Intraoperative cholangiography and bile duct flushing in 47 dogs receiving laparoscopic cholecystectomy for benign gallbladder disease: A retrospective analysis.

OBJECTIVE: To describe a laparoscopic approach for performing intraoperative cholangiography (IOC) and bile duct flushing (BDF) during laparoscopic cholecystectomy (LC) in dogs. To investigate the clinical outcomes of dogs undergoing these procedures for the treatment of benign gallbladder disease, ie gallbladder mucocele (GM) or cholecystitis. STUDY DESIGN: Retrospective study. ANIMALS: Forty-seven client-owned dogs. METHODS: Medical records of client-owned dogs with benign gallbladder diseases that underwent IOC and BDF during LC between September 2016 and December 2019 were reviewed. Of these dogs, only dogs with GM or cholecystitis were included in the study. The fundus dissection first method was used for LC. Intraoperative cholangiography and BDF procedures were performed laparoscopically using a catheter inserted into the cystic duct following dissection within the subserosal layer of the gallbladder. Videos recorded during each procedure were reviewed, and data on procedure duration, completion, outcome, and technical approach were recorded. RESULTS: Forty-seven dogs were included in the study. The median procedure time for BDF and IOC was 4 min (range, 2-48 min), and no intraoperative or postoperative complications occurred. CONCLUSION: During LC, BDF and IOC were performed safely and successfully. Intraoperative cholangiography identified obstructions and strictures in the common bile duct that were not detected using BDF alone. CLINICAL SIGNIFICANCE: Our findings suggest that BDF and IOC are both safe and time effective and should be considered for routine use by surgeons during LC.

Transanal minimally invasive surgery for rectal neuroendocrine tumors.

BACKGROUND: There is no literature that mainly searched for rectal neuroendocrine tumor (rNET) using transanal minimal invasive surgery (TAMIS). We report our clinical experiences of TAMIS for rectal neuroendocrine tumors to evaluate the feasibility and safety. METHODS: Between December 2010 and March 2020, the 25 consecutive patients with rectal neoplasma underwent the TAMIS procedure performed by single laparoscopic surgeon at the two hospitals. Of these, ten patients with rectal neuroendocrine tumors were reviewed retrospectively. The full-thickness excision down to the outer fatty tissues was completed using TAMIS technique. Clinicopathological findings, perioperative and postoperative complications were recorded. RESULTS: TAMIS for small rNET was successfully completed in all cases. There were seven cases with a tumor size of less than 10 mm, and three cases with a tumor size between 10 and 15 mm. Six patients underwent the primary tumor excision; the remaining four patients underwent resection for the scar after endoscopic procedure. The median surgical duration was 80.5 (53-124) minutes and the median blood loss was 1 (1-12) ml. All removed tumors in the 6 primary excisions were diagnosed as neuroendocrine tumor G1. The margins of specimens were completely free in all cases. Among the four patients after endoscopic procedure, all had no histological evidence of residual tumor. The median length of hospital stay was 7 days postoperatively. There was no post-operative mortality or severe complication. The median length of observation was 54 months. No recurrence, no local or distant metastasis and no mortality of all patients were observed. CONCLUSIONS: TAMIS is safety and feasible procedure for small rNET. Further experience and clinical trials are needed to fully define the advantages, disadvantages, and indications of TAMIS for rNET.

Urinary bladder fistula following laparoscopic inguinal hernioplasty: a case report.

BACKGROUND: Fistula formation due to mesh erosion into hollow viscera, such as the urinary bladder, is uncommon. To date, there have been no reports of fistula formation into the urinary bladder without evidence of mesh erosion after hernioplasty; herein, we report one such rare case, in which the clinical symptoms improved without any surgical intervention. CASE PRESENTATION: A 73-year-old man underwent a trans-abdominal preperitoneal repair for bilateral direct inguinal hernia. One month later, the patient experienced a painful induration in the right inguinal region, and computed tomography revealed fluid collection in this region. A culture of the aspirated fluid yielded no bacteria. Seven months later, he experienced another episode of painful induration in the same region. However, blood examination revealed a normal white blood cell count and C-reactive protein level. Moreover, no organisms were detected by aspirated fluid culture. Although the painful induration subsided after aspiration of the fluid collection, he developed gross hematuria and dysuria a month later. Cystoscopy revealed a fistula in the right wall of the urinary bladder that discharged a purulent fluid. Culture of the fluid revealed no bacteria, and there was no evidence of mesh erosion. Hematuria improved without therapeutic or surgical intervention. The patient's clinical symptoms improved without mesh removal. Moreover, cystoscopy revealed that the fistula was scarred 12 months after the initial appearance of urinary symptoms. No further complications were observed during a 42-month follow-up period. CONCLUSIONS: We report a rare case of a fistula in the urinary bladder without evidence of mesh erosion after laparoscopic hernioplasty. The patient's condition improved without mesh removal. Fluid collection due to foreign body reaction to meshes can cause fistula formation in the urinary bladder without direct mesh contact.

