Functional characterization of iPSC-derived arterial- and venous-like endothelial cells.

The current work reports the functional characterization of human induced pluripotent stem cells (iPSCs)- arterial and venous-like endothelial cells (ECs), derived in chemically defined conditions, either in monoculture or seeded in a scaffold with mechanical properties similar to blood vessels. iPSC-derived arterial- and venous-like endothelial cells were obtained in two steps: differentiation of iPSCs into endothelial precursor cells (CD31pos/KDRpos/VE-Cadmed/EphB2neg/COUP-TFneg) followed by their differentiation into arterial and venous-like ECs using a high and low vascular endothelial growth factor (VEGF) concentration. Cells were characterized at gene, protein and functional levels. Functionally, both arterial and venous-like iPSC-derived ECs responded to vasoactive agonists such as thrombin and prostaglandin E2 (PGE2), similar to somatic ECs; however, arterial-like iPSC-derived ECs produced higher nitric oxide (NO) and elongation to shear stress than venous-like iPSC-derived ECs. Both cells adhered, proliferated and prevented platelet activation when seeded in poly(caprolactone) scaffolds. Interestingly, both iPSC-derived ECs cultured in monoculture or in a scaffold showed a different inflammatory profile than somatic ECs. Although both somatic and iPSC-derived ECs responded to tumor necrosis factor-α (TNF-α) by an increase in the expression of intercellular adhesion molecule 1 (ICAM-1), only somatic ECs showed an upregulation in the expression of E-selectin or vascular cell adhesion molecule 1 (VCAM-1).

Flexible nanofilms coated with aligned piezoelectric microfibers preserve the contractility of cardiomyocytes.

The use of engineered cardiac tissue for high-throughput drug screening/toxicology assessment remains largely unexplored. Here we propose a scaffold that mimics aspects of cardiac extracellular matrix while preserving the contractility of cardiomyocytes. The scaffold is based on a poly(caprolactone) (PCL) nanofilm with magnetic properties (MNF, standing for magnetic nanofilm) coated with a layer of piezoelectric (PIEZO) microfibers of poly(vinylidene fluoride-trifluoroethylene) (MNF+PIEZO). The nanofilm creates a flexible support for cell contraction and the aligned PIEZO microfibers deposited on top of the nanofilm creates conditions for cell alignment and electrical stimulation of the seeded cells. Our results indicate that MNF+PIEZO scaffold promotes rat and human cardiac cell attachment and alignment, maintains the ratio of cell populations overtime, promotes cell-cell communication and metabolic maturation, and preserves cardiomyocyte (CM) contractility for at least 12 days. The engineered cardiac construct showed high toxicity against doxorubicin, a cardiotoxic molecule, and responded to compounds that modulate CM contraction such as epinephrine, propranolol and heptanol.

Sarcoma the great "masquerader" hematoma/deep vein thrombosis manifestation.

INTRODUCTION: The clinical presentation of patients with soft-tissue sarcoma is highly variable. Most patients present with a painless mass, typically one that is increasing in size, and few have systemic symptoms such as fever, weight loss, or malaise. Soft tissue sarcomas can initially present as, or even be misdiagnosed as, deep venous thrombosis (DVT), leading to a late diagnosis. CASE REPORT: A 51-year-old woman presented to the hospital with complaints of pain and swelling in her left thigh, interpreted as an infected hematoma with an associated deep vein thrombosis and treated accordingly. The patient presented to our emergency department two more times. In the last visit and due to an unresolving clinical scenario a MRI and surgical byopsies were made that confirmed a sarcoma diagnosis. DISCUSSION: When a patient presents with an expanding, nontraumatic mass simulating a haematoma, several other differential diagnoses should be considered including aneurysm, bleeding tendency, chronic expanding haematoma and soft-tissue sarcoma. The growth of the tumor undetected while being treated for the DVT and then posteriorly for the hematoma, was without a doubt dismal to the patient, so earlier diagnosis would have been preferable. CONCLUSION: When a patient presents with an unusual history of hematoma in the extremities, it is necessary to consider the possibility of a malignant soft tissue tumor.

