Enhancement of in vitro tumor—cell transcellular migration by tumor—cell—secreted endothelial—cell—retraction factor

To investigate the factors affecting endothelial-cell retraction, we have studied the interaction of tumor cells with endothelial cells in 2 human pancreatic cancer cell lines, PSN-1 and MiaPaca-2. The extent of endothelial-cell retraction measured by the amount of intercellular junctional transport of F1TC-dextran through an endothelial monolayer was increased by the addition of a conditioned medium (CM) from both cell lines, while CM from PSN-I cells was 2 to 3 times more potent than that from MiaPaca-2 cells. After the treatment of endothelial monolayer with CM of PSN-1 cells, the ability of both PSN-1 cells and MiaPaca cells to adhere to or invade the monolayer increased. The addition of CM from PSN-1 cells did not affect the growth rate of either the endothelial or the tumor cells. The activity in the CM was heat-stable and bound to heparin-Sepharose, but was inactivated when treated by 0.5% trypsin. Protease inhibitors did not influence the activity. Pre-treatment of PSN-I cells by an inhibitor of protein synthesis, cyclohexi-mide, or of protein processing, benzyl-N-acetyl-a-D-galactos-aminide, reduced endothelial-cell-retraction activity in the CM. The active substance in the CM fractionated in the molecular-weight range of 10,000 to 50,000. These results suggest that PSN-1 cells produce and secrete (a) soluble factor(s) that can induce endothelial-cell retraction, thus facilitating tumor-cell invasion.     

Exosomal miRNA profiles of triple-negative breast cancer in neoadjuvant treatment

Triple-negative breast cancer (TNBC) is characterized by aggressive clinicopathological features and is associated with a poor prognosis. Identifying patients that are non-responsive to chemotherapy remains a critical goal for effective personalized therapies. In the present study, the predictive value of exosomal microRNAs (miRNAs) was investigated in patients with TNBC. Exosomes were isolated from patients with TNBC undergoing neoadjuvant chemotherapy. Microarray-based miRNA profiles were compared between patients with pathological complete response (pCR; n=12) and non-pCR (n=12). Furthermore, the miRNA profiles of non-pCR patients with breast cancer recurrence were compared with those with no recurrence. A total of 16 differentially expressed exosomal miRNAs were identified between the patients with pCR and non-pCR by microarray analysis. Of these, a combined signature of four miRNAs (miR-4448, miR-2392, miR-2467-3p and miR-4800-3p) could be used to discriminate between pCR and non-pCR patients with TNBC with an area under the curve value of 0.7652. Furthermore, this study found 43 differentially expressed miRNAs between the patients with non-pCR and recurrence and non-pCR patients without recurrence. In network analysis, ‘pathway in cancer’, ‘focal adhesion’ and ‘cell cycle’ were identified as the crucial pathways in patients with non-pCR who also developed recurrence. Several exosomal miRNAs may be useful biomarkers to predict treatment efficacy for TNBC. The present study identified patients who were resistant to standard chemotherapy and therefore more likely to develop breast cancer recurrence.     

Effect of micro-environment maintenance on embryo culture after in-vitro fertilization: comparison of top-load mini incubator and conventional front-load incubator

Purpose: To investigate the effect of microenvironment maintenance on embryo culture and clinical results using two types of incubators. Methods: Temperature and oxygen concentration in a mini-incubator and a conventional incubator were compared following a 5-s door opening/closing procedure. Embryos of 30 in-vitro fertilization embryo transfer (IVF-ET) cases were randomly allocated to either one of the incubator, cultured, and the early-stage good embryo formation rate and the good blastocyst formation rate were compared, as indicators for micro-environment maintenance ability. Results: Temperature recovery after a 5-s door opening/closing procedure was approximately 5 min for the mini-incubator and 30 min for the conventional incubator. The oxygen concentration return was significantly improved in the mini-incubator (3.0 ± 0 min) compared with the conventional incubator (7.8 ± 0.9 min). Both the early-stage good embryo formation rate and the good blastocyst formation rate were significantly higher in the mini-incubator (39.5% and 15.1%) than the conventional incubator (28.4% and 7.8%). Conclusion: The microenvironment maintenance ability of incubators appears to significantly influence the formation of good embryos.     

