Alignment of CH3F in para-H2 crystal studied by IR quantum cascade laser polarization spectroscopy.

In order to investigate the alignment of CH3F in para-H2 crystals, high resolution polarization spectroscopy of the ν3 vibrational band is studied using a quantum cascade laser at 1040 cm(-1). It is found that the main and satellite series of peaks in the ν3 vibrational band of CH3F have the same polarization dependence. This result supports the previously proposed cluster model with ortho-H2 in first and second nearest neighbor sites. The observed polarization dependence function is well described by a simple six-axis void model in which CH3F is not aligned along the c-axis of the crystal but tilted to 64.9(3)° from it.

Molecular epidemiology of livestock rabies viruses isolated in the northeastern Brazilian states of Paraíba and Pernambuco from 2003 - 2009.

BACKGROUND: Limited or no epidemiological information has been reported for rabies viruses (RABVs) isolated from livestock in the northeastern Brazilian states of Paraíba (PB) and Pernambuco (PE). The aim of this study was to clarify the molecular epidemiology of RABVs circulating in livestock, especially cattle, in these areas between 2003 and 2009. FINDINGS: Phylogenetic analysis based on 890 nt of the nucleoprotein (N) gene revealed that the 52 livestock-derived RABV isolates characterized here belonged to a single lineage. These isolates clustered with a vampire bat-related RABV lineage previously identified in other states in Brazil; within PB and PE, this lineage was divided between the previously characterized main lineage and a novel sub-lineage. CONCLUSIONS: The occurrences of livestock rabies in PB and PE originated from vampire bat RABVs, and the causative RABV lineage has been circulating in this area of northeastern Brazil for at least 7 years. This distribution pattern may correlate to that of a vampire bat population isolated by geographic barriers.

Efficient transformation of small hepatocytes into insulin-expressing cells by forced expression of Pdx1.

BACKGROUND/PURPOSE: The expression of ectopic pancreatic and duodenal homeobox factor 1 (Pdx1) can transform hepatocytes into pancreatic endocrine cells. Small hepatocytes (SHs) have a high possibility to be a cellular source for islet cell transplantation. However, the efficacy of the transformation of SHs into pancreatic endocrine cells is not fully understood. The focus of our study was to compare the efficacy of the transformation into pancreatic endocrine cells of SHs and mature hepatocytes (MHs). METHODS: MHs and SHs were cultured for 3 and 10 days, respectively, before Adeno-Pdx1 gene transduction. Western blot analysis was performed for pancreatic transcription factors, and reverse-transcription polymerase chain reaction (RT-PCR) was performed for the gene expression of pancreatic hormones. Confocal laser microscanning analysis was used to observe insulin and glucagon expression. RESULTS: Although the pancreatic transcription factors Pdx1, Ngn3, NeuroD, and Pax6 were induced in both SHs and MHs after Adeno-Pdx1 gene expression, the pancreatic transcription factors Nkx2.2 and Nkx6.1 were induced in SHs more than in MHs. Glucagon mRNA expression was seen in both SHs and MHs, whereas insulin mRNA expression was higher in SHs than in MHs. Confocal laser microscanning analysis showed that SHs expressed both insulin and glucagon, whereas MHs predominantly expressed glucagon. CONCLUSIONS: SHs were transformed into both insulin-and glucagon-expressing cells, and the efficacy of the transformation into insulin-expressing cells of SHs was higher than that for MHs. Thus, SHs could be a more suitable source of future cell therapy than MHs.

[A case of advanced pancreatic cancer successfully treated by combined chemotherapy of S-1 and gemcitabine].

The patient was a 71-year-old woman. In March of 2007, she experienced abdominal pain, and consulted a physician at our hospital. She was followed on a outpatient basis, but the symptom did not improve, so she consulted our division. As a result of examinations, pancreatic head tumor, multiple liver metastasis, and para-aortic lymph node swelling were detected. She was then admitted for chemotherapy in our hospital. She was treated with combined chemotherapy of S-1 and GEM. Tumor marker(CEA, CA19-9)decreased and the size of the pancreatic head tumor and liver metastasis decreased. No toxic events were observed due to this therapy. She was treated as an outpatient, and was considered to have good quality of life.

