Jornal de Biomarcadores Moleculares e Diagnóstico

Biochemistry provides knowledge about commonality principles, explains particularities of individuals, and discloses targets for therapeutic approaches. Although common biochemical pathways have been conserved during evolution, and although molecules and pathways have been generated based on existing ones, a one-fitsall medicine is about to be more and more replaced by personalized medicine. The declared objective of personalized medicine is to either predict a person’s risks for developing a disease or to treat a patient according to his or her metabolic predisposition and capacity, genetic mutations, or polymorphisms. The genotype of a person can hint at imminent risks and prevent the outbreak of diseases if lifestyle or behavior is changed according to the risk profile. The phenotype does not only describe proteins, enzymes and metabolites from the expression of the person’s genes, but provides data to recognize patterns belonging to an existing or eventually silent disease which can be treated effectively.

Background: Although the contribution of platelets to the measured serum EGF levels was reported by Oka since 1983, the majority of reports in healthy donors or patients do not control clotting times during the collection of the sera. This results in a variation of the notified values, additionally to the one already caused by the functional EGF SNP of the gene, that modulates the expression of the molecule. Both issues -platelets and SNP- make the conventional stratification by absolute serum EGF levels not suitable. Within this study we evaluated serum EGF levels in a panel of 105 healthy Cuban donors, balanced by gender and age (from 18-78 years). As a result, a new stratification methodology for the comparison between individuals was proposed.

Methods: The EGF was estimated in sera collected at two different clotting times: 1h and 4h. Comparisons between groups were carried out. The estimations, normalized through the calculation of ratios from the two measured levels (r=[EGF]1h/[EGF]4h), were used for the stratification.

Results: Differences were found by age (4h, donors younger and older than 45 years, p=0.0083) and gender (1h, p=0.0167), and between 1h and 4h levels (p<0.0001, n=105). While 38 out of 105 individuals ranked different in 1h and 4h conventional stratifications, the methodology using ratios yielded a unique score for each individual.

Conclusions: The proposed methodology of stratification by ratios, in contrast to the conventional approach, allows for a proper comparison between EGF levels and individuals. Thus, it should have an impact on diseases for which the association of EGF with the illness has been established, aiding to clarify the connection of the molecule with the disease. This work might be of value to clinicians, scientists, and the healthcare community in general, conducting research regarding the role of EGF as a biomarker.

STEAP1 is one of six-transmembrane epithelial antigen of prostate (STEAP) highly expressed in human prostate cancer and is up-regulated in multiple cancers, including bladder, colon, breast, Ewing and lung cancers. Immunohistochemical analysis demonstrates significant STEAP1 expression at the intercellular communication between adjacent cells suggesting that this antigen must be a channel, or a transport protein indicating its potential role in tumor cell intercellular communication increasing the potential of STEAP1 as a diagnostic, prognostic, prophylactic and/or therapeutic target for new immunotherapeutic strategies. In prostate cancer, overexpression relates adenocarcinoma and prostatic intraepithelial neoplasia scores suggest that STEAP1 could be involved in tumor initiation and progression and may be of clinical usefulness in early disease diagnosis. Also, STEAP1 can serve as a biomarker of worse prognosis because of its association with higher Gleason score, seminal vesicle invasion, biochemical recurrence, and worse outcome. Therapeutically, STEAP1 is one of a few prostate cancer antigens that meet the appropriate criteria and represent an attractive target for antibody cancer therapy. It can be a target of anti-tumor CD8, or serve as a tool for vaccination as shown by xenograft models and phases I/II studies. STEAP1 could also serve as a new marker of tumor angiogenesis in lung cancer, indicate for occult residual tumor cells in patients with Ewing Sarcoma and also be used as a cell surface antigen for the development of breast cancer immunotherapy. The standardization of its evaluation as well as the validation through randomized trials would be necessary.

Philadelphia-positive (Ph+), BCR-ABL1, chronic myeloid leukemia (CML) is a model of leukemia driven by a single, specific, chromosome translocation, the t (9;22) (q22;q11). This translocation, leading to a new, hybrid, leukemia-specific gene (BCR-ABL1) encoding for a deregulated tyrosine-kinase protein (p210), drives the leukemic transformation of hematopoietic stem cells [1-6] and induces the progression of the disease from the early chronic phase (CP) to the late blastic phase BP) which close the natural history of the disease. In the 2000s, the introduction of Imatinib, the first tyrosinekinase inhibitor (TKI) able to target the protein p210, significantly changed the fate of CML to fatal disease in real chronic disease.

