist), chlorpromazine (dopamine-receptor antagonist) and haloperidol (dopamine-receptor and sigma-receptor antagonist) all did not produce anticonflict effects in this test using ICR mice. The results suggest that the Vogel conflict test is applicable to ICR mice and that this test in mice is appropriate as a screening method for drugs that have apparent anti-anxiety actions.

Circadian behavior and plasticity of light-induced c-fos expression in SCN of tau mutant hamsters.

In hamsters homozygous for the circadian clock mutation tau, the photic history dramatically affects the magnitude of light-induced circadian phase shifts. The maximum amplitude of phase shifts produced by 1-h light pulses presented at CT 14 was less than 2 h in animals that had been in DD for 2 days, whereas animals that had been kept in DD for 49 days could be shifted by more than 8 h. In this study, the authors compared the effect of previous light history on the amplitude of circadian phase shifts and on c-fos expression in the SCN of tau mutant hamsters. Although the maximum amplitude of behavioral phase shifts was drastically different between animals that had been held for either 2 or 49 days in DD, maximal fos induction was not significantly different in these two groups. However, photic thresholds for light-induced behavioral phase shifts, c-fos mRNA, and Fos immunoreactivity were closely correlated within both groups, and these thresholds were lower (more sensitive to light) after 49 than after 2 days in DD. The correlation between phase shifting and Fos induction thresholds, under conditions where both responses are dramatically altered by the previous light history, demonstrates an association between changes in circadian behavioral phase-shifting responses of tau mutant hamsters and plasticity of light-induced c-fos expression in SCN. However, because the maximum amplitudes of Fos induction and phase shifting were not correlated in animals that had been in DD for 2 days, we speculate that the level of c-fos expression does not directly determine phase shift amplitude.

Tris(2-chloroethyl)phosphate increases ambulatory activity in mice: pharmacological analyses of its neurochemical mechanism.

The present study was conducted to clarify the acute effect of tris(2-chloroethyl)phosphate (TRCP), an organophosphate flame-retardant, on spontaneous ambulatory activity (AA) in male ICR mice and to examine the neurochemical mechanism of this effect. Single dose administration of 200 mg/kg i.p. of TRCP increased AA in ICR mice. Neither the nicotinic cholinergic antagonist mecamylamine (MA) nor the muscarinic cholinergic antagonist scopolamine (SCP) affected the AA response to TRCP. On the other hand, the benzodiazepine agonist diazepam (DZ), the GABAA agonist muscimol (MUS) and the GABAB agonist baclofen (BAC) all attenuated the effect of TRCP. DZ and MUS blocked the increase of AA within the first 10 min after administration of TRCP. These drugs did not attenuate the AA-increasing effect of SCP, suggesting that the mechanism of TRCP action is distinct from that of SCP. MUS and BAC did, but DZ did not, inhibit the AA increasing effect of the dopaminergic agonist apomorphine (APO), suggesting that dopamine is involved in the control of AA, and that GABA can affect AA through interaction with dopaminergic neurons. These results suggest that TRCP acts as a GABA antagonist and not as a cholinergic agonist, and that TRCP increases AA in ICR mice through a GABAergic mechanism.

Behavioral effects of trichloroethylene and tetrachloroethylene in mice.

This study was performed to clarify the toxicological profiles of trichloroethylene (TRCE) and tetrachloroethylene (TECE) when they are administered intraperitoneally in mice. The ED50 for loss of righting reflex were 2596 mg/kg in TRCE and 4209 mg/kg in TECE. TRCE and TECE impaired bridge test performance at 500 and 2000 mg/kg, respectively. An operant behavior performance was also inhibited by TRCE at 1000 mg/kg and by TECE at 2000 mg/kg. Both TRCE and TECE exhibited anticonflict effects in a Vogel-type task at 500 mg/kg. This effect was confirmed by the finding that TRCE exhibited anticonflict action in a Geller-type paradigm at 250 mg/kg and more, as did TRCE did at 1000 mg/kg. These results show that TRCE and TECE affect various behaviors in mice and suggest that conflict behaviors are one of the most sensitive behavioral indicators of the effects of these substances. The toxicological profiles of TRCE and TECE with respect to behavioral effects were very similar, and they can be classified in a single category.

