Jornal de Biomarcadores Moleculares e Diagnóstico

A different number of modern molecular technologies, genomics and proteomics in applied areas of veterinary parasitology are increasing rapidly. To assess the molecular and genetic host parasitic interaction different advanced molecular tools are required and it is useful in the diagnosis of parasitic diseases. The important molecular tool for identification and diagnosis of parasites is polymerase chain reaction as well as there are other so many applications. In genomics, species-specific probes or primers are available along with conventional techniques. These techniques will give more information on the DNA sequences of parasites will reveal many more unique sequences which can be used for identification, diagnosis of parasitic infections. Genomics is crucial for the assessment of evolutionary biology, molecular epidemiology, physiology of parasites and vaccine development. These techniques may also help in the selection of genetically resistant hosts to parasite infection and discovery of new antiparasitic drugs, improvements to existing chemotherapeutic families and monitoring of drug resistance. From the last decade utilization of the aspects of molecular biology and their applications increased and it is essential to teach and train the veterinary parasitologists.…

Background: Substantial miss-rate of gastric cancer by conventianl endoscope was reported, indicating the importance of novel biomarkers for early detection of precancerous and cancerous lesions.

Objective: To identify potential biomarker molecules for early gastric cancer detection and evaluation of their application for molecular in vivo imaging.

Method: Besides the CEA-TAG+/- mouse model, 3 human and 2 murine gastric cancer cell lines were applied in this study. Cells lines were cultured at standard conditions. To determine the expression pattern of the targets, total RNA was extracted from cultured cells; cDNA was synthesised and qPCR was performed using spcefic primers. The murine stomach was removed and washed with PBS. The stomach was macroscopically categorized into tumor region and normal tissues. Histopathology and imunohistochemitry was performed in one half of the tumor tissues using specific antibodies. In the other half of the tumour tissue, RNA was extracted, reverse transcribed into cDNA and qPCR was performed. Histopathology, immhunohistochemistry and qPCR was performed for tumor-adjescent normal tissues to compare the difference in expression pattern of the target. Finally, a cathepsin activatable probe was injected into mice intravenously. Ex vivo fluorescent imaging was performed after 24 h of intravenous injection of a cathepsin activatable probe. Gene expression was quantified by standard method with normalizing CT values to the housekeeping gene (Cyclophilin A).

Results: Significantly elevated expression of cathepsins B, H and MMP2 in human and murine gastric cancer cell lines was detected. In addition, increased expression of cathepsins and MMPs was observed in murine primary gastric tumour specimens compared to the corresponding adjacent normal gastric mucosa (p<0.05, tumor versus normal mucosa). Accordingly, the NIRF probe was specifically activated in stomach tumor of CEA-TAG+/- mice and allowed ex vivo detection of the stomach tumor by Odyssey planar near-infrared scanner. Furthermore, immunohistochemical stainings with antibodies specific for cathepsins and MMPs indicated a stronger signal towards the tumor tissues compared to the adjacent normal mucosa.

Conclusion: Cathepsins and MMPs are potential biomarkers for detection of high grade dysplasia and early gastric cancer in mouse models, revealing the potential applicability of the biomarkers in tumor imaging and therapeutic monitoring.

Introduction: Kidney cancer is relatively rare when compered to other malignancies. It accounts for approximately 3% of adult malignancies. The renal clear cell carcinoma (RCCC) is the most common. In 70% of cases the disease is confined to the kidney and surgical approaches are generally curative. One third of cases is discovered in metastatic stage (30% of cases), specially bone and lung metastases; however, other rare metastases sites have also been described.

Case report: We report a fascinating case of kidney cancer with thyroid metastases, discovered fortuitously during the surgical management of multinodular goitre; in a 54 years-old-woman with a history of nephrectomy 9 years ago for renal neoplasm.

Conclusion: Thyroid metastases of kidney cancer are uncommon, it´s important for the surgeon and the oncologist to be able to recognize and differentiate thyroid metastases from primary tumours. The diagnosis can be suspected if the patient has a thyroid tumour and a past history of kidney cancer. These tumours, on the whole, tend to behave more aggressively and in most cases the use of multimodal therapy is recommended.

Background and objectives: Reactive oxygen species have been extensively studied in a wide range of human cancers, but are poorly studied in cutaneous malignant melanomas, particularly with respect to the lipid peroxidation product 4-hydroxy-2-nonenal. This was the reason why we chose to introduce this marker in this study, its use still being a novelty in these neoplasms. In addition, comparative analyzes of 4-hydroxy-2-nonenal expression in benign and malignant (primary or metastatic) human melanocytic lesions have not been performed so far.

Materials and methods: The immunohistochemical study was performed in a total of 55 patients, divided into a control group, which included 5 cases of simple nevi and 5 cases of dysplastic nevi, as well as a study group consisting of 35 cases of primary malignant melanoma and 10 metastases. The immunohistochemical method used to identify epitopes of interest was two-time, polymer-specific, with is characterized by high sensitivity, specificity and affinity.

