Patient-derived spheroids and patient-derived organoids simulate evolutions of lung cancer.

Cancer cells harbor many genetic mutations and gene expression profiles different from normal cells. Patient-derived cancer cells (PDCC) are preferred materials in cancer study. We established patient-derived spheroids (PDSs) and patient-derived organoids (PDOs) from PDCCs isolated from the malignant pleural effusion in 8 patients. The morphologies suggested that PDSs may be a model of local cancer extensions, while PDOs may be a model of distant cancer metastases. The gene expression profiles differed between PDSs and PDOs: Gene sets related to inflammatory responses and EMT were antithetically regulated in PDSs or in PDOs. PDSs demonstrated an attenuation of the pathways that contribute to the enhancement of transforming growth factor beta (TGF-ß) induced epithelial mesenchymal transition (EMT), while PDOs demonstrated an attenuation of it. Taken together, PDSs and PDOs have differences in both the interaction to the immune systems and to the stroma. PDSs and PDOs will provide a model system that enable intimate investigation of the behavior of cancer cells in the body.

Cytotoxicity against HL60 Cells of Ficifolidione Derivatives with Methyl, n-Pentyl, and n-Heptyl Groups.

Ficifolidione, a natural insecticidal compound isolated from the essential oils of Myetaceae species, is a spiro phloroglucinol with an isobutyl group at the C-4 position. We found that ficifolidione showed cytotoxicity against cancer cells via apoptosis. Replacement of the isobutyl group by n-propyl group did not influence the potency, but the effect of the replacement of this group by a shorter or longer alkyl group on the biological activity remains unknown. In this study, ficifolidione derivatives with alkyl groups such as methyl, n-pentyl, and n-heptyl group-instead of the isobutyl group at the C-4 position-were synthesized to evaluate their cytotoxicity against the human promyelocytic leukaemia cell line HL60 and their insecticidal activity against mosquito larvae. The biological activities of their corresponding 4-epimers were also evaluated. As a result, the conversion of the isobutyl group to another alkyl group did not significantly influence the cytotoxicity or insecticidal activity. In HL60 cells treated with the n-heptyl-ficifolidione derivative, the activation of caspase 3/7 and the early stages of apoptosis were detected by using immunofluorescence and flow cytometric techniques, respectively, suggesting that the cytotoxicity should be induced by apoptosis even though the alkyl group was changed.

Bilateral submandibular sialadenitis following influenza A virus infection.

A 48-year-old male presented to our hospital with pyrexia and general fatigue. The patient was diagnosed with influenza A virus infection, and then given laninamivir octanoate hydrate. Eleven days after onset, he noticed bilateral swollen submandibular glands. He visited our hospital again, and underwent blood examination and cervical computed tomography (CT). CT indicated that bilateral submandibular glands were swelling. We believe that this is the first report of bilateral submandibular sialadenitis following influenza A virus infection, and thus we presented this patient.

Revised stereochemistry of ficifolidione and its biological activities against insects and cells.

Ficifolidione (1), a moderately active insecticidal compound from two species of Myrtaceae, and its derivatives were synthesized to evaluate their insecticidal activity. X-ray crystallographic analyses and specific rotation values of ficifolidione and its C-4 (2) demonstrated that the structure of ficifolidione differs from the reported absolute structure; that is, the C-4 configuration of ficifolidione should have an S configuration. The reported insecticidal activity of ficifolidione (1) and its C-4 epimer (2) against adult houseflies (Musca domestica), mosquito larvae (Culex pipiens), and cutworms (Spodoptera litura) was not observed. The cytotoxicities of ficifolidione and its derivatives (1-4) against four cell lines, Sf9, Colon26, HL60, and Vero, were also measured because ficifolidione has a phloroglucinol-derived moiety, a motif that is often present in the structure of cytotoxic chemicals. Compound 1 exhibited IC50 values of ca. 32, 9, 3, and 12 µM for Sf9, Colon26, HL60, and Vero cells, respectively, indicating that ficifolidione possesses selective cytotoxicity against the four cell lines. In HL60 cells treated with 1, DNA fragmentation and the activation of procaspase 3 were observed, suggesting that the cytotoxicity is induced by apoptosis.

Three-dimensional relationship between the conus branch and the precordial leads confirmed by 64-multidetector-row computed tomography.

