RESUMO
BACKGROUND: Primary thymic mucinous adenocarcinoma is an extremely rare and aggressive tumor with poor prognosis. The tumor may present as a heterogeneous solid or cystic mass accompanied by calcifications. However, clinical and radiologic features of the tumor are not well known due to the rarity of the disease, which makes accurate diagnosis difficult. CASE PRESENTATION: Here we present a rare case of primary thymic mucinous adenocarcinoma in the anterior mediastinum, including computed tomography (CT) and magnetic resonance imaging (MRI) findings. Chest computed tomography revealed a large anterior mediastinal mass with extensive calcifications with poor enhancement. MRI showed that anterior mediastinal mass showed intermediate signal intensity on T1-weighted images (T1WI), high SI on T2-weighted images (T2WI), and heterogeneous enhancement. Biopsy was performed and the anterior mediastinal tumor was diagnosed as thymic mucinous adenocarcinoma by histopathologic examination and immunohistochemical staining. CONCLUSION: Thymic mucinous adenocarcinomas could be included in differential diagnoses of anterior mediastinal tumors showing extensive calcification, and common imaging findings of mucinous adenocarcinoma such as T2 high signal intensity and heterogeneous enhancement on MRI may be helpful to diagnose thymic mucinous adenocarcinoma.
Assuntos
Adenocarcinoma Mucinoso , Humanos , Adenocarcinoma Mucinoso/diagnóstico por imagem , Adenocarcinoma Mucinoso/patologia , Diagnóstico Diferencial , Tomografia Computadorizada por Raios X , Imageamento por Ressonância MagnéticaRESUMO
BACKGROUND: Palifermin (trade name Kepivance®) is an amino-terminally truncated recombinant human keratinocyte growth factor 1 (KGF-1) with 140 residues that has been produced using Escherichia coli to prevent and treat oral mucositis following radiation or chemotherapy. In this study, an amino-terminally shortened KGF-1 variant with 135 residues was produced and purified in E. coli, and its cell proliferation activity was evaluated. RESULTS: We expressed soluble KGF-1 fused to thioredoxin (TRX) in the cytoplasmic fraction of E. coli to improve its production yield. However, three N-truncated forms (KGF-1 with 140, 138, and 135 residues) were observed after the removal of the TRX protein from the fusion form by cleavage of the human enterokinase light chain C112S (hEKL C112S). The shortest KGF-1 variant, with 135 residues, was expressed by fusion with TRX via the hEKL cleavage site in E. coli and purified at high purity (> 99%). Circular dichroism spectroscopy shows that purified KGF-1135 had a structure similar to that of the KGF-1140 as a random coiled form, and MCF-7 cell proliferation assays demonstrate its biological activity. CONCLUSIONS: We identified variations in N-terminus-truncated KGF-1 and selected the most stable form. Furthermore, by a simple two-step purification, highly purified KGF-1135 was obtained that showed biological activity. These results demonstrate that KGF-1135 may be considered an alternative protein to KGF-1.
Assuntos
Escherichia coli , Fator 7 de Crescimento de Fibroblastos , Humanos , Fator 7 de Crescimento de Fibroblastos/genética , Fator 7 de Crescimento de Fibroblastos/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismoRESUMO
Recombinant human keratinocyte growth factor 2 (KGF-2), also known as repifermin, is used in various therapeutic applications. However, KGF-2 production has not been optimized for facilitating large-scale production. Therefore, we attempted to attain high-level production of bioactive KGF-2. KGF-2 was fused with 6HFh8 (6HFh8-KGF-2) at the tobacco etch virus protease cleavage site. The 6HFh8-KGF-2 was expressed in Escherichia coli with high expression levels of approximately 33% and 20% of soluble protein in flask culture and 5 L fermentation, respectively. 6HFh8-KGF-2 was purified via nickel affinity chromatography. To maintain a stable form of KGF-2, the conditions of the cleavage reaction were optimized based on the isoelectric point. KGF-2 was purified via ion-exchange chromatography to high purity (>99%) with an optimal purification yield (91%). Circular dichroism spectroscopy demonstrated that purified KGF-2 had a secondary structure and thermal stability similar to that of commercial KGF-2. Bioactivity assays indicated that purified KGF-2 could induce MCF-7 cell proliferation in the same manner as commercial KGF-2. These results demonstrate that bioactive KGF-2 was overexpressed in E. coli and purified to high quality. Our findings indicated that bioactive KGF-2 can be produced in large quantities in E. coli.
