Predictors of a difficult Pringle maneuver in laparoscopic liver resection and evaluation of alternative procedures to assist bleeding control.

PURPOSE: To evaluate the predictors of a difficult Pringle maneuver (PM) in laparoscopic liver resection (LLR) and to assess alternative procedures to PM. METHODS: Data from patients undergoing LLR between 2013 and 2020 were reviewed retrospectively. Univariate and multivariate analyses were performed and the outcomes of patients who underwent PM or alternative procedures were compared. RESULTS: Among 106 patients who underwent LLR, PM could not be performed in 18 (17.0%) because of abdominal adhesions in 14 (77.8%) and/or collateral flow around the hepatoduodenal ligament in 5 (27.8%). Multivariate analysis revealed that Child-Pugh classification B (p = 0.034) and previous liver resection (p < 0.001) were independently associated with difficulty in performing PM in LLR. We evaluated pre-coagulation of liver tissue using microwave tissue coagulators, saline irrigation monopolar, clamping of the hepatoduodenal ligament using an intestinal clip, and hand-assisted laparoscopic surgery as alternatives procedures to PM. There were no significant differences in blood loss (p = 0.391) or transfusion (p = 0.518) between the PM and alternative procedures. CONCLUSIONS: Child-Pugh classification B and previous liver resection were identified as predictors of a difficult PM in LLR. The alternative procedures were found to be effective.

Chromosomal Aberrations in Large Japanese Field Mice (Apodemus speciosus) Captured near Fukushima Dai-ichi Nuclear Power Plant.

Since the Fukushima Dai-ichi Nuclear Power Plant accident, radiation effects on nonhuman biota in the contaminated areas have been a major concern. Here, we analyzed the frequencies of chromosomal aberrations (translocations and dicentrics) in the splenic lymphocytes of large Japanese field mice (Apodemus speciosus) inhabiting Fukushima Prefecture. A. speciosus chromosomes 1, 2, and 5 were flow-sorted in order to develop A. speciosus chromosome-specific painting probes, and FISH (fluorescence in situ hybridization) was performed using these painting probes to detect the translocations and dicentrics. The average frequency of the translocations and dicentrics per cell in the heavily contaminated area was significantly higher than the frequencies in the case of the noncontaminated control area and the slightly and moderately contaminated areas, and this aberration frequency in individual mice tended to roughly increase with the estimated dose rates and accumulated doses. In all four sampling areas, the proportion of aberrations occurring in chromosome 2 was approximately >3 times higher than that in chromosomes 1 and 5, which suggests that A. speciosus chromosome 2 harbors a fragile site that is highly sensitive to chromosome breaks induced by cellular stress such as DNA replication. The elevated frequency of chromosomal aberrations in A. speciosus potentially resulting from the presence of a fragile site in chromosome 2 might make it challenging to observe the mild effect of chronic low-dose-rate irradiation on the induction of chromosomal aberrations in A. speciosus inhabiting the contaminated areas of Fukushima.

Serum p53 antibody as a potential tumor marker in extrahepatic cholangiocarcinoma.

PURPOSE: Only a few studies have evaluated the clinicopathological significance of the p53 protein expression and s-p53-Abs level in patients with cholangiocarcinoma. We therefore analyzed the clinicopathological and prognostic significance of s-p53-Abs in patients with extrahepatic cholangiocarcinoma. METHODS: We prospectively evaluated s-p53-Abs levels before and after surgery in 61 patients with extrahepatic cholangiocarcinoma to determine the relationship between clinicopathological factors and the prognostic significance of s-p53-Abs. RESULTS: Among a total of 61 primary extrahepatic cholangiocarcinoma cases, 23% were positive for s-p53-Abs. Combination of s-p53-Abs with the conventional serum markers carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) significantly increased the rate of positive extrahepatic cholangiocarcinoma cases (57% for CEA and/or CA19-9 vs. 75% for CEA and/or CA19-9 and/or s-p53-Abs, P = 0.035). There were no significant differences in clinicopathological factors between the p53-seropositive and p53-seronegative patients. An immunohistochemical analysis showed the presence of significant associations between the intensity (P = 0.003) and extent (P = 0.001) of p53 immunoreactivity and p53-seropositivitly. Although s-p53-Abs was not a significant prognostic factor for the survival in either univariate or multivariate analyses, p53 immunoreactivity was independently associated with a poor survival. Among patients positive for s-p53-Abs before surgery, the s-p53-Abs levels were reduced after surgery in most. CONCLUSION: These findings suggested that s-p53-Abs might be associated with p53 immunoreactivity. In addition, s-p53-Abs may be useful for a diagnosis, but was not useful for predicting tumor recurrence or the survival. This study was registered as UMIN000014530.

