Differences in post-operative complications after reconstruction for congenital biliary dilatation in a single institution-Roux-en-Y hepaticojejunostomy versus hepaticoduodenostomy.

BACKGROUND: The standard surgical procedure for congenital biliary dilatation (CBD) consists of excision of extrahepatic bile duct and Roux-en-Y hepaticojejunostomy (HJ). However, alternative reconstructive operations for CBD includes hepaticoduodenostomy (HD). We compared postoperative complications and therapeutic outcomes of these two operations at our institution. METHODS: From 1981 to 2009, there were 23 traceable patients who underwent operation for CBD, They were divided into an HJ Group (n = 15) and an HD Group (n = 8). Demographic and outcome data were compared. RESULTS: There were no significant differences in postoperative complications (cholangitis, pancreatitis, and anastomotic stenosis) and current blood test data (AST, ALT, Total Bilirubin, Direct Bilirubin, Amylase) between the two groups. Current abdominal pain and carcinogenesis were not observed in either group. Intrahepatic stones occurred in one patient in each group, both > 25 years post-operation. CONCLUSION: HD is considered to be an acceptable reconstruction method compared to HJ, based on our results. There has been a suggestion that inadequate diversion in HD might increase the risk of cholangiocarcinoma. To date, in this series, that has not happened.

Analysis of an acyl-CoA binding protein in Aspergillus oryzae that undergoes unconventional secretion.

Acyl-CoA binding protein (ACBP) plays important roles in the metabolism of lipids in eukaryotic cells. In the industrially important filamentous fungus Aspergillus oryzae, although we have previously demonstrated that the A. oryzae ACBP (AoACBP) localizes to punctate structures and exhibits long-range motility, which is dependent on autophagy-related proteins, the physiological role of AoACBP remains elusive. Here, we describe identification and characterization of another ACBP from A. oryzae; we named this ACBP as AoAcb2 and accordingly renamed AoACBP as AoAcb1. The deduced amino acid sequence of AoAcb2 lacked a signal peptide. Phylogenetic analysis classified AoAcb2 into a clade that was same as the ACBP Acb1 of the model yeast Saccharomyces cerevisiae, but was different from that of AoAcb1. In contrast to punctate localization of AoAcb1, AoAcb2 was found to be dispersedly distributed in the cytoplasm, as was previously observed for the S. cerevisiae Acb1. Since we could not generate an Aoacb2 disruptant, we created an Aoacb2 conditional mutant that exhibited less growth under Aoacb2-repressed condition, suggesting that Aoacb2 is an essential gene for growth. Moreover, we observed that A. oryzae AoAcb2, but not A. oryzae AoAcb1, was secreted under carbon-starved condition, suggesting that AoAcb2 might be secreted via the unconventional protein secretion (UPS) pathway, just like S. cerevisiae Acb1. We also demonstrated that the unconventional secretion of AoAcb2 was dependent on the t-SNARE AoSso1, but was independent of the autophagy-related protein AoAtg1, suggesting that the unconventional secretion of AoAcb2, unlike that of S. cerevisiae Acb1, via the UPS pathway, is not regulated by the autophagy machinery. Thus, the filamentous fungus A. oryzae harbors two types of ACBPs, one of which appears to be essential for growth and undergoes unconventional secretion.

Subcellular localization of acyl-CoA binding protein in Aspergillus oryzae is regulated by autophagy machinery.

In eukaryotic cells, acyl-CoA binding protein (ACBP) is important for cellular activities, such as in lipid metabolism. In the industrially important fungus Aspergillus oryzae, the ACBP, known as AoACBP, has been biochemically characterized, but its physiological function is not known. In the present study, although we could not find any phenotype of AoACBP disruptants in the normal growth conditions, we examined the subcellular localization of AoACBP to understand its physiological function. Using an enhanced green fluorescent protein (EGFP)-tagged AoACBP construct we showed that AoACBP localized to punctate structures in the cytoplasm, some of which moved inside the cells in a microtubule-dependent manner. Further microscopic analyses showed that AoACBP-EGFP co-localized with the autophagy marker protein AoAtg8 tagged with red fluorescent protein (mDsRed). Expression of AoACBP-EGFP in disruptants of autophagy-related genes revealed aggregation of AoACBP-EGFP fluorescence in the cytoplasm of Aoatg1, Aoatg4 and Aoatg8 disruptant cells. However, in cells harboring disruption of Aoatg15, which encodes a lipase for autophagic body, puncta of AoACBP-EGFP fluorescence accumulated in vacuoles, indicating that AoACBP is transported to vacuoles via the autophagy machinery. Collectively, these results suggest the existence of a regulatory mechanism between AoACBP localization and autophagy.