In vitro transformation of adult rat hepatic progenitor cells into pancreatic endocrine hormone-producing cells.

BACKGROUND/PURPOSE: Pancreatic and duodenal homeobox factor 1 (Pdx-1) plays an important role in initiating differentiation toward pancreatic endocrine cells. The transdifferentiation or transformation of hepatocytes into pancreatic endocrine cells could be feasible, due to their similar cellular origins. Our goal in this study was to see if small hepatocytes (SHs) could give rise to pancreatic endocrine cells via exogenous Pdx-1 gene expression. METHODS: SHs were cultured for 10 days before adenovirus (Adt)-mediated Pdx-1 gene transfection. We performed western blot analysis for pancreatic transcription factors in the nuclei and reverse-transcription polymerase chain reaction (RT-PCR) for the gene expression of pancreatic endocrine hormones. Confocal laser microscanning analysis was used to observe the transformation of SHs. RESULTS: Pancreatic transcription factors such as Pdx-1, Ngn3, NeuroD, Nkx2.2, and Pax6 were induced after Adt-Pdx-1 gene transfection. The mRNA expression of pancreatic endocrine hormones (insulin, glucagon, and somatostatin) was induced after the gene transfection. Pdx-1 was expressed in the nucleus, where the cells were positive for one or more of the hormones and cytokeratin (CK) 8. Some cells were positive for multiple hormones. The insulin level increased while the glucagon level decreased after the glucose loading test, depending on the glucose concentration. CONCLUSIONS: SHs are transformed into functional pancreatic endocrine cells after Pdx-1 gene transfection.

Tangent height registration method for the Version 1.4 data retrieval algorithm of the solar occultation sensor ILAS-II.

The Improved Limb Atmospheric Spectrometer-II (ILAS-II) is a satellite-borne solar occultation sensor onboard the Advanced Earth Observing Satellite-II (ADEOS-II). The ILAS-II succeeded the ILAS. The ILAS-II used four grating spectrometers to observe vertical profiles of gas volume mixing ratios of trace constituents and was also equipped with a Sun-edge sensor to determine tangent heights geometrically with high precision. The accuracy of gas volume mixing ratios depends on the accuracy of the tangent height determination. The combination method is a tangent height registration method that was developed to give appropriate tangent heights for the ILAS-II Version 1.4 data retrieval algorithm. This study describes the method used in the ILAS-II Version 1.4 retrieval algorithm to register tangent heights. The root-sum-square total random error is estimated to be 30 m, and the total systematic error is 180 m at an altitude of 30 km. The influence of the tangent height errors on the vertical profiles of gas volume mixing ratios in ILAS-II Version 1.4 is estimated by using the relative difference. The relative difference for each species is within 7% (20%) for an altitude shift of +/-100 m(+/-300 m).

Assessment of liver fibrosis by a noninvasive method of transient elastography and biochemical markers.

AIM: To assess the correlation between the fibrotic area (FA) as calculated by a digital image analysis (DIA), and to compare the diagnostic accuracy of FibroScan to the other existing Liver fibrosis (LF) markers using the receiver operating curve analysis. METHODS: We recruited 30 patients who underwent a liver resection for three different etiologies including normal liver, hepatitis B, and hepatitis C. Liver stiffness was measured by using a FibroScan. The FA was then calculated by DIA to evaluate LF in order to avoid any sampling bias. RESULTS: The FA negatively correlated with Prothrom-bin time (PT), platelet count, lecithin-cholesterol acyltransferase (LCAT), and pre-albumin (ALB). On the other hand, the findings of FibroScan correlated with similar markers. The FA positively correlated with FibroScan, serum hyaluronate level, and type IV collagen level, and aspartate transaminase to platelet ratio index (APRI). The area under the receiver operating curve for FibroScan was higher than that for the other markers, even though the statistical significance was minimal. CONCLUSION: Our findings suggest that FibroScan can initially be used to assess LF as an alternative to a liver biopsy (LB) and serum diagnosis, because it is a safe method with comparable diagnostic accuracy regarding the existing LF markers.