UBE4B protein couples ubiquitination and sorting machineries to enable epidermal growth factor receptor (EGFR) degradation.

The signaling of plasma membrane proteins is tuned by internalization and sorting in the endocytic pathway prior to recycling or degradation in lysosomes. Ubiquitin modification allows recognition and association of cargo with endosomally associated protein complexes, enabling sorting of proteins to be degraded from those to be recycled. The mechanism that provides coordination between the cellular machineries that mediate ubiquitination and endosomal sorting is unknown. We report that the ubiquitin ligase UBE4B is recruited to endosomes in response to epidermal growth factor receptor (EGFR) activation by binding to Hrs, a key component of endosomal sorting complex required for transport (ESCRT) 0. We identify the EGFR as a substrate for UBE4B, establish UBE4B as a regulator of EGFR degradation, and describe a mechanism by which UBE4B regulates endosomal sorting, affecting cellular levels of the EGFR and its downstream signaling. We propose a model in which the coordinated action of UBE4B, ESCRT-0, and the deubiquitinating enzyme USP8 enable the endosomal sorting and lysosomal degradation of the EGFR.

Chromosomal Aberrations in Wild Mice Captured in Areas Differentially Contaminated by the Fukushima Dai-Ichi Nuclear Power Plant Accident.

Following the Fukushima Dai-ichi Nuclear Power Plant accident, radiation effects on nonhuman biota in the contaminated areas have been a great concern. The induction of chromosomal aberrations in splenic lymphocytes of small Japanese field mice (Apodemus argenteus) and house mice (Mus musculus) inhabiting Fukushima Prefecture was investigated. In mice inhabiting the slightly contaminated area, the average frequency of dicentric chromosomes was similar to that seen in mice inhabiting a noncontaminated control area. In contrast, mice inhabiting the moderately and heavily contaminated areas showed a significant increase in the average frequencies of dicentric chromosomes. Total absorbed dose rate was estimated to be approximately 1 mGy d(-1) and 3 mGy d(-1) in the moderately and heavily contaminated areas, respectively. Chromosomal aberrations tended to roughly increase with dose rate. Although theoretically, the frequency of chromosomal aberrations was considered proportional to the absorbed dose, chromosomal aberrations in old mice (estimated median age 300 days) did not increase with radiation dose at the same rate as that observed in young mice (estimated median age 105 days).

(135)Cs/(137)Cs isotopic ratio as a new tracer of radiocesium released from the Fukushima nuclear accident.

Since the Fukushima Daiichi nuclear power plant (FDNPP) accident in 2011, intensive studies of the distribution of released fission products, in particular (134)Cs and (137)Cs, in the environment have been conducted. However, the release sources, that is, the damaged reactors or the spent fuel pools, have not been identified, which resulted in great variation in the estimated amounts of (137)Cs released. Here, we investigated heavily contaminated environmental samples (litter, lichen, and soil) collected from Fukushima forests for the long-lived (135)Cs (half-life of 2 × 10(6) years), which is usually difficult to measure using decay-counting techniques. Using a newly developed triple-quadrupole inductively coupled plasma tandem mass spectrometry method, we analyzed the (135)Cs/(137)Cs isotopic ratio of the FDNPP-released radiocesium in environmental samples. We demonstrated that radiocesium was mainly released from the Unit 2 reactor. Considering the fact that the widely used tracer for the released Fukushima accident-sourced radiocesium in the environment, the (134)Cs/(137)Cs activity ratio, will become unavailable in the near future because of the short half-life of (134)Cs (2.06 years), the (135)Cs/(137)Cs isotopic ratio can be considered as a new tracer for source identification and long-term estimation of the mobility of released radiocesium in the environment.

Efficacy of laparoscopic liver resection in colorectal liver metastases and the influence of preoperative chemotherapy.