Serum hepatocyte growth factor levels are increased in patients with congestive heart failure

BACKGROUND: Hepatocyte growth factor (HGF) is a potent endothelial cell-specific mitogen. We investigated the clinical importance of HGF in congestive heart failure (CHF). METHODS AND RESULTS: Thirty-five patients with acute exacerbation of CHF and 7 control subjects were examined. Serum and peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood on days 1, 7, and 14 after admission. PBMCs were cultured at a density of 1 x 10(7) cells/mL for 24 hours. HGF levels in serum and the PBMC culture medium and serum interleukin 6 (IL-6) levels were measured by enzyme-linked immunosorbent assay. Serum HGF levels in patients with CHF were markedly increased at admission compared with those in control subjects and gradually returned to control levels during hospitalization. HGF levels in the PBMC culture medium were also significantly increased in CHF patients compared with control subjects. There was a positive correlation between HGF levels in serum or those in the PBMC culture medium and serum IL-6 levels. HGF levels in serum and the culture medium were not notably different between CHF patients regularly treated with and without angiotensin-converting enzyme inhibitors. CONCLUSIONS: HGF levels in serum are increased in patients with acute exacerbation of CHF.     

Identification of biomarkers in ductal carcinoma <Emphasis Type="Italic">in situ</Emphasis> of the breast with microinvasion

Background  

Widespread use of mammography in breast cancer screening has led to the identification of increasing numbers of patients with ductal carcinoma in situ (DCIS). DCIS of the breast with an area of focal invasion 1 mm or less in diameter is defined as DCIS with microinvasion, DCIS-Mi. Identification of biological differences between DCIS and DCIS-Mi may aid in understanding of the nature and causes of the progression of DCIS to invasiveness.     

Long-term course of lipoprotein lipase (LPL) deficiency due to homozygous LPL(Arita) in a patient with recurrent pancreatitis, retained glucose tolerance, and atherosclerosis

CONTEXT: Lipoprotein lipase (LPL) deficiency is a rare autosomal recessive disorder caused by LPL gene mutation and is characterized by severe hyperchylomicronemia. Patients with LPL deficiency suffer from the frequent recurrence of acute pancreatitis, but the underlying mechanisms are not fully understood. CASE REPORT: A 22-yr-old male Japanese patient with severe hyperchylomicronemia was admitted to our hospital in 1973. He had no consanguinity and no family history of hyperlipidemia. He was genetically diagnosed as LPL deficiency (homozygous for LPL(Arita)) with no LPL mass or activity in postheparin plasma. He has experienced recurrent acute pancreatitis 22 times during our 31-yr clinical follow-up, but no pancreatic pseudocyst, irregularity of the pancreatic duct, or abnormal pancreatic calcification was observed in computed tomography. Moreover, his pancreatic endocrine function, as assessed by the oral glucose tolerance test, has preserved more than 30 yr. Although he was a current smoker, no clinically significant atherosclerotic lesion had been observed. CONCLUSIONS: From the long-term observation of this patient, we propose that LPL deficiency is not invariably associated with high mortality and that even with repeated episodes of acute pancreatitis, pancreatic function may be slow to decline.     

