The Effect of Enzymatic Digestion on Cultured Epithelial Autografts.

Severe burns are often treated by means of autologous skin grafts, preferably following early excision of the burnt tissue. In the case of, for example, a large surface trauma, autologous skin cells can be expanded in vitro prior to transplantation to facilitate the treatment when insufficient uninjured skin is a limitation. In this study we have analyzed the impact of the enzyme (trypsin or accutase) used for cell dissociation and the incubation time on cell viability and expansion potential, as well as expression of cell surface markers indicative of stemness. Skin was collected from five individuals undergoing abdominal reduction surgery and the epidermal compartment was digested in either trypsin or accutase. Trypsin generally generated more cells than accutase and with higher viability; however, after 7 days of subsequent culture, accutase-digested samples tended to have a higher cell count than trypsin, although the differences were not significant. No significant difference was found between the enzymes in median fluorescence intensity of the analyzed stem cell markers; however, accutase digestion generated significantly higher levels of CD117- and CD49f-positive cells, but only in the 5 h digestion group. In conclusion, digestion time appeared to affect the isolated cells more than the choice of enzyme.

Correction: W.C. Mak, et al. Controlled Delivery of Human Cells by Temperature Responsive Microcapsules. J. Funct. Biomater. 2015, 6, 439-453.

Recently, we found a mistake in Figure 4D in our previously published paper[...].

Auditoría de oxígenoterapia de pacientes hospitalizados en establecimientos del Servicio de Salud Talcahuano / Audit on oxygen therapy in hospitalized patients from Public Health Service of Talcahuano, Chile

El oxígeno (O2) es una de las drogas más utilizadas en clínica, su uso no está exento de riesgos. Existen guias internacionales para su uso, pero en nuestro medio no sabemos si se aplican. Nuestro objetivo fue describir la forma en que se utiliza y controla la oxígenoterapia en los 3 hospitales de la red del Servicio de Salud Talcahuano. Aplicamos una encuesta un día de agosto de 2016 a los pacientes hospitalizados en las instituciones de la red que recibían O2 en ese momento. Se recolectaron datos sobre la prescripción, administración y seguimiento de la terapia de O2. De los 381 pacientes auditados, un 13,7% recibía oxígenoterapia. Los diagnósticos más frecuentes fueron de causa respiratoria (46,15%) y cardiológica (25%). La indicación la dio un médico en 88,5% de los casos y en un 3,8% no había registro. En un 17,3% de los pacientes no había fundamento para la indicación. Se indicó una dosis fija en el 75% con una meta de SaO2 en el 50%, siendo naricera y máscara de Venturi los métodos de administración más frecuentes monitorizándose con oximetría de pulso en los hospitales menos complejos y gasometría arterial en el hospital terciario. La duración media de la oxígenoterapia fue de 7,8 días. Habiendo un buen fundamento y control de la oxígenoterapia aún no se indican metas a obtener. No hay un buen registro de la indicación ni de los cambios realizados. Creemos útil la realización periódica de este tipo de control para optimizar su uso evitando los potenciales efectos adversos en los pacientes.

Abstract Oxygen is a commonly used drug in clinics and its use must be judicious. There are guidelines for oxygen therapy but we ignore if these are respected in our country. We conducted an audit of oxygen therapy by applying a survey to 381 patients in the three hospitals of Talcahuano Public Health Service. The day of the audit 13.7% of the hospitalized patients were on oxygen, most of them with respiratory (46.15%) or cardiovascular (25%) diseases. Indication of O2 administration was given by a physician in 88.5% and there was not registry in 3.8% of the cases. There was not foundation for supplying O2 in 13.3% of patients. A fixed dose was indicated in 75% of cases and 50% had an oximetry value as a target. Oxygen was administered in most of the cases by nasal prongs and Venturi masks. Monitoring was based on pulse oximetry in the less complex hospitals and on arterial blood gases in the tertiary hospital. 100% of patients at urgency ward were receiving a different dose from that indicated at their admission time and none of them had a registry of the new dose. Mean duration of therapy was 7.8 days. We believe our results might represent what is going on with oxygen therapy in our country; having a good foundation and monitoring, we still don t use targets and there is a bad system of registry. We think that it would be advisably to carry out audits on oxygen therapy at national level on regular basis.

Controlled Delivery of Human Cells by Temperature Responsive Microcapsules.

Cell therapy is one of the most promising areas within regenerative medicine. However, its full potential is limited by the rapid loss of introduced therapeutic cells before their full effects can be exploited, due in part to anoikis, and in part to the adverse environments often found within the pathologic tissues that the cells have been grafted into. Encapsulation of individual cells has been proposed as a means of increasing cell viability. In this study, we developed a facile, high throughput method for creating temperature responsive microcapsules comprising agarose, gelatin and fibrinogen for delivery and subsequent controlled release of cells. We verified the hypothesis that composite capsules combining agarose and gelatin, which possess different phase transition temperatures from solid to liquid, facilitated the destabilization of the capsules for cell release. Cell encapsulation and controlled release was demonstrated using human fibroblasts as model cells, as well as a therapeutically relevant cell line-human umbilical vein endothelial cells (HUVECs). While such temperature responsive cell microcapsules promise effective, controlled release of potential therapeutic cells at physiological temperatures, further work will be needed to augment the composition of the microcapsules and optimize the numbers of cells per capsule prior to clinical evaluation.

The inverted CD4/CD8 ratio and associated parameters in 66-year-old individuals: the Swedish HEXA immune study.

