Pilot study of a novel vacuum-assisted method for decellularization of tracheae for clinical tissue engineering applications.

Tissue engineered tracheae have been successfully implanted to treat a small number of patients on compassionate grounds. The treatment has not become mainstream due to the time taken to produce the scaffold and the resultant financial costs. We have developed a method for decellularization (DC) based on vacuum technology, which when combined with an enzyme/detergent protocol significantly reduces the time required to create clinically suitable scaffolds. We have applied this technology to prepare porcine tracheal scaffolds and compared the results to scaffolds produced under normal atmospheric pressures. The principal outcome measures were the reduction in time (9 days to prepare the scaffold) followed by a reduction in residual DNA levels (DC no-vac: 137.8±48.82 ng/mg vs. DC vac 36.83±18.45 ng/mg, p<0.05.). Our approach did not impact on the collagen or glycosaminoglycan content or on the biomechanical properties of the scaffolds. We applied the vacuum technology to human tracheae, which, when implanted in vivo showed no significant adverse immunological response. The addition of a vacuum to a conventional decellularization protocol significantly reduces production time, whilst providing a suitable scaffold. This increases clinical utility and lowers production costs. To our knowledge this is the first time that vacuum assisted decellularization has been explored. Copyright © 2015 John Wiley & Sons, Ltd.

Urothelial cells undergo epithelial-to-mesenchymal transition after exposure to muscle invasive bladder cancer exosomes.

Bladder cancer, the fourth most common noncutaneous malignancy in the United States, is characterized by high recurrence rate, with a subset of these cancers progressing to a deadly muscle invasive form of disease. Exosomes are small secreted vesicles that contain proteins, mRNA and miRNA, thus potentially modulating signaling pathways in recipient cells. Epithelial-to-mesenchymal transition (EMT) is a process by which epithelial cells lose their cell polarity and cell-cell adhesion and gain migratory and invasive properties to become mesenchymal stem cells. EMT has been implicated in the initiation of metastasis for cancer progression. We investigated the ability of bladder cancer-shed exosomes to induce EMT in urothelial cells. Exosomes were isolated by ultracentrifugation from T24 or UMUC3 invasive bladder cancer cell conditioned media or from patient urine or bladder barbotage samples. Exosomes were then added to the urothelial cells and EMT was assessed. Urothelial cells treated with bladder cancer exosomes showed an increased expression in several mesenchymal markers, including α-smooth muscle actin, S100A4 and snail, as compared with phosphate-buffered saline (PBS)-treated cells. Moreover, treatment of urothelial cells with bladder cancer exosomes resulted in decreased expression of epithelial markers E-cadherin and ß-catenin, as compared with the control, PBS-treated cells. Bladder cancer exosomes also increased the migration and invasion of urothelial cells, and this was blocked by heparin pretreatment. We further showed that exosomes isolated from patient urine and bladder barbotage samples were able to induce the expression of several mesenchymal markers in recipient urothelial cells. In conclusion, the research presented here represents both a new insight into the role of exosomes in transition of bladder cancer into invasive disease, as well as an introduction to a new platform for exosome research in urothelial cells.

Characterisation of porcine dermis scaffolds decellularised using a novel non-enzymatic method for biomedical applications.

Off-the-shelf availability of tissue-engineered skin constructs, tailored by different combinations of reagents to produce a highly preserved biological matrix is often the only means to help patients suffering skin damage. This study assessed the effect of five different decellularisation methods on porcine dermal scaffolds with regard to matrix composition, biomechanical strength, and cytotoxicity using an in vitro biocompatibility assay. Results demonstrated that four out of the five tested decellularisation protocols were efficient in producing acellular scaffolds. Nevertheless, decellularisation method using osmotic shock without enzymatic digestion showed to be efficient not only in removing cellular material and debris from dermal scaffolds but was also beneficial in the preservation of extracellular matrix components (glycosaminoglycans and collagen). Histological assessment revealed that the dermal architecture of coarse collagen bundles was preserved. Examinations by scanning electron microscopy and transmission electron microscopy showed that the arrangement and ultrastructure of collagen fibrils in the scaffolds were retained following non-enzymatic method of decellularisation and also after collagen crosslinking using genipin. Moreover, this decellularised scaffold was not only shown to be biologically compatible when co-cultured with bone marrow-derived mesenchymal stem cells and fibroblasts, but also stimulated the cells to release trophic factors essential for tissue regeneration.