Both Intraoperative Medial and Lateral Joint Stabilities at Midflexion Influence Postoperative Patient-Reported Outcome Measures Following Bi-Cruciate Stabilized Total Knee Arthroplasty.

Bi-cruciate stabilized (BCS) total knee arthroplasty (TKA) was developed to approximate normal knee kinematics and is expected to improve clinical outcomes. However, the effects of soft tissue balance at the medial or lateral compartment on patient-reported outcome measures (PROMs) following BCS TKA are unclear. The purpose of this study was to clarify the relationship between the medial or lateral component gaps and PROMs following BCS TKA. One hundred seventeen knees with varus deformities which underwent BCS TKA were included in this study. They were divided into two groups according to the validated Japanese version of the Knee Injury and Osteoarthritis Outcome Score for each subscale of pain, symptoms, and activities of daily living (ADL) at 1 year postoperatively: group H consisted of patients with ≥90 points and group L consisted of patients with <90 points. Intraoperative medial and lateral joint laxities at 0°, 30°, 60°, 90°, and 120° flexion measured using a tensor device were compared between the two groups in each subscale. In the pain subscale, the medial joint laxities at 30° (p < 0.05) and 60° flexion (p < 0.05) in group H were significantly smaller than those in group L. In the ADL subscale, the medial joint laxity at 60° flexion in group H was significantly smaller than that in group L (p < 0.05). In the symptom subscale, the lateral joint laxity at 60° flexion in group H was significantly smaller than that in group L (p < 0.05). Surgeons should pay attention to the importance of both medial and lateral joint stabilities to achieve better postoperative PROMs following BCS TKA.

[Investigation of urinary management after removal of indwelling urethral catheters in a general hospital: associations between bedridden state and impaired bladder emptying].

In this study we investigated the influences of decreased levels of activities of daily living (ADL), especially in bedridden patients,on lower urinary tract dysfunction and urinary management during hospital care. All 1,106 non-urological inpatients (896 non-bedridden patients and 210 bedridden patients) with an indwelling urethral catheter treated at Noto General Hospital between April 2006 and October 2009 were retrospectively evaluated. Maximum bladder capacity and post-void residual urine volume (PVR) were evaluated with uroflowmetry or voiding cystourethrography at the time the catheter was removed. Clean intermittent catheterization (CIC) and drug administration were performed for patients who had a PVR of 100 ml or more. Bedridden patients required urinary interventions at a higher rate than did non-bedridden patients (bedridden : 29.0%,non-bedridden : 17.6%). Although indwelling urethral catheters were reinserted in 13 patients in the bedridden group and 16 patients in the non-bedridden group,many patients in both groups could be free from the catheter. Our results indicate that patients with low ADL are vulnerable to impaired bladder emptying,and early diagnosis of impaired bladder emptying and active urinary management are required to solve their urinary problems.

[Mixed epithelial and stromal tumor of the kidney: a case report].

We report a case of a mixed epithelial and stromal tumor of the kidney. A 64-year-old female who had no complaint was found to have a left renal tumor by computed tomography (CT) for an examination of a right breast tumor and was referred to our department. CT revealed a gradually enhancing 5 cm mass in the left kidney. The patient underwent left radical nephrectomy, and the tumor was histologically diagnosed as a mixed epithelial and stromal tumor. The patient has been followed up for 6 months with no evidence of local recurrence or metastasis.

The Relationship between Soft-Tissue Balance and Intraoperative Kinematics of Guided Motion Total Knee Arthroplasty.

A new design of the so-called "guided-motion" total knee arthroplasty (TKA) is expected to produce normal-like kinematics. The implant behaves strictly as a mechanically constraint-guided motion system. However, no previous reports have demonstrated the most appropriate surgical technique or soft-tissue balance that would reproduce ideal kinematics. The purpose of this study was to clarify the relationship between soft-tissue balance and the intraoperative kinematics of guided-motion TKA. In this study, intraoperative kinematics of 95 patients whose TKA was performed with a guided-motion prosthesis (Journey II BCS Smith and Nephew) were measured using the computed tomography (CT)-free navigation system. All procedures were performed via the same soft-tissue balancing technique, which focused on the medial compartment because guided-motion TKA must acquire medial stability to induce medial pivot motion. We measured the extension and flexion osteotomy gaps using a force-controlled compartment-specific ligament tensioner with a distraction force of 80 N for each compartment and divided patients into three groups based on the relationship between extension and flexion joint osteotomy gaps of the medial compartment: group1- loose flexion gap, group 2-equal joint gap, and group 3-tight flexion gap. We compared the preoperative demographic characteristics, implant alignment, and intraoperative kinematics among the three groups. There was no difference between the preoperative demographic characteristics and postoperative implant alignment in the three groups. The relative tibial internal rotational angles in groups 1 and 2 were significantly larger than that in group 3 at 60°, 90°, and maximum flexion (p < 0.05). The appropriate soft-tissue balance of the medial compartment for guided-motion TKA was an equal joint osteotomy gap or a larger flexion than extension gap. A tight flexion gap should be avoided.