The Kite Latissimus Dorsi Flap for Breast Reconstruction: An Attempt to Reduce Lateral Chest Wall Deformity and Axillary Bulking.

UNLABELLED: The latissimus dorsi flap is a commonly used tissue transfer for volume replacement in partial or total breast reconstruction. In this era of cosmetic awareness and oncoplastic breast surgery, two main defects are related to the conventional technique: the back scar and the bulkiness on the lateral chest wall, under the axilla. Axillary bulking, a disturbing defect for the majority of patients, is a persistent consequence, independent of the technique used, even when the proximal tendon is cut. We describe a new approach, the kite latissimus dorsi flap, consisting of harvesting the flap, partially or totally, with pedicle dissection from the muscle, extending dissection, perforator style if needed, until the external border of the breast (anterior axillary line) is reached. The muscle is then cut at that level, leaving no unnecessary volume under the axilla, which would cause bulkiness and chest wall deformity. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Thyroid metastasis from renal cell carcinoma-A case report after 9 years.

INTRODUCTION: The thyroid gland is a rare site of clinically detectable tumor metastasis. As thyroid tumors are usually assumed to be primary in origin, its recognition as a secondary is difficult. PRESENTATION OF CASE: We report a case of an 80-year old female who was referred to the Department of Surgery for a symptomatic thyroid nodule. Her medical history included a radical nephrectomy for renal cell carcinoma (RCC) nine years ago. During follow-up a pancreatic nodule was noted suggestive of a neuroendocrine tumor and the von Hippel-Lindau syndrome had to be ruled out. The fine-needle aspiration biopsy (FNAB) guided by ultrasound (US) of the thyroid nodule was inconclusive and a hemithyroidectomy and isthmectomy were performed. Histological examination revealed metastasis of a clear cell carcinoma. DISCUSSION: RCC disseminates in an unpredictable manner and can show late recurrences. Although secondary involvement of the thyroid gland by RCC is rare, it is still one of the more common neoplasms to metastasize to this site. There are no specific clinical features and few characteristic findings of metastatic thyroid carcinoma on imaging studies. FNAB is a useful procedure to diagnose metastatic thyroid cancer, but one should remain suspicious when the result for malignant cells is negative or indeterminate. After thyroidectomy the diagnosis of RCC is confirmed immunohistochemically. There is a clear survival benefit if a surgical approach to the thyroid metastasis is chosen. CONCLUSION: Thyroid metastasis should be considered in patients with a thyroid nodule and positive history for RCC.

Walking Clinic in ambulatory surgery--A patient based concept: A Portuguese pioneer project.

INTRODUCTION: Walking Clinic is an innovative, efficient and easily reproducible concept adapted to ambulatory surgery. It consists of a preoperative single day work-up, with a surgeon, an anesthetist and a nurse. The aim of this study was to evaluate patient satisfaction and its determinants. METHODS: A survey was applied to 171 patients (101 of the Walking Clinic group and 70 not engaged in this new concept). Patient satisfaction was assessed evaluating five major questionnaire items: secretariat (quality of the information and support given), physical space (overall comfort and cleanliness), nurses and medical staff (willingness and expertise), and patients (waiting time until pre-operative consults and exams, waiting time until being scheduled for surgery, surgery day waiting time and postoperative pain control). Furthermore, overall assessment of the received treatment, and probability of patient recommending or returning to our ambulatory unit were also analyzed. RESULTS: Walking Clinic group had overall better results in the five major questionnaire items assessed, with statistical significance, except for the physical space. It also showed better results regarding the sub-items postoperative pain control, waiting time until being scheduled for surgery and surgery day waiting time. DISCUSSION: The results confirm better patient satisfaction with this new concept. CONCLUSION: The Walking Clinic concept complements all the tenets of ambulatory surgery, in a more efficient manner.

Translating clinical trials to clinical practice: outcomes of men with metastatic castration resistant prostate cancer treated with docetaxel and prednisone in and out of clinical trials.

