Portal Region Research Articles

Salvage Ablative Radiation Therapy for Isolated Intraabdominal Recurrences of Hepatocellular Carcinoma

<h3>Purpose/Objective(s)</h3> The standard of care for extrahepatic hepatocellular carcinoma (HCC) is a systemic therapy. However, some patients present with limited intra-abdominal disease that might benefit from local ablative radiation therapy (RT) with, or instead of, systemic therapy. This study explores outcomes of ablative RT for isolated intra-abdominal recurrences from HCC. <h3>Materials/Methods</h3> A single-institution retrospective chart review was conducted from 2013 to 2021. Inclusion criteria were any patient with HCC that had limited disease in an abdominal lymph node (LN) or adrenal gland (AG) treated with salvage ablative RT. The primary outcome was local control (LC) in the target lesion. Secondary outcomes were progression-free survival (PFS) and overall survival (OS) from the time of recurrence after completing RT. <h3>Results</h3> 31 patients met inclusion criteria, of which 21 (67.7%) had a recurrence in a LN while 10 (32.2%) had recurrences in an AG. 19 (90%) of LN recurrences were in the portal region. The median time from initial HCC diagnosis to extrahepatic recurrence was 18 months (range 0 to 152 months). Among all patients, 21 patients received immunotherapy (prior, concurrent, or after) in addition to salvage RT. 27 (87.1%) patients received salvage SBRT, and 4 (12.9%) patients received fractionated RT. The median dose was 4500 in 5 fractions. LC, as defined as no progression in target lesion, was 100%. The median PFS, defined by recurrence or death, was 18 months. The median OS was 29 months. There was a statistical difference in OS in patients that received IO and RT vs. RT alone (p=0.038) in favor of RT alone. There was no significant difference in OS for a LN recurrence versus salvage RT for an AG recurrence (p=0.389). <h3>Conclusion</h3> Salvage ablative RT, with or without immunotherapy, can produce meaningful PFS and OS in well-selected patients with limited intra-abdominal extrahepatic recurrence of HCC. This benefit appears the same for LN recurrences and AG metastases and might be independent of immunotherapy. Prospective data are needed to verify these results.

Single-cell transcriptomics shows dose-dependent disruption of hepatic zonation by TCDD in mice.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) dose-dependently induces the development of hepatic fat accumulation and inflammation with fibrosis in mice initially in the portal region. Conversely, differential gene and protein expression is first detected in the central region. To further investigate cell-specific and spatially resolved dose-dependent changes in gene expression elicited by TCDD, single-nuclei RNA sequencing and spatial transcriptomics were used for livers of male mice gavaged with TCDD every 4 days for 28 days. The proportion of 11 cell (sub)types across 131 613 nuclei dose-dependently changed with 68% of all portal and central hepatocyte nuclei in control mice being overtaken by macrophages following TCDD treatment. We identified 368 (portal fibroblasts) to 1339 (macrophages) differentially expressed genes. Spatial analyses revealed initial loss of portal identity that eventually spanned the entire liver lobule with increasing dose. Induction of R-spondin 3 (Rspo3) and pericentral Apc, suggested dysregulation of the Wnt/β-catenin signaling cascade in zonally resolved steatosis. Collectively, the integrated results suggest disruption of zonation contributes to the pattern of TCDD-elicited NAFLD pathologies.

Multiple Timescale Dynamic Analysis of Functionally-Impairing Mutations in Human Ileal Bile Acid-Binding Protein.

