RESUMEN
The multi-attribute method (MAM) has garnered attention as a new quality control method of therapeutic monoclonal antibodies (mAbs). MAM analysis allows multiple relative quantifications of several structural attributes of therapeutic mAbs; however, some issues remain to be addressed in its procedures especially for sample preparation. The goal of this study was to optimize the sample preparation method for MAM analysis of mAbs. Using a model mAb, we compared five sample preparation methods based on sequence coverage, peptide redundancy, missed cleavage and chemical deamidation. It was found that low pH buffer and short digestion time reduced artificial deamidation. The desalting process after carboxymethylation was essential to obtaining high sequence coverage by a short digestion time. The generation of missed cleavage peptides was also improved by using a trypsin/lysyl endopeptidase (Lys-C) mixture. Next, we evaluated the usefulness of our method as a part of MAM analysis. Finally, 17 glycopeptides, 2 deamidated peptides and N- and C-terminal peptides of the heavy chain were successfully monitored with acceptable mass accuracy and coefï¬cient of variation (CV, %) of the relative peak area. On the other hand, 4 oxidated peptides indicated the unavoidable slightly higher inter-assay CV (%) of the peak area ratio due to the instability in the MS sample solution. Collectively, we demonstrated that our method was applicable as an easy and reliable sample preparation method for MAM analysis, and the variation in the relative peak area could be influenced by the modification type rather than by the amount of each peptide.
Asunto(s)
Anticuerpos Monoclonales/metabolismo , Serina Endopeptidasas/metabolismo , Tripsina/metabolismo , Anticuerpos Monoclonales/genética , Humanos , Concentración de Iones de Hidrógeno , Control de Calidad , Serina Endopeptidasas/genética , Tripsina/genéticaRESUMEN
BACKGROUND: Orthodontically induced root resorption (OIRR) is considered as an undesirable and unpredictable sequel of orthodontic treatment. Recent reports demonstrated that interleukin (IL)-17/IL-34, and T cells secrete inflammatory/osteoclastogenic cytokines, which might stimulate osteoclastogenesis/bone resorption. However, little is known about the role played by IL-17/IL-34 in OIRR. The present study was aimed at investigating the odontoclastic expression pattern of IL-17 and IL-34 in resorbed cementum during different experimental tooth movements in vivo. METHODS: Twenty-four 8-week-old male Wistar rats were divided into four groups: control group, optimal force group (10 g), heavy force group (50 g), and jiggling force group (compression and tension, repetition; 10 g). After 7, 14, and 21 days, the expression levels of IL-17 and IL-34 protein in the resorbed cementum were analyzed using immunohistochemical methods. RESULTS: On day 21, the immunoreactivity for IL-17 and IL-34 in resorbed roots in the jiggling force group was stronger than that in the heavy force and optimal force groups. Moreover, the number of IL-17-positive and IL-34-positive odontoclasts was significantly increased in the jiggling force group compared with those in the other groups on day 21. CONCLUSIONS: These results suggest that jiggling forces might exacerbate OIRR compared with heavy forces, as evidenced by the increased expression of IL-17 and IL-34 in odontoclasts obtained from resorbed roots.
Asunto(s)
Interleucina-17/metabolismo , Interleucinas/metabolismo , Resorción Radicular/etiología , Resorción Radicular/metabolismo , Técnicas de Movimiento Dental/efectos adversos , Animales , Peso Corporal , Cemento Dental/metabolismo , Inmunohistoquímica/métodos , Masculino , Osteoclastos/metabolismo , Osteogénesis , Ligamento Periodontal/patología , Ratas , Ratas Wistar , Resorción Radicular/fisiopatología , Linfocitos T/metabolismo , Fosfatasa Ácida TartratorresistenteRESUMEN
The palliative care team's roles are to provide a symptom relief to cancer patients, help them accept their medical conditions, and offer advice regarding the selection of appropriate medical treatments to suit their needs. Seeking the comfort of their homes, patients prefer a home care of superior medical care provided at hospitals. In 2008, 25 of the end-stage cancer patients at hospitals were expressed their desires to have a home medical care, and 10 of them were allowed to do so. We considered the following contributing factors that a patient should have for a smooth transition from hospital care to home medical care: (1) life expectancy of more than 2 months, (2) no progressive breathing difficulties experienced daily, (3) good awareness of medical condition among patients and families, (4) living with someone who has a good understanding of the condition, (5) availability of an appropriate hospital in case of a sudden change in medical requirements, and (6) good collaboration between emergency care hospitals, home physicians, and visiting nurses. To treat the end-stage cancer patients at home, there is a need for information sharing and a joint training of physicians specialized in cancer therapy, palliative care teams, home physicians, and visiting nurses. This would ensure a sustainable "face-to-face collaboration" in community health care.
Asunto(s)
Servicios de Atención de Salud a Domicilio , Neoplasias/terapia , Cuidados Paliativos , Grupo de Atención al Paciente , Alta del Paciente , Cuidado Terminal , Redes Comunitarias , Femenino , Hospitales Universitarios , Humanos , Masculino , Persona de Mediana EdadRESUMEN
In this article we illuminate the narratives of women living with a spinal cord injury (SCI) with regard to (a) learning how to live with a changed body and (b) exploring the factors that influence how they feel toward their new bodies. An SCI produces immediate physical impairments resulting in a changed body, which can then have physical, emotional, and social consequences to these women. Through its focus on enhancing the body, physical therapy can help to promote a positive view of the self within the changed body. Our analysis of these women's experiences resulted in a fluid, three-phase framework of learning to live with a changed body that generally moves from (a) discomfort, to (b) moving toward comfort, to (c) comfort. Physical therapy can potentially influence the process of women's gaining comfort with their changed bodies following an SCI. The framework provides a basis for future research on adaptation following SCI.