Zinc Deficiency via a Splice Switch in Zinc Importer ZIP2/SLC39A2 Causes Cystic Fibrosis-Associated MUC5AC Hypersecretion in Airway Epithelial Cells.

Airway mucus hyperproduction and fluid imbalance are important hallmarks of cystic fibrosis (CF), the most common life-shortening genetic disorder in Caucasians. Dysregulated expression and/or function of airway ion transporters, including cystic fibrosis transmembrane conductance regulator (CFTR) and epithelial sodium channel (ENaC), have been implicated as causes of CF-associated mucus hypersecretory phenotype. However, the contributory roles of other substances and transporters in the regulation of CF airway pathogenesis remain unelucidated. Here, we identified a novel connection between CFTR/ENaC expression and the intracellular Zn2+ concentration in the regulation of MUC5AC, a major secreted mucin that is highly expressed in CF airway. CFTR-defective and ENaC-hyperactive airway epithelial cells specifically and highly expressed a unique, alternative splice isoform of the zinc importer ZIP2/SLC39A2 (ΔC-ZIP2), which lacks the C-terminal domain. Importantly, ΔC-ZIP2 levels correlated inversely with wild-type ZIP2 and intracellular Zn2+ levels. Moreover, the splice switch to ΔC-ZIP2 as well as decreased expression of other ZIPs caused zinc deficiency, which is sufficient for induction of MUC5AC; while ΔC-ZIP2 expression per se induced ENaC expression and function. Thus, our findings demonstrate that the novel splicing switch contributes to CF lung pathology via the novel interplay of CFTR, ENaC, and ZIP2 transporters.

Pharmacological and genetic reappraisals of protease and oxidative stress pathways in a mouse model of obstructive lung diseases.

Protease-antiprotease imbalance and oxidative stress are considered to be major pathophysiological hallmarks of severe obstructive lung diseases including chronic obstructive pulmonary disease (COPD) and cystic fibrosis (CF), but limited information is available on their direct roles in the regulation of pulmonary phenotypes. Here, we utilized ßENaC-transgenic (Tg) mice, the previously established mouse model of severe obstructive lung diseases, to produce lower-mortality but pathophysiologically highly useful mouse model by backcrossing the original line with C57/BL6J mice. C57/BL6J-ßENaC-Tg mice showed higher survival rates and key pulmonary abnormalities of COPD/CF, including mucous hypersecretion, inflammatory and emphysematous phenotypes and pulmonary dysfunction. DNA microarray analysis confirmed that protease- and oxidative stress-dependent pathways are activated in the lung tissue of C57/BL6J-ßENaC-Tg mice. Treatments of C57/BL6J-ßENaC-Tg mice with a serine protease inhibitor ONO-3403, a derivative of camostat methylate (CM), but not CM, and with an anti-oxidant N-acetylcystein significantly improved pulmonary emphysema and dysfunction. Moreover, depletion of a murine endogenous antioxidant vitamin C (VC), by genetic disruption of VC-synthesizing enzyme SMP30 in C57/BL6J-ßENaC-Tg mice, exaggerated pulmonary phenotypes. Thus, these assessments clarified that protease-antiprotease imbalance and oxidative stress are critical pathways that exacerbate the pulmonary phenotypes of C57/BL6J-ßENaC-Tg mice, consistent with the characteristics of human COPD/CF.

Pharmacist blood pressure management programs using telemonitoring systems are useful for monitoring side effects of antihypertensive drugs in a community pharmacy.

We previously started pharmacist blood pressure (BP) management programs using telemonitoring systems for monitoring side effects of antihypertensive drugs in a community pharmacy. The present case demonstrates that pharmacist BP management programs using telemonitoring systems are useful for monitoring side effects of antihypertensive drugs in a community pharmacy.

Hepatocellular carcinoma incidentally detected at second hepatectomy for repeated colorectal liver metastasis in a patient with hepatitis C virus-related cirrhosis: a case report.