BACKGROUND: Multiple factors can influence outcomes of patients receiving identical interventions in clinical trials and in routine practice. Here, we compare outcomes of men with metastatic castrate-resistant prostate cancer (mCRPC) treated with docetaxel and prednisone in routine practice and in clinical trials. PATIENTS AND METHODS: We reviewed patients with mCRPC treated with docetaxel at Princess Margaret Cancer Centre. Primary outcomes were overall survival and PSA response rate. Secondary outcomes were reasons for discontinuation and febrile neutropenia. Outcomes were compared for men treated in routine practice and in clinical trials, and with data from the TAX 327 study. RESULTS: From 2001 to 2011, 438 men were treated, of whom 357 received 3-weekly docetaxel as first-line chemotherapy: 314 in routine practice and 43 in clinical trials. Trial patients were younger and had better performance status. Median survival was 13.6 months [95% confidence interval (95% CI) 12.1-15.1 months] in routine practice and 20.4 months (95% CI 17.4-23.4 months, P = 0.007) within clinical trials, compared with 19.3 months (95% CI 17.6-21.3 months, P < 0.001) in the TAX 327 study. PSA response rates were 45%, 54%, and 53%, respectively (P = NS). Reasons for treatment discontinuation were similar although trial patients received more cycles (median: 6 versus 8 versus 9.5, P < 0.001). Rates of febrile neutropenia were 9.6, 0, and 3% (P < 0.001) while rates of death within 30 days of last dose were 4%, 0%, and 3%, respectively (P = NS). CONCLUSIONS: Survival of patients with mCRPC treated with docetaxel in routine practice is shorter than for men included in trials and is associated with more toxicity.

A new level 1 oncoplastic technique for breast conserving surgery: rotation glandular flap.

BACKGROUND: When performing conservative surgery for breast cancer, breast reshaping can be a challenging procedure. Level 1 oncoplastic surgery (OPS) techniques, i.e., advancement or rotation of glandular flaps, should be performed when less than 20 per cent breast volume is excised. OBJECTIVE: A new Level 1 OPS technique is described. A wide centro-lateral glandular flap is created after extensive undermining of the skin and nipple-areolar complex, and rotated into the cavity. DISCUSSION: This rotation glandular flap is a new technique for use following a wide excision, in glandular, not fatty, breasts, and when standard closure of the cavity would not leave a satisfactory cosmetic result.

In vitro assessment of gentian violet anti- candida activity.

BACKGROUND/AIM: Recurrent vulvovaginal candidosis (RVVC) needs alternative therapeutic approaches. Gentian violet (GeV) has been traditionally used to treat mucocutaneous candidosis. The aim of the present study was to evaluate the in vitro activity of GeV against Candida spp. and contribute to clarify the mechanism of action, supporting its clinical therapeutic use. METHODS: Seventeen clinical Candida isolates from RVVC and one C. albicans type collection (ATCC 10231) were studied; the antifungal activity of GeV was evaluated according to the CLSI M27-A3 protocol. To elucidate its mechanism of action, cells were stained with propidium iodide and afterwards analyzed by flow cytometer. RESULTS: GeV showed a fungicidal activity against most Candida spp. C. albicans and C. tropicalis were the most susceptible species. Minimal lethal concentrations were similar to minimal inhibitory concentrations for most tested strains. The fungicidal effect was not related to a primary lesion of the cytoplasmic membrane. CONCLUSION: In accordance with our findings, GeV is a valuable potent fungicidal drug to be used topically, isolated or in combination with oral antifungal drugs, particularly in RVVC cases.

Severe malaria--clinical case.

We report a clinical case of severe malaria, where the rate of initial parasitaemia by Plasmodium falciparum was 43 %. Multiple organ dysfunction, including ARDS, forced admission in a close surveillance unit, with survival of the same. A brief review of the subject is made, focusing on severity and general conduct, alerting and awareness for this entity, whose expression, among us, could take on increasing importance.

Prevalence of human cytomegalovirus congenital infection in Portuguese newborns.

