Renal failure is a frequent and costly complication of many chronic diseases, including diabetes and hypertension. One common feature of renal failure is glomerulosclerosis, the pathobiology of which is unclear. To help elucidate this, we generated a mouse strain carrying the missense mutation Wt1 R394W, which predisposes humans to glomerulosclerosis and early-onset renal failure (Denys-Drash syndrome [DDS]). Kidney development was normal in Wt1؉/R394W heterozygotes. However, by 4 months of age 100% of male heterozygotes displayed proteinuria and glomerulosclerosis characteristic of DDS patients. This phenotype was observed in an MF1 background but not in a mixed B6/129 background, suggestive of the action of a strain-specific modifying gene(s). WT1 encodes a nuclear transcription factor, and the R394W mutation is known to impair this function. Therefore, to investigate the mechanism of Wt1 R394W-induced renal failure, the expression of genes whose deletion leads to glomerulosclerosis (NPHS1, NPHS2, and CD2AP) was quantitated. In mutant kidneys, NPHS1 and NPHS2 were only moderately downregulated (25 to 30%) at birth but not at 2 or 4 months. Expression of CD2AP was not changed at birth but was significantly upregulated at 2 and 4 months. Podocalyxin was downregulated by 20% in newborn kidneys but not in kidneys at later ages. Two other genes implicated in glomerulosclerosis, TGFB1 and IGF1, were upregulated at 2 months and at 2 and 4 months, respectively. It is not clear whether the significant alterations in gene expression are a cause or a consequence of the disease process. However, the data do suggest that Wt1 R394W-induced glomerulosclerosis may be independent of downregulation of the genes for NPHS1, NPHS2, CD2AP, and podocalyxin and may involve other genes yet to be implicated in renal failure. The Wt1 R394W mouse recapitulates the pathology and disease progression observed in patients carrying the same mutation, and the mutation is completely penetrant in male animals. Thus, it will be a powerful and biologically relevant model for investigating the pathobiology of the earliest events in glomerulosclerosis.Glomerulosclerosis, whether primary or secondary to other disease processes, is a key and common feature of progressive renal failure, which is a major cause of morbidity and mortality in the United States. Although several genetic and environmental insults are known to cause primary glomerulosclerosis, the cellular mechanism by which they initiate this process is still largely unknown. A knowledge of these mechanisms would greatly aid in identifying strategies to prevent or slow the development of glomerulosclerosis, regardless of its etiology.Glomeruli are complex and specialized structures responsible for blood filtration in the kidney and are targets of injury in a number of human diseases. The major functional features of the glomerulus are capillary loops lined with fenestrated endothelial cells, supporting mesangial cells, the glomerular basement membrane (GBM), and podocytes. The "octopuslike" ...

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Restoration of the tumor-suppression function by gene transfer of the melanoma differentiationassociated gene 7 (MDA7)/interleukin 24 (IL-24) successfully induces apoptosis in melanoma tumors in vivo. To address the molecular mechanisms involved, we previously revealed that MDA7/ IL-24 treatment of melanoma cells down-regulates interferon regulatory factor (IRF)-1 expression and concomitantly up-regulates IRF-2 expression, which competes with the activity of IRF-1 and reverses the induction of IRF-1-regulated inducible nitric oxide synthase (iNOS). Interferons (IFNs) influence melanoma cell survival by modulating apoptosis. A class I IFN (IFN alfa) has been approved for the treatment of advanced melanoma with some limited success. A class II IFN (IFN gamma), on the other hand, supports melanoma cell survival, possibly through constitutive activation of iNOS expression. We therefore conducted this study to explore the molecular pathways of MDA7/ IL-24 regulation of apoptosis via the intracellular induction of IFNs in melanoma. We hypothesized that the restoration of the MDA7/IL-24 axis leads to upregulation of Class I IFNs and induction of the apoptotic cascade. We found that MDA7/IL-24 induces the secretion of endogenous IFN beta, another class I IFN, leading to the arrest of melanoma cell growth and apoptosis. We also identified a series of apoptotic markers that play a role in this pathway, including the regulation of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and Fas-FasL. In summary, we described a novel pathway of MDA7/IL-24 regulation of apoptosis in melanoma tumors via endogenous IFN beta induction followed by IRF regulation and TRAIL/FasL system activation.