Human ileal bile acid-binding protein (hI-BABP) has a key role in the enterohepatic circulation of bile salts. Its two internal binding sites exhibit positive cooperativity accompanied by a site-selectivity of glycocholate (GCA) and glycochenodeoxycholate (GCDA), the two most abundant bile salts in humans. To improve our understanding of the role of dynamics in ligand binding, we introduced functionally impairing single-residue mutations at two key regions of the protein and subjected the mutants to NMR relaxation analysis and MD simulations. According to our results, mutation in both the vicinity of the C/D (Q51A) and the G/H (Q99A) turns results in a redistribution of motional freedom in apo hI-BABP. Mutation Q51A, deteriorating the site-selectivity of GCA and GCDA, results in the channeling of ms fluctuations into faster motions in the binding pocket hampering the realization of key side chain interactions. Mutation Q99A, abolishing positive binding cooperativity for GCDA, leaves ms motions in the C-terminal half unchanged but by decoupling βD from a dynamic cluster of the N-terminal half displays an increased flexibility in the vicinity of site 1. MD simulations of the variants indicate structural differences in the portal region and mutation-induced changes in dynamics, which depend on the protonation state of histidines. A dynamic coupling between the EFGH portal, the C/D-region, and the helical cap is evidenced highlighting the interplay of structural and dynamic effects in bile salt recognition in hI-BABP.

Artificial intelligence based liver portal tract region identification and quantification with transplant biopsy whole-slide images

Liver fibrosis staging is clinically important for liver disease progression prediction. As the portal tract fibrotic quantity and size in a liver biopsy correlate with the fibrosis stage, an accurate analysis of portal tract regions is clinically critical. Manual annotations of portal tract regions, however, are time-consuming and subject to large inter- and intra-observer variability. To address such a challenge, we develop a Multiple Up-sampling and Spatial Attention guided UNet model (MUSA-UNet) to segment liver portal tract regions in whole-slide images of liver tissue slides. To enhance the segmentation performance, we propose to use depth-wise separable convolution, the spatial attention mechanism, the residual connection, and multiple up-sampling paths in the developed model. This study includes 53 histopathology whole slide images from patients who received liver transplantation. In total, 6,012 patches derived from 30 images are used for our deep learning model training and validation. The remaining 23 whole slide images are utilized for the model testing. The average liver portal tract segmentation performance of the developed MUSA-UNet is 0.94 (Precision), 0.85 (Recall), 0.89 (F1 Score), 0.89 (Accuracy), 0.80 (Jaccard Index), and 0.91 (Fowlkes–Mallows Index), respectively. The clinical Scheuer fibrosis stage presents a strong correlation with the resulting average portal tract fibrotic area (R = 0.681, p<0.001) and portal tract percentage (R = 0.335, p = 0.02) computed from the MUSA-UNet segmentation results. In conclusion, our developed deep learning model MUSA-UNet can accurately segment portal tract regions from whole-slide images of liver tissue biopsies, presenting its promising potential to assist liver disease diagnosis in a computational manner.

333.2: Vascular Complications in Pediatric Liver Transplants And Their Management

Introduction: Unlike other organs, the blood supply of the liver occurs through two systems. One of these is the hepatic artery, and it supply approximately one-third to one-fifth of the liver. The rest of the liver is supplied with blood via the portal vein. The outflow of blood circulation in the liver is via the hepatic veins. Any disruption in this blood circulation results in deterioration in liver functions. In this study, we aimed to evaluate early vascular complications in pediatric liver transplants. Methods: From 8 November 1988 to 31 December 2021, we performed 701 LT procedures and 334 of them were pediatric.). We reviewed the medical records of these recipients for the following: primary cause of liver failure, age, and weight at the time of transplantation, type of graft, vascular complications and their management. One hundred and seventy six of the recipients were male and 158 were female. Mean age of 7.34 years (0.5 months – 17 years). Nineteen (5.7%) of the LT were deceased donor LT and 315 (94.3%) were living related liver transplant. Most cause of liver failure was biliary atresia (n=169). Mean weight of recipients was 23.3 kg. Most of graft types was left lateral graft (n=204) Results: Hepatic vein complications occurred in 3 patients. In all three patients, stenosis was detected in the portal vein anastomosis region and was successfully treated with interventional radiological methods by placing a stent in the anastomosis region. Portal vein complications occurred in 3 patients. In one of these patients, hemostasis was performed by surgical method due to bleeding from the portal vein anastomosis. In the second patient, the anastomosis was surgically revised due to thrombus formation in the portal vein. In the third patient, due to a stenosis of more than 50% in the portal vein anastomosis, a stent was placed in the anastomosis region after balloon dilation using interventional radiological methods, and blood flow was successfully maintained. Hepatic artery complications occurred in 54 patients. Hepatic artery thrombosis occurred in 31 patients, hepatic artery stenosis in 13 patients, bleeding from hepatic artery anastomosis in 7 patients, hepatic artery dissection in 2 patients, and pseudoaneurysm in the hepatic artery in 1 patient. 43 of these patients were successfully treated with interventional radiological methods and 11 of them surgically. Conclusion: Vascular complications seen in liver transplants can cause deterioration in hepatic functions and acute liver failure. Especially hepatic artery complications are one of the most important causes of biliary tract complications that will develop in the future. Vascular complications can be successfully treated at an early stage in experienced organ transplant centers.