Human cytomegalovirus (HCMV) is considered the most frequent cause of congenital infection, occurring in 0.2 to 2.2% of all live births. Since this is a wide range of prevalences observed in different studies, it would be desirable to investigate the prevalence of this infection at national level. The aim of this study was the evaluation of the national prevalence of HCMV congenital infection. We analysed a total of 3,600 Guthrie cards collected from Portuguese newborns during a period of 14 months (August 2003 to September 2004). The cards covered all regions of Portugal and were proportional to the number of births in each region. A heat DNA extraction method was used, followed by DNA amplification by nested PCR. Sensitivity and specificity of this method were evaluated as 93% and 100%, respectively, using 28 cards from HCMV-positive and 280 cards from HCMV-negative children. The national prevalence of congenital HCMV was determined as 1.05% (95% confidence interval: 0.748-1.446). This is the first study of the prevalence of HCMV congenital infection at national level in Portugal. It suggests that Portugal may have one of the highest prevalences of congenital HCMV infection in Europe.

Diagnóstico laboratorial pré-natal da infecçao congénita por cytomegalovirus: revisão da experiência de 10 años / Prenatal laboratory diagnosis of cytomegalovirus congenital infection: a ten year review

El virus humano del citomegalovirus (CMV) es el agente más frecuente de infección congénita, afectando aproximadamente al 0,2 a 2,2% de todos los recién nacidos. El Laboratorio de Microbiología del Hospital de Santa Cruz inició su colaboración con varias instituciones obstétricas portuguesas en 1994 en el área del diagnóstico prenatal de laboratorio para el CMV y en el año 2001 el Servicio de Patología Clínica del Centro Hospitalario Cova da Beira inició, igualmente, esta colaboración con algunas instituciones obstétricas del centro y norte del país. Dada la uniformidad de los métodos utilizados por los dos laboratorios, se ha efectuado una revisión conjunta de esta experiencia en esta área. De los 596 líquidos amnióticos estudiados, 30 fueron positivos para CMV. La sensíbilidad, la especificidad, el valor predictivo negativo y valor predictivo positivo del diagnóstico prenatal (evaluados en los casos en que se efectuó la comparación con los métodos de referencia, de la viruria en el recién nacido y la investigación de inclusiones en el feto) fueron, respectivamente del 90, 99, 98 y 95%. Los resultados presentados en esta revisión sugieren que el diagnóstico prenatal virológico para CMV, efectuado siguiendo la metodología que se ha descrito, constituye un arma diagnóstica fiable para la evaluación prenatal de esta infección congénita. La selección de los casos para amniocentesis debe obedecer a las indicaciones de «seroconversión para IgG», «IgM confirmada» (debiendo los métodos de confirmación ser la avidez de las IgG y, en algunos casos, el Western blot para IgM) y las alteraciones eco gráficas de etiología suspechosa para CMV

Human cytomegalovirus (CMV) is the most frequent cause of congenital infection, affecting 0.2 to 2.2% of all live births. In 1994, the Laboratory of Microbiology of the Hospital de Santa Cruz began working in collaboration with several Portuguese Institutions, in the field of prenatal CMV diagnosis. In 2001, the Department of Clinical Pathology of the Centro Hospitalar Cova da Beira also started this collaboration with several Obstetrical Institutions from the north and center regions of Portugal. Since both laboratories have adopted similar experimental methodologies used in CMV prenatal diagnosis, the present work represents a collection of results obtained from both institutions. From the 596 amniotic fluids collected, 30 were found positive for CMV. Sensitivity, specificity, negative predictive and positive predictive values were, 90, 99, 98 and 95%, respectively (when compared with the urine cultures in the newborns or histological examination of the fetuses). The results obtained suggest that virological diagnosis, performed as described above, can be very useful for prenatal diagnosis of CMV congenital infections. Selection for amniocentesis should be restricted to cases with «IgG seroconversion», «confirmed IgM» (by IgG avidity and, in some cases, by IgM Western blot) and ecographic abnormalities

[Implicit memory and its contribution to the rehabilitation of an amnesic patient: case report]. / Memória implícita e sua contribuição à reabilitação de um paciente amnéstico: relato de caso.

