Ion Conductance-Based Perfusability Assay of Vascular Vessel Models in Microfluidic Devices.

We present a novel methodology based on ion conductance to evaluate the perfusability of vascular vessels in microfluidic devices without microscopic imaging. The devices consisted of five channels, with the center channel filled with fibrin/collagen gel containing human umbilical vein endothelial cells (HUVECs). Fibroblasts were cultured in the other channels to improve the vascular network formation. To form vessel structures bridging the center channel, HUVEC monolayers were prepared on both side walls of the gel. During the culture, the HUVECs migrated from the monolayer and connected to the HUVECs in the gel, and vascular vessels formed, resulting in successful perfusion between the channels after culturing for 3-5 d. To evaluate perfusion without microscopic imaging, Ag/AgCl wires were inserted into the channels, and ion currents were obtained to measure the ion conductance between the channels separated by the HUVEC monolayers. As the HUVEC monolayers blocked the ion current flow, the ion currents were low before vessel formation. In contrast, ion currents increased after vessel formation because of creation of ion current paths. Thus, the observed ion currents were correlated with the perfusability of the vessels, indicating that they can be used as indicators of perfusion during vessel formation in microfluidic devices. The developed methodology will be used for drug screening using organs-on-a-chip containing vascular vessels.

LC-MS analyses of N-acetyl-p-benzoquinone imine-adducts of glutathione, cysteine, N-acetylcysteine, and albumin in a plasma sample: A case study from a patient with a rare acetaminophen-induced acute swelling rash.

Acetaminophen (Paracetamol, APAP) has been widely used for many decades as an analgesic and antipyretic agent but APAP overdose often causes acute adverse reactions, particularly liver damage. The metabolically oxidized form of APAP, N-acetyl-p-benzoquinone imine (NAPQI), is chemically reactive and binds covalently to proteins. Therefore, NAPQI is believed to be the key metabolite that causes hepatotoxicity, especially under conditions of glutathione depletion. Other APAP-induced adverse reactions, such as skin damage, are rare and remain poorly studied. Here, we report a case study of a male patient who presented with an acute swelling skin rash (without hepatotoxicity) caused by therapeutic doses of APAP. Plasma samples were collected at 17 hr after dosing (during the manifestation of symptoms) and at one month (after recovery) and were subjected to LC-MS analysis of NAPQI-adducts. A significant concentration of NAPQI-cysteine adduct (33 pmol/mL) was found together with low concentrations of NAPQI-N-acetylcysteine adduct (2.0 pmol/mL) and NAPQI-glutathione adduct (0.13 pmol/mL). However, the NAPQI-albumin adduct was below the detection limit (below 0.001% modification on albumin) despite a previous report of high concentrations of NAPQI-albumin adduct following acute liver injury. Therefore, the observed APAP-induced skin damage may have had a different cause from APAP-induced liver injury.

A study of poor responders for long-term, low-dose macrolide administration for chronic sinusitis.

PURPOSE: We investigated the clinical factors (CT images, endoscopic nasal findings and allergic factors) involved in resistance of chronic sinusitis to macrolide therapy (ME) retrospectively. METHODS: ME was administered for 8-20 weeks in 68 adults with chronic sinusitis cases. The effect was evaluated in each factor from radiographic findings (R0-R3 according to the severity of the images), nasal findings (N0: no polyp, N1: a single polyp and N2: multiple polyps), allergic factors (A0: no allergy, A1: nasal allergy, A2: bronchial asthma) and objective nasal symptoms. In addition, an effect after polypectomy and histological examination were assessed for N1 and N2 groups. RESULTS: ME was effective in 70.6% (48/68 patients). The efficacy of ME was significantly less in the polyp group compared with the polyp-free group (p < 0.05). Therapeutic efficacy was significantly different between R1 and R3 groups (p < 0.05) with a tendency for worse outcome from R1 to R3. The efficacy in asthma patients was significantly less compared with patients with allergic rhinitis or no allergy (p < 0.05). The efficacy after polypectomy was significantly improved in N2 group but not in N1 group. The number of eosinophil/total inflammatory cells (%) in nasal polyps of resistant cases was significantly higher than in marked improved cases. CONCLUSION: The efficacy of ME was less in patients with polyposis; CT scans indicating severe findings, bronchial asthma and polyps with increased eosinophil infiltrations. Polypectomy resulted in significant improvement in the efficacy of ME.