A 65-year-old man with effort angina pectoris underwent percutaneous coronary intervention of the proximal right coronary artery. The lesion was dilated with a bare-metal stent under wire protection of the conus branch (CB). However, the jailed CB was occluded. Electrocardiogram with conventional precordial leads (V(1) through V(6)) accompanied with the supplementary leads (V(1) through V(6)) positioned 1 intercostal space higher showed marked ST elevation in V(1) through V(3) that was more prominent in V(1) through V(3). The 64-multidetector-row computed tomographic coronary angiography showed recanalization of the CB located just in the center of the V(1), V(2), V(1), and V(2) electrodes.

Potent antioxidant and radical-scavenging activities of traditional Japanese cereal grains.

To estimate the preventive potential of Japanese traditional cereals against oxygen radical-related chronic diseases such as cardiovascular diseases and diabetes, antioxidant and radical-scavenging activities in the extracts of five Japanese traditional cereal grains were analyzed by using an assay system of lipid peroxidation and a radical compound, 1,1-diphenyl-2-picrylhydrazyl (DPPH). DPPH radical-scavenging activities in the extracts of Japanese cereal grains were divided into two groups. One group including Japanese sorghum, black rice and red rice showed strong radical-scavenging activities, but the other group including Japanese barnyard millet and foxtail millet did not exhibit significant radical-scavenging activities. The DPPH radical-scavenging activities of these extracts were closely correlated to the contents of phenolic compound in the extracts, but not to the sugar or protein content in the extracts. In contrast, all the methanol and water extracts of the cereal grains caused significant antioxidant activities against hydroperoxide generation in the peroxidation of linoleic acid, in which the water extracts of these cereal grains caused much higher antioxidant activities than the methanol extracts of the same cereals. These results suggest that Japanese traditional cereals contain qualitatively different principles associated with antioxidant and radical-scavenging activities, and possible principles responsible for the antioxidant and radical-scavenging activities in the cereal grains are discussed.

Outcome of percutaneous intrapericardial fibrin-glue injection therapy for left ventricular free wall rupture secondary to acute myocardial infarction.

Left ventricular free wall rupture (LVFWR) is a fetal complication of acute myocardial infarction. This study was conducted to test the feasibility of percutaneous intrapericardial fibrin-glue injection therapy (PIFIT) for LVFWR after acute myocardial infarction and to assess its clinical outcome. From January 2000 to December 2004, LVFWR was confirmed by echocardiography in 22 patients. Thirteen patients showing abrupt hemodynamic collapse failed to recover from resuscitation maneuvers and died <2 hours after LVFWR. The remaining 9 patients (5 women, mean age 73 +/- 10 years) underwent PIFIT. Pericardiocentesis was performed from the subxiphoid process, and a 6Fr pigtail catheter was introduced into the pericardial space. After bloody fluid was drained from the catheter, the fibrin glue was injected into the pericardial space. There were no complications relating to pericardiocentesis and PIFIT. One patient underwent surgical repair on the day of PIFIT because of uncontrollable bleeding from pericardial drainage. In-hospital death as a result of rerupture occurred in 2 patients on days 4 and 7 after PIFIT. Echocardiography during follow-up revealed no evidence of pseudoaneurysm or left ventricular restriction. On follow-up at a median of 4.0 years (interquartile range 3.1 to 4.8), 1 noncardiac death occurred at 3.3 months. The other 5 patients were free of cardiovascular events and in New York Heart Association functional class I. In conclusion, PIFIT is a simple, effective, and less invasive technique for the management of LVFWR and thus can be an alternative to surgical repair for LVFWR after acute myocardial infarction.

Responsiveness to dobutamine stimulation in patients with left ventricular apical ballooning syndrome.

To investigate the underlying mechanisms of the left ventricular (LV) apical ballooning syndrome, we evaluated the functional responsiveness to dobutamine stimulation in patients with the syndrome. Over a 22-month period, 11 consecutive patients with the apical ballooning syndrome were referred to our institution. All 11 patients were women and 57 to 85 years of age (mean 73 +/- 10). Among them, 10 patients underwent low-dose dobutamine echocardiography within 24 hours after admission (17 +/- 8 hours). Echocardiography was repeated in the convalescent phase (48 +/- 33 days) to assess functional outcome. In the resting state, all patients showed akinetic wall motion in the midportion of the left ventricle and apical left ventricle. After low-dose dobutamine infusion, akinetic wall motion detected at rest did not show any improvement despite the hypercontractile basal LV wall. In the convalescent phase, LV dysfunction was not observed on echocardiography in all 11 patients. The LV apical ballooning syndrome has a unique feature that reversible dysfunction lacks functional amelioration during dobutamine administration. In conclusion, this finding suggests that the pathophysiologic mechanisms of the syndrome appear to be distinct from those of myocardial stunning after transient ischemia, and catecholamine-mediated cardiac toxicity may play a role in the development of the syndrome.