Even severe amnesic patients usually are able to learn new information using implicit memory strategies. In the present study we describe our experience in the rehabilitation of an amnesic patient who had suffered cerebral anoxia. He was taught to use a computer text editor, enabling him to sheltered work. The training program lasted 14 weeks and was based on errorless learning and drilling practice techniques. The patient succeeded in all tasks, even though he could not remember accurately the training sessions. These results meet previous studies which point out that interaction between implicit and residual explicit memory can provide a basis new learning in amnesic patients. The domain-specific knowledge acquisition is demonstrating effectiveness in neuropsychological rehabilitation of brain injured patients.

Simple mucin-type carbohydrate antigens (Tn, sialosyl-Tn, T and sialosyl-T) and gp 230 mucin-like glycoprotein are candidate markers for neoplastic transformation of the human cervix.

Mucins and simple mucin-type carbohydrates are cancer-associated antigens in several human tumors. Expression of Tn, sialosyl-Tn, Thomsen-Friedenreich (T), sialosyl-T and of a recently identified mucin-like glycoprotein (gp230) has not yet been thoroughly investigated in human cervix carcinogenesis. In the present study sections from normal cervix (n=10), CIN III lesions (n=10), and invasive carcinomas (n=47) were evaluated immunohistochemically using monoclonal antibodies. In normal cervix there was: cytoplasmatic expression of Tn in 1 case (10%); membranous expression of STn in 8 cases (80%); no expression of T and cytoplasmatic expression of ST in 1 case (10%); gp 230 was expressed in all cases with a membranous pattern. In CIN III lesions there was cytoplasmatic and membranous expression of Tn in 3 cases (30%) and of STn in 9 cases (90%); T and ST were not expressed; gp 230 was expressed in 5 cases (50%) both in the cytoplasm and at the cell membrane. In invasive carcinomas we observed Tn expression in 30 cases (63.8%) and STn in 31 cases (66%); T antigen was not expressed; expression of both ST and gp 230 in 24 cases (51.1%); all antigens showed membranous and cytoplasmatic staining. Our results show that Tn and ST are good markers of invasive carcinomas of the human cervix. We have also shown that loss of expression of the mucin-like glycoprotein gp 230 is associated with malignant transformation at a preinvasive stage.

[Procedures to improve daily activities performance in individuals with cerebral anoxia: case report]. / O uso de procedimentos de estruturação de rotina em indivíduos com anóxia cerebral: relato de caso.

OBJECTIVE: To present a rehabilitation case study; procedures to improve the performance on daily activities were used by the caregiver. PATIENT: Cerebral anoxia due to cardiac arrest. He presented memory deficits for both visual and verbal material, temporal disorientation, abulia, psychomotor slowing and bradypsychism. METHODS: daily activities were developed in fixed hours; calendar was presented 3 times a day; daily walks were performed; self-care activities were performed by the patient with short verbal commands and cues from the caregiver. RESULTS: We observed improvement on initiative and autonomy for daily activities, attention, temporal orientation and bradypsychism. There was also reduction on response time. CONCLUSIONS: Behavioral control strategies result on improvement for patients and their family although limit their routine. Behavioral control strategy promotes learning through the preserved implicit memory mechanism and results on enhancement at patient's autonomy at home.

Reparacao de ferimento cardiaco:ocorrencia de dois episodios no mesmo paciente. / Repair of cardiac injury: ocurrence of two episodes in the same patient

Os autores relatam o caso de um paciente que por duas vezes foi vitima de ferimentos cardiacos com perfuracao de ventriculo esquerdo, tendo em ambas as ocasioes sido submetido a cirurgia de urgencia em um hospital de interior, sem equipamentos especializados ou medicos treinados em cirurgia cardiaca. Houve pleno sucesso em ambas as cirurgias e o paciente manteve-se vivendo normalmente apesar das sequelas remanescentes que, mais tarde, foram reveladas pela cineangiocoronariografia (obstrucao da coronaria descendente anterior esquerdo a aneurisma de ventriculo esquerdo)