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Revista de diagnóstico y biomarcadores moleculares

Volumen 8, Asunto 4 (2017)

Artículo de investigación

Stem Cell Organoids in Primary Cultures of Human Non-Malignant and Malignant Colon

Sahrish Tariq, Muhammad Tahseen, Mariam Hassan, Muhammad Adnan Masood, Shahid Khattak, Aamir Ali Syed, Asad Hayat Ahmad, Mudassar Hussain, Muhammed Aasim Yusuf, Chris Sutton, and Saira Saleem

Aims: A sub-population of cells named cancer stem cells (CSCs) that initiate and promote tumour growth have been demonstrated to exist in several malignancies including colon carcinoma. The objective of our pilot study was to isolate CD133+CD26+CD44+ CSCs from patient colon tumours, culture spheres or organoids and observe their proliferation in primary cultures. Parallel cultures of non-cancer controls from colon normal lining and nonadenomatous polyps were set up.

Methods: Magnetic activated cell sorting was used to isolate CD133+CD26+CD44+ cell populations followed by primary cell culturing under stem cell culture conditions. Number, cells/organoid and daughter generations of organoids were calculated using phase contrast microscope. Trypan blue exclusion method was used to test the viability of the cells.

Results: Both colon tumour and colon non-adenomatous polyp formed floating organoids in suspension; however non-adenomatous polyp cultures did not show self-renewal properties for more than 1 passage. Normal colon singlecell suspension did not create organoids. Metastatic colon tumours rapidly produce cancer cell organoids in less than 24 hours in larger numbers compared to non-metastatic colon tumours (1-3 weeks). Metastatic colon tumour organoids have the ability for proliferation for upto five daughter generations in primary culture compared to three generations for those grown from non-metastatic tumours.

Conclusions: This in vitro CSC organoid model will help study colon cancer biology, in particular providing a valuable source of primary cell-derived tissue for studying personalized molecular profiling using ‘omics strategies to direct therapeutic intervention.

Artículo de investigación

Study of Association Between the Polymorphism of MICA Gene (met 129 val) and Ankylosing Spondylitis in a Sample of the Population of Western Algeria

Noria Bouras, Ahmed Benzaoui, Ibtissem Messal, Nadjet Boushaba, and Abdallah Boudjema

Ankylosing spondylitis (AS) is a chronic inflammatory rheumatism characterized by a predominantly axial (spinal) localization, followed by joint damage and enthesis. The AS may be associated with other conditions such as reactive arthritis, psoriatic arthritis, chronic inflammation of the intestine and pulmonary manifestation. In this case, we speak of Spondylarthroparies (SPA). The etiology of AS is not well known, however, genetic factors as well as environmental factors can play a very important role in the onset of the disease. Although several genes appear to be associated with SA, the concept of genetic ground relies heavily on the association with HLA-B27 specificity. Recently, the MICA gene has aroused the interest of several studies of associations MICA and autoimmune diseases (MAI), in particular, the association MICA and SA. In our work, we are interested in the polymorphism existing at exon 3 and which encodes the α2 domain of the MICA protein. This polymorphism has the position 129 either the methionine (met) allele or the valine (val) allele. MICA proteins with methionine residue at position 129 react strongly with its NKG2D receptors found on the surface of NK and LT, thereby increasing the cytotoxicity threshold. In contrast, MICA proteins that have a valine residue at the same position have a low affinity with NKG2D. This weakens the threshold of cytotoxicity. The aim of our work is to look for the associations between the polymorphism MICA met129val and the SA in a sample of 90 cases suffering from the SA and 78 controls, within the population of western Algeria. We then tested the effect of this polymorphism on HLA-B27 status. Finally, we looked for a possible association between the MICA-129 genotyping and the early onset of SA. The results show that the MICA-129met allele is strongly associated with SA in patients compared to controls since it is found at an allelic frequency of 0.54 vs 0.30 (p=11.10-6). On the other hand, the MICA-129val allele is strongly found in the controls than in the cases, with a frequency of 0.70 vs 0.46 (p=11.10-4). However, the polymorphism MICA met129val showed no synergistic or independent effect on the distribution of HLA-B27 specificity in either cases or patients. It is interesting in this case to study other interactions between this polymorphism and other genes or alleles already associated with SA. In our work, we could not determine the effect of MICA-129 genotypes on the early onset of the disease, as reported by another similar study carried out in an Algerian population.