Correlation between follicular diameters and flushing versus no flushing on oocyte maturity, fertilization rate and embryo quality.

OBJECTIVE: To determine (a) the correlation between follicular sizes, oocyte maturity, normal fertilization rate, cleavage and embryo quality; and (b) to establish whether oocytes recovered with or without follicular flushing have different developmental competence. DESIGN: Prospective observational study. SETTING: Academic medical center. PATIENTS: Forty nine cycles (37 ICSI and 12 IVF). INTERVENTIONS: Measurement of 360 follicular diameters on the day of egg retrieval and classification into three groups Group A (mean diameter 12-14.5 mm.), group B (mean diameter 15-18 mm.) and group C (diameter >18.5 mm.). MAIN OUTCOME MEASURE: Correlation between follicular size at the time of retrieval and oocyte maturity, fertilization and cleavage rate in 226 oocytes (163 ICSI and 63 IVF). Developmental competence of oocytes retrieved with flushing versus non flushing. RESULTS: Almost all (99 %) of the oocytes recovered from follicles of group C were in metaphase II as opposed to 80 % in group A and 81 % in group B (p < 0.01). Overall there was a progressive and significant increase in fertilization rates from group A follicles to group C (47 % vs. 67 %, p 0.05). Overall 53 % of oocytes retrieved from group A follicles showed either no fertilization or abnormal fertilization versus 27 % in group C (p 0.05). The oocyte recovery rate with follicular flushing improved from group A to group B and to group C follicles (65 % vs. 49 % vs.37 % respectively p < 0.01). There were no differences in rates of immature oocyte, fertilization, abnormal or not fertilization and cleavage. CONCLUSIONS: The results of this study shows that: a) Follicles larger than 18 mm at retrieval have consistently mature oocytes with a higher rate of fertilization; b) Small size follicles are still capable of containing mature oocytes, but their rate of abnormal or no fertilization is high; c) Oocytes recovered with flushing are still able to produce embryos with full developmental competence.

High density micromass cultures of a human chondrocyte cell line: a reliable assay system to reveal the modulatory functions of pharmacological agents.

Osteoarthritis is a highly prevalent and disabling disease for which we do not have a cure. The identification of suitable molecular targets is hindered by the lack of standardized, reproducible and convenient screening assays. Following extensive comparisons of a number of chondrocytic cell lines, culture conditions, and readouts, we have optimized an assay utilizing C-28/I2, a chondrocytic cell line cultured in high-density micromasses. Utilizing molecules with known effects on cartilage (e.g. IL-1ß, TGFß1, BMP-2), we have exploited this improved protocol to (i) evoke responses characteristic of primary chondrocytes; (ii) assess the pharmacodynamics of gene over-expression using non-viral expression vectors; (iii) establish the response profiles of known pharmacological treatments; and (iv) investigate their mechanisms of action. These data indicate that we have established a medium-throughput methodology for studying chondrocyte-specific cellular and molecular responses (from gene expression to rapid quantitative measurement of sulfated glycosaminoglycans by Alcian blue staining) that may enable the discovery of novel therapeutics for pharmacological modulation of chondrocyte activation in osteoarthritis.

The role of combination medical therapy in benign prostatic hyperplasia.

