Jornal de Biomarcadores Moleculares e Diagnóstico

Background and purpose: Predicting the efficacy of anticancer therapy is the holy grail of drug development and treatment selection in the clinic. To achieve this goal, scientists require pre-clinical models that can reliably screen anticancer agents with robust clinical correlation. However, there is increasing challenge to develop models that can accurately capture the diversity of the tumor ecosystem, and therefore reliably predict how tumors respond or resistant to treatment. Indeed, tumors are made up of a heterogeneous landscape comprising malignant cells, normal and abnormal stroma, immune cells, and dynamic microenvironment containing chemokines, cytokines and growth factors. In this mini-review we present a focused, brief perspective on emerging preclinical models for anticancer therapy that attempt to address the challenge posed by tumor heterogeneity, highlighting biomarkers of response and resistance.

Recent findings: Starting from 2-dimensional and 3-dimensional in vitro models, we discuss how organoid cocultures have led to accelerated efforts in anti-cancer drug screening, and advanced our fundamental understanding for mechanisms of action using high-throughput platforms that interrogate various biomarkers of ‘clinical’ efficacy. Then, mentioning the limitations that exist, we focus on in vivo and human explant technologies and models, which build-in intrinsic tumor heterogeneity using the native microenvironment as a scaffold. Importantly, we will address how these models can be harnessed to understand cancer immunotherapy, an emerging therapeutic strategy that seeks to recalibrate the body’s own immune system to fight cancer.

Conclusion: Over the past several decades, numerous model systems have emerged to address the exploding market of drug development for cancer. While all of the present models have contributed critical information about tumor biology, each one carries limitations. Harnessing pre-clinical models that incorporate cell heterogeneity is beginning to address some of the underlying challenges associated with predicting clinical efficacy of novel anticancer agents.

Currently a wide range of biomarkers and potential biomarkers exist across a variety of fields for the management of colorectal cancer. These can be a specific molecule or a radiographical finding and can predict outcome or response to treatment. The field is being developed along several fronts with many new innovations happening in the last few years however few have made it into routine clinical practice with others requiring validation. The evolving markers of significance are; MicroRNA, epithelial-to-mesenchymal-transition, imaging and metabolic with several sub-divisions depending on the pathway effected. This review will provide a narrative appraisal of our current understanding and clinical application of these biomarkers.

Introduction: Spontaneous bacterial peritonitis (SBP) is the most frequent and life-threatening infection in patients with decompensated liver cirrhosis. Diagnosis should be prompt and treatment must not be delayed until the microbiology results are available.

Aims and Methods: The current study was conducted to assess the potential role that MPV may have in the diagnosis of SBP in cirrhotic patients with ascites Three groups were included in the study. Group I (SBP group) included 100 patients with liver cirrhosis and ascites complicated by SBP, group II (non-SBP group) included 98 patients with liver cirrhosis and ascites without SBP and group III included 50 healthy subjects to serve as a control group.

Results: ESR, CRP and total leucocytic count (TLC) in ascitic fluid were significantly higher in SBP group compared to non-SBP group (median 37.5 vs. 12, 12 vs. 5 and 530 vs. 60 respectively with p value<0.01). The MPV was significantly higher in SBP group vs. non-SBP group and healthy control group (8.5, 7.9 and 8.3 respectively and P value<0.0001). On constructing ROC curve for the MPV; at a cutoff value of 8.4 fl, MPV had 73% sensitivity and 85.7% specificity for detecting SBP with overall accuracy 79.3%, (AUC=0.84 with negative predictive value (NPV) and positive predictive value (PPV) for MPV of 75.7 and 83.9%, respectively). Regarding ESR; at a cutoff value of 20.

Conclusion: MPV is increased in SBP in cirrhotic patients with ascites than the other inflammatory markers.

Introduction: Hand assisted laparoscopic surgery is an updated highly advanced version of laparoscopic technique. Such technique bridges the gap between traditional surgery and total laparoscopic surgery. Introduction of the hand intracorporeally enhanced the degree of freedom, hence, a remarkable degree of precision and safety in task performance.Clinical and experimental studies confirmed safe use of the hand with insufflation pressure enhancing dexterity as well as a steep learning curve. Therefore, the author made an overview analysis to the factors related to safety; efficiency; dexterity; instrumentation and cost-effectiveness for the use of hand assisted laparoscopic surgery; with an emphasis on live donor nephrectomy.
Results and discussion: Prospective studies made by Kolvenbach on the use of hand assisted laparoscopic surgery in aortic aneurysm repair proved high degree of safety and efficiency as well as cost effectiveness. Several studies highlighted a multitude of factors significantly contributing into a high degree of precision and task performance; which reflected on uneventful enhanced recovery programme.The introduction of either hand intracorporeally enhanced the limited degree of freedom for the current laparoscopic tools. There are various hand port devices of which the pros and cons for each port will be discussed in detail. The author’s experimental studies confirmed that optimum safe insufflation pressure would be 10 mm Hg with no leak from the hand port and optimum dexterity and task performance.
Conclusion: Hand assisted laparoscopic surgery is a safe and efficient technique. It significantly enhances concept of Enhanced Recovery programme. Raising public awareness can provide a high impact in enhancing live donor nephrectomy; hence reducing the inexorable renal transplant waiting list for patients with end stage renal disease. Such patients are at progressive rise of mortality risk with prolonged waiting list.

