Isolation of cancer stem cells by selection for miR-302 expressing cells

Background Cancer stem cells are believed to be a major reason for long-term therapy failure because they are multi-drug resistant and able to rest mitotically inactive in the hypoxic center of tumors. Due to their variable number and their often low proliferation rate, cancer stem cells are difficult to purify in decent quantities and to grow in cell culture systems, where they are easily outcompeted by faster growing more ‘differentiated’, i.e., less stem cell-like tumor cells. Methods Here we present a proof of principle study based on the idea to select cancer stem cells by means of the expression of a stem cell-specific gene. A selectable egfp-neo coding sequence was inserted in the last exon of the non-coding murine miR-302 host gene. As a stem cell specific regulatory element, 2.1 kb of the genomic region immediately upstream of the miR-302 host gene transcription start site was used. Stable transgenic CJ7 embryonic stem cells were used to induce teratomas. Results After three weeks, tumors were removed for analysis and primary cultures were established. Stem cell-like cells were selected from these culture based on G418 selection. When the selection was removed, stem cell morphology and miR-302 expression were rapidly lost, indicating that it was not the original ES cells that had been isolated. Conclusions We show the possibility to use drug resistance expressed from a regulatory sequence of a stem cell-specific marker, to isolate and propagate cancer stem cells that otherwise might be hidden in the majority of tumor cells

A randomized, triple-blind, placebo-controlled clinical trial, evaluating the sesamin supplement effects on proteolytic enzymes, inflammatory markers, and clinical indices in women with rheumatoid arthritis

Inflammation is one of the main characteristics of rheumatoid arthritis. Based on the antiinflammatory properties of sesame, this study was conducted to evaluate the sesamin supplement effects on serum levels of some proteolytic enzymes, inflammatory biomarkers, and clinical indices in women with rheumatoid arthritis. In this randomized, triple‐blind, placebo‐controlled clinical trial, 44 patients were randomly divided in intervention and control groups. Patients received 200‐mg/day sesamin supplement or placebo in the intervention and control group for 6 weeks. Serum levels of proteolytic enzymes (hyaluronidase, aggrecanase, and matrix metalloproteinases‐3) and inflammatory biomarkers (hs‐CRP, IL‐1β, IL‐6, TNF‐α, and cyclooxygenase‐2) were measured with enzyme‐linked immunosorbent assay method at the beginning and end of the study. After intervention, serum levels of hyaluronidase and matrix metalloproteinases‐3 decreased significantly in sesamin group. Also, serum levels of hs‐CRP, TNF‐α, and cyclooxygenase‐2 in intervention group were significantly decreased in intervention group compared with placebo group. Sesamin supplementation also caused a significant reduction in the number of tender joints and severity of pain in these patients. According to the results, it seems that the sesamin by reducing inflammatory mediators can relieve clinical symptoms and pathological changes that caused by inflammatory impairment in patients with rheumatoid arthritis. KEYWORDS inflammatory factors, proteolytic enzymes, rheumatoid arthritis, sesami

A molecular analysis of desiccation tolerance mechanisms in the anhydrobiotic nematode Panagrolaimus superbus using expressed sequenced tags

<p>Abstract</p> <p>Background</p> <p>Some organisms can survive extreme desiccation by entering into a state of suspended animation known as anhydrobiosis. <it>Panagrolaimus superbus </it>is a free-living anhydrobiotic nematode that can survive rapid environmental desiccation. The mechanisms that <it>P. superbus </it>uses to combat the potentially lethal effects of cellular dehydration may include the constitutive and inducible expression of protective molecules, along with behavioural and/or morphological adaptations that slow the rate of cellular water loss. In addition, inducible repair and revival programmes may also be required for successful rehydration and recovery from anhydrobiosis.</p> <p>Results</p> <p>To identify constitutively expressed candidate anhydrobiotic genes we obtained 9,216 ESTs from an unstressed mixed stage population of <it>P. superbus</it>. We derived 4,009 unigenes from these ESTs. These unigene annotations and sequences can be accessed at <url>http://www.nematodes.org/nembase4/species_info.php?species=PSC</url>. We manually annotated a set of 187 constitutively expressed candidate anhydrobiotic genes from <it>P. superbus</it>. Notable among those is a putative lineage expansion of the <it>lea </it>(late embryogenesis abundant) gene family. The most abundantly expressed sequence was a member of the nematode specific <it>sxp/ral-2 </it>family that is highly expressed in parasitic nematodes and secreted onto the surface of the nematodes' cuticles. There were 2,059 novel unigenes (51.7% of the total), 149 of which are predicted to encode intrinsically disordered proteins lacking a fixed tertiary structure. One unigene may encode an exo-β-1,3-glucanase (GHF5 family), most similar to a sequence from <it>Phytophthora infestans</it>. GHF5 enzymes have been reported from several species of plant parasitic nematodes, with horizontal gene transfer (HGT) from bacteria proposed to explain their evolutionary origin. This <it>P. superbus </it>sequence represents another possible HGT event within the Nematoda. The expression of five of the 19 putative stress response genes tested was upregulated in response to desiccation. These were the antioxidants <it>glutathione peroxidase, dj-1 </it>and <it>1-Cys peroxiredoxin</it>, an <it>shsp </it>sequence and an <it>lea </it>gene.</p> <p>Conclusions</p> <p><it>P. superbus </it>appears to utilise a strategy of combined constitutive and inducible gene expression in preparation for entry into anhydrobiosis. The apparent lineage expansion of <it>lea </it>genes, together with their constitutive and inducible expression, suggests that LEA3 proteins are important components of the anhydrobiotic protection repertoire of <it>P. superbus</it>.</p