Aim: The study aims to assess the adequacy and appropriateness of the use of chemical prophylaxis/enoxaparin in total knee and/or hip replacement (TKR and THR) surgery patients at National University Hospital (NUH) with respect to the compliance/non-compliance to NUH Venous-Thromboembolism (VTE) Prophylaxis Guidelines. This is done with the objective to identify potential gaps in current prescribing patterns that may require interventions to improve clinical efficacy and safety outcomes.

Methodology: A retrospective drug utilization evaluation was performed for NUH patients aged≥18 years old who have undergone TKR and/or THR surgery from 1st January to 31st May 2013 and excluded foreigners not residing in Singapore. The study indicators included compliance of chemoprophylaxis/enoxaparin prescribing patterns to NUH guidelines. Efficacy and safety related clinical outcomes in terms of VTE and hemorrhagic events respectively in a 3-months follow-up period post-surgery were also measured.

Results: A total of 127 patients were available for evaluation but data for 82 patients were collected and analyzed. Chemoprophylaxis prescribing patterns for only 46 (56.1%) patients were compliant to NUH guidelines in terms of indication. The need for chemoprophylaxis exceeded bleeding risks for 55 (67.1%) patients but only 30 (36.6%) patients were given chemoprophylaxis (enoxaparin). When enoxaparin was prescribed, none of the dosing regimens were compliant to NUH guidelines in all aspects of dose and frequency, prophylaxis duration and time of first dose initiation. During the 3-months follow-up, no bleeding events due to enoxaparin occurred. 9 (11.0%) patients developed thrombosis, 2 of which considered as clinically significant by physicians.

Conclusion: The study revealed the baseline chemoprophylaxis and enoxaparin usage patterns in NUH TKR and THR patients. The adverse clinical outcomes that occurred identified potential safety gaps within the prescribing practices, for which recommendations were made to improve the safe and effective use of VTE chemoprophylaxis in NUH post-surgical orthopedic patients.

Antigen surface markers represent as the new prognostic tool for detection of acute leukemia. To investigate the prevalence expression of lymphoid and myeloid antigen lineage in acute leukemias. This study included 100 acute leukemias patients. Specimens were selected from consecutive patients who had sufficient material available. Among the 100 patients in which a detailed history, hematological, clinical and immunophenotyping analysis were performed. This study was showed distribution of immunophenotyping characters between studied AML and ALL cases. The most abundant immunophenotyping features in acute myeloid leukemia were cMPO, CD33, CD117, CD13, CD14 and CD64, while the most abundant immunophenotyping features in acute lymphoblastic leukemia were CD19, CD79a, TdT, CD20, CD10 and CD34. cMPO which act as independent prognostic factor for AML, CD10 and TdT can used as independent prognostic factor for differentiate between ALL and AML.

People often look for complex ways to solve our daily problems. We have visited many patients in clinic that resist to accept our simple advices and instead ask for more complex herbal or chemical medicines. On the other hand, there are many patients with chronic gastrointestinal (GI) and dermal problems who benefit from observing simple health advices. Chewing well, eating in peace, and avoiding simultaneous eating of some foods including meat, dairy products, fruits, and salads are considered as mainstay measures for maintaining health according to traditional Persian medicine (TPM). For instance, observing such recommendations is the initial stage in managing a serious health issue like infertility.