BACKGROUND: It has been reported that liver metastasis rarely occurs in a cirrhotic/hepatitic liver. Thus, coexistence of liver metastasis and hepatocellular carcinoma has been scarcely reported. To the best of our knowledge, there are no cases with hepatocellular carcinoma, which developed during an observational period after hepatectomy for colorectal liver metastasis, in the worldwide English literature. Here we present a case of hepatocellular carcinoma which occurred during a period between the first and second hepatectomy for repeated colorectal liver metastasis. CASE PRESENTATION: A 65-year-old Japanese woman underwent rectal resection for advanced rectal cancer. Hepatitis C cirrhosis was diagnosed at that time and antiviral therapy was offered but rejected because of socioeconomic reasons. At the age of 68, she developed two colorectal liver metastases originating from the rectal cancer, which were treated by local ablation and partial hepatectomy. At the age of 71, solitary recurrent colorectal liver metastasis was observed adjacent to the previously ablated lesion in segment 4, and thus segmentectomy 4 was performed. During surgery, a small tumor in segment 8 was incidentally identified. Taking into account her history, the tumor was considered to be recurrent colorectal liver metastasis and it was extirpated by partial hepatectomy. However, the segment 4 tumor was diagnosed as recurrent colorectal liver metastasis on the basis of histological findings and the segment 8 tumor was diagnosed as hepatocellular carcinoma. Although she had a cut surface abscess postoperatively, she was discharged from hospital 21 days after the surgery and is currently doing well 18 months after the second hepatectomy. She is currently receiving interferon and ribavirin therapy to eliminate hepatitis C virus. CONCLUSIONS: If antiviral therapy was performed earlier for the present case and viral elimination was achieved, hepatocellular carcinoma might not have developed. This case reemphasizes the importance of antiviral therapy for preventing carcinogenesis of hepatocellular carcinoma in patients with viral hepatitis even if they have other cancers.

Use of pharmacist blood pressure telemonitoring systems in diagnosis of nocturnal hypertension in a young healthy male.

Blood pressure (BP) telemonitoring systems and pharmacist management programs were introduced into Haruka Community Pharmacy. A 22-year-old healthy male came to the community pharmacy, although he was not in a diseased state, he had been informed previously that he had a moderately high BP during a routine examination. He continued home BP telemonitoring for 28 days. A pharmacist intervention was conducted at 2 week intervals. His average nighttime systolic BP was higher than the daytime systolic BP. The pharmacist consulted a doctor based on the BP telemonitoring results, and ambulatory blood pressure monitoring (ABPM) was initiated. The doctor detected nocturnal hypertension based on the results of ABPM monitoring. BP telemonitoring systems have been introduced into a small percentage of pharmacies in Japan, and this is the first case report for the usefulness of these systems in a community pharmacy.

Blunt penetration technique for treatment of a completely obstructed anastomosis after rectal resection: a case report.

INTRODUCTION: We present a case of completely obstructed anastomosis after rectal resection which was nonsurgically and successfully treated with a blunt penetration technique using a commonly used device for transanal ileus drainage. The technique we used in this case has not been previously reported. CASE PRESENTATION: A 79-year-old Japanese man underwent redo rectal resection for completely separated anastomosis which was caused by anastomotic leakage after a sigmoidectomy performed 3 years previously that was remedied by diverging ileostomy. Immediately after the redo surgery, fluoroscopy showed good passage through the colorectal anastomosis but no anastomotic leakage. However, fluoroscopy and colonoscopy prior to the ileostomy takedown showed complete obstruction of the anastomosis. Unlike usual anastomotic strictures, the lumen between colon oral and rectum anal to the anastomosis was completely discontinued by a membranous structure. Therefore, a conventional balloon dilatation technique was unsuitable for this condition. We applied a blunt penetration technique using a commercially available device designed as a transanal drainage system for obstructing colorectal cancer to restore the continuity between the colon oral and rectum anal to the anastomosis. After restoring the continuity, we performed conventional balloon dilatation for the anastomosis and successfully treated the anastomotic obstruction. Subsequently, the patient underwent ileostomy takedown and is currently doing well 12 months after the ileostomy takedown. CONCLUSIONS: The penetration technique we applied is easy and less stressful to adopt because it does not require usage of materials specialized for other particular purposes. Furthermore, we believe that this technique is superior in safety to other reported methods for this condition even if applied in the wrong direction because this technique does not utilize electrocision or sharp needle puncture.

