ABSTRACT
Delivery of small interfering RNAs (siRNAs) into primary T cells is quite challenging because they are non-proliferating cells and are difficult to transfect with non-viral approaches. Because sonoporation is independent of the proliferation status of cells and siRNA acts in the cell cytoplasm, we investigated whether sonoporation could be used to deliver siRNA into mouse and human T cells. Cells mixed with Definity microbubbles and siRNA were sonicated with a non-focused transducer of center frequency 2.20 MHz producing ultrasound at a 10% duty cycle, pulse repetition frequency of 2.20 kHz and spatial average temporal average ultrasound intensity of 1.29 W/cm2 for 5 s and then examined for siRNA fluorescence by flow cytometry analysis. These sonoporation conditions resulted in high-efficiency transfection of siRNA in mouse and human T cells. Further, the efficacy of siRNA delivery by sonoporation was illustrated by the successful visualization of decreased methylation-controlled J protein expression in mouse and human CD8 T cells via Western blot analysis. The results provide the first evidence that sonoporation is a novel approach to delivery of siRNA into fresh isolated mouse and human T cells in vitro, and might be used for in vivo studies in the future.
Subject(s)
RNA, Small Interfering/administration & dosage , Sonication/methods , T-Lymphocytes , Animals , Blotting, Western , Flow Cytometry , Humans , In Vitro Techniques , Mice , Models, AnimalABSTRACT
BACKGROUND: Pigmented lesions of the eye represent a challenging field from both clinical and pathologic approaches. The importance of a correct diagnosis in this case is enhanced by the delicate structures involved and their difficult accessibility. CASE: We report a case of a 51-year-old man with acute vision loss and the feeling of a foreign body in the left eye diagnosed with vitreous hemorrhage. A small, pigmented lesion in the proximity of optic nerve head was detected. After a 9-year follow-up, a definitive diagnosis of melanocytoma was achieved in vitrectomy cytology. Smears showed a population of plump, cohesive melanocytes and elongated cells with cytoplasmic melanin and uniform rounded nuclei. No change in the lesion has occurred 3 years after vitrectomy. CONCLUSION: Fine needle aspiration proved to be helpful and reliable for the correct diagnosis of melanocytoma, which does not require additional treatment.
Subject(s)
Nevus, Pigmented/diagnosis , Optic Nerve Neoplasms/diagnosis , Biopsy, Fine-Needle , Humans , Male , Middle AgedABSTRACT
OBJECTIVE: To compare sensitivity an effectiveness of ThinPrep vs. Pap Test and determine whether the Hybrid Capture System II (HCII) for detection of human papillomavirus (HPV) could increase the sensitivity and effectiveness of cervical screening in a low risk population. STUDY DESIGN: We present a comparative observer, blind, "split sample" study of ThinPrep and Papanicolaou staining in a population of 2026 consecutive women. RESULTS: Conventional Papanicolaou stain detected lesions in 62 cases (3.06%), including 1 atypical glandular cell of undetermined significance (AGUS), 34 atypical squamous cells of undetermined significance (ASCUS) (1.73% of atypias), 20 (0.99%) low grade squamous intraepithelial lesions (LSIL), and 7 (0.35%) high grade squamous intraepithelial lesions (HSIL). ThinPrep detected lesions in 63 cases (3.11%), including 26 (1.28%) ASCUS, 27 (1.33%) LSIL and 10 (0.49%) HSIL. Despite this overall similarity, only 35 had a coincident cytopathologic result by both methods. The kappa concordance index between ThinPrep and Pap was 0.546. Sensitivity and specificity were 74.6 and 45.0 in ThinPrep and 69.5 and 25.0 in Papanicolaou-stained smears. High risk HPV (HR-HP 17) was positive in 13 of ZOASCUS and in 21 of 22 SIL. CONCLUSION: Technologies now widely available such as Thin Prep and viral detection aim to improve accuracy of screening.
Subject(s)
Carcinoma, Squamous Cell/prevention & control , Papanicolaou Test , Papillomaviridae/isolation & purification , Papillomavirus Infections/pathology , Tumor Virus Infections/pathology , Uterine Cervical Neoplasms/prevention & control , Vaginal Smears/methods , Carcinoma, Squamous Cell/epidemiology , Carcinoma, Squamous Cell/pathology , Colposcopy , Female , Humans , Mass Screening/methods , Papillomavirus Infections/epidemiology , Risk Factors , Sensitivity and Specificity , Tumor Virus Infections/epidemiology , Uterine Cervical Neoplasms/epidemiology , Uterine Cervical Neoplasms/pathology , Uterine Cervical Dysplasia/epidemiology , Uterine Cervical Dysplasia/pathology , Uterine Cervical Dysplasia/prevention & controlABSTRACT
OBJECTIVE: To compare the cytologic features of histologically proven lymphocytic (Hashimoto's) thyroiditis (Hashimoto's thyroitidis) and primary thyroid lymphomas (TL). STUDY DESIGN: Clinical histories, smears (stained with Diff-Quik, Papanicolaou stain or hematoxylin and eosin [HE]) and surgical specimens (HE slides) were reviewed in 25 cases of lymphocytic thyroiditis and 12 of thyroid lymphomas. RESULTS: Surgical specimens of thyroiditis were obtained for other medical reasons: goiter and compressive symptomatology in 21 cases and neoplasms in 4 (2 papillary carcinomas, 1 follicular carcinoma and 1 oncocytic adenoma). Seven cases were primary lymphomas, and 5 were secondary. Histologically there were 6 large B-cell lymphomas, 2 mantle cell lymphomas, 1 Burkitt lymphoma, 2 mucosal-associated lymphoid tissue lymphomas in blastic transformation and 1 of unknown type. Sensitivity for the diagnosis was 67.5% for HT and 92.3% for lymphoma. CONCLUSION: A heterogeneous population of small and large lymphocytes was the most frequent pattern in both diseases. The presence of a monotonous population of large lymphocytes or, more rarely, of small cells indicates a probable TL. Plasma cells favor HT. Other techniques are mandatory for the differentiation of cases with inconclusive diagnoses.
