Duct Lumen Research Articles

ThP5.13 - Lessons learnt from a neuroendocrine tumour presenting as choledocholithiasis

Abstract A 79 year old male presented to A&E with obstructive jaundice. A computerised tomography (CT) scan of his abdomen and pelvis showed intra- and extra-hepatic duct dilation, a small calculus within the lumen of the common bile duct (CBD), and gallbladder distension. His liver function tests were deranged, with an alkaline phosphatase of 280IU/L. Successful stone retrieval was achieved by endoscopic retrograde cholangiopancreatography (ERCP), with an interim CBD stent inserted. Within the same admission, the patient underwent an uncomplicated emergency cholecystectomy. Histological analysis highlighted moderate chronic inflammation and fibromuscular hyperplasia, cholesterolosis, and ruled out dysplasia or malignancy. Four weeks later, the patient re-presented with septic obstructive jaundice. A repeat CT scan identified a dilated CBD of 2.2cm, suggesting an occluded stent. On ERCP for stent removal, ampulla narrowing was identified, hence concurrent brush cytology was performed. Due to clinical suspicion, a CT Positron emission tomography (PET) was requested, which identified metabolically active disseminated malignancy, likely pancreatic in origin, and with nodal, liver, and likely peritoneal metastases. A Ca 19-9 subsequently came back raised at 157 kIU/L. Liver biopsy confirmed a poorly differentiated neuroendocrine carcinoma (NEC) with intermediate features between small and large cell NEC. In such cases with discrepancies between treatment success and lack of biochemical improvement, a low threshold of suspicion for further investigation and oncological consideration is imperative.

The spermatheca ultrastructure of the ground beetle Clinidium canaliculatum (Costa) (Carabidae, Rhysodinae)

The ground beetle Clinidium canaliculatum is a member of Rhysodinae, a taxon with still discussed systematic position. The spermatheca of this species is a small cylindrical structure connected to the common oviduct by a thin duct. The ultrastructure of the organ has revealed that the apical receptacle is provided with an epithelium lined by a thick cuticle from the deeper region of which several finger-like cuticular structures extend into the cytoplasm. On these structures adhere microtubule bundles that cross the whole cytoplasm to anchor on short densities along the basal plasma membrane. These specializations are strongly reminiscent of the hemidesmosomes, possibly playing a mechanical role enabling the cells to resist to the muscle contractions pushing the sperm towards the spermathecal duct. The cells are rich in mitochondria and glycogen granules and they are possibly involved in fluid uptake from the spermathecal lumen. The spermathecal duct has a simple epithelium lined by a soft cuticle. The sperm present in the apical receptacle and in the duct lumen maintain the structure described in the male genital apparatuses. They are generally free and embedded in a homogeneous electron-dense material. Occasionally, a sperm bundle, still with an apical cap, was visible in the spermathecal receptacle.

Proteomics analysis of carcinogenesis in a rat model of mammary cancer induced by DMBA (7,12-dimethylbenz[a]anthracene)

Background: Mammary cancer, called breast cancer in humans, results from the abnormal growth of cells in the mammary glands that attack the surrounding tissue. The process of carcinogenesis at the molecular level can be monitored through the production of proteins as biomarkers for carcinogenesis. 7,12-Dimethylbenz[a]anthracene (DMBA) is a known carcinogenic compound. This study aimed to analyze the proteomic profile as critical data regarding DMBA-induced carcinogenesis in Sprague‒Dawley rats. Methods: Experimental animals were divided into two groups: a treatment group given DMBA at a dose of 10 mg/kg (intramammary) at intervals of 48 hours for a total of 10 doses, and a negative control group that was not given any treatment. Measurement of the total protein concentration was carried out using a spectrophotometer, and the data were analyzed using a t-test, while the characterization of protein profiles was carried out based on molecular weight data using SDS‒PAGE. Mammary gland histopathology was evaluated by hematoxylin and eosin (H&E) staining. Results: The results showed a significant (p<0.05) increase of 27% in the total protein concentration in the rat mammary cancer model. The results of proteomic characterization showed a protein profile containing proteins of 187, 169, 68, 64, 53, 41, 24, 18, and 14 kDa, which were suspected to be HER-2, Nischarin, COX-2, Albumine, Vimentin, ACTB, TNF, p16, and fatty acid binding protein 3 (FABP3), respectively. Histopathology of the mammary glands showed an irregular and indistinct arrangement of the alveoli and extensive epithelial cell proliferation from the surface to the lumen of the mammary ducts, and the mammary stroma showed the formation of new epithelial cells, which were cancer cells that spread to surrounding tissue. Conclusions: The proteomic profile was strongly associated with morphological alterations in mammary carcinogenesis in a rat model of DMBA-induced mammary cancer.