Efficacy of combined androgen blockade with zoledronic acid treatment in prostate cancer with bone metastasis: the ZABTON-PC (zoledronic acid/androgen blockade trial on prostate cancer) study.

AIM: Zoledronic acid (ZA) reduces the risk of skeletal-related events (SREs) in castration-resistant prostate cancer (CRPC) with bone metastasis and improves quality of life. It remains unclear when clinicians should initiate ZA treatment. PATIENTS AND METHODS: Hormone-naïve patients were randomized to a combined androgen blockade (CAB) group or CAB with ZA group (CAB-ZA) based on Gleason score (GS) or extent of disease. The primary end-point of the study was progression-free survival (PFS) and the secondary end-point was incidence of SREs and bone pain. RESULTS: Thirty-one and 29 patients among 60 enrolled patients were assigned to the CAB group and the CAB-ZA group, respectively. There was no significant difference in PFS between the two groups. Subgroup analyses revealed better PFS in the CAB-ZA group with GS ≥8 (p=0.021). Moreover, incidence of SREs, including bone pain, was lower in the CAB-ZA group (p=0.019). CONCLUSION: CAB-ZA treatment was found to improve PFS for patients with prostate cancer with high GS. CAB-ZA treatment could be recommended for treatment of patients with prostate cancer.

Environmental chlamydiae alter the growth speed and motility of host acanthamoebae.

Symbiosis between living beings is an important driver of evolutionary novelty and ecological diversity; however, understanding the mechanisms underlying obligate mutualism remains a significant challenge. Regarding this, we have previously isolated two different Acanthamoeba strains harboring endosymbiotic bacteria, Protochlamydia (R18 symbiotic amoebae: R18WT) or Neochlamydia (S13 symbiotic amoebae; S13WT). In this study, we treated the symbiotic amoebae R18WT and S13WT with doxycycline (DOX) and rifampicin (RFP), respectively, to establish the aposymbiotic amoebae R18DOX and S13RFP, respectively. Subsequently, we compared the growth speed, motility, phagocytosis, pinocytosis, and morphology of the symbiotic and aposymbiotic amoebae. The growth speed of R18DOX was decreased, although that of S13RFP was increased. A marked change in motility was observed only for R18DOX amoebae. There was no difference in phagocytic and pinocytic activities between the symbiotic and aposymbiotic amoebae. Meanwhile, we observed a significant change in the phalloidin staining pattern and morphological changes in R18DOX (but not S13RFP) aposymbiotic amoebae, indicating a change in actin accumulation upon removal of the Protochlamydia. Infection of C3 (a reference strain) or S13RFP amoebae with Protochlamydia had a harmful effect on the host amoebae, but R18DOX amoebae re-infected with Protochlamydia showed recovery in both growth speed and motility. Taken together, we conclude that endosymbiont environmental chlamydiae alter the growth speed and/or motility of their host Acanthamoeba, possibly implying an close mutual relationship between amoebae and environmental chlamydiae.

Survival and transfer ability of phylogenetically diverse bacterial endosymbionts in environmental Acanthamoeba isolates.

Obligate intracellular bacteria are commonly found as endosymbionts of acanthamoebae; however, their survival in and ability to transfer to amoebae are currently uncharacterized. In this study, six bacterial endosymbionts, found in five environmental Acanthamoeba isolates (S13, R18, S23, S31, S40) from different locations of Sapporo city, Japan, were characterized. Phylogenetic analysis revealed that three bacterial endosymbionts (eS23, eS31, eS40a) belonged to α- and ß-Proteobacteria phyla and the remaining endosymbionts (eS13, eR18, eS40b) belonged to the order Chlamydiales. The Acanthamoeba isolate (S40) contained two phylogenetically different bacterial endosymbionts (eS40a, eS40b). Fluorescent in situ hybridization analysis showed that all bacterial endosymbionts were diffusely localized within amoebae. Transmission electron microscopy also showed that the endosymbionts were rod-shaped (eS23, eS31, eS40a) or sphere- or crescent-shaped (eS13, eR18, eS40b). No successful culture of these bacteria was achieved using conventional culture methods, but the viability of endosymbionts was confirmed by live/dead staining and RT-PCR methods. However, endosymbionts (except eR18) derived from original host cells lost the ability to be transferred to another Acanthamoebae strains [ATCC strain (C3), environmental strains (S14, R23, S24)]. Thus, our data demonstrate that phylogenetically diverse bacterial endosymbionts found in amoebae maintain a stable interaction with amoebae, but the transferability is limited.