To review key trials of monotherapy and combination therapy of alpha(1)-adrenergic receptor antagonists (alpha(1)-ARAs), 5alpha-reductase inhibitors (5alphaRIs) and anti-muscarinic agents in the treatment of lower urinary tract symptoms (LUTS) associated with benign prostatic hyperplasia (BPH). To assess the safety and efficacy of combination therapies for LUTS associated with BPH, a search of the MEDLINE and Cochrane databases (1976-2008) was conducted for relevant trials and reviews using the terms benign prostatic hyperplasia, lower urinary tract symptoms, alpha(1)-adrenergic receptor antagonists, 5alpha-reductase inhibitors, anti-muscarinics, anticholinergics, combination therapy, alfuzosin, doxazosin, tamsulosin, terazosin, dutasteride, finasteride, tolterodine, flavoxate, propiverine, oxybutynin, erectile dysfunction, sildenafil, vardenafil and tadalafil. Data from the Medical Therapy of Prostatic Symptoms (MTOPS) study indicated a role for long-term use of alpha(1)-ARAs and 5alphaRIs in combination. In the MTOPS study, combination therapy with the alpha(1)-ARA doxazosin and the 5alphaRI finasteride was significantly more effective than either component alone in reducing symptoms (P=0.006 vs doxazosin monotherapy; P<0.001 vs finasteride monotherapy) and in lowering the rate of clinical progression (P<0.001 vs either monotherapy). These findings were confirmed by the 2-year preliminary results of the Combination of Avodart and Tamsulosin study. In this study, combination therapy of the alpha(1)-ARA tamsulosin and the 5alphaRI dutasteride resulted in a significantly greater decrease in International Prostate Symptom Score (IPSS) when compared with either monotherapy. Several recent trials have studied the efficacy of combining alpha(1)-ARAs and anti-muscarinic agents in the treatment of BPH. These studies have found this combination to result in statistically significant benefits in quality of life scores, patient satisfaction, urinary frequency, storage symptoms and IPSS scores. Studies have not shown an increased risk of urinary retention associated with the use of anti-muscarinics in a highly select cohort of men with BPH. The available data suggest that combination therapy can be beneficial in the treatment of BPH and associated LUTS. The greatest efficacy for the alpha(1)-ARA and 5alphaRI combination was shown in patients with larger prostate size and more severe symptoms. The combination of alpha(1)-ARAs and 5alphaRIs appears to prevent disease progression in these patients. The combination of alpha(1)-ARAs with anti-muscarinic agents is useful for relieving symptoms of bladder outlet obstruction and detrusor overactivity. Theoretic concerns regarding the risk of acute urinary retention have been refuted in several recent clinical trials; however, it must be noted that the patients in these trials were a highly select cohort of men. Men with overactive bladder and BPH who are not receiving adequate alleviation of symptoms from the first-line alpha(1)-ARAs may benefit from the addition of an anti-muscarinic agent.

A retrospective study of the safety of intramuscular ziprasidone in agitated elderly patients.

OBJECTIVE: Authors evaluated the safety of intramuscular ziprasidone for use in acute agitation in an elderly population. METHOD: Medical records were reviewed retrospectively to identify consecutive patients who were admitted to our neuropsychiatry service with the presenting complaint of dementia (DSM-IV) with agitation and who were given intramuscular ziprasidone and then administered an electrocardiogram (ECG) (N = 23). Some patients also had a baseline ECG (N = 14). QTc intervals were recorded, and significance was defined as a QTc of > or =450 ms or a 10% prolongation from baseline. A paired-samples t test was performed to compare the baseline and postmedication QTc intervals. Confounding factors were examined, and cardiac events (torsades de pointes, cardiac arrest) were recorded. RESULTS: There was no significant difference in the QTc interval between the baseline and the post-ziprasidone values. One patient had a QTc greater than 500 ms and 25% over baseline, and therefore the medication was discontinued. The mean prolongation of the QTc interval was only 0.5 ms. There were no episodes of torsades de pointes. Other medications that the patients were taking did not appear to affect the QTc interval in an expected manner. CONCLUSION: Larger studies need to be done to evaluate the safety of intramuscular ziprasidone in agitated elderly patients, a population with an increased risk of QT prolongation and torsades de pointes because of their age, comorbid conditions, and concomitant use of multiple medications.

Lymphatic system changes in diabetes mellitus: role of insulin and hyperglycemia.