Effects of body-color mutations on vitality: an attempt to establish easy-to-breed see-through medaka strains by outcrossing.

"See-through" strains of medaka are unique tools for experiments: their skin is transparent, and their internal organs can be externally monitored throughout life. However, see-through fish are less vital than normally pigmented wild-type fish, which allows only skilled researchers to make the most of their advantages. Expecting that hybrid vigor (heterosis) would increase the vitality, we outcrossed two see-through strains (SK(2) and STIII) with a genetically distant wild-type strain (HNI). Fish with the see-through phenotypes were successfully restored in the F2 generation and maintained as closed colonies. We verified that genomes of these hybrid see-through strains actually consisted of approximately 50% HNI and approximately 50% SK(2) or STIII alleles, but we could not obtain evidence supporting improved survival of larvae or fecundity of adults, at least under our breeding conditions. We also found that four of the five see-through mutations (b(g8), i-3, gu, and il-1 but not lf) additively decrease viability. Given that heterosis could not overwhelm the viability-reducing effects of the see-through mutations, easy-to-breed see-through strains will only be established by other methods such as conditional gene targeting or screening of new body-color mutations that do not reduce viability.

Synergism between interleukin (IL)-17 and Toll-like receptor 2 and 4 signals to induce IL-8 expression in cystic fibrosis airway epithelial cells.

Cystic fibrosis (CF) is the most common lethal inherited disorder and is caused by mutations in the gene encoding the CF transmembrane regulator (CFTR). The CF lung expresses a profound proinflammatory phenotype that appears to be related to a constitutive hypersecretion of interleukin (IL)-8 from airway epithelial cells in response to microbial infection. Since overproduction of IL-8 in CF contributes to massive bronchial infiltrates of neutrophils, identification of the pathways underlying IL-8 induction could provide novel drug targets for treatment of neutrophil-dominated inflammatory diseases such as CF. Here, we show that IL-17A synergistically increases IL-8 production induced by a toll-like receptor (TLR) 2 agonist, peptidoglycan (PGN), or TLR4 agonist, lipopolysaccharide (LPS), in a human CF bronchial epithelial cell line (CFBE41o-). A strong synergism was also observed in primary human CF bronchial epithelial cells, but not in human non-CF cell lines and primary cells. Notably, despite the induction of nuclear factor-κB and MAP kinases during TLR2 or TLR4 activation in CFBE41o-, IL-17A-dependent synergism appears to be the result of enhanced PGN- or LPS-induced phosphorylation of p38. Taken together, these studies provide evidence that IL-17A is a critical factor in increasing IL-8 expression in bacteria-infected CF airways via a pathway that regulates p38 phosphorylation.

Rb/E2F1 regulates the innate immune receptor Toll-like receptor 3 in epithelial cells.

Tumor suppressor genes regulate the antiviral host defense through molecular mechanisms that are not yet well explored. Here, we show that the tumor suppressor retinoblastoma (Rb) protein positively regulates Toll-like receptor 3 (TLR3) expression, the sensing receptor for viral double-stranded RNA and poly(I · C). TLR3 expression was lower in Rb knockout (Rb(-/-)) mouse embryonic fibroblasts (MEF) and in mammalian epithelial cells transfected with Rb small-interfering RNA (siRNA) than in control cells. Consequently, induction of cytokines interleukin-8 and beta interferon after poly(I · C) stimulation was impaired in Rb(-/-) MEF and Rb siRNA-transfected cells compared to controls. TLR3 promoter analysis showed that Rb modulates the transcription factor E2F1, which directly binds to the proximal promoter of TLR3. Exogenous addition of E2F1 decreased TLR3 promoter activity, while Rb dose dependently curbed the effect of E2F1. Interestingly, poly(I · C) increased the Rb expression, and the poly(I · C)-induced TLR3 expression was impaired in Rb-depleted cells, suggesting the importance of Rb in TLR3 induction by poly(I · C). Together, these data indicated that E2F1 suppresses TLR3 transcription, but during immune stimulation, Rb is upregulated to block the inhibitory effect of E2F1 on TLR3, highlighting a role of Rb-E2F1 axis in the innate immune response in epithelial cells.

