Determination of Secondary Bile Acids in the Mice Feces. Controversies on their Involvment in the Pathogenesis of Colorectal Cancer

Abstract Objectives: The aim of the study was to determine the level of secondary bile acids (SBA) in the diets and feces of mice and the variation of amount ingested/excreted if these SBA are administered as monotherapy or in 1:1 dose. Methods: The mice were divided into 4 groups and fed for 140 days with different diets. The control lot received a normal diet and the others received diets supplemented with 0.25% deoxycholic acid (DCA), 0.25% lithocholic acid (LCA) and 0.125% DCA+0.125% LCA. After 140 days, the mice feces were collected and homogenized to obtain a mixture for each lot from which the determinations of the studied SBA were performed. For the mice food evaluation, portions of 10 g from each of the 4 diets were subjected to the SBA determination. Results: The daily ingestion over more than 4 months of DCA or LCA added to the diet and administered as monotherapy determine a significantly increase of the SBA eliminated into the feces (the DCA level was 11x higher, and of the LCA 233x higher). If half of the LCA dose is replaced with DCA, the level of LCA in the feces gets comparable with that of the DCA (their combined amounts represents only 13x higher increase of these two bile acids in feces). Conclusions: The simultaneous ingestion and excretion of DCA and LCA can be considered as a particular situation ruled by endogenous mechanisms. This behavior represents an important observation, knowing that the bile acids effects in the colorectal cancer are dose dependent.</jats:p

Colorectal cancer: an update upon the diagnostic and therapeutic transdiciplinary approach

Scop: Prezentarea unor date actualizate referitoare la rolul ONCOTEAM în diagnosticul și terapia pacientului cu cancerului colorectal (CRC). Material și metode: În perioada 2018-2023, 147 de pacienți cu CRC au beneficiat de un abord individualizat. Evaluarea imagistică preoperatorie cu CT-scan/MRI cu difuzie a fost urmată de o descriere de tip ”hartă” a informațiilor pe baza cărora s-a efectuat intervenția chirurgicală și procesarea histopatologică conform metodologiei descrise anterior de echipa noastră în jurnalelel Diagnostics (DOI: 10.3390/diagnostics11020314) și Journal of the Belgian Society of Radiology (DOI: 10.5334/jbsr.3186). Examinările genetice au fost efectuate la indicațiile oncologului iar terapia post-operatorie a luat în considerare profilul molecular al celulelor tumorale. Rezultate: Utilizând acest protocol adaptat, am obținut un număr mediu de 15±2.23 limfonoduli prelevați per caz. Numărul depozitelor tumorale a fost, de asemenea, crescut și a dus la o supra-stadializare a 15% din cazuri. Utilizând o valoare a ”lymph node ratio” de 0.15, am obținut valori superioare celor obținute la abordarea clasică a 120 cazuri examinate anterior (p=0.002). Determinările genetice efectuate în timp au dus la o începere rapidă a terapiei oncologice individualizate și, deși profilul genei BRAF V600E este dificil a fi evaluat în țesuturi incluse în parafină, extracția ADN și determinările PCR au fost adecvate în toate cazurile examinate. Concluzii: Abordarea transdiciplinară a CRC poate fi efectuată doar dacă fiecare membru al echipei este implicat conștiincios în fiecare pas al diagnosticului sau terapiei. Costurile determinărilor au fost parțial acoperite în cadrul proiectelor PCCF 20/2018 și 10127/13/2021.Aim: To present an update regarding the role of the ONCOTEAM in the diagnosis and therapy of colorectal cancer (CRC). Materials and methods: During 2018-2023, 147 patients with CRC have benefited by an individualized approach. Preoperatively evaluation was done with CT-scan/diffusion-weighted MRI and a lymph node station map was typed. The next step consisted on surgical removal, based on the indications included in the map. Histopathological examination was based on the methods described by our team previously (DOI: 10.3390/diagnostics11020314; DOI: 10.5334/jbsr.3186). Genetic examinations were done based on the indications of the oncologist and the post-operative therapy was performed according to the molecular profile. Results: Based on the in-house adapted protocol, the median number of harvested lymph nodes per case was 15±2.23. The number of identified deposits was also significant and up-staged the tumors in 15% of the cases. The lymph node ratio value, using a cut-off of 0.15, was also superior to the classic approach of other 120 cases (p=0.002). The genetic examinations proved to be useful for an earlier start of post-operative therapy, without any cost for the patients. As regarding pre-analytical factors, although BRAF V600E gene profile is hard to be detected from paraffin-embedded tissues, the DNA extraction and PCR examinations were succesful in all of the cases. Conclusions: A proper transdiciplinary approach can be done only if any member of the team is attentively involved in each step of the diagnosis and therapy. The costs were partially supported by the projects PCCF 20/2018, and 10127/13/2021