Results: Immunostaining of the control and study groups with anti-4-HNE antibody included the analysis of the percentage of positive cells (with membranous/cytoplasmic staining and staining intensity for each type of melanocytic lesion analyzed (benign, dysplastic and malignant).

Conclusion: Oxidative stress-induced expression of the lipid peroxidation product 4-hydroxy-2-nonenal is significantly increased in dysplastic nevi versus benign nevi and is maintained at a level comparable to that in dysplastic lesions in cutaneous malignant melanomas. 4-hydroxy-2-nonenal intervenes early in the tumorigenic process of cutaneous malignant melanomas, being highly overexpressed once melanocytes have undergone dysplastic changes. Metastases lose the lipid peroxidation product, aspect correlated with the increased proliferative activity detected in metastases from cutaneous malignant melanoma.

Study background: In our environment, oral cancer is one of the most common lethal diseases encountered in dental practice. It is frequently diagnosed in the late stages because most patients present late in the course of the disease. This may be attributed to their low socioeconomic status, illiteracy, and some traditional beliefs in alternative native therapies. Some authors have reported on oral cancers specifically in their individual geographic settings; however, there is a paucity of reviews on Oral cancers generally in our environment. This study aims to review the prevalence, awareness and clinicopathologic patterns of oral cancers across the different geographic zones in Nigeria.

Methods: Information was sourced from journals, electronic data base such as Medline, Pubmed, Elsevier Science Direct and personal research work.

Result: Several prevalence rates have been reported in different geopolitical locations in our environment. Orofacial carcinomas were reported mostly in the older age groups while the Orofacial sarcomas were found in the slightly younger age groups. Squamous cell carcinoma was the predominant histopathological type. There is a low level of awareness of these lesions among the low socio-economic groups which makes them present late in our health care facilities hence a poor prognosis.

Conclusion: There is a need for increased awareness, advocacy and preventive care and early detection.

Memory and fear are two overlapping CNS processes in which mechanism of synaptic plasticity plays a key role. Fear increases likelihood fixation in memory behaviorally important information associated with a traumatic event. For healthy psychological functioning is crucial the ability to update the content and emotional charge of memories (cognitive plasticity) contributing to fading and extinction of fear memories once this event has been passed. Memory performance and its plasticity are associated with ability of synapses to change of number α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid glutamate receptors (AMPAR) on the post-synaptic membrane surface. AMPAR undergo exocytosis/endocytosis (AMPAR trafficking), thus changing the strength of neural connections (synaptic plasticity) and contributing to formation, maintaining and transformation of memories. Impairment of AMPAR trafficking may become a cause of deficiency of fear extinction, and development of posttraumatic stress disorder (PTSD). AMPAR trafficking is controlled by the nervous and the immune systems jointly. Kidney and brain expressed protein (KIBRA) and interleukin (IL) -1β are important regulators of AMPAR trafficking. Data show that genes encoding of these proteins (KIBRA-WWC1 gene and IL-1β gene) and their alleles mediates of stress impact on synaptic plasticity and are a bridge between memory and diseases. Disease susceptibility in humans is most commonly associated with single nucleotide polymorphisms (SNPs), when corresponding sequences of DNA from different individuals differ at one DNA base. PTSD is polygenic disorder, so can be identified the combination of allelic variations of SNPs, associated with strength and plasticity of memory that distinguish PTSD patients from the control. Our preliminary results showed that the combination of the certain allelic variants of WWC1, SNP rs17070145 (T/C), and the IL-1β gene, SNP rs16944 (A/G), is characteristic for PTSD compared to depressive and healthy groups. We assume that this combination might be used for pre-clinical diagnosis PTSD and in clinical practice.

Fishes that belong to the family Pangasiidae are widely recognized to have good potential for aquaculture and are highly valued as flesh food in the markets of Vietnam. However, there is much debate on the identification and phylogeny of the available species of Pangasiidae in the Asia. In the present study, nine species of two genera (Pangasianodon and Pangasius) of Pangasiidae were investigated using two partial sequences of the COI and cytochrome b mitochondrial genes to differentiate among them and study their phylogenetic relationships. A total of 42 haplotypes were identified (21 haplotssypes of each gene). The highest interspecies genetic distance was between Pangasius larnaudii and Pangasius bocourti (0.189) for COI and was between Pangasius macronema and Pangasianodon hypophthalmus (0.179) for cyt b. Whereas the lowest genetic distance was between Pangasius macronema and Pangasius conchophilus for both genes (0.65 for COI and 0.92 for cyt b, respectively). The phylogenetic tree analyses of two genes showed two major clusters that are genetically distant from the two genera. The results obtained in this study also show that beside COI gene, the cyt b gene region can be successfully used for differentiating between species and accepted as a standard region for DNA barcoding.