Comparative susceptibility of newborn and young rats to six industrial chemicals

To elucidate the comparative susceptibility of newborn rats to chemicals, newborn and young animals were administered six industrial chemicals by gavage from postnatal days (PND) 4 to 21, and for 28 days starting at 5-6 weeks of age respectively, under the same experimental conditions as far as possible. As two new toxicity endpoints specific to this comparative analysis, presumed no-observed-adverse-effect-levels (pNOAELs) were estimated based on results of both main and dose-finding studies, and presumed unequivocally toxic levels (pUETLs) were also decided. pNOAELs for newborn and young rats were 40 and 200 for 2-chlorophenol, 100 and 100 for 4-chlorophenol, 30 and 100 for p-(alpha,alpha-dimethylbenzyl) phenol, 100 and 40 for (hydroxyphenyl)methyl phenol, 60 and 12 for trityl chloride, and 100 and 300 mg/kg/day for 1,3,5-trihydroxybenezene, respectively. To determine pUETLs, dose ranges were adopted in several cases because of the limited results of experimental doses. Values for newborn and young rats were thus estimated as 200-250 and 1000 for 2-chlorophenol, 300 and 500 for 4-chlorophenol, 300 and 700-800 for p-(alpha,alpha-dimethylbenzyl) phenol, 140-160 and 1000 for (hydroxyphenyl)methyl phenol, 400-500 and 300 for trityl chloride, and 500 and 1000 mg/kg/day for 1,3,5-trihydroxybenzene, respectively. In most cases, newborn rats were 2-5 times more susceptible than young rats in terms of both the pNOAEL and the pUETL. An exception was that young rats were clearly more susceptible than their newborn counterparts for trityl chloride.     

Toxicological analyses over the past five years at a single institution

We investigated 352 patients with acute poisoning, 145 males and 207 females, who were sent to the Department of Emergency and Critical Care Center between June 1999 and March 2004. The insurance application rate averaged 72.5%. The causative agents included 98 prescribed drugs, 23 pesticides and herbicides, cyanide, arsenic and boric acid. Hypnotic and psychotropic drugs accounted for 192 cases (47.5%); antipyretics and analgesics, 93 (23.0%); pesticides and herbicides, 65 (16.1%); amphetamine-like drugs, 6 (1.5%); boric acid, 4 (1.0%); cyanide, 3 (0.7%); arsenic, 1 (0.2%); and other agents, 40 (9.9%). In young people, the most prevalent cause of poisoning was hypnotic,or psychotropic drugs prescribed in psychiatry clinics. In people older than 30, poisoning due to pesticides and herbicides was increased, reaching a peak among those in their 40s. Of 23 patients who died, 20 took Paraquat and Diquat, 2 took organophosphates and 1 took an overdose of antidepressants. The other 329 were transferred to different medical departments or were discharged from our hospital after recovery.     

Cardiovascular and intravesical pressure responses during natural micturition in conscious rats

Urinary bladder distension is known to influence the cardiovascular system under a pathophysiological condition such as spinal cord injury, hypertension, and arteriosclerosis. A reflex due to bladder distension and/or contraction is considered as one reason for the cardiovascular disturbance associated with micturition. However, it has remained unknown how much intravesical pressure (IVP) rises during micturition in daily life and to what extent mean arterial blood pressure (MAP) and heart rate (HR) respond at that time. To answer these questions, we attempted to examine the direct changes in IVP, MAP, and HR during natural micturition in freely moving conscious rats. IVP increased from the baseline value of 4 +/- 0.2 mmHg to 14 +/- 0.5 mmHg during natural micturition. Although MAP and HR began to increase before micturition, the increases in MAP and HR became significant 1-4 s before its onset. The peak increases in MAP and HR (7 +/- 0.8 mmHg and 14 +/- 3 beats/min, respectively) were delayed by 2 s from the peak IVP. Following an administration of xylocaine into the urinary bladder, the increases in MAP and HR during micturition were significantly blunted to 5 +/- 2 mmHg and 8 +/- 3 beats/min, although IVP increased the same as it did during micturition without xylocaine. Moreover, the relationship between IVP and MAP or HR during natural micturition resembled that between IVP and the vesico-cardiovascular reflex responses during isovolumic bladder contraction in anesthetized rats. Therefore it is concluded that natural micturition in freely moving conscious rats accompanies the significant cardiovascular responses despite a limited increase in intravesical pressure, to which a reflex from the urinary bladder may substantially contribute.