Canine idiopathic chylothorax: Anatomic characterization of the pre- and postoperative thoracic duct using computed tomography lymphography.

Surgical treatment has improved the prognosis of canine idiopathic chylothorax, although a recurrence of the disease occurs occasionally after the procedure. An improved understanding of possible causes for this recurrence would be helpful for prognosis and treatment planning in affected patients. In this retrospective case series study, we described the detailed pre- and postoperative computed tomographic lymphography (CTLG) imaging characteristics for a group of dogs with surgically confirmed idiopathic chylothorax. Preoperative CTLG was performed in 12 of 14 dogs diagnosed with idiopathic chylothorax. Thoracic ducts were present on the right side in 10 dogs, left side in one dog, and bilaterally in one dog. All the 14 dogs received a combination therapy of pericardiectomy and thoracic duct ligation (TDL) by video-assisted thoracoscopic surgery. One week after surgery, a postoperative CTLG was performed, and the thoracic ducts were apparent in seven of 14 dogs. Three dogs had an unchanged course of the thoracic duct, which could have resulted from a missed duct. Four dogs were identified as having a bypass formation: the oblique duct originated at the ligation site and connected to the duct on the other side. Our findings indicated that one of the possible causes for postoperative recurrence of chylothorax in dogs could be "invisible or sleeping" fine ducts that are collapsed and not visible in preoperative CTLG scans. After TDL causes a change in the pressure of lymphatic flow, these fine thoracic ducts may become apparent using postoperative CTLG.

Efficacy of en bloc thoracic duct ligation in combination with pericardiectomy by video-assisted thoracoscopic surgery for canine idiopathic chylothorax.

OBJECTIVE: To compare the outcomes of pericardiectomy performed with conventional clipping thoracic duct ligation (C-TDL) to those with en bloc thoracic duct ligation (EB-TDL) using video-assisted thoracoscopic surgery (VATS) for canine idiopathic chylothorax. STUDY DESIGN: Retrospective consecutive case series. ANIMALS: Thirteen client-owned dogs with idiopathic chylothorax. METHODS: Medical records of dogs treated with pericardiectomy in combination with TDL by VATS without intraoperative contrast were reviewed. Five and seven dogs underwent C-TDL and EB-TDL, respectively, and 11 dogs were evaluated by preoperative and 7- to 10-days-postoperative computed tomography-lymphography (CTLG). No clinical symptoms with absent or minimal pleural effusion was defined as clinical improvement. Long-term remission (LTR) was defined as rapid resolution of pleural effusion and no recurrence for more than 1 year. Anesthesia time, operation time, the duration of hospitalization, and time until pleural effusion resolution were compared. RESULTS: Clinical improvement was achieved in 91.7% of the cases (C-TDL, 4/5; EB-TDL, 7/7), excluding one case of intraoperative death. The LTR rate was significantly higher with EB-TDL (6/7 [85.7%]) than with C-TDL (1/5 [20%]). Anesthesia time, operation time, and time until pleural effusion resolution were significantly better with EB-TDL than with C-TDL. The rates of thoracic ducts visualization by postoperative CTLG were 100% (5/5) with C-TDL and 42.9% (3/7) with EB-TDL. CONCLUSION: En bloc TDL was an effective treatment for canine idiopathic chylothorax in this patient population. It compared favorably to C-TDL, although missed branches at the time of surgery may explain the difference between C-TDL and EB-TDL in this small population of cases. CLINICAL SIGNIFICANCE: En bloc TDL by VATS was an effective minimally invasive treatment for canine idiopathic chylothorax. Computed tomography-lymphography can be used for surgical planning and postoperative evaluation.