New insight in secretory structures and secretion composition in Rhus typhina L. – Anatomical, histochemical, and ultrastructural studies

Rhus typhina is a valuable plant used in the pharmaceutical, cosmetic, and food industries due to the presence of biologically active substances accumulated in its organs, especially in secretory structures, i.e. trichomes and secretory ducts. Light microscopy, scanning electron microscopy, and transmission electron microscopy were used to examine the structure of glandular and non-glandular trichomes, as well as secretory ducts present in inflorescence peduncles of R. typhina. The chemical composition of the secretion produced by trichomes and ducts was assessed using histochemical techniques, including observations under brightfield and fluorescence microscopes. Two types of capitate glandular trichomes producing secretions with a similar composition and non-glandular trichomes exhibiting secretory activity were identified. The secretion of glandular trichomes was dominated by acidic and neutral lipids, essential oil, sesquiterpenes, and steroid-containing terpenes. The schizogenic secretory ducts located in the phloem produced a viscous milky substance with acidic polysaccharides, acidic lipids, phenolic compounds, and proteins. The secretion was released into the duct lumen through notches in the walls of the secretory epithelial cell facing the duct lumen. The location, type, and traits of the non-glandular trichomes and secretory structures, as well as the composition of the secreted products are considered important taxonomic features in the family Anacardiaceae and the Rhus genus. Additionally, these characters are important diagnostic markers for the pharmacobotanical identification of the species in medicinal and cosmetic raw materials. The various compounds present in the secretory structures of R. typhina may contribute to plant protection against pathogens or herbivory and probably play a role as attractants for pollinators and seed dispersers.

Manajemant Breast Engogerment Kompres Daun Kubis Dingin (Brassica Oleracea Var. Capitata) dan Breastcare terhadap Pembengkakan Payudara pada Ibu Nifas

Breast milk duct dams and swelling of the mammary glands are one of the lactation problems that occur after childbirth and during breastfeeding. Breast swelling occurs when the milk ducts do not drain completely and narrow. Milk thickens and the lumen of the milk ducts is blocked. From 3 to 6 days after birth until the milk is expelled normally, symptoms such as warmth, pain to the touch, tenderness, and swelling associated with breast swelling will appear. From 3 to 6 days after birth until the milk is expelled normally, symptoms such as warmth, pain to the touch, tenderness, and swelling associated with breast swelling will appear. The use of cabbage leaves for the treatment and prevention of breast swelling is very simple, the inside of cabbage leaves can be put in the freezer for approximately 20-30 minutes before the procedure. Place cold cabbage leaves inside your bra for 30 minutes and do it twice a day for 3 days. This study aims to determine the difference in effectiveness before and after cabbage leaves (Brassica Oleracea Var.Capitata) and Breast Care in Reducing Breast Swelling. This research is a quasi-experimental design research with one group pre and post design research methods. The population in this study was 34 postpartum mothers who experienced breast swelling on day 3 to day 14. The results of the Wilcoxon test found differences in breast swelling scores before and after applying cold cabbage leaf compresses and breastcare, so it can be concluded that there is a statistically significant average difference between breast swelling scores before and after treatment, where the mean after treatment decreased by 1.15 compared to the mean before treatment 2.79. This means that giving cold cabbage leaf compresses and breastcare can effectively reduce the problem of breast swelling in puerperal mothers. Applying cabbage leaf compresses (Brassica Oleracea var. Capitata) cold and Breast Care combined with very effective in reducing breast swelling in puerperal mothers.