MORPHOLOGICAL CHARACTERISTICS OF THE LIVER IN DIFFERENT PHASES OF THE INVASION OPISTHORCHIS FELINEUS IN RABBITS ORYCTOLAGUS CUNICULUS

Morphological changes in the liver, as the main organ in which Opisthorchis felineus is localized, according to the literature, are characterized by significant structural differences in the acute and chronic phases of the disease. Comparative characteristics of the morphological changes in the liver in different phases of opisthorchiasis in the experiment in rabbits, such as Oryctolagus cuniculus, was not carried out. To simulate the acute and chronic phases of invasion, the animals were divided into two observation groups. The histological material was processed by standard methods, liver tissue sections were stained with hematoxylin and eosin and according to Van Gieson to identify connective tissue structures. Histological preparations were studied by the methods of light-optical microscopy with immersion, video microscopy with morphometry. As a result of the study, it was found that in the acute phase of opisthorchiasis, the general infiltration of the portal tract area prevails compared to the infiltration of the bile duct wall, in the chronic phase, the thickness of the infiltrate of the bile duct wall approaches the total values. The cellular composition of infiltrates is characterized by the presence of lymphocytes, fibroblasts, epithelioid cells and cells of foreign bodies in both phases, if macrophages predominate in the acute phase of invasion, then fibrocytes and mast cells predominate in the chronic phase. Hepatocytes in the acute phase have disturbances in the tinctorial properties of the karyolemma and cytoplasm, mainly due to grain segregation. In the chronic phase of opisthorchiasis, polyploid hepatocytes are found. The connective tissue of the portal tract region in both phases quantitatively predominates in comparison with the connective tissue of the region of the boundary plates. The maximum amount of connective tissue in the area of ​​the portal tract in the chronic phase increases by 4.4 times compared with the acute phase of the parasitic disease. Thus, morphological changes in the liver of rabbits in different phases are characterized by both commonality and significant differences, consisting in different cellular composition of the invasion zone, the amount of connective tissue formed in different areas of the organ, and changes in hepatocytes.

Characteristics of SOX9-positive progenitor-like cells during cholestatic liver regeneration in biliary atresia