[Preoperative Chemoradiotherapy for Stage II or III Esophageal Squamous Cell Carcinoma].

BACKGROUND: The goal of this retrospective study was to investigate the efficacy and safety of preoperative chemoradiotherapy( NACR)in patients with Stage II or Stage III esophageal squamous cell carcinoma(SCC). METHODS: Between 2004 and 2014, a total of 86 patients underwent surgical resection in conjunction with NACR for esophageal SCC at our institute. Thirty-one patients(36.0%)had Stage II disease and 55 patients(64.0%)had Stage III disease. RESULTS: The median age was 64(43-81)years. A total of 78 patients received the full NACR regimen. The most common major Grade 3 hematologic toxic effects of NACR were leukopenia and neutropenia(48 cases), while the most common major Grade 3 non-hematologic toxic effect was anorexia(12 cases). One patient died in the hospital and no patients died within 30 days after surgery. A pathological complete response was achieved in 23 cases. Pathological staging(number of cases)was Stage 0(23), Stage I (8), Stage II (28), Stage III (25), and Stage IV (2). The 5-year overall survival rate(OS)was 51.0%, and was 83.2% in Stage II patients and 29.9% in Stage III patients. CONCLUSION: Preoperative NACR is safe and may improve OS and downstaging rates in patients with esophageal SCC.

Evaluation of three types of platforms in single-incision laparoscopic surgery for performing transanal endoscopic microsurgery (SILSTEM).

BACKGROUND/AIMS: Transanal endoscopic microsurgery using a platform for single-incision laparoscopic surgery (SILSTEM) is safe for excising rectal lesions. We tested three types of platforms. METHODOLOGY: Nine patients underwent SILSTEM by one surgeon. Tumors located 5­15 cm from the anal verge were eligible. After measuring their dimensions, length, and trocar channels, the platforms tested were the SILSTM port (SP), EZTM access (EA), and GelPOINTTM Path (GP). Clinicopathology, intraoperative parameters, and postoperative outcomes were recorded. RESULTS: Six men and three women (median age 63 years) underwent SILSTEM using platform SP in three patients, EA in four, GP in two. Median operation time was 128 min (range 71­313). Median blood loss was 3 ml (range 1­71). Pathology confirmed adenocarcinoma in five patients, adenoma in three, and carcinoid in one. Patients were discharged within 2­13 days postoperatively. There was no postoperative fecal incontinence or soiling. Overall median follow-up was 13.3 months (range 1.3­27.2). There were no recurrences. CONCLUSION: SILSTEM can effectively resect rectal tumors using any of three platforms. Large prospective trials are needed to define the advantages, disadvantages, and indications for each platform and to draw conclusions regarding operation time, anorectal function, and costs.

Radiochemotherapy for esophageal squamous cell carcinoma in elderly patients.

BACKGROUND/AIMS: The goal of this retrospective study was to investigate the efficacy and safety of curative radiochemotherapy (CRT) in elderly patients with esophageal squamous cell cancer (SCC). METHODOLOGY: Between 1986 and 2011, 38 consecutive patients aged over 75 years with esophageal SCC were initially treated with a CRT regimen comprising radiation therapy at a dose of 60 Gy and two cycles of chemotherapy with 5-fluorouracil/cisplatin. RESULTS: Fourteen patients were aged over 80 and 24 under 79 years. Complete treatment compliance was obtained in 30 (78.9%) of the patients. Chemotherapy was terminated and the dose of radiation reduced in 7 and 1 patients, respectively. The overall incidence of CRT-related complications was 86.8% (33 patients). Grade 3 leukocytopenia was observed in 6 (15.8%) patients, all of whom recovered promptly. A complete response was observed in 17 patients (44.7%). The overall 3-year survival rate was 32.0%. CONCLUSIONS: Curative CRT is safe and may improve overall and progression-free 3-year survival rates in elderly patients aged more than 75 years with esophageal SCC.

Mouse prion protein (PrP) segment 100 to 104 regulates conversion of PrP(C) to PrP(Sc) in prion-infected neuroblastoma cells.