A better understanding of novel and emerging longitudinal data collection strategies will ultimately improve longitudinal data collection as well as foster research efforts. Nurse researchers, along with all researchers, must be aware of and consider implementing novel and emerging strategies to ensure future healthcare research success.

This study examined the prevalence of self-reported depressive symptoms and the self reported somatic depressive symptoms as measured by the Beck Depression Inventory-II (BDI-II) among patients hospitalized for acute coronary syndrome (ACS), and explored the impact of gender on both. A convenience sample of 789 adults (248 women and 541 men) was recruited for the study during hospital admission for ACS and participants were screened for self-reported depressive symptoms. BDI-II scores of ≥14 indicate a moderate level of depressive symptoms and this cut-off score was used to categorize patients into depressed and non-depressed groups. Pearson chi-square tests for independence (categorical variables) and t tests for independent samples (continuous variables) were used for gender comparisons. Results showed that depressive symptoms during ACS episodes were different between women and men. Women reported greater overall depressive symptoms (BDI-II mean = 11.89, S.D. = 9.68) than men (BDI-II mean = 9.00, S.D. = 7.93) (P < 0.000). Significantly more women (7.66%) were identified positive for somatic depressive symptoms (sleep and appetite disturbances and fatigue) than men (2.22%) (P = 0.0003). Findings support that there are gender differences in depressive symptoms experienced by patients hospitalized for ACS. Somatic symptoms of depression may be important indicators of depression especially among female ACS patients.

Preservation of biospecimens for biobanking applications traditionally involves freezing while maintaining the integrity of the product throughout multiple freeze-thaw cycles. The protection and stabilization of DNA at room temperature (RT) may eliminate the costs associated with freezer storage and reduce the maintenance costs for biobanks. However, there is a paucity of information describing the yield, purity, and integrity of DNA extracted from biospecimens stored at RT. To evaluate the yield, purity, and integrity of DNA extracted from whole blood samples stored at RT (18°C), low (-20°C), and ultra-low (-80°C) temperatures, whole blood samples from sheep and human subjects were collected, and aliquots were stored at RT (18°C), -20°C, and -80°C. Blood samples at RT were stored utilizing biostabilization technology designed to protect genomic DNA in whole blood. The quantification of the extracted DNA was determined by spectrophotometry, and the integrity was assessed following gel electrophoresis. Storage temperature did not influence the DNA yield (p=0.52); DNA yield averaged 13.6 ± 1.2 ng/μL across all storage temperatures. However, DNA yield was influenced (p=0.04) by species. The DNA yield was not influenced by a species × storage temperature interaction (p=0.84). Among the samples stored at RT, the species, type of technology utilized, and the interaction did not influence (p>0.13) DNA yield for both DNAgard and DNAstable. The 260/280 ratio was influenced by a species × storage temperature interaction (p=0.01). Generally, the 260/280 ratios were higher (p<0.05) for human samples stored at low and ultra-low temperatures compared to sheep samples stored at similar temperatures. Ambient temperature-based technologies offer an alternative to low temperature biospecimen preservation for blood that can be utilized by biobanks to reduce freezer storage costs while maintaining the quality of the biospecimen.

Background Despite the availability of established guidelines for measuring platelet serotonin, these guidelines may be difficult to follow in a hospital setting where time to processing may vary from sample to sample. Purpose The purpose of this study was to evaluate the effect of the time to processing of human blood samples on the stability of the enzyme-linked immunosorbent assay (ELISA) for the determination of platelet serotonin levels in human plasma. Method Human blood samples collected from a convenience sample of eight healthy volunteers were analyzed to determine platelet serotonin levels from plasma collected in ethylene diamine tetra acetic acid (EDTA) tubes and stored at 4°C for 3 hr, 5 hr, 8 hr, and 12 hr. Results Refrigeration storage at 4°C for 3 hr, 5 hr, 8 hr, and 12 hr altered the platelet serotonin measurement when compared to immediate processing. The bias for the samples stored at 4°C for 3 hr was 102.3 (±217.39 ng/109 platelets), for 5 hr was 200.1 (±132.76 ng/109 platelets), for 8 hr was 146.9 (±221.41 ng/109 platelets), and for 12 hr was –67.6 (±349.60 ng/109 platelets). Discussion Results from this study show that accurate measurement of platelet serotonin levels is dependent on time to processing. Researchers should therefore follow a standardized laboratory guideline for obtaining immediate platelet serotonin levels after blood sample collection.

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