BackgroundThe progression of Biliary Atresia (BA) is associated with the number of reactive ductular cells (RDCs) whose heterogeneity in origin and evolution in humans remains unknown. SOX9-positive liver progenitor-like cells (LPLCs) have been shown to participate in RDCs and new hepatocyte formation during cholestatic liver regeneration in an animal model, which implies the possibility that hepatocyte-reprogrammed LPLCs could be a source of RDCs in BA. The present study aimed to elucidate the characteristics of SOX9-positive LPLCs in BA for exploring new possible therapeutic targets by manipulating the bi-differentiation process of LPLCs to prevent disease progression.MethodsTwenty-eight patients, including 24 patients with BA and 4 patients with Congenital Choledochal Cyst as the control group, were retrospectively recruited. Liver biopsy samples were classified histologically using a 4-point scale based on fibrosis severity. LPLCs were detected by SOX9 and HNF4A double positive staining. Single immunohistochemistry, double immunohistochemistry, and multiple immunofluorescence staining were used to determine the different cell types and characteristics of LPLCs.ResultsThe prognostic predictors of BA, namely total bile acid (TBA), RDCs, and fibrosis, were correlated to the emergence of LPLCs. SOX9 and HNF4A double-positive LPLCs co-stained rarely with relevant markers of portal hepatic progenitor cells (portal-HPCs), including CK19, CK7, EPCAM, PROM1 (CD133), TROP2, and AFP. Under cholestasis conditions, LPLCs acquired superior proliferation and anti-senescence ability among hepatocytes. Moreover, LPLCs arranged as a pseudo-rosette structure appeared from the periportal parenchyma to the portal region, which implied the differentiation from hepatocyte-reprogrammed LPLCs to RDCs with the progression of cholestasis.ConclusionsLPLCs are associated with disease progression and prognostic factors of BA. The bipotent characteristics of LPLCs are different from those of portal-HPCs. As cholestasis progresses, LPLCs appear to gain superior proliferation and anti-senescence ability and continually differentiate to RDCs.

High-fidelity biosensing of dNTPs and nucleic acids by controllable subnanometer channel PaMscS

Current tools for dNTP analysis mainly rely on expensive fluorescent labeling, mass spectrometry or electrochemistry. Single-molecule assay by protein nanopores with an internal diameter of ca. 1–3.6 nm provides a useful tool for dNTP sensing. However, the most commonly used protein nanopores require additional modifications to enable dNTP detection. In this study, the PaMscS channel (mechanosensitive channel of small conductance from Pseudomonas aeruginosa) embedded in the bilayer lipid membrane (BLM) of E. coli polar lipid extract was applied as a nanopore for single molecular sensing. Two mutants of PaMscS nanopores on the side portal region (PaMscS W130A and PaMscS K180R) were selected for direct dNTP or pyrophosphoric acid (PPi) detection without aptamer or protein modification. Notably, the PaMscS mutant pore can be adjusted by regulation of osmolarity differences, which is crucial for the optimal detection of specific molecules. In addition, we established a PaMscS-based diagnosis method for the rapid sensing of disease-associated nucleic acids by monitoring the consumption of dNTPs, with 86% specificity and 100% sensitivity among 22 clinical samples. This protein nanopore, without aptamer or modification, paves a new way for dNTPs, PPi direct sensing and nucleic acid detection with low cost but high versatility.

Clinical and oncological benefits of left hepatectomy for Bismuth type I/II perihilar cholangiocarcinoma.

This retrospective study aimed to clarify whether the postoperative prognosis differs between right and left hepatectomy for Bismuth type I/II perihilar cholangiocarcinoma. Preoperative images of 195 patients with perihilar cholangiocarcinoma were reexamined. Patients with Bismuth type I/II perihilar cholangiocarcinoma without a difference in extraductal tumor invasion between the right and left sides of the hepatic portal region were classified into those undergoing left (L group) or right (R group) hepatectomy. Twenty-three patients (11.8%) were classified into the L group and 33 (16.9%) into the R group. All eight patients with pTis/1 belonged to the L group. The L group had significantly less liver failure than the R group (p = 0.001). One patient (4.3%) in the L group and four patients (12.1%) in the R group died from postoperative complications. Among 48 patients with pT2, the L group tended to have better overall survival (median, 12.2 vs. 5.6years; p = 0.072), but not recurrence-free survival (median, 9.1 vs. 3.6years; p = 0.477), in comparison to the R group. Postoperative survival after left hepatectomy for Bismuth type I/II perihilar cholangiocarcinoma is expected to be as long as that after right hepatectomy.