Deficiency of adenosine deaminase 2: a challenging differential diagnosis of polyarteritis nodosa

Deficiency of adenosine deaminase 2 (DADA2) is an autosomal recessive that was first described in 2014. It is a monogenic disease that is caused by loss-of-function variants in the ADA2 gene. DADA2 involves small- and medium-sized vessels and its clinical presentations include polyarteritis nodosa (PAN)-like features such as livedoid rash, early-onset stroke, hypogammaglobulinemia, hematological abnormalities, and systemic inflammation. Early diagnosis and treatment of DADA2 are crucial as the clinical features could be potentially life-threatening but might be treatable. The first-line treatment of choice in DADA2 is tumor necrosis factor (TNF)-inhibitors. We aimed to give an overview of the known pathophysiology, clinical presentations, diagnosis, and treatment of DADA2. A clearer knowledge of DADA2 may help to better diagnose, manage, and improve the clinical outcome of DADA2 patients. However, further studies are required to investigate the genotype-phenotype associations and exact pathophysiology of DADA2

The impact of intra-sacroiliac joint methylprednisolone injection in the recovery of patients with spondyloarthropathy: a randomized controlled trial

Introduction Spondyloarthropathies are a group of chronic inflammatory diseases with specific clinical symptoms in rheumatic diseases. These patients suffer from pain in the joints. Physicians have tried several ways to decrease the pain in these patients. This study aimed to evaluate the effect of intra-sacroiliac joint methylprednisolone injection under the guidance of ultra- sonography in spondyloarthropathy patients. Material and methods In this randomized control trial we studied 60 patients with spondyloarthropathy (30 patients in the intervention group and 30 patients in the control group) from January 2020 to December 2020. The intervention group patients received 40 mg of intra-sacroiliac joint (SIJ) methylprednisolone injection at the beginning in addition to treatment with nonsteroidal anti-inflammatory drugs (inflammatory dose) and sulfasalazine (2 to 3 g/day). Patients’ pain intensity and symptoms were assessed in the 2nd, 4th, 6th, and 8th weeks after glucocorticosteroid injection. Quantitative factors were compared by independent Student’s t-test. Data analysis was performed using SPSS version 22.0 software. A p-value < 0.005 was considered significant. Results There were no statistically significant differences in Visual Analogue Scale (VAS) and Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) criteria and finger-to-floor (FTF) levels in the intervention and control groups. There were significant differences in VAS and BASDAI criteria and FTF levels 2 weeks after the injection, and this difference remained the same until the end of the 8th week. The p-value was significant (p-value < 0.0001). Conclusions The sacroiliac joint methylprednisolone injection approach with ultrasound guidance seems to be effective in pain relief and function, and patient satisfaction scores. Additionally using the guidance of ultrasonography in this approach is without the risk of radiation exposure

Association of erythrocyte sedimentation rate and C-reactive protein and clinical findings with HLA-DQ8 allele in Rheumatoid Arthritis patients

Background ― Rheumatoid arthritis (RA) is an inflammatory, autoimmune disease induced by certain auto-antigens. HLA-DRB1*0401 allele has a significant relationship with RA incident. Additionally, DQβ1*0301, *302(DQ8), *303, and *304 can increase RA risk especially in DQA1*0301 and *302 coincident. Recent studies suggest that distribution of this allele is different in various populations Material and Methods ― 70 patients and 70 healthy controls were analyzed for human leukocyte antigen (HLA) allele by specific primer-polymerase chain reaction (SSP-PCR) method. Patients were evaluated in terms of ESR and CRP. Data analysis was performed in SPSS V.17. Results ― HLA-DQ8 allele was significantly more frequent in RA patients compared to control (P<0.0001). However, no significant relationship was observed between increased ESR (P=0.527), CRP (P=0.505), and mean counts of arthritic (P=0.691) and tender joints (P=0.669) among the patients who were carriers of HLA-DQ8. Conclusion ― There is a significant association between RA and HLA-DQ8 allele, this allele can increase susceptibility to RA. These findings might relate to the ethnical variations of RA patients but we couldn’t find a significant association between CRP and ESR with HLA-DQ8. We recommend to add specific inflammatory markers to CRP as well as assess ESR in larger sample sizes to obtain accurate results