BACKGROUND: Since 1993, we have performed minimally invasive laparoscopic liver resection (LLR) to treat malignant liver cancer, including colorectal liver metastases (CLM). However, further studies are needed to accumulate sufficient evidence on the oncological outcome of LLR for CLM. METHODS: To elucidate the efficacy of LLR for CLM, this study comparatively analyzed the invasiveness and short-term prognosis of LLR (n=43 cases) and open liver resection (OR) (n=62 cases) performed for CLM after 2006 and also investigated the safety of LLR following chemotherapy. RESULTS: Compared with the OR group, the LLR group had significantly less blood loss (P<0.001) and a shorter hospital stay (P<0.001). The E-PASS scoring system was used to compare surgical invasiveness, and although the preoperative risk score did not differ between the groups, the surgical stress score and comprehensive risk score were significantly lower in the LLR group (P<0.001). Concerning the survival rate and disease-free survival rate, there were no significant differences between procedures. However, more clinical cases and longer follow-up periods are needed to reach a definitive conclusion.Preoperative hemanalysis, intraoperative bleeding, complications, and postoperative length of stay did not differ significantly between LLR patients with preoperative chemotherapy and those with surgery alone, indicating no adverse effects of chemotherapy. CONCLUSIONS: LLR can be an effective minimally invasive surgery in CLM patients receiving both perioperative chemotherapy and surgery. Because LLR is comparable with OR with regard to short-term oncological outcome, LLR may be a valuable option for CLM.

[A case of pancreatic cancer with distant metastasis successfully resected after neoadjuvant chemotherapy with a combination of S-1 and gemcitabine].

The patient was a 70-year-old man with a chief complaint of cough.After careful examination, he was diagnosed with pancreatic body cancer with metastasis to the liver and right pleura and with early gastric cancer.He was treated with S-1 and gemcitabine combination chemotherapy.After completing 3 courses, the distant metastasis could no longer be observed. After completing 4 courses, the tumor marker level in the serum was normalized.The pancreatic lesion was restricted by the end of 10 courses, and the pancreas body and tail were resected.After additional chemotherapy with S-1, he was switched to weekly paclitaxel therapy because of peritoneal dissemination.The patient survived for 15 months after surgery.In cases of unresectable pancreatic cancer with distant metastasis, it may be possible to consider the surgical option when chemotherapy is effective.

[An analysis of cases with malignant pyloroduodenal obstruction from unresectable pancreatic cancer who underwent palliative gastrojejunostomy].

The aims of this study were to examine the effectiveness of palliative gastrojejunostomy in terms of the postoperative food intake and hospital stay in patients with advanced pancreatic cancer. Retrospective data from 17 consecutive patients who underwent gastrojejunostomy from 2008 to 2013 in Toho University Omori Medical Center were analyzed. In our analysis, patients without peritoneal dissemination and distant metastasis were associated with poor oral intake and patients with lower neutrophil to lymphocyte ratio (NLR) were associated with rate of discharge.

Effectiveness of laparoscopic subtotal cholecystectomy: perioperative and long-term postoperative results.

BACKGROUND/AIMS: We analyzed perioperative and long-term postoperative data to determine the effectiveness of laparoscopic subtotal cholecystectomy (LSC). METHODOLOGY: 89 LSCs done at our center were classified into 3 groups according to the operative procedure required to treat severe cholecystitis. Perioperative and long-term postoperative results were then compared with standard laparoscopic cholecystectomy (s-LC). RESULTS: Operative time was longer and postoperative CRP level was higher for LSC. Significantly more bleeding and longer operative times were seen only in the LSC-II subgroup, and average postoperative hospital stay was longer only in the LSC-III subgroup. As for long-term (1.5 to 8 years) postoperative results, 3 of 26 LSC-III cases had a relapse of cholecystolithiasis in remnant gallbladder tissue 2 years or later after initial surgery. CONCLUSIONS: There were no serious intraoperative complications such as the bile duct injury, and a smaller proportion of procedures were intraoperatively converted to open laparotomy. It is considered that LSC is a safe, useful surgical procedure to the patients in whom the neck of the gallbladder is anatomically unclear due to cholecystitis or fibrosis although patients undergoing LSC-III should be monitored for cholecystolithiasis in remnant gallbladder tissue. LSC is often the procedure of choice for patients.

[Laparoscopic surgery for pancreatic cancer].

Laparoscopic distal pancreatectomy is currently in widespread use because of its technical simplicity, although laparoscopic pancreaticoduodenectomy is practiced in only a limited number of surgical institutions due to the associated technical difficulties especially at reconstruction. The indication for the treatment of pancreatic cancer by a laparoscopic procedure remains controversial. Because pancreatic cancer is already advanced at the time of diagnosis in many cases, it requires a difficult radical operation in terms of lymphadenectomy with nerves plexus in laparoscopic surgery. The oncologic clearance of pancreatic resection may be achievable by laparoscopic surgery, although longer follow-up and larger series of treatment are still needed.