Advances in acute care medicine have increased the chances of survival for patients with severe illness or trauma. The major causes of modifiable and non-modifiable mortality among patients treated in medical or surgical intensive care units (ICUs) are trauma, sepsis, complications of diabetes mellitus type 2 and hypertension, respiratory support, CVA, electrolyte imbalance, poisoning and snake bite. Such analysis will give an insight into the various factors which led to death. The pre-hospital co-morbidities will reinforce the internist to anticipate and take appropriate measures to mitigate their onslaught. In this way, the mortality in the ICU may be curtailed.

Background: Atherosclerosis is leading to mortal diseases such as Cerebral Infarction (CI), and Cardiovascular Disease (CVD), and closely related to Diabetes Mellitus (DM) and Chronic Kidney Disease (CKD). Biomarkers are useful but not sufficient for early detection. In the present study, we aimed to identify novel antibody markers for atherosclerosis-related diseases.

Methods: The protein array method was used for the initial screening, and a peptide containing a possible epitope domain was used to evaluate serum antibody levels using the Amplified Luminescent Proximity Homogeneous Assay (AlphaLISA) method.

Results: The protein array identified prolylcarboxypeptidase (PRCP) as one of the target antigens recognized by IgG antibodies in the sera of patients with atherosclerosis. We then prepared a possible antigenic peptide of amino acids 214-227 of PRCP. Results of AlphaLISA showed significantly higher serum antibody levels against the PRCP peptide in patients with DM, CVD, acute-phase cerebral infarction, transient ischemic attack or CKD, than in healthy donors. Furthermore, areas under the receiver operating characteristic curves of these antibodies were higher in patients with DM or CKD than in other patients. Spearman correlation analysis revealed that the serum anti-PRCP antibody levels were associated with hypertension, artery stenosis, and smoking habit..

Conclusion: The serum anti-PRCP antibody may be useful for early detection of atherosclerosis-related diseases, and may have a pathogenic role in the development of atherosclerosis.

In spite of the interferon (INF) redundancy in treating HCV, ribavirin (RBV) is still included with the new direct antiviral therapies. Ribavirin-induced hematological alterations had been referred to ITPA gene polymorphisms.

Objective: Evaluate ITPA gene polymorphisms (rs1127354 and rs7270101) with development of anemia in chronic hepatitis C (CHC) Egyptian patients during treatment with pegylated-interferon (Peg-IFN) plus ribavirin (RBV).

Methods: 100 Egyptian CHC patients treated with PEG-IFN/RBV were recruited, 55 patients developed anemia (Hb decline>2 g/dl), and other 45 would not developed anemia (Hb decline ≤ 2 g/dl) at week 12 throughout the treatment course. Routine laboratory investigations were done for all participates (HCV-Abs, HBs Ag, HCV-RNA levels, complete blood picture, liver and kidney function tests. Single nucleotide polymorphism (SNP) was done by ABI TaqMan allelic discrimination kit for ITPA polymorphisms (rs1127354 and rs7270101).

Results: CC and AA were the most prevalent genotypes of SNPs rs1127354 and rs7270101 respectively among two studied groups. rs1127354 polymorphism was associated with Hb-decline at week 12 of treatment and with rs7270101 polymorphism for predicting platelet decline during treatment. Lower levels of platelet decline were detected with CC rs1127354 and AA rs7270101.

Conclusion: While ITPA polymorphisms rs1127354 CC genotype carried a predilection of anemia occurrence in PEG-IFN/RBV HCV treated subjects, the minor allele rs1127354 AA plays a crucial role in their protection. Platelet decline was reported in both ITPA rs1127354 and rs7270101 polymorphisms. Screening for ITPA polymorphisms in Egyptian HCV patients would be of value in avoiding hematological disturbances and dose modulations in RBVbased therapies.

Background: One of the strongest prognostic factor for breast cancer is the regional lymph node status, which can be evaluated intraoperatively by sentinel lymph node biopsy. Many methods (e.g. frozen section, touch imprint cytology (TIC) and one-step nucleic-acid amplification) are available to detect metastatic cells in axillary lymph nodes. The aim of this study is to evaluate the feasibility of using TIC to detect metastatic cells in sentinel lymph nodes.

Methods: This is a retrospective single center cohort analysis conducted on prospectively recorded data. The study included all patients admitted to the Department of General Surgery of Trieste University Hospital for invasive clinically node-negative breast cancer who underwent sentinel lymph node assessment by means of TIC.

Results: Between January 2015 and December 2016, 343 patients (338 females and 5 males) underwent breast surgery and sentinel lymph node TIC. Patient’s median age was 66 (33-92) years.

The sensitivity of TIC was 54% (95% C.I. 39% to 69%), whereas its specificity and accuracy were 99% (95% C.I. 98% to 100%) and 90%, respectively. The median time required to obtain the result was 20 (15-45) min. The overall cost per each TIC analysis was about 20.50€.

Conclusion: Touch imprint cytology appears to be a fast, cost-effective and reliable technique to intraoperatively detect breast cancer lymph node metastasis.