Hypoxic and nitrosative stress conditions modulate expression of myoglobin genes in a carcinogenic hepatobiliary trematode, Clonorchis sinensis

Despite recent evidence suggesting that adult trematodes require oxygen for the generation of bioenergy and eggshells, information on the molecular mechanism by which the parasites acquire oxygen remains largely elusive. In this study, the structural and expressional features of globin genes identified in Clonorchis sinensis, a carcinogenic trematode parasite that invades the hypoxic biliary tracts of mammalian hosts, were investigated to gain insight into the molecules that enable oxygen metabolism. The number of globin paralogs substantially differed among parasitic platyhelminths, ranging from one to five genes, and the C. sinensis genome encoded at least five globin genes. The expression of these Clonorchis genes, named CsMb (CsMb1—CsMb3), CsNgb, and CsGbX, according to their preferential similarity patterns toward respective globin subfamilies, exponentially increased in the worms coinciding with their sexual maturation, after being downregulated in early juveniles compared to those in metacercariae. The CsMb1 protein was detected throughout the parenchymal region of adult worms as well as in excretory-secretory products, whereas the other proteins were localized exclusively in the sexual organs and intrauterine eggs. Stimuli generated by exogenous oxygen, nitric oxide (NO), and nitrite as well as co-incubation with human cholangiocytes variously affected globin gene expression in live C. sinensis adults. Together with the specific histological distributions, these hypoxia-induced patterns may suggest that oxygen molecules transported by CsMb1 from host environments are provided to cells in the parenchyma and intrauterine eggs/sex organs of the worms for energy metabolism and/or, more importantly, eggshell formation by CsMb1 and CsMb3, respectively. Other globin homologs are likely to perform non-respiratory functions. Based on the responsive expression profile against nitrosative stress, an oxygenated form of secreted CsMb1 is suggested to play a pivotal role in parasite survival by scavenging NO generated by host immune cells via its NO dioxygenase activity.

First Report and 3D Reconstruction of a Presumptive Microscopic Liver Lipoma in a Black Barbel (Barbus balcanicus) from the River Bregalnica in the Republic of North Macedonia

A lipoma is a benign tumour of mature adipocytes which may appear in various species, including marine and freshwater fish. It usually occurs in isolated locations, such as a superficial or deep mass, mainly in the skin and seldom in other organs. In non-mammalian vertebrates, there is no agreed minimal size for the mass to be considered a lipoma. This study histologically describes a case proposed to be a microlipoma in the liver of Barbus balcanicus. The structure was an oval-shaped mass of well-differentiated adipocytes, surrounded by hepatic parenchyma. The adipocyte cluster did not contact with major vascular or biliary tracts, the liver capsule, or the hilum. The cell mass reached a maximal linear length and width of ~0.5 mm and ~0.4 mm. A three-dimensional and software-assisted reconstruction of the adipocytic mass showed that it had the shape of a flattened prolate spheroid (~0.01 mm3). Given the histological criteria currently used in the literature, we consider the mass as a lipoma, or, better, a microlipoma because it was tiny. We interpret this structure as an early growing lipoma. This work is the second description of a liver lipoma in a fish to the best of our knowledge.

Overexpression of Cancer-Associated Stem Cell Gene OLFM4 in the Colonic Epithelium of Patients With Primary Sclerosing Cholangitis

Background To examine immune-epithelial interactions and their impact on epithelial transformation in primary sclerosing cholangitis–associated ulcerative colitis (PSC-UC) using patient-derived colonic epithelial organoid cultures (EpOCs). Methods The EpOCs were originated from colonic biopsies from patients with PSC-UC (n = 12), patients with UC (n = 14), and control patients (n = 10) and stimulated with cytokines previously associated with intestinal inflammation (interferon (IFN) γ and interleukin (IL)-22). Markers of cytokine downstream pathways, stemness, and pluripotency were analyzed by real-time quantitative polymerase chain reaction and immunofluorescence. The OLFM4 expression in situ was assessed by RNAscope and immunohistochemistry. Results A distinct expression of stem cell–associated genes was observed in EpOCs derived from patients with PSC-UC, with lower expression of the classical stem-cell marker LGR5 and overexpression of OLFM4, previously associated with pluripotency and early stages of neoplastic transformation in the gastrointestinal and biliary tracts. High levels of OLFM4 were also found ex vivo in colonic biopsies from patients with PSC-UC. In addition, IFNγ stimulation resulted in the downregulation of LGR5 in EpOCs, whereas higher expression of OLFM4 was observed after IL-22 stimulation. Interestingly, expression of the IL-22 receptor, IL22RA1, was induced by IFNγ, suggesting that a complex interplay between these cytokines may contribute to carcinogenesis in PSC-UC. Conclusions Higher expression of OLFM4, a cancer stemness gene induced by IL-22, is present in PSC-UC, suggesting that IL-22 responses may result in alterations of the intestinal stem-cell niche in these patients.