Chromosomal Aberrations in Large Japanese Field Mice (Apodemus speciosus) Captured in Various Periods after Fukushima Daiichi Nuclear Power Plant Accident.

After the Fukushima Daiichi Nuclear Power Plant accident, we studied the chromosomal aberrations (dicentrics and translocations) in the splenic lymphocytes of wild mice inhabiting Fukushima prefecture. Here, we report the frequencies of chromosomal aberrations in large Japanese field mice (Apodemus speciosus) captured from 2012 to 2016 in a heavily contaminated area. The chromosomal aberrations were detected using newly developed 4-color FISH (fluorescence in situ hybridization) with A. speciosus chromosome 1-, 3-, 4- and 5-specific painting probes. The frequencies of chromosomal aberrations in mice captured in July 2012 and October 2014 were significantly higher than that in the mice inhabiting the non-contaminated control area; however, the frequency of chromosomal aberrations in mice captured in January 2016 was not. The frequency of chromosomal aberrations in individual mice tended to increase with certain dose rates and accumulated doses. Regression tree analyses suggested increasing chromosomal aberration rate in mice exposed to chronic radiation at dose rates of more than 1.1 mGy day-1 and at accumulated doses of more than 200 mGy. It is concluded that ambient dose rates in the most severely contaminated area of Fukushima prefecture and radiation doses to wild mice inhabiting this area decrease with time; consequently, chromosomal aberrations induced by radiation have not been detected 5 years after the accident.

The impacts of geometry and binding on CaMKII diffusion and retention in dendritic spines.

We used a particle-based Monte Carlo simulation to dissect the regulatory mechanism of molecular translocation of CaMKII, a key regulator of neuronal synaptic function. Geometry was based upon measurements from EM reconstructions of dendrites in CA1 hippocampal pyramidal neurons. Three types of simulations were performed to investigate the effects of geometry and other mechanisms that control CaMKII translocation in and out of dendritic spines. First, the diffusional escape rate of CaMKII from model spines of varied morphologies was examined. Second, a postsynaptic density (PSD) was added to study the impact of binding sites on this escape rate. Third, translocation of CaMKII from dendrites and trapping in spines was investigated using a simulated dendrite. Based on diffusion alone, a spine of average dimensions had the ability to retain CaMKII for duration of ~4 s. However, binding sites mimicking those in the PSD controlled the residence time of CaMKII in a highly nonlinear manner. In addition, we observed that F-actin at the spine head/neck junction had a significant impact on CaMKII trapping in dendritic spines. We discuss these results in the context of possible mechanisms that may explain the experimental results that have shown extended accumulation of CaMKII in dendritic spines during synaptic plasticity and LTP induction.

Lobe specific Ca2+-calmodulin nano-domain in neuronal spines: a single molecule level analysis.

Calmodulin (CaM) is a ubiquitous Ca(2+) buffer and second messenger that affects cellular function as diverse as cardiac excitability, synaptic plasticity, and gene transcription. In CA1 pyramidal neurons, CaM regulates two opposing Ca(2+)-dependent processes that underlie memory formation: long-term potentiation (LTP) and long-term depression (LTD). Induction of LTP and LTD require activation of Ca(2+)-CaM-dependent enzymes: Ca(2+)/CaM-dependent kinase II (CaMKII) and calcineurin, respectively. Yet, it remains unclear as to how Ca(2+) and CaM produce these two opposing effects, LTP and LTD. CaM binds 4 Ca(2+) ions: two in its N-terminal lobe and two in its C-terminal lobe. Experimental studies have shown that the N- and C-terminal lobes of CaM have different binding kinetics toward Ca(2+) and its downstream targets. This may suggest that each lobe of CaM differentially responds to Ca(2+) signal patterns. Here, we use a novel event-driven particle-based Monte Carlo simulation and statistical point pattern analysis to explore the spatial and temporal dynamics of lobe-specific Ca(2+)-CaM interaction at the single molecule level. We show that the N-lobe of CaM, but not the C-lobe, exhibits a nano-scale domain of activation that is highly sensitive to the location of Ca(2+) channels, and to the microscopic injection rate of Ca(2+) ions. We also demonstrate that Ca(2+) saturation takes place via two different pathways depending on the Ca(2+) injection rate, one dominated by the N-terminal lobe, and the other one by the C-terminal lobe. Taken together, these results suggest that the two lobes of CaM function as distinct Ca(2+) sensors that can differentially transduce Ca(2+) influx to downstream targets. We discuss a possible role of the N-terminal lobe-specific Ca(2+)-CaM nano-domain in CaMKII activation required for the induction of synaptic plasticity.