Endosymbiotic bacterium Protochlamydia can survive in acanthamoebae following encystation.

Obligate intracellular bacteria are commonly seen as endosymbionts of acanthamoebae. However, whether endosymbionts can survive amoebal encystations remains a significant challenge in cellular biology. The survival of the endosymbiotic bacteria Protochlamydia belonging to environmental chlamydiae found in an amoebal isolate that we have previously reported (Environmental Microbiology Reports, DOI: 10.1111/j.1758-2229.2009.00094.x, 2009) following encystation was therefore assessed. The bacteria were observed in cysts and trophozoites reverted from cysts by analysis with transmission electron microscope, and the bacterial 16S rRNA transcripts were detected in amoeba cultures following encystations by reverse transcription polymerase chain reaction method. Furthermore, the bacterial growth was also confirmed, by fluorescent in situ hybridization analysis and the AIU assay that we have previously established (Applied Environmental Microbiology, 74: 6397-6404, 2008), in trophozoites reverted from cysts stored at 4°C for up to a month after encystation. Thus, these results demonstrated that Protochlamydia could survive in acanthamoebae following encystation. Our findings suggest that amoeba cysts might be further studied in order to understand their role in the environmental survival of endosymbionts.

Ciliates rapidly enhance the frequency of conjugation between Escherichia coli strains through bacterial accumulation in vesicles.

The mechanism underlying bacterial conjugation through protozoa was investigated. Kanamycin-resistant Escherichia coli SM10λ+ carrying pRT733 with TnphoA was used as donor bacteria and introduced by conjugation into ciprofloxacin-resistant E. coli clinical isolate recipient bacteria. Equal amounts of donor and recipient bacteria were mixed together in the presence or absence of protozoa (ciliates, free-living amoebae, myxamoebae) in Page's amoeba saline for 24 h. Transconjugants were selected with Luria broth agar containing kanamycin and ciprofloxacin. The frequency of conjugation was estimated as the number of transconjugants for each recipient. Conjugation frequency in the presence of ciliates was estimated to be approximately 10⁻6, but in the absence of ciliates, or in the presence of other protozoa, it was approximately 10⁻8. Conjugation also occurred in culture of ciliates at least 2 h after incubation. Successful conjugation was confirmed by the polymerase chain reaction. Addition of cycloheximide or latrunculin B resulted in suppression of conjugation. Heat killing the ciliates or bacteria had no effect on conjugation frequency. Co-localization of green fluorescent protein-expressing E. coli and PKH-67-vital-stained E. coli was observed in the same ciliate vesicles, suggesting that both donor and recipient bacteria had accumulated in the same vesicle. In this study, the conjugation frequency of bacteria was found to be significantly higher in vesicles purified from ciliates than those in culture suspension. We conclude that ciliates rapidly enhance the conjugation of E. coli strains through bacterial accumulation in vesicles.

Acute urinary retention associated with fractured osteophyte.

A 24-year-old man presented to our hospital twice with acute urinary retention in March 2002. Myelographic computed tomography demonstrated a posterior osteophyte. The preoperative urodynamic study indicated detrusor overactivity and detrusor sphincter dyssynergia 30 days after the first occurrence of urinary retention. During osteophyte resection and laminoplasty, the osteophyte, which was found to be fractured and to press on the cauda equina, was removed. The postoperative urodynamic study showed that detrusor overactivity remained and detrusor sphincter dyssynergia had disappeared. The patient was regularly followed up as an outpatient, and no dysuria had been noted by January 2003.

Case of leiomyosarcoma of the renal pelvis.

A 54-year-old man presented at the Mibayashi clinic with bowel discomfort. Ultrasonography showed a left renal mass and the patient was referred to Noto General Hospital for urological evaluation. Results of the physical examination were unremarkable, but computed tomography and magnetic resonance imaging demonstrated a large tumor in the middle of the left kidney. The patient underwent left nephrectomy. On hemisection of the kidney, a firm tumor, measuring 8 x 7.5 cm, was seen occupying the renal pelvis. The histological diagnosis was leiomyosarcoma arising from the right renal pelvis. No treatment was provided after surgery and no recurrence was observed 6 months postoperatively.