Objective: The skin pathergy test (SPT) is an important tool in the diagnosis of Behçet disease (BD), but its decreasing sensitivity over years has resulted in less frequent use in the clinical practice. This study aimed to improve the sensitivity of the SPT without compromising its specificity. Methods: BD patients, patients with other inflammatory diseases, recurrent aphthous stomatitis, and healthy controls comprised the study group. The SPT was conducted using 20G needle and 21G lancet pricks, or with additional application of 23-valent polysaccharide pneumococcal vaccine (PS-23), Alum, or ATP to the prick site. Development of erythema and induration at 24 h/48 h were evaluated by the same observer. Induration (≥2 mm) with erythema at 48 h was accepted as a positive reaction. Proinflammatory cytokine production following stimulation with lipopolysaccharide or PS-23 was investigated by whole-blood assay (WBA) in a subgroup. Results: Stimulation of the forearm skin by PS-23 and a 20G needle prick showed the highest sensitivity and specificity in BD (64.3% and 100%, respectively), especially in patients with active disease (80.3% and 100%, respectively), compared with a sensitivity of 4.8% in all and 6.1% in active patients using a single 20G prick. A positive result was associated with active disease and no use of immunosuppressives. In WBA, increased IL-1β and IL-1Ra production in response to PS-23 was observed in the group with active BD, while the cytokine response to lipopolysaccharide was similar in all groups. Conclusions: The SPT conducted using a 20G needle prick and PS-23 antigens was shown to be a promising tool for the diagnosis of BD owing to its improved sensitivity compared with the standard approach.

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive disease resulting from the accumulation of genetic changes that affect the development of T-cells. The precise role of lymphoid enhancer-binding factor 1 (LEF1) in T-ALL has been controversial since both overexpression and inactivating LEF1 mutations have been reported to date. Here, we investigate the potential gene targets of LEF1 in the Jurkat human T-cell leukemia cell line. We used small interfering RNA (siRNA) technology to knock down LEF1 in Jurkat cells and then compared the gene expression levels in the LEF1 knockdown cells with nontargeting siRNA-transfected and non-transfected cells by employing microarray analysis. We identified DHRS2, a tumor suppressor gene, as the most significantly downregulated gene in LEF1 knockdown cells, and we further confirmed its downregulation by real-time quantitative polymerase chain reaction (qRT-PCR) in mRNA and at protein level by western blotting. Our results revealed that DHRS2 is positively regulated by LEF1 in Jurkat cells, which indicates the capability of LEF1 as a tumor suppressor and, together with previous reports, suggests that LEF1 exhibits a regulatory role in T-ALL via not only its oncogenic targets but also tumor suppressor genes.

Objectives: The role of neutrophil extracellular traps (NETs) in IgA vasculitis (IgAV) pathogenesis is emerging, with NETosis-associated markers potentially linked to disease activity. This study aimed to explore the relationship between NETosis biomarkers and IgAV disease phases. Methods: A longitudinal study involving 33 paediatric IgAV patients and 26 healthy controls was conducted. Blood and urine samples were collected from healthy controls and patients during active and inactive disease phases. NETosis markers, including cell-free DNA (cf-DNA), neutrophil elastase, MPO and citrullinated histone H3 (cit-H3) were measured using ELISA kits. Statistical analyses were conducted to compare differences for NETosis markers between groups and to evaluate correlations among variables using appropriate statistical tests. Results: There was no significant difference in gender and age between the patient and control groups. The serum cf-DNA level was significantly higher in the active patient group compared with the control and inactive patient groups (P = 0.04; P = 0.04, respectively). In urine, MPO levels were significantly lower in the active phase of patients than controls (P = 0.009), while cit-H3 levels were higher in both active and inactive phases compared with controls (P = 0.01 and P = 0.03, respectively). A cf-DNA threshold of 935 ng/ml was identified, which achieved a sensitivity of 93% (correctly identifying 93% of active patients) and a specificity of 72% (correctly identifying 72% of healthy controls). Conclusion: Elevated serum cf-DNA and urine cit-H3 suggest a potential role for NETosis in IgAV activity, highlighting these markers as potential indicators for disease monitoring. Further studies are warranted to establish standardized protocols for NETosis marker assessment in IgAV.