Comunicación corta

Favourable/Intermediate ELN-Risk Acute Myeloid Leukemia to Transplant or Not to Transplant First-Line?

Michele Malagola, Federica Cattina, Valeria Cancelli, Benedetta Rambaldi, Enrico Morello, Nicola Polverelli, Alesssandro Turra, Simona Bernardi and Domenico Russo

Favourable/intermediate ELN-risk acute myeloid leukemias (AMLs) (e.g. those harboring t(8;21) or inv(16) or NPM1A mutations or CEBP-alpha bi-allelic mutations) account for 30% to 50% of all newly diagnosed AMLs [1,2]. In this setting, conventional induction treatments may induce complete remission (CR) in up to 70% to 80%, but relapses still occur in 40% to 50% of cases and, at the end, no more than 30% to 40% of patients can be cured. Therefore, the optimization of post- remission therapy represents the greatest challenge in the treatment of favourable/intermediate-I ELN-risk AML.

Artículo de investigación

Effect of High-Intensity Laser Treatments on Chronic Pain Related to Osteoarthritis in Former Professional Athletes: A Case Series

Paul F White, Xuezhao Cao, Loani Elvir-Lazo and Hector Hernandez

Background: We evaluated the use of a new higher intensity 42 Watt cold laser for treating chronic pain related to osteoarthritis (OA) in former NFL football players.

Methods: 39 consenting former NFL football players with OA underwent 1-3 treatment sessions lasting 10-20 min with a 42 Watt FDA-approved high-intensity cold laser (Phoenix thera-lase, Dallas, TX) at a wavelength of 1275 nm. We recorded their pain verbal rating scale (VRS) score at rest and with activity before and after each treatment using an 11-point VRS with 0=no pain to 10=worst pain imaginable. In addition, we assessed the duration of the painrelieving effect produced by each laser treatment, as well as its effects on other OA-related symptoms.

Results: The chronic pain scores were significantly reduced both at rest and with activity after each treatment. Baseline VRS pain scores were 3.5 ± 2.9 at rest and 6.0 ± 2.6 with activity. After the initial treatment, the pain scores were reduced to 1.2 ± 1.8 (p<0.01) at rest and to 2.0 ± 2.0 (p<0.01) with activity. The overall beneficial effect was 7.2 ± 1.8 on a scale from 0=no relief to 10=complete relief, and the duration of the beneficial effect lasted 1-3 weeks in 64% of the players treated. Finally, 90% of the players would recommend the laser treatment to their colleagues.

Conclusion: High-intensity cold laser treatments reduced chronic OA-related pain in former NFL football players by ~67% at rest and with activity and the beneficial effect typically persisted for 1 week or longer after 1-3 treatments in the majority of these chronic pain patients.

Artículo de investigación

Comparative Immunohistochemical Study of P63, SMA, CD10 and Calponin in Distinguishing In Situ from Invasive Breast Carcinoma

Dina M Abdallah, and Nevine MF El Deeb

Background: Loss of the outer myoepithelial layer is the hallmark of invasive carcinoma, and demonstration of this loss can be documented by immunohistochemical techniques. The purpose of this study was to compare the specificity and sensitivity of four of the most commonly used-markers of myoepithelial cells: P63, SMA, CD10 and Calponin in distinguishing in situ from invasive breast carcinoma.

Material and methods: Immunostaining using antibodies against P63, SMA, CD10 and Calponin was performed on representative paraffin sections from 40 cases of breast masses examined at the Department of Pathology, Alexandria Faculty of Medicine, and diagnosed as ductal carcinoma in situ ± an invasive ductal or lobular carcinoma.