The Swedish OCTO and NONA immune longitudinal studies were able to identify and confirm an immune risk profile (IRP) predictive of an increased 2-year mortality in very old individuals, 86-94 years of age. The IRP, was associated with persistent cytomegalovirus infection and characterized by inverted CD4/CD8 ratio and related to expansion of terminally differentiated effector memory T cells (TEMRA phenotype). In the present HEXA immune longitudinal study, we have examined a younger group of elderly individuals (n = 424, 66 years of age) in a population-based sample in the community of Jönköping, Sweden, to examine the relevance of findings previously demonstrated in the very old. Immunological monitoring that was conducted included T cell subsets and CMV-IgG and CMV-IgM serology. The result showed a prevalence of 15 % of individuals with an inverted CD4/CD8 ratio, which was associated with seropositivity to cytomegalovirus and increases in the level of TEMRA cells. The proportion of individuals with an inverted CD4/CD8 ratio was significantly higher in men whereas the numbers of CD3+CD4+ cells were significantly higher in women. In conclusion, these findings are very similar to those previously found by us in the Swedish longitudinal studies, suggesting that an immune profile previously identified in the very old also exists in the present sample of hexagenerians. Therefore, it will be important to examine clinical parameters, including morbidity and mortality, to assess whether the immune profile also is a risk profile associated with higher mortality in this sample of hexagenerians.

Expression and prognostic value of transcription factor PROX1 in colorectal cancer.

BACKGROUND: PROX1 is a specific target of the ß-catenin/TCF pathway in the intestinal epithelium. It acts as a regulator of progression from a benign to a highly dysplastic phenotype in colorectal tumours. However, the clinical significance of PROX1 expression is not known. METHODS: We studied the prognostic value of immunohistochemical expression of PROX1 in a series of 517 patients with colorectal cancer (CRC). RESULTS: The majority of the tumour samples expressed PROX1 (91%, 471 out of 517). High PROX1 expression was associated with a poor grade of tumour differentiation (P<0.0001). In the subgroup of patients with colon cancer, high PROX1 expression was associated with unfavourable colorectal cancer-specific survival (CCSS) as compared with low PROX1 expression (CCSS 47% vs 62%; P=0.045; RR 1.47). The association between high PROX1 and poor outcome was further strengthened in female colon cancer patients (CCSS 38% vs 63%; P=0.007; RR 2.02). Nonetheless, in multivariate survival analysis PROX1 expression was not retained as an independent prognostic factor. CONCLUSION: High PROX1 expression is associated with a poor grade of tumour differentiation, and, in colon cancer patients, also with less favourable patient outcome. Our results strengthen the previous preclinical observations that PROX1 has a role in tumour progression in CRC.

Learning dementia care in three contexts: practical training in day-care, group dwelling and nursing home.

During the period 1996-1999, 18 licensed practical nurses (LPNs) received specialized training to become caregivers and mentors in the field of dementia care at the Silvia Home Foundation in Stockholm, Sweden. The aim of the study was to illuminate how the trainees utilized their practical training to learn about dementia care. The trainees gained practical training within three care models for elderly persons with dementia. The three forms of care and the context for practical training included the school's integrated day-care, a group dwelling and a nursing home. The findings show that the trainees made use of each training context in a similar fashion but there were differences between the contexts. A perspective of human dignity characterized the day-care. This was an opportunity for the nursing philosophy taught by the programme to be put to practical use, and for reflection and experiences pertaining to the individual patient to be developed. In the group dwelling, the trainees encountered patients with different forms of dementia and studied how the care-giving could be adapted to the individual patient's symptoms - the disease perspective. In the nursing home, the trainees chose a staff perspective in which they focused on organization, management and working conditions as well as staff attitudes and the effects of these factors on patient care.

The patient as 'teacher': learning in the care of elderly persons with dementia.

In 1996 HM Queen Silvia of Sweden started a non-governmental education programme with an integrated day-care unit devoted to elderly persons with dementia. A total of 18 Licensed Practical Nurses (LPN) from various parts of Sweden took part in the year theoretical and practical education in dementia care. The purpose was to develop specialized skills in the particular field and more generally to develop mentor capabilities. The aim of the study was to examine the experiences of the trainees, gained from following a single patient during their entire practical training period in the school's integrated day-care unit. The study was based on a combination of participant observations, interviews, diaries and recorded data and used an ethnographic approach. The results showed that themes such as 'personal guide', 'creating a relationship', 'reducing the working pace' and 'investigative mealtimes' were of major importance for the trainees' learning. In their role as 'personal guides', the trainees fell into six categories with different educational focuses. In their relationships with the patients, the trainees were able to form their own impressions of the patients' present and former lives. By training their ability to adapt the pace at which they worked, they had time to observe symptoms and be aware of the patients' needs, as well as the patients' signs of appreciation. The results also indicated that 'investigative mealtimes' can be an important element in the trainee's education in dementia care.

Change of outlook on elderly persons with dementia: a study of trainees during a year of special education.

In 1996 the Silvia Home Foundation started a non-governmental education programme with an integrated day-care unit devoted to patients with dementia: Working chairwoman of the foundation is HM Queen Silvia of Sweden. This study aims to describe the trainees' changed outlook on elderly people with dementia during a year of special education. Data were collected by interviews, informal discussions and participant observations during the lessons. The investigation focused on two questions: the trainees' outlook on the patients and their outlook on the work with the patients. At the beginning of their education, the trainees looked at the patients from a staff's perspective. During their education, this was gradually toned down and they changed to a disease perspective, and eventually to a human dignity perspective. After initially seeing dementia patients as a homogeneous group, the trainees went on to see them as unique human beings. Their outlook on their work also changed, from being task-oriented to a more humanitarian approach.