Resumen: Objectivos: La leucemia mieloide aguda (AML, por sus siglas en inglés) es una enfermedad muy heterogénea. Aunque se puede clasificar a los pacientes en grupos de riesgo según sus mutaciones genéticas, el pronóstico dentro de cada categoría varía sustancialmente. Es perentorio identificar nuevos marcadores moleculares de la AML. Recientemente, se ha descrito la elevación del inhibidor de la serina peptidasa Kazal tipo 2 (SPINK2) en la AML, habiendo sido relacionada con peores resultados clínicos en metaanálisis, así como en un número limitado de pacientes con AML. Métodos: Analizamos la expresión de SPINK2 en 62 pacientes (45 adultos y 17 niños) con AML y en 11 líneas celulares mediante PCR cuantitativa (qRT-PCR). Los niveles de la proteína SPINK2 se determinaron en líneas celulares mediante ELISA. Resultados: Observamos un aumento de expresión del ARNm de SPINK2 y de los niveles de la proteína en las líneas celulares de AML (HL60 y NB4), frente a otras líneas celulares (K562, Jurkat y NALM6, MCF7, HeLa, HUVEC, hFOB, 293T, U87). Los pacientes con AML mostraron una expresión elevada de ARNm de SPINK2 frente a los controles (p=0,004) y esta fue significativamente menor en los pacientes t(8;21) positivos, frente a los pacientes negativos (p=0,0006). Conclusions: Estos resultados sugieren que el gen SPINK2 tiene un papel relevante en el desarrollo de la AML. Son necesarios más estudios para evaluar la expresión de SPINK2 en los pacientes con AML con la mutación t(8.21) e investigar su valor pronóstico en varios subgrupos de pacientes con AML.

Amaç: Bortezomib, multipl miyelom tedavisinde yaygın olarak kullanılan önemli bir proteazom inhibitörüdür; ancak kazanılmış direnç, ilacın uzun dönem klinik etkinliğini sınırlandırmaktadır. MikroRNA’lar apoptoz, stres yanıtı, hücresel adaptasyon ve ilaç direnci süreçlerinde rol oynayan önemli post-transkripsiyonel düzenleyicilerdir. Bu çalışmada, multipl miyelom hücre modellerinde bortezomib direnci ile ilişkili olabilecek seçilmiş mikroRNA’ların ekspresyon profillerinin araştırılması amaçlandı. Yöntem: Bortezomib dirençli U266 ve RPMI-8226 multipl miyelom hücreleri, artan konsantrasyonlarda bortezomibe sürekli maruz bırakılarak oluşturuldu. Dirençli ve kontrol hücrelerinde miR-25, miR-30c, miR-34a, miR-34c, miR-200b, miR-200c, miR-326, miR-429, miR-449a, miR-449b, miR-499a, miR-582 ve miR-708 ekspresyon düzeyleri kantitatif gerçek zamanlı PCR yöntemi ile analiz edildi. Endojen kontrol olarak U6 snRNA kullanıldı ve relatif ekspresyon düzeyleri 2⁻ΔΔCt yöntemi ile hesaplandı. Bulgular: Bortezomib dirençli U266 ve RPMI-8226 hücrelerinde hem ortak hem de hücre hattına özgü mikroRNA ekspresyon değişiklikleri gözlendi. miR-34a, miR-34c, miR-326, miR-429, miR-449a ve miR-449b her iki dirençli hücre hattında da anlamlı olarak yüksek eksprese edildi. miR-25, miR-200c ve miR-708 ekspresyonları yalnızca dirençli RPMI-8226 hücrelerinde arttı. Buna karşılık, miR-200b, miR-499a ve miR-582 iki dirençli model arasında zıt ekspresyon paternleri sergilerken, miR-30c ekspresyonunda değişiklik saptanmadı. Sonuç: Multipl miyelomda kazanılmış bortezomib direnci, hem ortak hem de hücre hattına özgü mikroRNA değişiklikleri ile ilişkilidir. miR-34, miR-326 ve miR-449 aile üyelerinin her iki dirençli hücre hattında tutarlı şekilde artmış olması, bu mikroRNA’ların proteazom inhibisyonuna karşı gelişen adaptif yanıtlarda rol oynayabileceğini düşündürmektedir. Diğer mikroRNA’larda gözlenen farklı ekspresyon paternleri ise direnç mekanizmalarının heterojen yapısına işaret etmektedir. Bu mikroRNA’ların bortezomib direncindeki fonksiyonel rollerinin aydınlatılması için ileri çalışmalara ihtiyaç vardır.

Breast cancer remains a leading cause of cancer-related mortality worldwide, with epigenetic mechanisms like N⁶ methyladenosine (m⁶A) modification playing a crucial role in tumorigenesis. The interaction between microRNAs and m⁶A regulators, such as the methyltransferase METTL14, is increasingly recognized as a key pathway in oncogenesis. This study investigated whether miR-30c-2-3p regulates METTL14 expression to influence global m⁶A levels and cell migration in breast epithelial (MCF12A) and breast cancer (MCF7) cell lines. Following transfection with miR-30c-2-3p mimics, successful overexpression was confirmed in both cell lines. Subsequent RT-qPCR and Western blotting analyses demonstrated that METTL14 mRNA and protein levels were significantly reduced at 24 and 48 h post-transfection (p < 0.05). Concurrently, global m⁶A RNA methylation levels decreased, with a more pronounced reduction observed in MCF12A cells (p < 0.001). Functionally, wound healing assays revealed that miR-30c-2-3p significantly inhibited migration, reducing wound closure by 30–44% in MCF7 cells and by 66–72% in MCF12A cells. These findings reveal a novel regulatory axis involving miR-30c-2-3p, METTL14, and m⁶A, suggesting that miR-30c-2-3p functions as a tumor suppressor and represents a promising biomarker and therapeutic target in breast cancer.