Technique of right hemihepatectomy preserving ventral right anterior section guided by area of hepatic venous drainage.

BACKGROUND: Although the consequences of partial venous outflow interruption have attracted only limited attention in liver surgery, maximal preservation of liver function after hepatic resection requires preservation of circulation in the remnant liver, especially hepatic vein drainage. METHODS: Data from 30 patients undergoing 3-dimensional imaging were analyzed to clarify the relationship between the area of the ventral right anterior section (RAS) and that drained by regional hepatic vein tributaries. The feasibility of our preliminary technique of right hemihepatectomy preserving the ventral RAS also was evaluated. RESULTS: The median estimated volume of the ventral RAS was 230 mL (range, 88-391). The average ratio of this estimated volume of the ventral RAS to total estimated liver volume was 18.0 +/- 4.9%. The median volume of the territory served by middle hepatic vein (MHV) tributaries draining the ventral RAS, expressed as a percentage of the whole volume of the ventral RAS, was 82.5%. Findings in fusion images of portal and hepatic vein territories demonstrated an area of MHV tributaries comparable with the ventral RAS area in 73.3% of all cases. As for the results of right hemihepatectomy with the ventral RAS preserved, no tumor was exposed on transection surfaces, and no recurrence took place within the preserved ventral RAS of the remnant liver. CONCLUSION: Procedures considering the importance of regional venous drainage offer the possibility of reducing the extent of surgery without loss of effectiveness.

Feasibility of AC/EC followed by weekly paclitaxel in node-positive breast cancer in Japan.

UNLABELLED: The feasibility and efficacy of adriamycin or epirubicin in combination with cyclophosphamide followed by weekly paclitaxel (AC/EC-weekly PAC) as adjuvant chemotherapy for breast cancer was investigated. PATIENTS AND METHODS: Node-positive breast cancer was treated with AC/ EC-weekly PAC, namely AC at 60/600 mg/m(2) or EC at 90/600 mg/m(2) x4 at three-week intervals, followed by weekly PAC (80 mg/m(2)) x 12, namely four cycles of single weekly administration for three weeks followed by a one-week rest (3 x 4 PAC) or single weekly administration for 12 consecutive weeks (12 PAC). RESULTS: One hundred and three of 109 consecutive patients enrolled were analyzed, of whom 96 (93.2%) completed the regimen. Grade 3/4 neutropenia occurred in 52.4% receiving AC/EC, and 10.9% of 55 receiving 12 PAC but only 2.1% of 48 receiving 3 x 4 PAC. Neuropathy disorders occurred in more than half receiving PAC, which did not improve after one-week rest in 3 x 4 PAC. CONCLUSION: AC/EC-weekly PAC is feasible and without serious complications.

Impact of the degree of liver resection on survival for patients with multiple liver metastases from colorectal cancer.

BACKGROUND: Although the degree of hepatic resection has been found to be a key aspect of tumor stimulation, the differences in clinical outcome between a massive liver resection and a less extensive resection for multiple colorectal metastases have not been well studied. The purpose of this study was to clarify the impact of the extent of liver resection on survival outcome. METHODS: Clinicopathologic data were available for 85 patients who were surgically treated for four or more liver metastases. Forty-nine patients who underwent a major hepatic resection were compared with patients who underwent minor hepatic resections (n = 36). RESULTS: As the patients undergoing major resection were more likely to have multiple (p = 0.014) and large tumors (p = 0.021) compared to the minor-resection patients, their overall survival was worse (p = 0.046) and the disease-free rate tended to be poorer. By multivariate analysis of the cohorts, the only independent factor affecting survival was the number of liver tumors (< or =5 or > or =6; relative risk [RR] = 0.427; p = 0.014). When patients with six or more metastases were selected and analyzed, the overall survival of patients who had a major resection was significantly poorer than those who had minor resections (p = 0.028), although the clinical characteristics were comparable between the two groups. CONCLUSION: Although the extent of hepatectomy was not an independent prognosticator, minor resections for multiple colorectal metastases may offer a long-term survival advantage compared to a major resection.