Serum lipid and lipoprotein alterations represent recovery of liver function after hepatectomy.

BACKGROUND: The assessment of liver function during human liver regeneration is necessary to prevent unexpected liver failure and to prepare for further treatment. We selected patients prospectively and measured serum lipid and lipoprotein levels to identify which lipids and lipoproteins could represent recovery of liver function in human liver regeneration. METHODS: Thirty selected patients who underwent hepatectomy were divided into three groups depending on the serum hyaluronate (HA) level and the type of liver resection. RESULTS: We found three patterns of lipid and lipoprotein alterations after hepatectomy. Among the lipids and lipoproteins examined, the serum beta-lipoprotein and low-density lipoprotein (LDL) levels were significantly different among the groups at 7 days after hepatectomy. The alteration of the apolipoprotein (Apo) B level was similar to that of LDL. The LDL level was correlated with both beta-lipoprotein and Apo B before hepatectomy (r=0.653 and 0.894, respectively) and at 7 days after hepatectomy (r=0.841 and 0.943, respectively). CONCLUSION: Serum HA before hepatectomy can reflect postoperative liver function depending on the type of liver resection. Recovery of the beta-lipoprotein and LDL levels can reflect the recovery of liver function in human liver regeneration within the early period in association with the Apo B level.

Liver repopulation and long-term function of rat small hepatocyte transplantation as an alternative cell source for hepatocyte transplantation.

Hepatocyte transplantation (HT) is an attractive therapeutic modality for liver disease as an alternative for liver organ transplantation. Primary fresh hepatocytes (FHs) are the exclusive cell source that has been used for clinical HT. However, the use of FHs is limited due to a shortage of donor cells. Small hepatocytes (SHs) are hepatic progenitor cells and can be isolated not only from rodents but also from humans. SHs can proliferate in vitro and express liver functions, although conventional hepatocytes lose them within a short period after culture. SH functions in vivo have never been studied. We therefore investigated HT using SHs to evaluate cell engraftment and function compared to HT using FHs. The donor cell number in the SH group was smaller than that in the FH group at HT. The cell engraftment in the SH group was smaller in the liver and larger in the spleen than in the FH group. The cell engraftment in the liver increased after HT; however, that in the spleen decreased after HT in both groups. HT using SHs supported the serum albumin level in the NAR experiment as well as that using FH, and albumin mRNA was detectable in the recipients' tissues at 12 weeks after HT. In conclusion, HT using SHs showed hepatic repopulation similar to that using FHs. This suggests that both SHs and FHs can repopulate the liver as if they were hepatic stem cells. In addition, HT using SHs supported liver functions such as albumin correction at the same level as that using FHs. These observations strongly support the idea that SHs could be an alternative to primary FHs as a novel cell source for future HT.

Hyperbaric oxygen stimulates cell proliferation and normalizes multidrug resistance protein-2 protein localization in primary rat hepatocytes.

Hyperbaric oxygen therapy (HBO) has been used for many clinical treatments, including primary liver non-function. However, the cellular mechanism by which HBO treatment ameliorates liver function is not understood. Therefore, the purpose of this study was to elucidate this cellular mechanism using primary cultured rat hepatocytes in in vitro studies. Hepatocytes were treated with HBO at 1 day after plating, and the morphological and functional characteristics of bile canaliculi formed in cultured hepatocytes were observed by time-lapse microscopy. Multidrug resistance protein-2 localization was observed by confocal laser microscopy. In cultured hepatocytes, the labeling index in the HBO group at 2 days after treatment was significantly higher than that in the control group. In addition, the proliferating cellular nuclear antigen level in the HBO group was significantly higher than that in the control group. The contraction of the bile canaliculi in the HBO group was slower than in the control group and the dilatation of bile canaliculi in the HBO group was much larger than in the control group. Multidrug resistance protein-2 in the HBO group was localized at the apical membrane. These results show that HBO stimulates hepatocytes to proliferate and HBO normalizes multidrug resistance protein-2 localization to the apical membrane, which could dilate bile canaliculi.