Hypersensitivity reactions to cisplatin following multiple uncomplicated courses: a report on two cases.

Two patients with ovarian cancer who had received multiple courses of cisplatin without complications experienced hypersensitivity reactions to cisplatin: one, involving intrahepatic artery infusion, manifested general erythema, dyspnea, and hypotension; the other, involving intravenous infusion, manifested abdominal pain, general erythema, and fever. When platinum-containing agents are administered, patients should be closely monitored for hypersensitivity reactions, even in previously uncomplicated cases.

[Assessment of anxiolytics (5)--Vogel-type conflict task in mice].

The Vogel-type conflict task, in which drinking of water by animals is punished by electric shock, is well known as a simple conflict procedure, and has come into wide use. This task has almost been established in rats. On the other hand, there are only few reports about application of the task to mice, and such studies have not proceeded smoothly. This report presents technical problems in the application of the task to mice based on various experiments, in which experimental conditions such as procedures, intensity of electric shock and mouse strain differed, in order to determine the anti-conflict action of diazepam (DZ). These experiments showed that successful detection of DZ action depended upon the experimental conditions. Therefore, experimental designs are important in evaluating anti-conflict action of chemicals in the Vogel-type task in mice.

Long-continuous observation of the effects of methamphetamine on wheel-running and drinking in mice.

1. Effects of methamphetamine (MAP) on wheel-running and drinking in mice, housed under 12-hr light-dark schedule (light period; 06:00-18:00), were investigated through long-continuous observation. 2. MAP (1, 2 and 4 mg/kg, s.c.) acutely increased the wheel-running and drinking for 2-4 hr in a dose-dependent manner after the administration at 11:00, midpoint of light period. 3. MAP administered at 11:00 sub-acutely suppressed the spontaneous increment during dark period (18:00-06:00) in both the behaviors. 4. Many factors, such as the time-of-day and interval of the administration as well as the dose administered, affected the behavioral suppression induced by MAP. 5. In addition to these findings, the wheel-running and drinking during the light period increased even on the days without MAP administration. 6. These results suggest that MAP have not only acute stimulant and sub-acute depressant effects, but also long-lasting effects.

Effects of ozone and nitrogen dioxide on drinking and eating behaviors in mice.

Male ICR mice were exposed continuously to ozone (O3) and nitrogen dioxide (NO2) for 7 days to examine the effects on drinking and eating behaviors. Ozone at 0.1 ppm did not affect drinking and eating activities, whereas drinking activity decreased in a concentration-dependent manner to 47.7, 12.8, and 3.0% of the control value with 2-day exposures to 0.2, 0.4, and 0.8 ppm O3, respectively, and eating activity decreased to 35.2 and 8.7% of the control value at 0.4 and 0.8 ppm O3, respectively. Body weight also decreased markedly by 2.0, 4.6, and 7.5 g at 0.2, 0.4, and 0.8 ppm O3, respectively. These decrements reached a maximum on the second day of exposure. However, alterations in drinking and eating activities and body weight were transient, leading to recovery during the continuous O3 exposures. The recovery processes were dependent on the concentrations of O3. Nitrogen dioxide at 4 ppm did not affect drinking and eating activities, whereas drinking activity decreased in a concentration-dependent manner to 56.8, 8.3, and 18.7% of the control value with 2-day exposures to 6, 8, and 12 ppm NO2, respectively, and eating activity decreased markedly to 21.8 and 16.4% at 8 and 12 ppm NO2, respectively. Body weight also decreased by 2.5, 5.5, and 6.1 g at 6, 8, and 12 ppm NO2, respectively. These decrements reached a maximum on the second day of exposure. As in the O3 exposures, the decrements in drinking and eating activities and body weight were transient and recovered during the continuous exposures to NO2 depending on the concentrations of NO2. Drinking and eating activities and body weights of mice that had been previously exposed to 12 ppm NO2 for 7 days did not show changes when the mice were exposed to 0.4 ppm O3 9 days after NO2 exposure. The present study demonstrates that photochemical oxidants suppress drinking and eating behaviors in mice and that they recover thereafter under the continuous exposure conditions.

Malignant mixed Müllerian tumor of the fallopian tube: report of two cases and review of literature.