Prion diseases are characterized by the replicative propagation of disease-associated forms of prion protein (PrP(Sc); PrP refers to prion protein). The propagation is believed to proceed via two steps; the initial binding of the normal form of PrP (PrP(C)) to PrP(Sc) and the subsequent conversion of PrP(C) to PrP(Sc). We have explored the two-step model in prion-infected mouse neuroblastoma (ScN2a) cells by focusing on the mouse PrP (MoPrP) segment 92-GGTHNQWNKPSKPKTN-107, which is within a region previously suggested to be part of the binding interface or shown to differ in its accessibility to anti-PrP antibodies between PrP(C) and PrP(Sc). Exchanging the MoPrP segment with the corresponding chicken PrP segment (106-GGSYHNQKPWKPPKTN-121) revealed the necessity of MoPrP residues 99 to 104 for the chimeras to achieve the PrP(Sc) state, while segment 95 to 98 was replaceable with the chicken sequence. An alanine substitution at position 100, 102, 103, or 104 of MoPrP gave rise to nonconvertible mutants that associated with MoPrP(Sc) and interfered with the conversion of endogenous MoPrP(C). The interference was not evoked by a chimera (designated MCM2) in which MoPrP segment 95 to 104 was changed to the chicken sequence, though MCM2 associated with MoPrP(Sc). Incubation of the cells with a synthetic peptide composed of MoPrP residues 93 to 107 or alanine-substituted cognates did not inhibit the conversion, whereas an anti-P8 antibody recognizing the above sequence in PrP(C) reduced the accumulation of PrP(Sc) after 10 days of incubation of the cells. These results suggest the segment 100 to 104 of MoPrP(C) plays a key role in conversion after binding to MoPrP(Sc).

Prevalence of preoperative asymptomatic deep vein thrombosis in patients undergoing elective general surgery for benign disease.

Background: The systemic inflammatory response following surgery as well as that of malignant disease itself is associated with a hypercoagulable state, and thromboprophylaxis is thus recommended during postoperative management of cancer patients. However, limited information is available on the prevalence of preoperative deep vein thrombosis (DVT) and its risk factors in surgical candidates, especially those receiving operations for benign diseases. Methods: This is a retrospective observational study with data of all patients scheduled for elective general surgery between January 2011 and September 2020, undergoing lower extremity venous ultrasonography as preoperative screening for DVT. The prevalence of preoperative asymptomatic DVT was estimated and its associations with clinical variables were evaluated. Results: Among 1512 patients included in the study, 161 (10.6%) had asymptomatic DVT before surgery. DVT prevalence was 13.7% in patients with malignant disease, while it was 8.6% in those with benign disease. The site of the thrombus was distal type in 141 (87.6%) patients, most commonly in the soleal vein. Advanced age (>70 years), female sex, and decreased hemoglobin level were significantly associated with preoperative asymptomatic DVT by multivariate analysis. The odds ratio for advanced age was the highest and rose as age increased. Malignant disease was not an independent risk factor for preoperative DVT. Conclusion: This study showed the prevalence of asymptomatic DVT to be equal in patients with and without malignant disease undergoing elective general surgery. Preoperative DVT assessment is necessary regardless of the disease indicated for surgery, especially in patients with the risk factors identified in this study.

Accumulation of L-type bovine prions in peripheral nerve tissues.

We recently reported the intraspecies transmission of L-type atypical bovine spongiform encephalopathy (BSE). To clarify the peripheral pathogenesis of L-type BSE, we studied prion distribution in nerve and lymphoid tissues obtained from experimentally challenged cattle. As with classical BSE prions, L-type BSE prions accumulated in central and peripheral nerve tissues.

An improved method for cell-to-cell transmission of infectious prion.

Prion diseases are characterized by the accumulation of a pathological form of prion protein (PrP(Sc)), which behaves as an infectious agent. Here we developed an in vitro co-culture system to analyze the PrP(Sc) transmission from ScN2a cell, which persistently retains PrP(Sc), to naïve N2a cell. In this cell-to-cell transmission system, PrP(Sc) transmitted to recipient N2a cell was able to be detected within 5-7days. Further characterization showed that higher cell density greatly facilitated the transmission of PrP(Sc). This improved in vitro transmission method may become a useful tool for unveiling the molecular mechanism of PrP(Sc) transmission.

Identification and structural analysis of C-terminally truncated collapsin response mediator protein-2 in a murine model of prion diseases.