Progression and regression of biloma after laparoscopic cholecystectomy evaluated by computed tomography.

The natural course of biloma after laparoscopic cholecystectomy (LC) was successfully followed and evaluated by computed tomography (CT). Biloma is one of the major complications after LC. However, the processes of biloma progression and regression have not yet been reported in detail. We encountered a case of minor leakage of bile juice after LC. Unexpected injury of the Luschka duct (accessory bile duct) was suspected. We examined this patient periodically by CT, which is very reliable and easy to use for postoperative evaluation. Biloma gradually developed within 6 months as we suspected. We successfully detected the presence of the Luschka duct by bilomagraphy. The patient was symptomatically silent for the next 6 months as the size of the biloma spontaneously decreased. Therefore, it is not always necessary to carry out treatment for biloma after LC if the patient has no symptoms. This is the first report that shows the progression and regression of biloma evaluated precisely by CT.

Serum hyaluronate level for predicting subclinical liver dysfunction after hepatectomy.

The serum hyaluronate (HA) level reflects sinusoidal endothelial cell function correlated with liver function. We have reviewed multiple liver function indicators from 37 patients who underwent hepatectomy for various liver diseases. The serum HA level was well correlated with the indocyanine green retention rate at 15 minutes (ICGR15), lectin-cholesterol (LCAT), hepatocyte growth factor (HGF), liver uptake ratio of technetium-99m galactosyl human serum albumin (99mTc-GSA) at 15 minutes (HH15), prealbumin, and hepatic uptake ratio of 99mTc-GSA at 15 minutes (LHL15). In addition, the model for end-stage liver disease (MELD) score at 7 days after operation was well correlated with serum HA, ICGR15, HH15, and LHL15. In patients who showed serum an HA level of = 100 ng/ml before hepatectomy, the MELD score had significantly deteriorated by 7 days after hepatectomy. Of the 20 patients who showed a serum HA level < 100 ng/ml before hepatectomy, 11 had high serum HA after hepatectomy. The bilirubin level 7 days after operation in this group was much higher than that for patients who maintained a serum HA level < 100 ng/ml after hepatectomy. In addition, the serum HGF level before hepatectomy in this group was significantly lower. We concluded that the serum HA level is a reliable indicator when evaluating liver function and predicting liver dysfunction after hepatectomy. Furthermore, patients with a significantly low HGF level who have a normal HA level are susceptible to liver dysfunction after hepatectomy.

Use of a randomized hybrid ribozyme library for identification of genes involved in muscle differentiation.

We have employed the hybrid hammerhead ribozyme-based gene discovery system for identification of genes functionally involved in muscle differentiation using in vitro myoblast differentiation assay. The major muscle regulatory genes (MyoD1, Mylk, myosin, myogenin, and Myf5) were identified endorsing the validity of this method. Other gene targets included tumor suppressors and cell cycle regulators (p19ARF and p21WAF1), FGFR-4, fibronectin, Prkg2, Pdk4, fem, and six novel proteins. Functional involvement of three of the identified targets in myoblast differentiation was confirmed by their specific knockdown using ribozymes and siRNA. Besides demonstrating a simple and an effective method of isolation of gene functions involved in muscle differentiation, we report for the first time that overexpression of Fem, a member of the sex-determining family of proteins, caused accelerated myotube formation, and its targeting deferred myoblast differentiation. This functional gene screening is not only helpful in understanding the molecular pathways of muscle differentiation but also to design molecular strategies for myopathologic therapies.