Autopsy findings of patients with urological neoplasms.

BACKGROUND: A study was carried out to gain extensive understanding of the disease status in patients who had suffered and died of urological neoplasms. METHODS: The subjects were 524 patients who had died at the Department of Urology of Gunma Cancer Center. The autopsy rate for each urological, neoplasm, the extent of the disease, and incidental diseases were analyzed. RESULTS: Autopsies were performed in 27.1% (142/524) of the patients. Frequent metastatic sites were the lymph nodes, bone, and lung in prostate neoplasms; the lymph nodes, liver, and lung in bladder neoplasms; and the lymph nodes, lung, and bone in kidney neoplasms. In the 116 patients with these three major urological neoplasms, the autopsy findings of all patients were compatible with progression of the disease, except for 5 cases (acute myocardial infarction in 2 and liver failure in 3). Multiple primary cancers were seen in 21.6% (25/116), and other cancers that caused death, apart from those at urological sites, were confirmed in 7 patients (pancreas in 2, and esophagus, lung, gallbladder, liver, and uterus in 1 patient each). CONCLUSION: Autopsies revealed the macroscopic and microscopic extent of the disease and the presence of incidental disease beyond the diagnosis made in the patient's lifetime.

Tumor-associated tissue eosinophilia of penile cancer.

BACKGROUND: Tumor-associated tissue eosinophilia (TATE) occurs in many kinds of cancer. This study analyzed whether TATE improves the survival of penile cancer patients. METHODS: In 17 patients with penile cancer, survival was compared between the TATE-positive group and the TATE-negative group. Tissue eosinophils were observed by transmission electron microscopy. RESULTS: In all cases, 5-year survival was 72.9% in the TATE-positive group (n = 8) and 38.9% in the TATE-negative group (n = 9; P = 0.131). In stage III and IV, it was 60% in the TATE-positive group (n = 5) and 0% in the TATE-negative group (n = 5; P = 0.058). Transmission electron microscopy images revealed the vigorous infiltration of tissue eosinophils. CONCLUSIONS: It is suggested that TATE improves the survival of patients with advanced penile cancer. However, a greater number of subjects is needed to prove statistical significance.

Database and analysis system for cDNA clones obtained from full-length enriched cDNA libraries.

We have developed an efficient sequence-analysis system and a database system for clones obtained from full-length enriched cDNA libraries made by using the oligo-capping method. We developed a semi-automatic analysis system for 5'- and 3'-end sequences. It pre-processes raw sequences (vector cut and accurate-sequence region extraction), clusters the sequences, searches for similarities through public databases, annotates completeness of clones and analyzes the ORFs in the sequences. Newly developed or improved programs are used in each step. A new program, ESTiMateFull is used to evaluate and to predict the sequence-fullness based on comparisons with mRNA and EST sequences, respectively. The ATGpr program is used to predict sequence-fullness based on statistical information. The combination of full-length enriched cDNA clones and ATGpr fullness prediction resulted in 70% accuracy in the specificity and the sensitivity of the fullness predictions. For the ORFs predicted by the ATGpr, the signal peptides are predicted and a motif search is performed by our new system. We also developed a program that assembles our sequences with dbEST sequences and developed a system to retrieve clones by the characteristics of the ORFs. As keywords, combination of various results of the analyses can be used for retrieval. And various results such as ORF features and database search results can be shown on the same screen by multiple displays. Full-length clones having interesting functions can thus be retrieved efficiently by using this system.