Peroral Cholangioscopy-Guided Forceps Mapping Biopsy for Evaluation of the Lateral Extension of Biliary Tract Cancer

Background: Peroral cholangioscopy (POCS)-guided forceps mapping biopsy (FMB) is a method for the accurate preoperative identification of the extent of the disease of biliary tract cancer (BTC). However, the diagnostic value of POCS-FMB is still uncertain. Objectives: We evaluated the diagnostic utility of POCS-FMB for the identification of lateral extension—superficial intraductal spread longitudinally and continuously from the main lesion—of BTC. Methods: In the retrospective study, patients who received POCS-FMB and surgery for curative resection of BTC between September 2016 and August 2019 at our medical institution were enrolled. The diagnostic accuracy of POCS-FMB for the identification of lateral extension of BTC was evaluated. Furthermore, we also evaluated the factors affecting the diagnostic accuracy of POCS-FMB. Results: A total of 23 patients with BTC were enrolled, and 24 procedures of POCS-FMB from 96 sites of biliary tracts were performed. The sensitivity, specificity, and accuracy of POCS-FMB were 53.8%, 63.9%, and 63.1%, respectively. In the multivariate logistic regression analyses, the biopsy from the bifurcation of biliary tracts was a significant factor affecting the diagnostic accuracy of POCS-FMB (odds ratio 3.538, 95%; confidence interval 1.151–10.875, p = 0.027). Conclusions: The diagnostic accuracy of POCS-FMB for the identification of lateral extension of BTC was insufficient. The biopsy from the bifurcation of biliary tracts was a positive factor affecting the diagnostic accuracy of POCS-FMB.

Acute Pancreatitis as an Uncommon Complication of Hydatid Cyst of the Liver: A Case Report and Systematic Literature Review .

BackgroundHydatid disease is a major health problem worldwide. The liver is the most frequent location of hydatid disease. Acute pancreatitis secondary to liver hydatid cyst ruptured in the biliary tract is scarce and fewly described in literature. The management of this pancreatic complication of liver hydatid disease is challenging and includes a combination of surgical and endoscopic approaches. We report herein a rare case of hydatid cyst of the liver with cysto-biliary communication revealed by acute pancreatitis. A systematic literature review of similar cases reported was provided to compare surgical and endoscopic techniques. Case presentationA thirty-year-old woman was referred to our emergency unit for acute epigastric pain evolving for four days. We found fever associated with epigastric guarding on physical examination. Laboratory tests showed inflammatory biological syndrome with frankly high serum lipase level and abnormal liver tests. The diagnosis of acute pancreatitis was set. We decided then to perform an abdominal ultrasound that showed dilatation of intra and extrahepatic biliary tracts without gallbladder stones. A cystic mass of the left liver was additionally found. On CT-scan, we found a C grade acute pancreatitis. Furthermore, a hydatid cyst of the left liver ruptured in the biliary tracts and daughter vesicles within were found. We decided then to perform an emergency surgery through a bisoucostal incision. We performed a cholecystectomy and a peroperative that showed the communication between the cyst and left biliary tracts and the presence of daughter vesicle within the common bile duct. We conducted an exploration of the common bile duct with extraction of vesicle daughters. We left behind a T-tube in the common bile duct and we sutured the cysto-biliary fistula. Drainage was left in the remnant cavity after unroofing the cyst. Postoperative course was uneventful. Six months follow-up showed no recurrence.ConclusionCysto-biliary communication of liver hydatid disease revealed by acute pancreatitis is uncommon. We chose to perform emergency open surgery. However, through a systematic literature review, we noticed that endoscopic treatment is an efficient therapeutic and diagnostic tool to delay a morbid surgery of the liver and the common bile duct.