S2761 Recurrent Oriental Cholangiohepatitis Despite Partial Hepatic Resection

Introduction: Recurrent pyogenic cholangiohepatitis (RPC), also known as “Oriental Cholangiohepatitis”, is commonly found in people of Southeast Asian origin. RPC is characterized by recurrent cholangitis secondary to bile stasis and stone formation proximal to biliary strictures. We present a case of recurrent pyogenic cholangiohepatitis with hepatic abscess in a Vietnamese male with history of partial hepatic resection. Case Description/Methods: A 42 year old Vietnamese male with a past medical history of recurrent pyogenic cholangitis and hepatolithiasis status post hepaticojejunostomy in 2009, followed by abscess resection with culture positive Klebsiella, segment 3 resection, and biliary stenting in 2013 presented with two days of RUQ pain, fevers and jaundice. On arrival, the patient was afebrile, tachycardic and mildly hypotensive. Laboratory evaluation demonstrated alkaline phosphatase 360 IU/L, total bilirubin 14.4 mg/dL, ALT 93 IU/L, AST 113 IU/L, WBC 24,000, INR 1.3. Findings on CT scan of the abdomen included 10 cm multiloculated liver abscess in segment 4A, dilated biliary tract with small pneumobilia and dense material, and a 3.9 x 2.8 cm mass-like hyperdense region in the portal triad region visualized in arterial phase. The patient was started on broad spectrum antibiotics and underwent percutaneous abscess drainage with 350 mL of purulent material removed. MRCP the following day redemonstrated a liver abscess measuring 9.5 cm, RUQ drain in place, diffuse dilatation of the intrahepatic and extrahepatic bile ducts with multiple filling defects consistent with calculi, and a 14mm common bile duct. Cultures from the abscess grew ESBL E.coli and Provotella, and the patient was transitioned to Ertapenem with plan to continue antibiotics for four weeks. The patient was discharged home with abscess drain in place. Discussion: RPC is a recurrent syndrome of cholangitis triggered by a parasitic infection which causes biliary fibrosis leading to strictures and stone formation within intrahepatic ducts, most often on the left. Treatment includes antibiotics, stricture dilation, biliary drainage, stone removal, and liver resection. Although liver resection of the segment decreases the risk of recurrent cholangitis, abscess, and eliminates the risk of carcinoma in that segment, resection does not resolve the biliary fibrosis throughout the remaining segments leaving the patient at risk for recurrence. Liver transplant can be considered for complicated cases that do not respond to conventional therapies.Figure 1.: 10 cm multiloculated liver abscess in segment 4A.

Downregulation of Programmed Death-1 Pathway Promoting CD8 + T Cell Cytotoxicity in Primary Biliary Cholangitis

Primary biliary cholangitis (PBC) is an autoimmune disease. CD8 + T cell (CTLs) cytotoxicity played a crucial rule in of PBC with unclear detailed pathogenesis. The role of the programmed death-1 (PD-1) pathway in CD8 + T cell cytotoxicity in patients with PBC was determined. We recruited 69 patients with PBC and 57 healthy controls (HCs). PD-1 pathway in peripheral CD8 + T cells and related cytokines were detected, and gene expression levels were detected. Immunofluorescence staining of PD-1/PD-L1 was performed on liver tissue. PD-1 ± CTLs were cocultured with human intrahepatic biliary epithelial cells (HiBECs) to measure CTL cytotoxicity, proliferation and cytokine levels and HiBEC apoptosis. The upstream signaling pathway of PD-1 was detected. PBC patients exhibited Tbet gene upregulation and PD-1 downregulation in CTLs, with PD-1 expression reduced in CTLs and PD-L1 reduced in the liver portal region relative to HCs. Higher plasma IL-10, interferon-γ and transforming growth factor-β concentrations were observed in the PBC group than the HC group. In CTL and HiBEC coculture experiment, compared with PD-1- CTLs, PD-1 + CTLs exhibited weaker cytotoxicity, less proliferation and lower cytokine production. When the system was blocked by anti-PD-1 antibodies, these effects were antagonized. PD-1 expression in CD8 + T cells decreased, and PD-1 pathway-related cytokines changed in patients with PBC. PD-1/PD-L1 pathway silencing increased CD8 + T cell proliferation, related cytokine production and CTL cytotoxic effects on HiBECs in coculture experiment. The PD-1/PD-L1 pathway might represent an important pathway in the immunological mechanism underlying PBC.