Malignant mixed Müllerian tumors are usually found in the endometrium, vagina, cervix, and ovary. It is extremely rare for this tumor to arise in the fallopian tube, and to date only 37 tubal cases have been reported. We recently experienced 2 such cases. The clinical features, pathologic findings, diagnosis, therapy, and outcome of these 39 cases were reviewed. The clinical features and diagnosis were similar to those of primary carcinoma of the fallopian tube. Correct preoperative diagnosis was difficult. Histologically, 18 patients had homologous elements and 21 had heterologous elements in the sarcomatous components. The most common type of heterologous element was cartilage, followed by striated muscle and bone. The clinical stage (FIGO staging of ovarian carcinoma) was stage I in 15 cases, stage II in 11 cases, stage III in 8 cases, stage IV in 3 cases, and unknown in 2 cases. In all the patients except 1, the tumor was surgically removed. Postoperatively, radiotherapy was given to 9 patients, chemotherapy to 9 patients, and both to 2 patients. Sixteen patients died of the disease, after a mean period of 16.1 months. Of the 15 stage I patients, 10 survived more than 12 months. The most important prognostic factor was spread of the tumor at diagnosis.

[Effects of nicotine on circadian rhythm of ambulatory activity and drinking in rats].

Effects of the intake of nicotine solution and/or forced administration of nicotine at a fixed time-of-day on circadian rhythm of ambulatory activity and drinking in rats were observed under various conditions. Rats showed nocturnal pattern under the light-dark cycle (L: D = 12:12, light period 6:00-18:00), and the patterns scarcely altered while the rats were given 150 micrograms/ml of nicotine solution. When a restricted feeding only for 1 h per day (10:00-11:00) was loaded to the rats, the ambulatory activity and drinking appeared in the light period, especially around the feeding time. While rats were given 15-150 micrograms/ml of nicotine solution under the restricted feeding condition, they demonstrated a marked phase-shift in the circadian rhythm in the concentration-dependent manner. Rats showed free-running rhythms with tau value of longer than 24 h under the constant red dim light condition (CRDL), and such rhythm was scarcely modified by the intake of nicotine solution of 50-150 micrograms/ml. However, when nicotine (1.5 mg/kg po), but not tap water, was administered at 10:00 every day under the CRDL condition, it also produced phase-shift which was similar to that induced by the nicotine ingestion under the redose of nicotine is administered or taken at a fixed time-of-day.

Early gastric cancer following gastrojejunostomy without gastrectomy.

Two rare cases involving distal gastrectomies for gastric cancer after an earlier gastrojejunostomies without gastrectomy for duodenal or gastric stricture due to a duodenal or pre-pyloric ulcer are reported. The time interval between the initial gastrojejunostomy and the resection of the distal stomach with gastrojejunal stoma were 17 and 36 years. One patient underwent reconstruction by Billroth II and the other by Roux-en-Y end-to-side gastrojejunostomies, both retrocolically without any peri or postoperative complications. The histological classification was confirmed for both as type IIc mucosal early gastric cancer. Only 6 other primary early gastric cancers following gastrojejunostomies without gastrectomy have been reported in the Japanese literature.

Postoperative subphrenic abscess following gastrectomy for gastric cancer.

From 1985 to 1989, a total of 598 gastric cancer patients underwent gastrectomies in the First Department of Surgery at Kurume University School of Medicine. Of these, 24 patients (4.0%) incurred a subphrenic abscess, postoperatively. Subphrenic abscesses occurred most frequently after total gastrectomies at a rate of 17 of 197 (8.8%) cases. Leaking gastrointestinal anastomoses were responsible for 12 of the 17 (70.5%) abscesses. In 384 cases of distal gastrectomy, only 6 (1.6%) cases with a subphrenic abscess occurred, and only 3 were due to anastomotic leakage. Among the total of 24 cases with subphrenic abscesses, 16 (67%) were related to an anastomotic leakage including a pancreatic fistula, while the other 8 (33%) showed no leakage by contrast roentgenography. When chest roentgenography shows a pleural effusion and an elevation of the diaphragm, then a subphrenic abscess must be suspected. The majority of patients were treated by replacing the intraabdominal drainage catheter inserted during the gastrectomy with a larger catheter. The overall operative mortality rate was 4.1% due to a death in only one (MOF) patient.