Background: Hepatic progenitor cells (HPC) as a hepatic regeneration reservoir, are now signified as one of the promising therapeutics. However, connection to cells resistant to apoptosis in pathogenesis of chronic hepatitis C virus (HCV) is still evolving.

Aim: title relationship between HPC and cells resistant to apoptosis in HCV along with liver disease severity and fibrosis progression. Methods: liver biopsies of 91 chronic HCV patients were immunohistochemically examined. Both demographic and clinical characteristics were sourced from the data registries. METAVIR scoring was unified for both Grading and staging. Immunostaining with CK7, Ki67, and bcl2 antibodies was done.

Results: Transaminases, platelets and prothrombin time exhibited significant relation with Ki67, CK7 both isolated and ductular and bcl2 both LPT and LAH. CK7 ductular showed association with fibrosis and necroinflammatory activity (P< 0.05), while non-significant relation was noticed with the CK7 isolated form and Ki67 (P> 0.05). Moreover, bcl2 both (LPT) and (LAH) demonstrated association with fibrosis and necroinflammatory activity (P< 0.05). Positive correlation between immunoexpression of HPCs both isolated (r=0.547, <0.001) and ductular with bcl2 LAH (r=0.476, p<0.001) was reported. bcl2 H showed positive correlation with the CK7 isolated form (r=0.476, p <0.001), with no correlation between it and the CK7 ductular (r=0.298, p= 0.003).

Conclusion: Hepatic progenitor cells and cells resistant to apoptosis are conversely interrelated to HCV pathogenesis with a pivotal role in disease severity and progression. It is a notion to be considered in development new therapies of HCV-related chronic liver disease.

Objective: Although Helicobacter pylori infection is endemic in Nigeria, the specific genotypes that influence treatment responses are scarcely known. We aimed to determine the specific genotypes of H. pylori virulence genes, vacA and cagA in dyspeptic patients.
Subjects and methods: Gastric biopsy samples were obtained from 80 dyspeptic patients referred for endoscopy. Genomic DNA was extracted from biopsies using the ReliaPrep DNA kit. H. pylori DNA was detected by a singleplex PCR based on the genus specific 16s rRNA gene. The vacA subtypes for the s1 and s2 regions and the m1 and m2 alleles were detected by allele specific multiplex PCR. The cagA gene was amplified by a singleplex PCR.
Results: Of the 80 samples, 30 (37.5%) had abnormal mucosa which were chronic gastritis. The rest (62.5%) had normal mucosa lining. Of the 30 chronic gastritis cases, 22 (73%) had H. pylori infection as detected by 16s rRNA PCR. Only 2 (4%) of the normal mucosa cases had H. pylori infection. For the vacA genotypes, 79% of the H. pylori infections were s1c/m2 genotype, followed by 8% s1b/m2 genotype. Three different genotypes: s1c/m1/m2, s1c/s2/m2 and s1c/m1 occurred at 4% each. The most virulent vacA genotype, s1/m1 was only 8% while the least virulent vacA genotype s2/m2 was 4%. The moderate virulent vacA genotype, s1/m2 was the most prevalent (83%) in the Nigerian patients. The most prevalent subtype was the s1c/m2. Only 7 cases (29%) were cagA positive.
Conclusion: The pathogenicity-associated virulence genes present in Nigerian dyspeptic patients were moderate types. The endemicity of the disease may not necessarily lead to high rate of fatal outcomes or treatment failures as reported in other parts of the world.

Purpose of review: The vision and strategy for the 21st century treatment of cancer calls for a personalized approach in which therapy selection is designed for each individual patient. While genomics has led the field of personalized cancer medicine over the past several decades by connecting patient-specific DNA mutations with kinase-targeted drugs, the recent discovery that tumors evade immune surveillance has created unique challenges to personalize cancer immunotherapy. In this mini-review we will discuss how personalized medicine has evolved recently to accommodate the emerging era of cancer immunotherapy. Moreover, we will discuss novel platform technologies that have been engineered to address some of the persisting limitations.

Recent finding: Beginning with early evidence in personalized medicine, we discuss how biomarker-driven approaches to predict clinical success have evolved to account for the heterogeneous tumor ecosystem. In the emerging field of cancer immunotherapy, this challenge requires the use of a novel set of tools, distinct from the classic approach of next-generation genomic sequencing-based strategies. We will introduce new techniques that seek to tailor immunotherapy by re-programming patient-autologous T-cells, and new technologies that are emerging to predict clinical efficacy by mapping infiltration of lymphocytes, and harnessing fully humanized platforms that reconstruct and interrogate immune checkpoint blockade, ex-vivo.