Repercussions of the COVID-19 Response in Pregnant Women in Western Uganda: Knowledge, Behavior, and Emotional State after the First Lockdown in 2020.

Limited research exists on pregnant women’s knowledge, attitudes, and behavior concerning COVID-19 in sub-Saharan Africa. We performed a cross-sectional study among 648 pregnant women in Fort Portal, Uganda, after the first lockdown starting in June 2020. Structured interviews were conducted at three different facilities during routine antenatal care, assessing sociodemographic background, knowledge of COVID-19, prevention behavior adherence, and psycho-emotional stress levels. We performed descriptive analyses and examined associated factors using multivariable logistic regression. In Fort Portal Region, 32.8% of pregnant women had a higher knowledge regarding the COVID-19 pandemic, while all women at least heard of COVID-19. 88.6% of the women showed low self-reported prevention behavior adherence. More than one third of the pregnant women experienced high psycho-emotional stress related to the pandemic (39.8%). The odds for psycho-emotional stress were increased among the age group 21–30 years (AOR 1.97; 95% CI 1.18–3.35) compared to women under the age of 21, and decreased in single or divorced women compared to women in partnerships (AOR 0.42; 0.22–0.77) and in women having less COVID-19-related knowledge (AOR 0.40; 0.27–0.58). In conclusion, prevention behavior adherence seemed challenging, and psycho-emotional stress was ubiquitous among our cohort. To avoid adverse consequences in maternal and neonatal health, campaigns for hygiene but also women’s emotional state should be a major focus of community healthcare in exceptional times such as the SARS-CoV-2 pandemic.

Effect of high fat diet on structure of liver and gallbladder of adult male mice – an experimental study

Introduction. High fat diet (HFD) intake induces obesity and adversely affects different body organs including liver and gallbladder. Aim. It was to clarify the effects of HFD on the liver and gallbladder structure using light microscopic (LM) examination. Material and methods. 16 healthy adult male mice were equally divided into 2 groups. Control group mice were fed normal diet. HFD group was fed using HFD. At the end of the 8-week experiment, mice were anesthetized. Liver and gallbladder were removed and prepared to histological processing. Sections were stained with hematoxylin and eosin (H&amp;E) and immunostaining for cyclooxygenase-2 (COX-2) cellular localization. Oil Red O (ORO)-stained frozen liver sections were prepared. Results. H&amp;E-stained sections of HFD group revealed rounded swollen hepatic cells with pale cytoplasm suggesting cellular ballooning. Dilated congested sinusoids and portal vein, cellular degeneration and collection of inflammatory cells were observed between hepatic cells and in portal region. Gallbladder sections showed epithelial stratification and cellular vacuolation. Strong immunoexpression of COX-2 was observed in Kupffer and hepatic cells of the liver and gallbladder mucosal epithelial cells. Conclusion. HFD is suggested to alter the normal histological features of liver and gallbladder represented by fatty liver and gallbladder epithelial hyperplasia and inflammatory reaction.

Immunoglobulin G4-associated autoimmune hepatitis with peripheral blood eosinophilia: a case report

BackgroundImmunoglobulin G4 (IgG4) associated autoimmune hepatitis (AIH) has been recognized as a type of autoimmune disease that responds to corticosteroid. The diagnosis is based on elevation of the serum IgG4 level, abundance of IgG4 enhanced plasma cell infiltration in the portal region of the liver, and satisfaction of the criteria for “definite AIH” under the revised International Autoimmune Hepatitis Group (IAIHG) scoring system. However, the clinical course of the disease is unclear.Case presentationA 65-year-old man with jaundice and peripheral blood eosinophilia.His IAIHG and simplified score was compatible with definite AIH and his IgG4 level was elevated. Magnetic resonance imaging did not reveal abnormalities in the hepatobiliary system or pancreas. A liver biopsy revealed interface hepatitis with IgG4 positive plasma cell infiltration in the portal region, without evidence of bile duct injury. He responded to 4-week period of induction prednisolone therapy and had no recurring symptoms under maintenance therapy of 5 mg prednisolone during the 3-year follow up.ConclusionsThis was a rare case that demonstrated an association between IgG4 associated AIH and the presence of peripheral blood eosinophilia.