Fixed parotid sialoliths: surgical treatment via transfacial approach using sialendoscopy and ultrasound guidance—retrospective cohort study

The most common cause of symptomatic parotitis is the obstruction of its duct, most commonly by a stone. Despite the development of minimally invasive endoscopic techniques, some of these obstructions cannot be treated entirely endoscopically, requiring combined approaches. This study reviewed the outcomes and surgical technique of ultrasound-guided transfacial parotid sialolithotomy following a failed endoscopic approach. Conducted as a case series with retrospective chart review at an academic tertiary care center, the study evaluated patients who underwent this combined transfacial-endoscopic operation for symptomatic parotid sialolithiasis from April 2022 through January 2023. Key outcomes included operative technique, stone size, stone location, complications, and symptom relief. A total of four male patients with a mean age of 66 years underwent the ultrasound-guided transfacial operation for symptomatic parotid sialolithiasis. Needle localization facilitated transfacial stone retrieval in all cases. The follow-up period ranged from 6 to 15 months. Stone locations included the proximal one-third of the ductal lumen (1 patient), the parotid hilum (1 patient), and within the gland parenchyma (2 patients). The average sialolith length was 8 ± 1.4 mm, and the width was 3 ± 0.8 mm. The average surgical time was 113 ± 16.5 minutes. All cases (100%) achieved successful stone retrieval and symptomatic improvement, with complete resolution of symptoms in 3 cases (75%). No major complications were reported. Ultrasound-guided transfacial parotid sialolithotomy is a safe and effective alternative to parotidectomy for patients who have failed a purely endoscopic approach. A novel transfacial surgical dissection method, based on the middle premasseter space, is proposed for accessing the main parotid duct when obstructions are located in the middle portion of the duct.

Structural Changes in Liver Histology in Common Kilka <i>Clupeonella caspia</i> Svetovidov, 1941 under Anthropogenic Pressure

Histopathological changes are widely used as biomarkers of the health status of fish exposed to chemical compounds. Individuals of the common kilka Clupeonella caspia Svetovidov, 1941 caught from the Middle Caspian Sea exhibited significant changes in the cellular structure of the liver. The more common were signs of hyperemia: an edema of the liver parenchyma, vascular congestion, lympho-macrophage infiltration, small hemorrhages, and a narrowing of the bile duct lumen.

Differential Expression of Mucin in Salivary Gland Tumours.

Background and Objectives: Mucin has been implicated via various mechanisms in the development and growth of tumour cells. However, mucin expression studies in salivary gland tumours are limited, especially with samples from minor salivary glands. This study aims to investigate and compare mucin expression in benign and malignant salivary gland tumours of minor and major salivary gland origins. Materials and Methods: Special stains were used to stain neutral mucin (Periodic acid Schiff), sialomucin (Alcian Blue) and sulfomucin (Aldehyde Fuschin) within tissues from six normal salivary glands and 73 salivary gland tumours including 31 pleomorphic adenomas, 27 mucoepidermoid carcinomas, and 15 adenoid cystic carcinomas. A semi-quantitative approach was used to evaluate mucin expression within ductal lumens. Sialomucin was the most expressed mucin in all salivary gland tumours, regardless of origin. Results: A significant difference was observed in the mucin expression between benign and malignant salivary gland tumours, as pleomorphic adenoma showed three times significantly higher expression of sialomucin compared to mucoepidermoid carcinoma and adenoid cystic carcinoma (p = 0.028). Pleomorphic adenomas of major glands showed 42 times significantly higher expression of sialomucin compared to those of minor glands (p = 0.000). Conclusions: Sialomucin content in pleomorphic adenomas of major glands was vastly increased compared to that in minor glands. Differential sialomucin expression in benign and malignant salivary gland tumours suggests a role in diagnosing of borderline salivary gland tumours.

A pediatric case of Burkitt’s lymphoma with bile duct obstruction requiring surgical biliary reconstruction

BackgroundBiliary obstruction due to compression by a B-cell solid tumor occurs rarely. A few reports have described biliary reconstruction surgery for obstructive jaundice caused by Burkitt’s lymphoma. However, there are no detailed reports on pediatric cases. We report a pediatric case of obstructive jaundice due to malignant lymphoma treated with biliary reconstruction surgery.Case presentationA 5-year-old girl presented to our hospital with a massive abdominal tumor that caused biliary stricture. Chemotherapy was initiated after an open tumor biopsy. However, endoscopic biliary stent placement was performed owing to elevated bilirubin levels. We treated the patient with chemotherapy for 9 months while endoscopically replacing the biliary stent every few months. She achieved complete tumor remission. However, sclerotic lymph nodes were persistent on the dorsal side of the cholecystic duct junction, and biliary stricture at the same site had changed to stent-dependent biliary obstruction. Therefore, we performed choledochojejunostomy and retrocolic Roux-en-Y reconstruction 15 months after initial admission. There were no postoperative complications or tumor recurrences, and the bilirubin level remained low. Histopathologically, the resected bile duct wall was fibrotic and thick, and the bile duct lumen narrowed.ConclusionsBiliary reconstruction is effective to achieve long-term biliary patency in pediatric patients with stent-dependent biliary obstruction due to malignant lymphoma. However, the decision on when to stop biliary stent replacement and proceed to biliary reconstruction surgery is a matter of debate. Further case studies are required to address this issue.