BACKGROUND: Diabetic alterations of blood vessels have been well studied, but much less is known about the lymphatic system, which plays an important role in the transport of particles and defensive responses. Accordingly, we investigated lymphatic changes in diabetic rats. METHODS: Ten, 30 or 60 days after alloxan-induced diabetes (40 mg/kg; i.v.), we studied thoracic duct lymph flow and lymphocyte output, thoracic duct lymph transport of radiotracer particles ((99m)Tc-dextran 500), lymph node uptake and scintigraphic visualization of subcutaneously injected radiotracer particles, as well as the effect of insulin administration and food deprivation. RESULTS: Diabetes significantly increased thoracic duct lymph flow and the transport of dextran from the footpad subcutaneous tissue. Abnormal lymphocyte output from the thoracic duct occurred in the first 10 days. Uptake of dextran into regional lymph nodes was decreased in diabetes. Insulin per se, although not normalizing blood sugar levels, appeared to recover thoracic duct lymphocyte output and lymph node uptake of (99m)Tc-dextran 500 without affecting the thoracic duct lymph flow or the amount of radiotracer recovered therein. Normalization of glycemia (by food deprivation) restored the lymph flow to control levels without modifying the lymphocyte output. On the other hand, under insulin-restored normoglycemic conditions, both the thoracic duct lymph flow and the lymphocyte output were normalized. CONCLUSIONS: These findings suggest that variables related to defensive mechanisms, such as lymphocyte recirculation and particles uptake into the lymph nodes can benefit from insulin treatment, whereas glycemic control can benefit transport mechanisms in the lymphatic system, such as lymph flow and lymphatic transport of particles.

Psychoactive medications and actigraphically scored sleep quality in frail nursing home patients.

OBJECTIVE: The objective of this study was to examine the association between psychoactive medications and sleep quality in a sample of nursing home patients. DESIGN: We studied a baseline data collection for a clinical trial of a nonpharmacologic sleep intervention program. SETTING: This study was conducted at six community nursing homes. PARTICIPANTS: We studied 168 nursing home patients. METHODS: Sleep was recorded by wrist actigraphy for three to five nights under usual care conditions. Demographic and clinical data were collected by medical record reviews and patient assessments. RESULT: One or more routine psychoactive medications were being taken by 109 (65%) of the patients. Number of minutes of sleep, percent of time in bed asleep, and number of awakenings did not differ between those receiving and not receiving a psychoactive medication. Neither the use of antidepressants nor the use of only psychoactive medications reported to cause sedation was associated with significantly better sleep quality. CONCLUSION: Psychoactive medications as a general class of drugs were not associated with better or worse sleep quality in this very frail nursing home population. The effect of individual classes of psychoactive drugs on sleep quality in this patient population requires further study.

Cancer genetics nursing: impact of the double helix.

PURPOSE/OBJECTIVES: To describe the impact of genetic information on oncology nursing practice and to identify roles for oncology nurses in the field of cancer genetics. DATA SOURCES: Published articles, abstracts, books, and clinical experience. DATA SYNTHESIS: Oncology nurses in all areas of practice are affected by the recent explosion of genetic information. The identification of genetic mutations associated with increased risk for certain cancers and subsequent development of cancer predisposition testing have created a tremendous need for health care professionals who can explain and interpret genetic information. CONCLUSIONS: Oncology nurses already have many basic skills essential in the management of genetic information. An area in which oncology nurses may have the most to offer is in helping patients who carry a genetic predisposition to cancer understand the cancer surveillance and risk-management options available to them. Oncology nurses will be on the forefront in helping patients understand what this information means to them and how to apply it to their lives. IMPLICATIONS FOR NURSING PRACTICE: As we enter the 21st century, the advances in genetic information and the impact of the Human Genome Project will change oncology nursing as we know it. Oncology nurses will be expected to assess and interpret genetic and nongenetic cancer risk as an integrated whole, including genetic risk factors, environmental and lifestyle risk factors, and the interaction of the two. The challenge will be to make this complex information meaningful to patients as they make choices to manage their own cancer risk.

Heart stopper genes: would you recognize a high risk patient?

Coronary artery disease (CAD) is one of the leading causes of death in the United States. Eighty percent of people having heart attacks have normal cholesterol levels. A quarter of the population have a condition called low-density lipoprotein (LDL) pattern B that has been associated with a threefold risk of myocardial infarction. Although early intervention can often prevent an otherwise fatal event, these patients often go unrecognized until after a myocardial infarction has occurred because they may not have the usual risk factors associated with cardiovascular disease. In patients with LDL pattern B, the standard lipid panel may be normal and inadequate for diagnosis and treatment of the condition. This article discusses how to identify a potentially high risk patient, available laboratory tests, management options, and the role of nurses in identifying high risk patients. The second author tells his personal story of surviving multiple cardiac arrests at a young age before being diagnosed with this condition.