BACKGROUND: Prion diseases are fatal neurodegenerative disorders that accompany an accumulation of the disease-associated form(s) of prion protein (PrPSc) in the central nervous system. The neuropathological changes in the brain begin with focal deposits of PrPSc, followed by pathomorphological abnormalities of axon terminal degeneration, synaptic loss, atrophy of dendritic trees, and eventual neuronal cell death in the lesions. However, the underlying molecular basis for these neuropathogenic abnormalities is not fully understood. RESULTS: In a proteomic analysis of soluble proteins in the brains of mice challenged intracerebrally with scrapie prion (Obihiro I strain), we found that the amount of the full-length form of collapsin response mediator protein-2 (CRMP-2; 61 kDa) decreased in the late stages of the disease, while the amount of its truncated form (56 kDa) increased to comparable levels observed for the full-length form. Detailed analysis by liquid chromatography-electrospray ionization-tandem mass spectrometry showed that the 56-kDa form (named CRMP-2-ΔC) lacked the sequence from serine518 to the C-terminus, including the C-terminal phosphorylation sites important for the regulation of axonal growth and axon-dendrite specification in developing neurons. The invariable size of the mRNA transcript in Northern blot analysis suggested that the truncation was due to post-translational proteolysis. By overexpression of CRMP-2-ΔC in primary cultured neurons, we observed the augmentation of the development of neurite branch tips to the same levels as for CRMP-2T514A/T555A, a non-phosphorylated mimic of the full-length protein. This suggests that the increased level of CRMP-2-ΔC in the brain modulates the integrity of neurons, and may be involved in the pathogenesis of the neuronal abnormalities observed in the late stages of the disease. CONCLUSIONS: We identified the presence of CRMP-2-ΔC in the brain of a murine model of prion disease. Of note, C-terminal truncations of CRMP-2 have been recently observed in models for neurodegenerative disorders such as ischemia, traumatic brain injury, and Wallerian degeneration. While the structural identity of CRMP-2-ΔC in those models remains unknown, the present study should provide clues to the molecular pathology of degenerating neurons in prion diseases in connection with other neurodegenerative disorders.

Comparison of Short-Term Outcomes Between Hand-assisted Laparoscopic Distal Gastrectomy and Laparoscopy-assisted Distal Gastrectomy in Gastric Cancer.

Hand-assisted laparoscopic surgery is considered to provide the benefits of laparoscopic surgery in various diseases. However, limited information is available regarding the feasibility of hand-assisted laparoscopic distal gastrectomy (HALDG)-a subset of hand-assisted laparoscopic surgery-as a gastric cancer treatment. This study aimed to evaluate the usefulness of HALDG compared with laparoscopy-assisted distal gastrectomy (LADG). Consecutive patients who underwent HALDG (n=58) or LADG (n=90) for stage I gastric cancer between 2005 and 2016 were eligible. Operative time was significantly shorter and blood loss was significantly higher in HALDG than in LADG (P<0.001, both). Postoperative aminotransferase levels were significantly lower in HALDG than in LADG (P<0.001). There was no significant difference in perioperative complications, a number of analgesics, postoperative C-reactive protein levels, and 3-year relapse-free and overall survival rates between the groups. This study suggests that HALDG is a safe and feasible approach and could become an effective option for stage I gastric cancer treatment.

Irinotecan plus S-1 for liver metastases of gastric cancer.

BACKGROUND/AIMS: This retrospective study evaluated the efficacy of and compliance with combined irinotecan hydrochloride (CPT-11) and S-1 chemotherapy in patients with liver metastases of gastric cancer. METHODOLOGY: A total of 28 gastric cancer patients with liver metastases received first-line chemotherapy. The response rate, overall survival, and toxicity were evaluated. Fourteen patients were treated with CPT-11+S-1 and they were compared with 14 patients who received cis-diamminedichloroplatinum (CDDP)+S-1. RESULTS: The CPT+S-1 group showed a higher response rate than the CDDP+S-1 group (57.1% [95% CI 31.2-83.1%] vs. 42.9% [95% CI 16.9-68.8%]; p < 0.44). The median survival time of the CPT-11+S-1 group was significantly longer than the CDDP+S-1 group (16.1 months [95% CI 10.5-21.2] vs. 7.3 months [95% CI 2.2-14.7]; hazard ratio for death, 0.35 [95% CI 0.14-0.83]; p < 0.02). By multivariate analysis for the treatment with CPT-11+S-1 was identified as an independent prognostic factor. The most common adverse effect of CPT-11+S-1 therapy was leukopenia (57.1%), which was Grade 3 in 3 patients (21.4%). However, all patients recovered rapidly and there were no significant differences of toxicity between the two regimens. CONCLUSIONS: CPT-11+S-1 therapy will achieve significantly longer survival than CDDP+S-1 without severe toxicity in gastric cancer patients with liver metastases.