Histopathological changes in salivary glands after radioactive iodine treatment for thyroid diseases

ObjectivesWe aimed to evaluate the histopathological alterations in human salivary glands after radioactive iodine (RAI) treatment for thyroid diseases. Study DesignWe retrospectively selected patients with a history of RAI treatment for thyroid diseases from a database of patients who underwent surgery for oral and maxillofacial diseases and had specimens of salivary glands at Peking University School of Stomatology between December 2012 and July 2023. The patients’ clinical records and histopathological slides of the salivary glands were carefully reviewed. ResultsSixteen patients were included. Three symptomatic patients showed duct cell cytoplasmic vacuolization and increased numbers of disordered duct cell layers (3/3), severe duct stenosis and dilation (2/3), and exfoliated epithelial cells in the duct lumen (1/3). The glandular parenchyma showed severe acinar atrophy (2/2), fat content enhancement (2/2), and severe periductal fibrosis (3/3). Thirteen asymptomatic patients showed duct cell cytoplasmic vacuolization (5/13), acinar atrophy and increased fat content in the parenchyma (5/13), and periductal fibrosis (5/13). ConclusionMain histopathologic changes in the salivary glands after RAI treatment for thyroid diseases are cytoplasmic vacuolization of duct cells, acinar atrophy, fat content enhancement, and periductal fibrosis. These changes were evident in symptomatic cases, and were also seen in some asymptomatic patients.

The ultrastructure of the spermatheca of Mordellistena brevicauda (Coleoptera, Tenebrionoidea) and the associated bacterial cells

The ultrastructural study on the female reproductive system of the beetle M. brevicauda (Mordellidae) confirmed the positive correlation between the length of the sperm and the size of the female seminal receptacle (Spermatheca).The spermatheca of the species is characterized by an apical bulb-like structure where the spermathecal duct forms numerous folds filled with sperm. At this level many bacterial cells are present intermingled with the duct folds. Some are organized in large structures, such as bacteriomes, while other are single bacteriocytes. The latter are often found near the basal lamina of duct epithelium. In addition, some bacteria are visible in the cytoplasm of the duct epithelial cells. Interestingly, bacterial cells have never been observed in the duct lumen. The possible function of the bacterial cells is discussed.

Restoration of Bile Outflow and Liver Revascularization After Cholecystectomy with Damage to the Hepaticocholedochus and Thrombosis of the Right Hepatic Artery

The problem of iatrogenic damage to the extrahepatic bile ducts during cholecystectomy remains relevant. Insufficient experience of the surgical team, limited knowledge of the topographic anatomy of the subhepatic space, especially in conditions of perivesical inflammatory infiltrate, poorly timed access conversion are some of the main reasons.The results of treatment of these patients depend on the timing of recognition of the injury, the type of the injury, the characteristics of reintervention, and timely reconstructive surgery performed in a specialized center. Up to 30 % of damage occurs due to combined injury to the extrahepatic bile ducts and afferent vessels of the liver, which aggravates the course of the disease.We report a clinical case of a female patient. The fragment of the hepaticocholedochus was excised during a planned cholecystectomy. In the same clinic, the primary reconstructive operation was performed (drainage of the hepaticocholedochus using a Kehr’s drain). Afterwards, an external fistula of the common hepatic duct was formed. The reconstructive operation was performed 4 days after the initial operation, in a specialized department of the regional clinical hospital. Relaparotomy was performed. In a dense inflammatory infiltrate during mobilization of the suspected hepaticocholedochus, a Kehr’s drain was identified, which distal branch was not located in the lumen of the bile duct. Thrombosis of the right hepatic artery was diagnosed. The Kehr’s drain was removed, the lobar ducts were identified to their confluence, and the confluence was preserved. The distal stump was sutured. Next, arteriotomy and thrombectomy were performed, and adequate retrograde and antegrade blood flow was recoverd. A hepaticoenteroanastomosis has been formed. The patient was discharged with recovery. When examined after 2 months, the condition was satisfactory. Timely diagnosis of thrombosis of the right hepatic artery, adequate revascularization and biliary reconstruction made it possible to avoid liver necrosis and achieve a good treatment outcome.