Role of Chemokines in the Biology of Cholangiocarcinoma

Cholangiocarcinoma (CCA), a heterogeneous tumor with poor prognosis, can arise at any level in the biliary tree. It may derive from epithelial cells in the biliary tracts and peribiliary glands and possibly from progenitor cells or even hepatocytes. Several risk factors are responsible for CCA onset, however an inflammatory milieu nearby the biliary tree represents the most common condition favoring CCA development. Chemokines play a key role in driving the immunological response upon liver injury and may sustain tumor initiation and development. Chemokine receptor-dependent pathways influence the interplay among various cellular components, resulting in remodeling of the hepatic microenvironment towards a pro-inflammatory, pro-fibrogenic, pro-angiogenic and pre-neoplastic setting. Moreover, once tumor develops, chemokine signaling may influence its progression. Here we review the role of chemokines in the regulation of CCA development and progression, and the modulation of angiogenesis, metastasis and immune control. The potential role of chemokines and their receptors as possible biomarkers and/or therapeutic targets for hepatobiliary cancer is also discussed.

Professor Theodor Kocher (1841–1917) – one of the founders of european scientific surgery (on the 100th anniversary of death)

The main milestones in the life and work of T. Kocher are presented. Illuminated little-known interesting facts from the life of a professor. T. Kocher studied the problems of asepsis and antisepsis, abdominal surgery, thyroid surgery, traumatology. He deepened his teaching of the antisepsis and asepsis, introduced the concept of Implant Infection. It is noted that Kocher has developed sterilization methods suture material. He suggested operative access to the liver and biliary tracts, an original method of gastrectomy, methods for mobilizing the duodenum, surgical treatment of inguinal hernias, which are now described in textbooks. He also developed gentle surgical approaches for the procedure on large joints (hook-shaped incisions). The reduction of shoulder dislocations according to Kocher became a part of the daily practice of surgeons and traumatologists. T. Kocher's research on the problems of diseases of the thyroid gland is world famous. T. Kocher was awarded the 1909 Nobel Prize for his work on the physiology, pathology and surgery of the thyroid gland. He proposed some surgical instruments (hemostatic clamp, grooved probe, gastric clamp, etc.), a gastric tube was put into practice. The published manual on operative surgery has survived 5 reprints (1892–1907) and has been translated into many world's languages.

Gallstone Disease and Microbiome

Gallstone disease (GSD) has, for many years, remained a high-cost, socially significant public health problem. Over the past decade, a number of studies have been carried out—both in humans and in animal models—confirming the role of the microbiota in various sections of the gastrointestinal tract as a new link in the etiopathogenesis of GSD. The microbiome of bile correlates with the bacterial composition of saliva, and the microbiome of the biliary tract has a high similarity with the microbiota of the duodenum. Pathogenic microflora of the oral cavity, through mechanisms of immunomodulation, can affect the motility of the gallbladder and the expression of mucin genes (MUC1, Muc3, MUC4), and represent one of the promoters of stone formation in the gallbladder. The presence of H. pylori infection contributes to the formation of gallstones and affects the occurrence of complications of GSD, including acute and chronic cholecystitis, cholangitis, pancreatitis. Intestinal bacteria (Clostridium, Bifidobacterium, Peptostreptococcus, Bacteroides, Eubacterium, and Escherichia coli) participating in the oxidation and epimerization of bile acids can disrupt enterohepatic circulation and lead to the formation of gallstones. At the same time, cholecystectomy due to GSD leads to the further transformation of the composition of the microbiota in various parts of the gastrointestinal tract, increasing the risk of developing stomach cancer and colorectal cancer. Further research is required to determine the possibility of using the evaluation of the composition of the microbiota of the gastrointestinal and biliary tracts as an early diagnostic marker of various gastroenterological diseases.