[Acquired human prion diseases--past and present issues].

Transmissible spongiform encephalopathies, or prion diseases, are fatal neurodegenerative disorders. In aetiological viewpoint, human prion diseases are classified into 1) sporadic Creutzfeldt-Jakob disease (CJD) which comprises 80-90% of the total population of human prion disaeses, 2) inherited forms, and 3) acquired types by prion-contaminated surgical instruments, biopharmaceuticals or foodstuffs. The diseases cause an accumulation of the disease-associated form(s) of prion protein (PrP(Sc)) in the central nervous system. PrP(Sc) is regarded as the entity of prion agents and generally exerts infectivity, irrespective of its origin being from the sporadic cases or the inherited cases. Variant CJD (vCJD), first identified in the United Kingdom (UK) in 1996, is an acquired type of human CJD by oral intake of BSE prion. Cumulative numbers of 215 patients in the world have been reported for definite or probable vCJD cases according to the UK National Creutzfeldt-Jakob Disease Surveillance Unit by September, 2009. Different from sporadic CJD cases, vCJD patients show an accumulation of PrP(Sc) in spleen and tonsils. Such distribution of PrP(Sc) in lymphoid tissues raised clinical concern about the potential infectivity in the blood or blood components used for blood transfusion. To date, five instances of probable transfusion-mediated transmission of vCJD prion have been found in UK. Here we review the past and the present issues about the acquired human prion diseases.

Tracking and clarifying differential traits of classical- and atypical L-type bovine spongiform encephalopathy prions after transmission from cattle to cynomolgus monkeys.

Classical- (C-) and atypical L-type bovine spongiform encephalopathy (BSE) prions cause different pathological phenotypes in cattle brains, and the disease-associated forms of each prion protein (PrPSc) has a dissimilar biochemical signature. Bovine C-BSE prions are the causative agent of variant Creutzfeldt-Jakob disease. To date, human infection with L-BSE prions has not been reported, but they can be transmitted experimentally from cows to cynomolgus monkeys (Macaca fascicularis), a non-human primate model. When transmitted to monkeys, C- and L-BSE prions induce different pathological phenotypes in the brain. However, when isolated from infected brains, the two prion proteins (PrPSc) have similar biochemical signatures (i.e., electrophoretic mobility, glycoforms, and resistance to proteinase K). Such similarities suggest the possibility that L-BSE prions alter their virulence to that of C-BSE prions during propagation in monkeys. To clarify this possibility, we conducted bioassays using inbred mice. C-BSE prions with or without propagation in monkeys were pathogenic to mice, and exhibited comparable incubation periods in secondary passage in mice. By contrast, L-BSE prions, either with or without propagation in monkeys, did not cause the disease in mice, indicating that the pathogenicity of L-BSE prions does not converge towards a C-BSE prion type in this primate model. These results suggest that, although C- and L-BSE prions propagated in cynomolgus monkeys exhibit similar biochemical PrPSc signatures and consist of the monkey amino acid sequence, the two prions maintain strain-specific conformations of PrPSc in which they encipher and retain unique pathogenic traits.

Interacting targets of the farnesyl of transducin gamma-subunit.

