Biliary tract cancer or extrahepatic cholangiocarcinoma (ECCA) represents the sixth commonest cause of cancer in the gastrointestinal tract in western countries. We aimed to characterize the microbiota and its predicted associated functions in the biliary tract of ECCA and benign biliary pathology (BBP). Samples were taken from 100 patients with ECCA and 100 patients with BBP by endoscopic cholangio-pancreatography for DNA extraction. Ten patients with ECCA and ten with BBP were selected for microbiota studies using the V4-16S rRNA gene and sequenced in Illumina platform. Microbiota analyses included sample-to-sample distance metrics, ordination/clustering and prediction of functions. Presence of Nesterenkonia sp. and Helicobacter pylori cagA and vacA genes were tested in the 100 ECCA and 100 BBP samples. Phylum Proteobacteria dominated all samples (60.4% average). Ordination multicomponent analyses showed significant microbiota separation between ECCA and BBP (p 0.010). Analyses of 4002 operational taxonomic units with presence variation in at least one category probed a separation of ECCA from BBP. Among these, Nesterenkonia decreased, whereas Methylophilaceae, Fusobacterium, Prevotella, Actinomyces, Novosphingobium and H. pylori increased in ECCA. Predicted associated functions showed increased abundance of H. pylori virulence genes in ECCA. cagA and vacA genes were confirmed by PCR in ECCA and BBP samples. This is the first microbiota report in ECCA and BBP to show significant changes in microbial composition. Bacterial species unusual for human flora were found: Methylophilaceae and Nesterenkonia are reported in hypersaline soils, and Mesorhizobium is a nitrogen-fixing bacterium. Enrichment of virulence genes confirms previous studies suggesting that H. pylori might be associated with ECCA.

Several experimental and clinical studies have shown that citrates are useful in dissolving calcifications and proteic plug in pancreatic ducts both of alcoholic etiology and in patients with chronic pancreatitis. Until now, using citrates to dissolve stones in clinical studies was performed orally with satisfactory medium-term results, including control of abdominal pain and eradication of shadows on X-rays. Laboratory studies have shown that these concretions dissolve quickly when such compounds are applied directly. This paper reports 2 women aged 27 and 40 with histories of chronic abdominal pain, and who, by abdominal X-ray and endoscopic retrograde cholangiopancreatography (ERCP), were shown to have multiple calcifications in the main and accessory pancreatic ducts. In both patients, endoscopic sphincterotomy of the bile and pancreatic segments of the sphincter of Oddi and introduction of a nasopancreatic catheter and intraductal infusion of citrates were performed. Radiological controls showed fragmentation and disappearance of calcifications. Clinically, there was complete absence of abdominal pain in the first week following the procedure. This is the first human study of intraductal administration of citrates to dissolve pancreatic lithiasis with highly favorable results.

Two young women with clinically established pseudocyesis were studied by endometrial biopsy, basal hormonal serum levels and dynamic pituitary testing. Basal serum levels of PRL and TSH were in the normal range; estradiol - 17 beta, progesterone and FSH were in the follicular phase range, but LH was in the follicular phase range in one patient and in the climateric range in the other one. The histologic assessment of the endometrial biopsies disclosed a proliferative endometrium in both patients. A group of six patients with hypothalamic amenorrhea were subjected to dynamic pituitary testing to compare results with those obtained in the two patients with pseudocyesis. The dynamic pituitary response to GnRH, TRH and metoclopramide was normal in the two patients with pseudocyesis and in the group with hypothalamic amenorrhea; moreover, challenge with estradiol benzoate (EB) in the two patients with pseudocyesis disclosed a normal positive feedback of LH. These observations and the analysis of data already published suggest that the amenorrhea of pseudocyesis is associated neither with a persistent corpus luteum nor chronic hyperprolactinemia. We suggest that an abnormality in neurotransmitter pathways results in alterations of pituitary hormone secretion. However, additional patients must be studied to prove or disprove this hypothesis.

Professor Ralph Kirsch for getting me involved with primary sclerosing cholangitis in the first place. Professor Solly Benatar for his ongoing support during my training at Groote Schuur Hospital. Professors Flip Bornman and Jake Krige for their surgical expertise and for teaching me to be brief.