Summary: While cancer immunotherapy is now leading to durable outcomes in difficult-to-treat cancers, success is highly variable. Developing novel approaches to study cancer immunotherapy, personalize treatment to each patient, and achieve greater outcomes is penultimate to developing sustainable cures in the future. Numerous techniques are now emerging to help guide treatment decisions, which go beyond simple biomarker-driven strategies, and are now we are seeking to interrogate the entirety of the dynamic tumor ecosystem.

Background: Aim of this study was to analyse our results of oriented parathyroidectomy (minimally-invasive parathyroidectomy (MIP) minimally-invasive video-assisted parathyroidectomy (MIVAP); and to confirm the usefulness of intra-operative parathyroid hormone (PTH) monitoring (ioPTH) when using minimally invasive techniques for treatment of sporadic primary hyperparathyroidism (pHTP). Material and methods: From June 2005 to May 2017, in the Division of General Surgery of University of Trieste, we operated 137 patients with pHPT. Minimally invasive video-assisted parathyroidectomy (MIVAP) by an anterior approach was proposed for 60 (43.8%) patients with sporadic pHPT and one unequivocally enlarged parathyroid gland on pre-operative ultrasound and 99mTc-SestaMIBI scintigraphy without previous neck surgery. Intraoperatively, a quick parathyroid assay was used during the last 107 (78.1%) surgical procedures. All patients underwent preoperative investigations of calcemia, phoshoremia and PTH levels and vocal cord function. Age, gender, operative times, pathologic findings, calcemia, PTH, length of hospital stay and complications were retrospectively analyzed. Results: MIVAP was successfully accomplished in 60 patients. Mean operative time was 87 minutes. Postoperative complications included 12/137 (8.75%) transient hypocalcemia. No laryngeal nerve palsies, no definitive hypocalcemias, no persistent pHPT and no recurrent pHPT were observed. Pre-operative 99mTc-SestaMIBI scintigraphy revealed the correct position of parathyroid adenoma in 91 cases (66.4%). Conclusion: Our decade experience in a single center demonstrated that minimally invasive parathyroidectomy for localized single-gland adenoma seems to have significant advantages, especially in terms of cosmetic results and post-operative pain, if performed by dedicated team, with an adequate activity volume. Moreover, the ioPTH determinations are very important during minimally invasive parathyroidectomy, as it allows avoiding bilateral neck exploration.

Background: Giant cell hepatitis is rarely described in adults; referred to as post infantile giant cell hepatitis (PIGCH). Most reports have mentioned PIGCH's association with systemic lupus erythromatosis, autoimmune hepatitis, lymphoma, and leukemia. However, links with other medical disorders are still evolving.

Reports of primary sclerosing cholangitis (PSC) presenting as IgG4-SC has been typically described in association with other IgG4-related disorders, most frequently autoimmune pancreatitis. However, some cases of isolated IgG4-SC have been reported. Herein; we report a case of IgG4-SC presented by PIGCH.

Case description: A 29-year-old gentleman presented with two month jaundice and biochemical evidence of acute hepatitis. He reported no history of drug exposure, had no gall bladder or pancreatic disease, nor prior similar attacks. The work up revealed negative serology for viral hepatitis. Markers of autoimmune liver disease were negative except for pANCA, and serum levels of pancreatic enzymes, copper and ceruplasmin were normal, and urinary copper was normal.

Results: Abdominal sonography and MRCP showed normal pancreas and biliary tract. ERCP showed that the common bile duct had a single short narrowed segment with thickened walls. Histological examination of colonoscopic biopsies taken from the terminal ileum and colon demonstrated no pathological alterations. Liver histology showed evidence of parenchymal extinction with extensive giant cell transformation, ductular proliferation, cholestasis, and positive IgG4 staining, a picture suggestive of PSC and PIGCH.

Discussion: In this case, we did not test for serum levels of IgG4, and resorted to immune-staining of liver tissue, as this is the hallmark for diagnosing IgG4-SC. Histopathological features and, more definitely, the positive IgG4 immunostaining were present in the liver tissue, which were crucial in diagnosing this case as IgG4-SC (IAC).

Conclusion: This case presented with both IgG4-SC and biopsy proven PIGCH, and had a favourable outcome with biliary drainage and immuno-suppression therapy

Epidermal inclusion cyst is a benign lesion that commonly occurs due to inclusion of epidermal cells into the dermal or deeper layers in a trauma event. Percutaneous release is a minimally invasive technique and good surgical outcomes can be achieved. However, the percutaneous procedure is a puncture injury and the epidermal inclusion cyst is reasonable to become a possible complication. In this article, we presented a case of trigger finger in left middle finger. She underwent percutaneous release as the treatment and a second percutaneous release 5 months later due to recurrence of the symptoms. An epidermal inclusion cyst was noted 5 months thereafter and it was treated with excision. With this case, we need to be more aware of this possibility if a mass lesion without infection signs occurs in the released area.