G protein gamma-subunits are isoprenylated and carboxyl methylated at the C-terminal cysteine residue, and the set of the posttranslational modifications is indispensable for the function of the photoreceptor G protein transducin (Talpha/Tbetagamma). To explore farnesyl-mediated molecular interactions, we investigated molecular targets of a Tbetagamma analogue that was engineered to have a photoreactive farnesyl analogue, (3-azidophenoxy)geranyl (POG), covalently bound to the C-terminal cysteine of Tgamma. POG-modified Tbetagamma was further subjected to modification by methylation at the C-terminal carboxyl group, which copies a complete set of the known posttranscriptional modifications of Tbetagamma. Photoaffinity labeling experiment with the photoreactive Tbetagamma analogue in its free form indicated that the POG moiety of Tgamma interacted with Tbeta. In the trimeric Talpha/Tbetagamma complex, the POG moiety was cross-linked with Talpha in addition to concurrent affinity labeling of Tbeta. When photoreactive Tbetagamma was reconstituted with Talpha and light-activated rhodopsin (Rh*) in rod outer segment (ROS) membranes, the POG moiety interacted with not only Talpha and Tbeta but also Rh* and membrane phospholipids. The cross-linked phospholipid species was analyzed by ELISA employing a variety of lipid-binding probes, which revealed phosphatidylethanolamine (PE) and phosphatidylserine (PS) as selective phospholipids for POG interaction in the ROS membranes. These results demonstrate that the modifying group of Tgamma plays an active role in protein-protein and protein-membrane interactions and suggest that the farnesyl-PE/PS interaction may support the efficient formation of the signaling ternary complex between transducin and Rh*.

Short-term outcome of laparoscopic cholecystectomy for benign gall bladder diseases in 76 dogs.

Laparoscopic cholecystectomy (LC) is widely accepted as the standard treatment for benign gall bladder diseases in humans because it has proven to be less invasive and safer than are traditional methods. However, the efficacy of LC in dogs remains unclear. The present study aimed to examine the short-term outcome of LC for benign gall bladder diseases in dogs. We enrolled 76 consecutive dogs that underwent LC for benign gall bladder diseases at our hospital between April 2008 and October 2016. Dogs with jaundice, gall bladder ruptures, abdominal effusion, or extrahepatic biliary obstruction were not excluded from the indication. Factors including age, body weight, sex, clinical sign, disease, operative time, conversion to open surgery, perioperative complications, and postoperative hospital stay were investigated. The median age of the dogs was 11 years, and the median body weight was 5.4 kg. Fifty percent of the dogs exhibited no symptoms at the initial visit. Preoperative elevation of total bilirubin levels was observed in 16 dogs (21%). LC was successfully completed in 71 dogs (93%); the median operative time was 124 min. Although gall bladder ruptures were observed in 2 (2.6%) dogs, the operations were completed successfully. Three dogs (4.1%) had to be converted to open cholecystectomy and 2 (2.6%) underwent reoperation. Two dogs (2.6%) died intraoperatively and 2 (2.6%) died postoperatively. LC was a feasible, safe, and appropriate procedure considering the current operative indications for benign gall bladder diseases in dogs.

Thiol-reactive reagents inhibits intracellular trafficking of human papillomavirus type 16 pseudovirions by binding to cysteine residues of major capsid protein L1.

BACKGROUND: A human papillomavirus (HPV) virion is composed of capsid proteins L1 and L2. Several cysteine residues are located on L1 of various HPVs at markedly similar relative positions, suggesting their important functions. Although the authentic virions cannot be studied with cultured cells, surrogate pseudovirions consisting of capsid and reporter plasmid are available for studies dealing with infectivity. RESULTS: HPV type16-pseudovirions (16PVs) were found to lose their infectivity after incubation with thiol-reactive reagents [biotin polyethyleneoxide iodoacetamide (BPEOIA), 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), N-ethylmaleimide (NEM), 4-(N-maleimido)benzyl-trimethylammonium iodide (MBTA), and [2-(trimethylammonium)ethyl] methanethiosulfonate bromide (MTSET)]. A labelled streptavidin was detected to bind to the complex of BPEOIA and L1 of the 16PVs incubated with BPEOIA. The analysis of molecular mass of trypsin-fragments derived from the complex of the BPEOIA and L1 indicated that BPEOIA bound to at least C146, C225, and C229. No appreciable change of the 16PVs carrying DTNB or NEM was detected by sedimentation analysis or electron microscopy. The 16PVs carrying DTNB or NEM were able to bind to and enter HeLa cells but degraded before they reached the perinuclear region. CONCLUSION: HPV16 L1 C146, C225, and C229 have free thiol, which are accessible to BPEOIA, DTNB, NEM, MBTA, and MTSET. Binding of DTNB or NEM to the thiols may cause conformational changes that result in the inhibition of the entry and trafficking of the 16PVs.