Three-dimensional ultrastructural and anatomical analysis of prostatic neuroendocrine cells in mice.

A few studies have examined the ultrastructure of prostatic neuroendocrine cells (NECs), and no study has focused on their ultrastructure in three dimensions. In this study, three-dimensional ultrastructural analysis of mouse prostatic NECs was performed to clarify their anatomical characteristics. Three 13-week-old male C57BL/6 mice were deeply anesthetized, perfused with physiological saline and 2% paraformaldehyde, and then placed in 2.5% glutaraldehyde in 0.1 M cacodylate (pH 7.3) buffer for electron microscopy. After perfusion, the lower urinary tract, which included the bladder, prostate, coagulation gland, seminal vesicle, upper vas deferens, and urethra, was removed, and the specimen was cut into small cubes and subjected to postfixation and en bloc staining. Three-dimensional ultrastructural analysis was performed on NECs, the surrounding cells, tissues, and nerves using focused ion beam/scanning electron microscope tomography. Twenty-seven serial sections were used in the present study, and 32 mouse prostatic NECs were analyzed. Morphologically, the NECs could be classified into three types: flask, flat, and closed. Closed-shaped NECs were always adjacent to flask-shaped cells. The flask-shaped and flat NECs were in direct contact with the ductal lumen and always had microvilli at their contact points. Many of the NECs had accompanying nerves, some of which terminated on the surface in contact with the NEC. Three-dimensional ultrastructural analysis of mouse prostatic NECs was performed. These cells can be classified into three types based on shape. Novel findings include the presence of microvilli at their points of contact with the ductal lumen and the presence of accompanying nerves.

Living tape worms in the duodenal lumen: A rare case report

Fascioliasis is an infection caused by a trematode of the liver, Fasciola hepatica, which affects sheep, goats and cattle. Humans become accidental hosts through drinking contaminated water or ingesting raw green vegetables that has been contaminated with encysted metacercariae, which is the infective form. Fascioliasis has a hepatic phase and a biliary phase, each displaying different clinical signs and symptoms. Common signs and symptoms of the hepatic phase are abdominal pain, fever, eosinophilia, and abnormal liver function tests. The biliary phase of the disease usually presents with intermittent right upper quadrant pain with or without cholangitis or cholestasis. However, it is very rare to find a living flukes in the common bile duct (CBD) and duodenal lumen as it usually lives in gall-bladder and smaller biliary tracts. Here we report a case of human infestation of Fasciola hepatica, who presented with respiratory tract infection with abdominal pain & dyspeptic symptoms and eventually during evaluation, an upper gastro-intestinal endoscopy revealed several flat worms at the second part of the duodenum. The worms were sent to microbiology department for identification. The organisms were confirmed to be Fasciola hepatica. The patient was given tablet Nitazoxanide 500mg 12 hourly for 7 days. The symptoms reduced in intensity after two days of nitazoxanide. The patient was then discharged with an advice of a stool routine examination after one month of completion of nitazoxanide. As fasciolosis is non endemic in our country, a high degree of suspicion is required to ensure early detection and management of such cases. Patients with abdominal pain, altered liver function tests & eosinophilia should always be evaluated for a suspected Fasciola hepatica infection and should be kept as a differential in these cases. Bangladesh Crit Care J March 2024; 12 (1): 54-57