Histomorphological investigation of intrahepatic connective tissue for surgical anatomy based on modern computer imaging analysis.

Background/PurposeComputer‐assisted tissue imaging and analytical techniques were used to clarify the histomorphological structure of hepatic connective tissue as a practical guide for surgeons.MethodsApproximately 5000 histological slides were prepared from liver specimens of five autopsied patients. Three‐dimensional (3D) reconstruction was performed and subjected to computer imaging analysis. Scanning electron microscopy was also performed on the liver specimens.ResultsThe 3D reconstructed images revealed the running form of the vasculature and the relationship between the hepatic lobule and connective tissue. The hepatic capsule or portal pedicle was consistently located at the periphery of the hepatic lobules. An artificial intelligence random forest approach clearly segmented hepatic cells, type I collagen (CF), type III collagen (RF), and other cells. The hepatic lobule, portal region, and hepatic capsule were significantly distinguished based on CF and RF occupancy. The capsule directly covering the liver lobule with an RF concentration up to 87% was provisionally named the proper hepatic capsule. The existence of a proper hepatic ligament with distinct occupation rates of CF and RF was also suggested.ConclusionsThe identified proper hepatic capsule and ligament can be important markers for demarcating the dissecting layer during surgical procedures.

Cryo-EM, X-ray diffraction, and atomistic simulations reveal determinants for the formation of a supramolecular myelin-like proteolipid lattice

Myelin protein P2 is a peripheral membrane protein of the fatty acid-binding protein family that functions in the formation and maintenance of the peripheral nerve myelin sheath. Several P2 gene mutations cause human Charcot-Marie-Tooth neuropathy, but the mature myelin sheath assembly mechanism is unclear. Here, cryo-EM of myelin-like proteolipid multilayers revealed an ordered three-dimensional (3D) lattice of P2 molecules between stacked lipid bilayers, visualizing supramolecular assembly at the myelin major dense line. The data disclosed that a single P2 layer is inserted between two bilayers in a tight intermembrane space of ∼3 nm, implying direct interactions between P2 and two membrane surfaces. X-ray diffraction from P2-stacked bicelle multilayers revealed lateral protein organization, and surface mutagenesis of P2 coupled with structure-function experiments revealed a role for both the portal region of P2 and its opposite face in membrane interactions. Atomistic molecular dynamics simulations of P2 on model membrane surfaces suggested that Arg-88 is critical for P2-membrane interactions, in addition to the helical lid domain. Negatively charged lipid headgroups stably anchored P2 on the myelin-like bilayer surface. Membrane binding may be accompanied by opening of the P2 β-barrel structure and ligand exchange with the apposing bilayer. Our results provide an unprecedented view into an ordered, multilayered biomolecular membrane system induced by the presence of a peripheral membrane protein from human myelin. This is an important step toward deciphering the 3D assembly of a mature myelin sheath at the molecular level.

Effect of Carcinogenic Substance (7,12 Dimethylbenz [a] anthracene (DMBA)) on Tissue, Hematology Character and Enzyme Activity in Rat