Results: Calponin yielded a slightly higher sensitivity than each of P63, CD10 and SMA (65% vs 54%, 19% and 17%, respectively). The results of both semiquantitative assessment and computerized image analysis of immunohistochemically-stained sections were statistically correlated, statistically p63 showed the highest specificity for myoepithelium and the least expression in non-myoepithelial layer of all antibodies tested. In contrast, SMA showed the least specificity and highest non-myoepithelial expression especially in stromal myofibroblasts and in vascular smooth muscle cells.

Conclusions: Calponin and P63 are more sensitive myoepithelial markers as compared to CD10 and SMA; with Calponin slightly more sensitive than P63. SMA should not be used alone as a myoepithelial marker due to its low specificity.

Artículo de investigación

Influence of Chronic Recurrent Multifocal Osteomyelitis (CRMO) On Densitometry Measurements Obtained by Dual X-Ray Absorptiometry

Danny L Costantini, Reza Vali Eman Marie, Mandy Kohli, Holly Convery, Martin Charron, Amer Shammas and Ronald M Laxer

The incidence of detecting focal chronic recurrent multifocal osteomyelitis (CRMO) lesions on dual-energy x-ray absorptiometry (DXA), and the effect of these lesions on DXA bone mineral density (BMD), bone mineral content (BMC), and their associated Z-scores were retrospectively reviewed. Materials and Methods. The study included 22 patients (14 females, 8 males; median age of 13 years) with CRMO and in whom a total body less head (TBLH) and lumbar spine DXA scan had been obtained. Whole-body bone scintigraphy and MRI were used as the reference standards. Sites involved with CRMO were subsequently detected and DXA measurements were re-measured after removing the sclerotic lesions from the analysis. Results. In total, sclerotic CRMO lesions were detected in 15 of the 22 patients (68%) by DXA, although the number of lesions detected (on a per-lesion analysis) was much less (i.e. 29 of 129 lesions; 19.4%) when compared to MRI and/or bone scintigraphy. Larger lesions had a greater impact on the derived BMD/BMC measurements, and changed the diagnosis in one patient from having normal to abnormal DXA results based on the final Z-score. Discussion. CRMO lesions detected on DXA examinations should be regarded as a potential source of error. Careful inspection and re-quantification of the BMD, BMC and associated Z-score after applying an appropriate correction should be considered in patients with large CRMO lesions identified on DXA examinations.

Reporte de un caso

Slurred Speech Spells Trouble

Hilary Wyllie, Carlo Canepa and Titilopemi Oladosu

An 86-year-old lady was referred to the TIA clinic with recurrent episodes of slurred speech, disorientation and flashing lights in her vision. She was known to have difficult to control hypertension and atrial fibrillation, but had been stable on warfarin for many years with no previous vascular events or bleeding episodes. MRI imaging showed a large number of cerebral microbleeds in a lobar distribution, and also 2 small acute subcortical infarcts in the right frontal lobe. The appearances overall were suggestive of cerebral amyloid angiopathy. Her recurrent and stereotyped symptoms were felt to be more in keeping with transient focal neurological events (TFNE) related to cerebral amyloid angiopathy rather than TIAs. However, this left us with a dilemma as to whether to stop or continue warfarin treatment in the light of her acute infarcts and established atrial fibrillation.

Reporte de un caso

Paratesticular Myxoid Liposarcoma: A Case Report of Unusual Age of Presentation

Khaleel Al-Obaidy, Eman Abdulfatah*, Fulvio Lonardo and Rafic Beydoun

Paratesticular liposarcomas are relatively common sarcomas in the paratesticular region, however, the myxoid variant; myxoid liposarcoma (MLS) is considered very rare. MLS peaks around the fourth-fifth decade. Herein, we describe a case of paratesticular MLS occurring in a 19-year-old African American male who underwent resection of a slowly growing testicular mass over a period of one and a half years that was considered initially by his primary care physician to be a benign lesion. The resection margins were positive for tumor and the patient developed local recurrence one year after the initial resection. An extremely unusual age of presentation along with the importance of margin status in tumor recurrence is highlighted.

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