Механизм повреждения мелких холангиоцитов при первичном билиарном холангите

Primary biliary cholangitis (PBC) is a chronic cholestatic progressive liver disease and one of the most important progressive cholangiopathies in adults. Damage to cholangiocytes triggers the development of intrahepatic cholestasis, which progresses to cirrhosis in the terminal stage of the disease. Accumulating data indicate that damage to biliary epithelial cells [(BECs), cholangiocytes] is most likely associated with the intracellular accumulation of bile acids, which have potent detergent properties and damaging effects on cell membranes. The mechanisms underlying uncontrolled bile acid intake into BECs in PBC are associated with pH change in the bile duct lumen, which is controlled by the bicarbonate (HCO3-) buffer system "biliary HCO3- umbrella". The impaired production and entry of HCO3- from BECs into the bile duct lumen is due to epigenetic changes in expression of the X-linked microRNA 506. Based on the growing body of knowledge on the molecular mechanisms of cholangiocyte damage in patients with PBC, we propose a hypothesis explaining the pathogenesis of the first morphologic (ductulopenia), immunologic (antimitochondrial autoantibodies) and clinical (weakness, malaise, rapid fatigue) signs of the disease in the asymptomatic stage. This review focuses on the consideration of these mechanisms.

3D-printed versatile biliary stents with nanoengineered surface for anti-hyperplasia and antibiofilm formation

Biliary strictures are characterized by the narrowing of the bile duct lumen, usually caused by surgical biliary injury, cancer, inflammation, and scarring from gallstones. Endoscopic stent placement is a well-established method for the management of biliary strictures. However, maintaining optimal mechanical properties of stents and designing surfaces that can prevent stent-induced tissue hyperplasia and biofilm formation are challenges in the fabrication of biodegradable biliary stents (BBSs) for customized treatment. This study proposes a novel approach to fabricating functionalized polymer BBSs with nanoengineered surfaces using 3D printing. The 3D printed stents, fabricated from bioactive silica poly(ε-carprolactone) (PCL) via a sol–gel method, exhibited tunable mechanical properties suitable for supporting the bile duct while ensuring biocompatibility. Furthermore, a nanoengineered surface layer was successfully created on a sirolimus (SRL)-coated functionalized PCL (fPCL) stent using Zn ion sputtering-based plasma immersion ion implantation (S-PIII) treatment to enhance the performance of the stent. The nanoengineered surface of the SRL-coated fPCL stent effectively reduced bacterial responses and remarkably inhibited fibroblast proliferation and initial burst release of SRL in vitro systems. The physicochemical properties and biological behaviors, including in vitro biocompatibility and in vivo therapeutic efficacy in the rabbit bile duct, of the Zn-SRL@fPCL stent demonstrated its potential as a versatile platform for clinical applications in bile duct tissue engineering.

Hepatobiliary Ascariasis in a Piglet

PurposeAscariasis caused by the helminth Ascaris suum is the most common parasitosis of swine worldwide and it may involve all age categories of pigs. The present study reports an unusual localization of A.suum worms in the biliary system of a piglet slaughtered for human consumption.MethodsThe liver was subjected to ultrasound scan and pathological examination. The isolated worms were morphologically examined and the DNA was extracted for the molecular identification of the species involved.ResultsA total of 43 preadult nematodes were found within the gallbladder and the bile ducts. Parasites were morphologically identified as belonging to the genus Ascaris and molecularly as A. suum. At gross examination, the liver was moderately enlarged, with the bile ducts severely dilated. A chronic inflammatory infiltrate was noted, often centered around ectatic bile ducts (up to 5 mm in diameter), lined by hyperplastic epithelium and filled with sections of nematodes. The worm sections showed smooth cuticle, coelomyarian musculature, and an intestinal tract lined by columnar, uninucleated cells within a pseudocoelom. The ex vivo ultrasonographic examination of the liver allowed the visualization of several nematodes in the bile duct lumen and could be suggested for in vivo diagnosis. Unfortunately, the absence of the intestine did not allow to define the pathogenesis of the infection.ConclusionAlthough, given the unusual nature of this finding, it is difficult to identify predisposing factors for this A. suum localization, it suggests that ascariasis should be considered in the differential diagnosis of pigs with hepatobiliary disease.

Hepatic and renal lesions in sheep intoxicated with Urochloa hybrid Mulato II in Argentina.