[Hepatic arterial infusion chemotherapy with oxaliplatin in unresectable liver metastases from colorectal cancer after systemic chemotherapy failure].

UNLABELLED: We report 5 cases of colorectal liver metastases (CRLM) with hepatic arterial infusion (HAI) oxaliplatin after systemic infusion chemotherapy failure. Patients with unresectable CRLM and history of systemic chemotherapy failure were treated with HAI oxaliplatin (L-OHP 100 mg/body, 2 hours) combined with intravenous (iv) levofolinate calcium (175 mg/body, 2 hours) and iv bolus 5-FU (500 mg/body) every 2 weeks. RESULT: An average age was 58 years. All patients had previously received FOLFOX. Lung metastases had already existence before HAI oxaliplatin in 4 patients. A median of 10 treatments were administered (range 5-14). Serum level of CEA was decreased in 4 cases. In 2 patients, lung metastasis developed while a PR was obtained in the liver metastasis. Progress disease (PD) was confirmed in other 3 patients. No major toxicity was presented. The median time to progression free survival was 3.0 months and the median overall survival was 7.1 months. CONCLUSION: HAI oxaliplatin might be beneficial as a salvage therapy for CRLM without extrahepatic metastasis, which demonstrated an acceptable tolerability and maintenance of QOL.

Dissecting cooperative calmodulin binding to CaM kinase II: a detailed stochastic model.

Calmodulin (CaM) is a major Ca(2+) binding protein involved in two opposing processes of synaptic plasticity of CA1 pyramidal neurons: long-term potentiation (LTP) and depression (LTD). The N- and C-terminal lobes of CaM bind to its target separately but cooperatively and introduce complex dynamics that cannot be well understood by experimental measurement. Using a detailed stochastic model constructed upon experimental data, we have studied the interaction between CaM and Ca(2+)-CaM-dependent protein kinase II (CaMKII), a key enzyme underlying LTP. The model suggests that the accelerated binding of one lobe of CaM to CaMKII, when the opposing lobe is already bound to CaMKII, is a critical determinant of the cooperative interaction between Ca(2+), CaM, and CaMKII. The model indicates that the target-bound Ca(2+) free N-lobe has an extended lifetime and may regulate the Ca(2+) response of CaMKII during LTP induction. The model also reveals multiple kinetic pathways which have not been previously predicted for CaM-dissociation from CaMKII.

Laparoscopic hepatectomy for liver tumors: proposals for standardization.

BACKGROUND/PURPOSE: We draw on our experience with laparoscopic hepatectomy (LH) to present recommendations for standardization of LH for the treatment of liver tumors. METHODS: At our center, 90 LHs were performed from April 1993 to January 2008. These were divided equally into early cases and late cases, and short-term postoperative results were compared. Forty-nine of the LH procedures were total-laparoscopic procedures, 16 were hand-assisted procedures, and 25 were laparoscopy-assisted procedures. The tumors were malignant in 76 cases and benign in 14 cases. RESULTS: Among late cases, the numbers of malignant tumors and tumors located in the posterosuperior region of the liver (Segments VII, VIII, and IVb) were significantly higher than among early cases; however, operative blood loss and postoperative hospital stay were significantly lower in the late cases (158.9 +/- 213.4 vs. 377.6 +/- 421.2 cc, P = 0.007; and 8.7 +/- 3.6 vs. 15.3 +/- 8.7 days, P = 0.0001, respectively). No operative deaths occurred in either group. CONCLUSIONS: Although LH does have a steep learning curve, we believe that it can be standardized and provide a less invasive surgical option--with no reduction in disease curability--for the treatment of liver tumors in selected patients.

Genome-wide gene expression profiling in gamma-irradiated green alga, Pseudokirchneriella subcapitata, by HiCEP.

Transcriptome was analyzed in gamma-irradiated green alga Pseudokirchneriella subcapitata by high coverage gene expression profiling (HiCEP). Approximately 7,800 expressed genes were detected. Expression levels of 623-707 genes were affected at 100-300 Gy. Nucleotide sequences of 41 up-regulated genes were determined. The quantitative reverse transcription polymerase chain reaction validated the up-regulation. Two genes had homology to genes related to ionizing radiation. These results indicate usefulness of HiCEP for screening of stress-responsive genes in species that are ecotoxicologically important but for which genomic sequence information is lacking.