Assuntos
Escherichia coli , Humanos , Escherichia coli/genética , Escherichia coli/metabolismo , Fator 10 de Crescimento de Fibroblastos/metabolismo , Células MCF-7 , FermentaçãoRESUMO
BACKGROUND/AIM: Poorly differentiated thyroid carcinoma (PDTC), anaplastic thyroid carcinoma (ATC), and advanced DTC have poor outcomes. MATERIALS AND METHODS: We performed next-generation sequencing in nine selected aggressive thyroid cancers. RESULTS: Among the nine patients, the driver gene mutations BRAF V600E (3/9) and NRAS Q61K (1/9) were detected. Other oncogenic mutations included ERBB2 (1/9) and CDK4 (1/9). Telomerase reverse transcriptase (TERT) promoter mutation was found in five cases. Among tumor suppressor genes, mutations in TP53 (3/9), ARID1A (1/9), APC (1/9), MEN1 (1/9), DICER1 (1/9), and MED12 (1/9) were identified. RET fusions were found in two cases, one with PTDC and the other with ATC. The ATC with RET fusion also harbored TP53 and TERT promoter mutations. None of the PDTC cases had BRAF or RAS gene alterations. CONCLUSION: Since genetic alterations with therapeutic and prognostic implications were detected using next-generation sequencing, this technique is recommended to be performed for patients with aggressive thyroid cancer.
Assuntos
Telomerase , Carcinoma Anaplásico da Tireoide , Neoplasias da Glândula Tireoide , RNA Helicases DEAD-box , Genômica , Humanos , Mutação , Proteínas Proto-Oncogênicas B-raf/genética , Ribonuclease III , Telomerase/genética , Carcinoma Anaplásico da Tireoide/genética , Neoplasias da Glândula Tireoide/genéticaRESUMO
BACKGROUND: To evaluate the short-term outcomes of switching to ranibizumab in aflibercept-resistant polypoidal choroidal vasculopathy (PCV). METHODS: This retrospective study included 18 eyes diagnosed with aflibercept-resistant PCV. All patients were treated with two to four consecutive ranibizumab injections at 4-5-week intervals. The best-corrected visual acuity (BCVA), and central retinal thickness (CRT) values before and after switching to ranibizumab were compared. The proportion of eyes showing ≥100 µm decrease in retinal thickness and/or complete resolution of fluid after switching was identified. RESULTS: The mean number of aflibercept injections before switching was 5.7 ± 3.3. After switching, a mean of 2.8 ± 0.6 consecutive ranibizumab injections was performed. The mean logarithm of minimal angle of resolution (logMAR) BCVA was 0.41 ± 0.26 (Snellen equivalents = 20/51) before switching, and 0.40 ± 0.30 (20/50) after switching (p = 0.574). The mean CRT was 422.2 ± 152.4 µm before switching, and 400.7 ± 182.0 µm after switching (p = 0.236). A decrease in CRT of ≥100 µm, and/or complete resolution of fluid was noted in three eyes (16.7%). CONCLUSIONS: Switching to ranibizumab in aflibercept-resistant polypoidal choroidal vasculopathy was not effective in most patients, suggesting the need for further investigation to seek more effective treatment options for this condition.
RESUMO
BACKGROUND: This study aimed to investigate the trend of selecting ranibizumab and aflibercept for the initial treatment of neovascular age-related macular degeneration (AMD) and polypoidal choroidal vasculopathy (PCV). METHODS: This was a retrospective study that included 460 patients who were diagnosed with treatment-naïve neovascular AMD and PCV and were initially treated with either ranibizumab or aflibercept. The patients were divided into two groups: the ranibizumab group (n = 96) and the aflibercept group (n = 324). The patients' characteristics and the proportion of the subtypes of macular neovascularization (MNV) were compared between the two groups. RESULTS: Patients in the ranibizumab group were significantly older (mean 74.3 ± 8.4 years) than those in the aflibercept group (mean 70.4 ± 8.8 years; p < 0.001). In the ranibizumab group, the proportions of type 1 or 2 MNV, type 3 MNV, and PCV were 50.0%, 27.1%, and 22.9%, respectively. In the aflibercept group, the proportions were 35.2%, 6.8%, and 58.0%, respectively. There was a significant difference in the proportion of MNV subtypes between the ranibizumab and aflibercept groups (p < 0.001). Ranibizumab was used in 54.2% of patients with type 3 MNVs. However, in patients with PCV, aflibercept was used in 89.5% of patients. CONCLUSIONS: Ranibizumab was preferred as an initial treatment agent in older patients and those with type 3 MNV, whereas aflibercept was highly preferred in patients with PCV. The different characteristics and efficacy of the two agents may have partially contributed to this trend.