A flock of 48 sheep in Argentina grazing on a pasture of hybrid Urochloa (formerly Brachiaria) Mulato II (Urochloa ruziziensis × Urochloa decumbens × Urochloa brizantha) developed facial dermatitis, severe jaundice, and weakness after brief physical activity. Blood biochemistry of 3 animals revealed azotemia, elevated aspartate aminotransferase activity, and increased direct, indirect, and total bilirubin concentrations. The urine was markedly turbid and contained large concentrations of bile pigments and protein. At autopsy of 2 animals, there was severe jaundice and subcutaneous submandibular edema. The livers were enlarged, intensely yellow, and had a marked acinar pattern. Gallbladders were distended, and the kidneys were diffusely dark in one animal and yellow-green in the other. Microscopically, there was lymphoplasmacytic and histiocytic cholangiohepatitis with abundant crystals in the lumen of bile ducts and in the cytoplasm of macrophages. The proximal and distal convoluted renal tubules had protein casts in their lumens, and crystals were observed in the lumen and epithelial cells. Lectin histochemistry showed strong affinity for Arachis hypogaea agglutinin in hepatic macrophages. In the one sheep that was tested for heavy metals, copper concentrations in the liver and kidney were within the RIs. Despite the immediate change of pasture, morbidity and mortality were 100% within 3 mo. The association between the consumption of this pasture, and the clinical, biochemical, pathology, and lectin histochemistry findings confirmed intoxication with Urochloa hybrid Mulato II. To our knowledge, intoxication by this hybrid of Urochloa has not been reported previously.

Novel Screening System for Biliary Excretion of Drugs Using Human Cholangiocyte Organoid Monolayers with Directional Drug Transport.

It has recently been reported that cholangiocyte organoids can be established from primary human hepatocytes. The purpose of this study was to culture the organoids in monolayers on inserts to investigate the biliary excretory capacity of drugs. Cholangiocyte organoids prepared from hepatocytes had significantly higher mRNA expression of CK19, a bile duct epithelial marker, compared to hepatocytes. The organoids also expressed mRNA for efflux transporters involved in biliary excretion of drugs, P-glycoprotein (P-gp), multidrug resistance-associated protein 2 (MRP2), and breast cancer resistance protein (BCRP). The subcellular localization of each protein was observed. These results suggest that the membrane-cultured cholangiocyte organoids are oriented with the upper side being the apical membrane side (A side, bile duct lumen side) and the lower side being the basolateral membrane side (B side, hepatocyte side), and that each efflux transporter is localized to the apical membrane side. Transport studies showed that the permeation rate from the B side to the A side was faster than from the A side to the B side for the substrates of each efflux transporter, but this directionality disappeared in the presence of inhibitor of each transporter. In conclusion, the cholangiocyte organoid monolayer system has the potential to quantitatively evaluate the biliary excretion of drugs. The results of the present study represent an unprecedented system using human cholangiocyte organoids, which may be useful as a screening model to directly quantify the contribution of biliary excretion to the clearance of drugs.

Morphology of the epididymal duct of the domestic pig (Sus scrofa domesticus).

The study provides a general overview of the morphology of the epididymal duct in pigs. Four epididymides from two sexually mature boars were dissected into 32 segments and examined histologically. Duct lumen and wall thickness were measured and relative surface area of different components was assessed by Chalkley's random hit method. The epithelial lining was characterized at X1000. Lumen diameter and wall thickness of efferent ductules averaged 177 and 30 µm, respectively. Of the epididymal duct from caput to distal corpus the luminal diameter was 332 µm, with a narrower section in the proximal corpus. Wall thickness averaged 70 µm. In the cauda, luminal diameter and wall thickness increased to 717and 751 µm, respectively. The epithelial lining of the efferent ductules consists of a single layer of columnar cells with average height 21 µm. The lining of the epididymal duct consists of ciliated, pseudo-stratified columnar epithelium composed of "basal cells" and "principal cells." Particularly tall principal cells (96 µm) were found in the proximal caput. Height decreased to 40 µm at the distal cauda. Microvilli from principal cells were 14-17 µm long in the distal caput but decreased to 5 µm in the distal cauda. The epithelial lining was folded in the proximal caput and more so in the distal cauda. Secretory granules (epididymosomes) were present in small amounts in efferent ductules and epididymal duct; the largest quantities occurred in the distal cauda. Leukocytes were present throughout the duct, albeit in insignificant numbers. Chalkley's random hit method showed rapid spermatozoan transport through efferent ductules and proximal caput in large amounts of fluid. Sperm concentration increased due to fluid resorption in the proximal caput, was highest from caput flexure to proximal cauda and decreased at the caudal flexure, indicating secretory activity.