Background: Bladder urothelial carcinoma has high recurrence rate so patients with bladder urothelial carcinoma have to be monitored thoroughly for disease recurrence, this makes bladder cancer one of the most expensive cancer types for the health care system. Voided urine cytology is the most commonly used noninvasive follow-up diagnostic tool. The purpose of this study was to assess the role of voided urine cytology for the follow-up of patients with non-muscle invasive urothelial carcinoma (NMIUC). Materials and methods: In total, 200 patients were enrolled in this study. Both urine cytology and cystoscopy were done for each patient every three months and end-point of the surveillance was at 9 months after the first collection of urine and first cystoscopy done. In each time we correlate the findings of urine cytology and cystoscopy. Results: The sensitivity of voided urine cytology was low while the specificity of voided urine cytology was high after their estimation every three months for nine months. The positive predictive value of voided urine cytology was 0.867, while the negative predictive value was 0.919 at the end point of surveillance. None of the 180 patients with negative results of both cytology and cystoscopy developed recurrence for 6 months after initial diagnosis. Conclusion: Cytology has higher specificity than sensitivity especially for high grade urothelial carcinoma and high predictive value but it still was not sufficiently good to be a substitute for cystoscopy. However, follow-up cystoscopy may be temporarily postponed for a period of 6 month if urine cytology is negative.

The prognostic utility of the proliferation marker Ki67 to decide breast cancer treatment has been widely investigated and continues to be a source of much controversy. In this study, we evaluated: i) the role of Ki67 as a prognostic and predictive tool in patients with hormone receptor positive (ER+/PR+) carcinoma of the breast and ii) analyzed its correlation with two commonly used clinicopathological parameters, viz node status and tumor grade to predict clinical outcomes. To determine the clinical utility of Ki67 in assessment of breast cancer prognosis, we examined its expression by immunohistochemistry (IHC) in a series of 160 hormone receptor positive patients in a retrospective cohort of Indian patients. Patients were stratified based on Ki67 expression and analyzed for 5-year distant metastasis free survival (DMFS). Amongst the baseline clinicopathologic variables, we found node status, tumor grade, and age correlated significantly with outcome. However no significant correlation was found between Ki67 based risk stratification and patient outcome. Interestingly, increased Ki67 expression was found to correlate significantly with higher tumor grade but not with worse DMFS. In our study, conducted in an Indian cohort comprising 160 patients, Ki67 was not found to be significantly prognostic or predictive in patients with hormone receptor positive breast cancer.

PUFAs, from the stem bark extract of Alstonia boonei (SBEAB) was hypothesized to possess anti-inflammatory, antioxidant, anti-diabetic, pro-spermatogenic and hepatoprotective activities. The present study investigated the possible biochemical and molecular mechanisms underlying the hepatoprotective and testoprotective effects of SBEAB in diabetic rat. Biomarkers of hepatic and testicular damage, histological and immunohistochemical techniques were used. The expression of cyclooxygenase (COX-2) and inducible nitric oxide synthase (iNOS) were also estimated. SBEAB administered orally at dose of 100 mg/kg for 14 days significantly lowered the activities of serum transaminases and MDA levels induced by single intraperitoneal administration of streptozotoxin (STREP) (80 mg/kg) and preserved the integrity of both hepatocytes and spermatocytes. Also, SBEAB elevated the STREPinduced reduced activities of Δ5-17β-HSD and Δ5-17β-HSD with corresponding decrease in the activity of CAT. SBEAB inhibited the STREP induced expression of COX-2 and iNOS. The protective effect of SBEAB was compared to that of metaglomide (METAG), an established anti-diabetic drug. METAG treatment on hepatic damage was most efficacious in diabetic rats; followed by post and pre-treatment respectively while pre-and post-treatment were more efficacious on testicular damage than anti-diabetic drug. Furthermore, pre and post-treatment were more efficacious in preventing pro-inflammation and testicular cancer in diabetic rats than METAG-administration. We therefore concluded that the repression of genes encoding COX-2 and iNOS proteins by SBEAB validates the molecular basis of testicular protection and further suggests the links between the hepatocellular damage and male reproductive dysfunctions in diabetic individuals.

We describe two unique cases of fulminant mycosis fungoides with remarkably similar and aggressive clinical courses resulting in death. Both cases demonstrated ulcerated palmar and periorbital plaques and marked tissue eosinophilia, which was confirmed by T-cell receptor γ chain gene rearrangement studies to display identical monoclonality at temporally and anatomically distinct sites. Dense eosinophilic infiltrates on biopsy led to misdiagnosis of inflammatory dermatoses in both instances.

While mycosis fungoides may be challenging to diagnose histologically, the presence of eosinophils in progressive disease may herald a poor prognosis and should not exclude the diagnosis.