Challenges and controversies in open pancreatoduodenectomies

Duodenopancreatectomia cefalopancreatica este o procedura intricata, ce necesita o inalta precizie datorita proximitatii structurilor vitale, sustinuta prin expertiza. O data cu evolutia chirurgiei si aparitia de instrumente inovatoare, mortalitatea a scazut sub 5%, dar morbiditatea a ramas la 30%, in special datorita fistulelor de anastomoza si a evenimentelor hemoragice. In acest sens, chirurgia deschisa confera rezultate mai bune, cu o curba de invatare rezonabila. Cu atat mai mult, cu cat efectuarea de trialuri clinice este dificila in acest domeniu, atat in chirurgia deschisa, cat si minim invaziva, pentru a obtine evidente valoroase, astfel subiectul ramane unul de dezbatere.Whipple procedure is intricate and demands high precision due to the proximity of critical structures, which requires an achieved expertise. With the innovative instruments and evolution of surgery, the perioperative morbidity still stands at 30% with a mortality lower than 5%, primary because of anastomotic leaks and haemorrhagic events. Therefore open surgery provides better outcomes with a decent learning curve. Furthermore, it is challenging to conduct clinical trials in the field of pancreatic surgery both open or minimally-invasive to obtain high-level evidence, remaining a subject open to debate

Emergency Treatment of Transverse Colon Cancer

This chapter deals with the emergency treatment of transverse colon cancer. The main complications that classify transverse colon cancer in an emergency setting are obstruction, perforation accompanied by localized or generalized peritonitis, and hemorrhage which may be occult or cataclysmic with hemorrhagic shock. We present the technical principles of radical surgical resection using embryological, anatomical, and oncological concepts. In this chapter we also discuss the principles of lymphadenectomy associated with complete excision of the mesocolon with high vascular ligation, in particular with T3 or T4 cancers requiring D2/D3 lymphadenectomy. The use of infrapyloric, gastro-epiploic, and prepancreatic lymphadenectomy is recommended due to the frequent metastases in these regional lymph nodes

Analysis of microRNA signatures using size-coded ligation-mediated PCR

The expression pattern and regulatory functions of microRNAs (miRNAs) are intensively investigated in various tissues, cell types and disorders. Differential miRNA expression signatures have been revealed in healthy and unhealthy tissues using high-throughput profiling methods. For further analyses of miRNA signatures in biological samples, we describe here a simple and efficient method to detect multiple miRNAs simultaneously in total RNA. The size-coded ligation-mediated polymerase chain reaction (SL-PCR) method is based on size-coded DNA probe hybridization in solution, followed-by ligation, PCR amplification and gel fractionation. The new method shows quantitative and specific detection of miRNAs. We profiled miRNAs of the let-7 family in a number of organisms, tissues and cell types and the results correspond with their incidence in the genome and reported expression levels. Finally, SL-PCR detected let-7 expression changes in human embryonic stem cells as they differentiate to neuron and also in young and aged mice brain and bone marrow. We conclude that the method can efficiently reveal miRNA signatures in a range of biological samples