Rapid separation and concentration of food-borne pathogens in food samples prior to quantification by viable-cell counting and real-time PCR.

Buoyant density gradient centrifugation has been used to separate bacteria from complex food matrices, as well as to remove compounds that inhibit rapid detection methods, such as PCR, and to prevent false-positive results due to DNA originating from dead cells. Applying a principle of buoyant density gradient centrifugation, we developed a method for rapid separation and concentration following filtration and low- and high-speed centrifugation, as well as flotation and sedimentation buoyant density centrifugation, for 12 food-borne pathogens (Salmonella enterica, Escherichia coli, Yersinia enterocolitica, Campylobacter jejuni, Vibrio cholerae O139, Vibrio parahaemolyticus O3K6, Vibrio vulnificus, Providencia alcalifaciens, Aeromonas hydrophila, Bacillus cereus, Staphylococcus aureus, and Clostridium perfringens) in 13 different food homogenates. This method can be used prior to real-time quantitative PCR (RTi-qPCR) and viable-cell counting. Using this combined method, the target organisms in the food samples theoretically could be concentrated 250-fold and detected at cell concentrations as low as 10(1) to 10(3) CFU/g using the RTi-qPCR assay, and amounts as small as 10(0) to 10(1) CFU/g could be isolated using plate counting. The combined separation and concentration methods and RTi-qPCR confirmed within 3 h the presence of 10(1) to 10(2) CFU/g of Salmonella and C. jejuni directly in naturally contaminated chicken and the presence of S. aureus directly in remaining food items in a poisoning outbreak. These results illustrated the feasibility of using these assays for rapid inspection of bacterial food contamination during a real-world outbreak.

Clinical relationship of myocardial sympathetic nervous activity to cardiovascular functions in chronic heart failure: assessment by myocardial scintigraphy with 123I-metaiodobenzylguanidine.

The aim of this study was to clarify the relationship between cardiac sympathetic nervous activity (SNA) assessed by radioiodinated metaiodobenzylguanidine (123I-MIBG), an analogue of norepinephrine and cardiovascular functions in patients with chronic heart failure (CHF). Subjects were 17 patients with CHF. A dose of 111 MBq of 123I-MIBG was administered intravenously, and 5-minute anterior planar images were obtained 15 minutes (early image) and 3 hours (delayed image) after the injection. The heart/mediastinum (H/M) count ratio was defined to quantify cardiac 123I-MIBG uptake. The washout ratio (WR) of 123I-MIBG from the heart was calculated as follows: (early counts-delayed counts)/early counts x 100 (%). Echocardiography was performed on all patients within 1 week of 123I-MIBG scintigraphy to measure stroke volume index (SVI). Blood pressure and heart rate (HR) in the resting state were also recorded to calculate cardiovascular functions including cardiac output, pulse pressure (PP), and mean blood pressure. Significant linear correlations were found between the early H/M ratio of 123I-MIBG and SVI, and between the delayed H/M ratio of 123I-MIBG and SVI, respectively. WR of 123I-MIBG was correlated with HR, and was inversely correlated with SVI and with PP, respectively. It is likely that a decrease in SVI is associated with enhanced cardiac SNA in severe CHF. 123I-MIBG scintigraphy is effective in assessing the cardiac functional status and SNA in patients with CHF in vivo. Moreover, changes in PP and HR indicate well alteration in SNA.

A new miniature catheter with side-holes for percutaneous transradial or transbrachial coronary angiography.

The percutaneous arm approach through the radial or brachial artery for diagnostic cardiac catheterization has advantages, such as a lower incidence of access-site complications and decreased patient discomfort, particularly when smaller diameter catheters are employed. However, the pressure produced by high-flow jets of contrast material exiting from an end-hole against the vascular wall can cause coronary dissection or myocardial blushing. To avoid this type of complication, we designed and developed a new miniature (4 French) catheter with two side-holes for coronary angiography. Under conditions similar to those of a clinical situation, we obtained the relationship between the pressure produced by the jets exiting from an end-hole and the force with which the tip of the catheter was pushed against the vascular wall. The presence of the side-holes substantially decreased the pressure at forces ranging between 0 and 15 gf, and at all injection rates (2.0 3.5 ml/second), but their location did not affect the pressure. In a preliminary clinical study, the new catheters with side-holes were feasible and effective for coronary angiography through the radial artery. Furthermore, the distribution of contrast material decreased the incidence of dislodgement of the catheter from the coronary ostia and enabled good visualization of the coronary ostia and proximal branches.