Anatomic versus limited nonanatomic resection for solitary hepatocellular carcinoma.

BACKGROUND: Although anatomic liver resection is preferred when treating hepatocellular carcinoma (HCC), evidence that it improves survival when compared with an adequate nonanatomic resection is lacking. The purpose of this study was to compare the survival impact of anatomic versus nonanatomic resection in patients with solitary HCC. PATIENTS AND METHODS: Clinicopathologic data were available for 125 patients who underwent hepatectomy for a solitary HCC confined to 1 or 2 Couinaud's segments. These patients were divided into 2 groups based on the hepatectomy procedure: anatomic (n = 83) and nonanatomic (n = 42) resection. RESULTS: No differences were detected either in the hepatic recurrence rates (P = .38) or in the overall survival rates (P = .34) between the anatomic group and the nonanatomic group. The hepatectomy procedure (anatomic vs nonanatomic resection) did not affect survival in either univariate (P = 0.34) or multivariate analysis (relative risk, 1.574; P = .22). The proportion of patients who survived after recurrence was greater in the nonanatomic (15/42) than the anatomic group (13/83; P = .049), and the median survival time after recurrence was greater in patients who underwent nonanatomic resection (991 days; range, 131-4073 days) than in patients with anatomic resection (310 days; range, 48-1887 days; P = .045). CONCLUSIONS: No superiority was seen in survival when HCC was treated by anatomic resection. Maintaining adequate liver function regardless of whether the resection is anatomic or not may be of greater importance.

Perioperative infection control and its effectiveness in hepatectomy patients.

BACKGROUND AND AIM: Post-hepatectomy infections require careful attention, because they may deteriorate into liver failure. We retrospectively reviewed such infections in cases without biliary or intestinal reconstruction. METHODS: This retrospective study involved 535 patients with liver tumors who underwent hepatectomy at the Department of Gastroenterological Surgery of Yokohama City University Hospital between April 1992 and March 2005. After classification into four groups depending on changes in infection countermeasures used during different periods, the treatment outcomes were examined. No such anti-infection measures were taken during the first period; a closed suction drainage system and early enteral nutrition after surgery were introduced between the first and second periods; thorough management of bile leakage and prevention of nosocomial infection were added between the second and third periods; and surgical site infection (SSI) surveillance together with absorbable sutures instead of silk sutures between the third and fourth periods. RESULTS: The incidence of postoperative infection decreased significantly with additional countermeasures: first period 44.7%; second period 24.1%; third period 15.0%; and fourth period 9.2%. The incidence of both SSI and remote infection were similarly reduced. Postoperative infection risk factors were age, presence of diabetes mellitus, the use of silk sutures and bile leakage, while those for SSI were the use of silk sutures and bile leakage. CONCLUSION: The incidence of postoperative infection and SSI were significantly reduced by our infection countermeasures, especially by bile leakage management and the use of absorbable sutures.

[A case of advanced breast cancer with multiple liver metastases resistant to chemotherapy responding to high-dose toremifene].

A 47-year-old woman was admitted to our hospital for advanced breast cancer with multiple liver metastases and liver dysfunction in January 2004. EC (EPI combined with CPA), weekly PTX, and AI were performed but were not effective for that tumor. Therefore, high-dose TOR was started. Liver dysfunction recovered after administration of TOR, and primary tumor and liver metastases were evaluated as a partial response (PR). The same therapy has been performed for six months with no evidence of deterioration.

Acute disseminated encephalomyelitis and poststreptococcal acute glomerulonephritis.

We experienced the case of a boy suffering from acute disseminated encephalomyelitis and concomitant acute glomerulonephritis. The multiple lesions observed on MR images, which located mainly in the cortical gray matter, quickly responded to methyl prednisolone pulse therapy. Renal biopsy confirmed the diagnosis of poststreptococcal acute glomerulonephritis. Streptococcus pyogenes was identified by pharyngeal culture, and the infection was serologically confirmed. We speculated that S. pyogenes infection was coincidentally involved in both diseases.