Development and testing of the symptom experience scale.

The Symptom Experience Scale (SES) was designed to measure women's experience of symptoms associated with treatment for breast cancer. The SES, a modification of McCorkle's Symptom Distress Scale, was developed and tested in a sample of 252 women with breast cancer. Exploratory factor analysis yielded six factors, which used all 24 SES items and accounted for 83.2% of the variance. The factors were nausea and appetite, fatigue and sleep, concentration, appearance, bowel pattern, and pain. Cronbach's alpha internal consistency reliability coefficients ranged from 0.92 to 0.96; the alpha for the total SES was 0.94. Subscale to subscale correlations ranged from 0.21 to 0.56. Additional research is recommended with samples large enough to permit confirmatory factor analysis and determine the stability of the factor structure identified in the present study. Additional research also is recommended to determine the applicability of the SES for men and women of diverse ethnic groups with various types of cancer and other chronic illnesses.

Regulation of advanced nursing practice: Part One--Second licensure.

PURPOSE: To present an overview of the issues relating to second licensure and advanced nursing practice in oncology and to summarize recommendations regarding second licensure from the State-of-the-Knowledge Conference on Advanced Practice in Oncology Nursing. DATA SOURCES: Published journal articles, a position paper and survey prepared by the National Council of State Boards of Nursing, and papers presented at the conference. DATA SYNTHESIS: The use of second licensure to regulate advanced nursing practice is controversial and has advantages and disadvantages. CONCLUSIONS: The Oncology Nursing Society should not adopt a position on second licensure until more is known about its advantages and disadvantages. IMPLICATIONS FOR NURSING PRACTICE: Advanced practice nurses must understand how licensure laws affect their scope of practice, their prescriptive authority, reimbursement, and other practice issues.

Prostate cancer prevention: risk reduction through life-style, diet, and chemoprevention.

PURPOSE/OBJECTIVES: To provide an overview of factors related to life-style, diet, and chemoprevention agents that may help lower the incidence of prostate cancer in the future. DATA SOURCES: Analysis of literature from MEDLINE (National Library of Medicine, Bethesda, MD), published journal articles, papers from journal bibliographies, and protocol of the Southwest Oncology Group Prostate Cancer Prevention Trial. DATA SYNTHESIS: Information related to prostate cancer prevention is accumulating, which may lead to a decreased disease incidence. Promising areas include use of chemoprevention, identification of life-style and dietary factors that may affect prostate cancer risk, and identification of genetic factors involved in prostatic carcinogenesis. CONCLUSIONS: Increased understanding of hormonal and genetic changes that occur during prostatic carcinogenesis may lead to effective methods of preventing this disease. Prostate cancer risk may be reduced by altering life-style and dietary factors and by using chemoprevention agents. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses play an important role in prostate cancer prevention through cancer risk assessment and patient and public education related to risk-reduction strategies.

Prostate cancer screening: getting your patients to follow-up.

Nurses have the clinical knowledge and organizational ability to carry out a quality prostate cancer screening program. In addition, nurses possess the communication and patient education skills necessary for effective follow-up. With the dramatic increase in prostate screening programs, nurses are likely to be among the leaders in this effort. Motivating patients to follow up when they have an abnormal DRE or serum PSA can increase the early detection of prostate cancer and potentially cure the disease. After all, this is the primary purpose of the screening.

Prostate-specific antigen: the new early detection test for prostate cancer.

Prostate cancer is the second leading cause of cancer deaths in men, affecting one in 11 men in the United States. Early detection in the preclinical stage offers the best chance of increased survival, decreased morbidity and possible cure. Prostate-specific-antigen assay is rapidly joining the digital rectal exam as the standard early detection test for prostate cancer because digital rectal exam alone is no longer considered adequate. The American Cancer Society now recommends an annual serum prostate-specific-antigen test for every man age 50 or older, and current evidence strongly suggests that combining the serum prostate-specific-antigen test with the digital rectal exam increases the early detection rate for prostate cancer. This article describes prostate-cancer screening guidelines, serum prostate-specific-antigen test interpretation and an algorithm for abnormal serum prostate-specific-antigen test follow-up.