This study aimed to investigate the effects of 7,12 dimethylbenz [a] anthracene (DMBA) on some blood parameters and hepatic histopathology in rats and antioxidant enzyme. twenty female Wistar albino rats, weighing 180–200 g, were randomly divided into two group. DMBA group (positive group) who received 20 mg DMBA/kg body weight/ (single Douse). Control group (negative group) don’t received any thing. The animals in the groups were sacrificed at the end of the 90 days: The histological structure of the liver tissues in the control group was normal. the liver exhibited hydropic degeneration and coagulation necrosis in hepatocytes, severe dilation in the sinusoids, congestion in the central and portal regions , DMBA groups, was degenerative and necrotic changes were detected. MDA levels increased in the DMBA group , compared to the control group, Antioxidant activity CAT, SOD and GPX, the results showed increased enzyme activity in positive control compared with negative groups , ,the hematological parameter was increase in WBC, PLT, GRA, and LYM and decrease in RBC value .Keywords: 7,12-dimethylbenz[a]anthracene (DMBA); Blood parameters; Antioxidant enzyme ; MDA ; Rat

Influence of Carcinogenic Substance (7, 12 Dimethylbenz [a] Anthracene (DMBA) on Tissue, Hematology Character and Enzyme Activity in Rat

This study designed to investigate the effects of 7,12 dimethylbenz [a] anthracene (DMBA)on some blood parameters and hepatic histopathology in rats and antioxidant enzyme. twenty female Wistar albino rats, weighing 180–200 g, were randomly divided into two group 1- DMBA group (positive group) who received 20 mg DMBA/kg body weight/ (single Douse) 2-Control group (negative group) doesn’t receive any item. The animals in these groups were sacrificed at the end of the 90 days:The histological structure of the liver tissues in the control group was normal. the liver exhibited hydropic degeneration and coagulation necrosis in hepatocytes, severe dilation in the sinusoids, congestion in the central and portal regions , DMBA groups, was degenerative and necrotic changes were detected.MDA levels increased in the DMBAgroup , compared to the control group, Antioxidant activity CAT, SOD and GPX, the results showed increased enzyme activity in positive control compared with negative groups , ,the hematological parameter was increase in WBC, PLT, GRA, and LYM and decrease in RBC value .

Effect of Carcinogenic Substance (7,12 Dimethylbenz [a] Anthracene (DMBA)) on Tissue, Hematology Character and Enzyme Activity in Rat

This study aimed to investigate the effects of 7,12 dimethylbenz [a] anthracene (DMBA) on some blood parameters and hepatic histopathology in rats and antioxidant enzyme. twenty female Wistar albino rats, weighing 180–200 g, were randomly divided into two group. DMBA group (positive group) who received 20 mg DMBA/kg body weight/ (single Douse). Control group (negative group) don’t received any thing. The animals in the groups were sacrificed at the end of the 90 days: The histological structure of the liver tissues in the control group was normal. the liver exhibited hydropic degeneration and coagulation necrosis in hepatocytes, severe dilation in the sinusoids, congestion in the central and portal regions , DMBA groups, was degenerative and necrotic changes were detected. MDA levels increased in the DMBA group , compared to the control group, Antioxidant activity CAT, SOD and GPX, the results showed increased enzyme activity in positive control compared with negative groups, ,the hematological parameter was increase in WBC, PLT, GRA, and LYM and decrease in RBC value .

Using 3D Numerical Analysis for the Assessment of Tunnel–Landslide Relationship: Bahce–Nurdag Tunnel (South of Turkey)

Owing to the increase in population, transportation has become one of the major research topics. Among existing transportation systems, railway is one of the most economic and fast systems, and therefore, there has been an increase in new railway constructions in recent years. In Turkey, the longest railway tunnels, the Bahce–Nurdag high-speed rail tunnels, are under construction in the southern part of the country. This twin tubes tunnels are approximately 10 km in length, and they are excavated using tunnel boring machines (TBM). The tunnel route locates in the East Anatolian Fault Zone and the portal region is composed of debris and old landslide deposits. For this reason, mitigating the potential landslide hazard is critical for the tunnel. Therefore, the present study investigates the TBM tunnelling–landslide relationship by employing 3D numerical analyses. The analyses show that the last 600 m of the tunnel has a potential landslide hazard and some additional measures should be considered along this part to ensure tunnel safety. The tunnel conditions are highly complex, and thus it is an interesting case for tunnel engineers.