Stage Two Malignant Melanoma of the Clitoris: A Case Report.
Mucosal melanomas are rare in comparison to cutaneous melanomas, accounting for approximately 1% of all melanoma cases. Vulvovaginal melanoma is the least common mucosal melanoma subtype. For patients without distant metastases at presentation, regional lymph node involvement is the most important prognostic indicator. Lymphoscintigraphy is a method used to identify the sentinel lymph node (SLN), directing subsequent biopsies to the lymph node at highest risk for cancer spread. We present a 67-year-old Hispanic female with stage II B (T4aN0M0) melanoma of the clitoris. The patient initially sought medical treatment for a pigmented vulvar lesion over her clitoris that occasionally bleeds. She stated that over the past month the lesion began to grow peripherally and then acquired a very dark color. The patient underwent local excision of the mass and subsequent pathology revealed malignant melanoma. The patient underwent lymphoscintigraphy which allowed localization and subsequent biopsy of the left inguinal sentinel node. The left inguinal sentinel node was negative for metastatic melanoma. It was opted to keep her under observation. Compared to cutaneous melanoma, patients with mucosal melanoma usually present with more advanced disease, and thus efficacious imaging practices play a significant role in the management of the disease. Lymphoscintigraphy is a well-tolerated, validated, cost-effective, and reliable method of detecting sentinel nodes with minimal radiation exposure to the patient. In our case, early clinical detection allowed for prompt surgical intervention, pathologic diagnosis, and reliable staging via lymphoscintigraphy of a rare form of malignant melanoma.


Parametric imaging of clear cell and papillary renal cell carcinoma using contrast-enhanced ultrasound (CEUS).
The aim of this study was to analyse clear cell and papillary renal cell carcinoma (RCC) examined with contrast-enhanced ultrasound (CEUS) and a second generation blood pool agent (SonoVue®, Bracco, Milan, Italy) before clinical intervention. A total of 41 patients with histologically proven subtypes of RCC were examined. 29 patients had a clear cell RCC and 12 patients showed a papillary RCC. Average size in the clear cell RCC group was 6.07 cm and 1.88 cm in the papillary RCC group. An experienced radiologist examined all patients with CEUS. The following parameters were analysed: maximum signal intensity (PEAK), time elapsed until PEAK is reached (MTT), local blood flow (RBF), area under the time intensity curve (AUC) and the signal intensity (SI) during the course of time. For both groups all comparisons were made based on healthy renal parenchyma. In the clear cell RCC significant differences (significance level p < 0.05) between cancerous tissue and the healthy renal parenchyma were noticed in all four parameters. The clear cell RCC showed a significant reduced blood volume. It reached the PEAK reading relatively rapidly and its signal intensity was always lower than that of the healthy renal parenchyma. In the arterial phase retarded absorption of the contrast agent was observed, followed by fast washing out of the contrast agent bubbles.In the papillary RCC group, significant findings as to PEAK and RBF as well as a slightly significant difference as to AUC were recorded. The papillary RCC had a lower blood supply and reached its PEAK reading later. Its signal intensity was also reduced. The signal intensity of papillary NCC was significantly lower compared with clear cell RCC; absorption and washing out of the contrast agent was delayed. CEUS seems to be an useful additional method to clinically differentiate between clear cell and papillary RCC. In daily clinical use, patients with contraindication for other imaging methods, especially the magnetic resonance imaging, might particularly benefit from this method.


Effects of delta opioid antagonists on enkephalin-induced seizures.
We examined the effect of opioid receptor antagonists on the seizure phenomena induced by specific delta opioid receptor agonist [D-Ser2,Leu5] enkephalyl-Thr (DSLET). The experiments have been performed in the anesthetized rats, and the DSLET-induced seizure phenomena were registered by electrocorticogram and electromyogram. It was demonstrated that two selective delta opioid receptor antagonists, ICI 152,129 and ICI 174,864 inhibited DSLET-induced epileptiform ECoG pattern and myoclonic contractions in a dose-related manner. An equimolar concentration of naloxone failed to antagonize the epileptiform effects of DSLET. It is concluded that delta opioid receptor agonist-induced seizure is mediated by delta receptors, since it can be blocked by delta opioid receptor antagonists. Evidently, delta opioid antagonists can be used as a good tool, in order to demonstrate a delta component in the seizure phenomena induced by other endogenous opioid peptides or their derivatives.


[Effect of benzo(a)pyrene on the expression of AhR-regulated microRNA in female and male rat lungs].
Smoking is the main risk factor for lung cancer, mainly due to presence of nitrosamines and polycyclic aromatic hydrocarbons, including benzo[a]pyrene (BP) in tobacco smoke composition. The genotoxic effect of BP is based on the high DNA-binding ability of its metabolites, while the epigenetic effects are mediated by a change in the expression of cancer related genes or regulatory RNAs. It has been shown that women have a higher risk to develop lung cancer upon smoking rather than men. We hypothesized that crosstalk between signaling pathways activated by BP and estrogens could underlie the sex-dependent differences in miRNAs expression. To test this hypothesis, male and female rats were subjected to short-term or long-term BP exposure. Using in silico analysis, miRNAs containing the ER- and AhR-binding sites in the promoters of the genes (or host genes) were selected. During chronic exposure of BP the expression of miR-22-3p, -29a-3p, -126a-3p, -193b-5p in the lungs of male rats were significantly increased, while the level of miRNA-483-3p were decreased. Expression of miRNA-483-3p was up-regulated during chronic BP exposure in the lungs of female rats and the levels of other studied miRNAs were unchanged. In turn, changes in the expression of miRNAs were followed by changes in the expression of their target genes, including PTEN, EMP2, IGF1, ITGA6, SLC34A2, and the observed changes in female and male rat lungs were varied. Thus, our results suggest that sex-dependent epigenetic effects of BP may be based on different expression of AhR- and ER- regulated miRNAs.


A new dump system design for stray light reduction of Thomson scattering diagnostic system on EAST.
Thomson scattering (TS) diagnostic is an important diagnostic for measuring electron temperature and density during plasma discharge. However, the measurement of Thomson scattering signal is disturbed by the stray light easily. The stray light sources in the Experimental Advanced Superconducting Tokamak (EAST) TS diagnostic system were analyzed by a simulation model of the diagnostic system, and simulation results show that the dump system is the primary stray light source. Based on the optics theory and the simulation analysis, a novel dump system including an improved beam trap was proposed and installed. The measurement results indicate that the new dump system can reduce more than 60% of the stray light for the diagnostic system, and the influence of stray light on the error of measured density decreases.


The stimuli releasing histamine from murine bone marrow-derived mast cells. 1. The presence of P2-purinoceptors.
We have examined the histamine-releasing effects of 4 different stimuli on murine bone marrow-derived mast cells (BMMC). We obtained 10(9) BMMC from one femur of a CBA/J mouse when it was cultured in the presence of conditioned medium from WEHI-3 cells for 4 weeks. When these cells were sensitized with monoclonal anti-DNP IgE antibodies, DNP35HSA antigen, ionomycin, thrombin and ATP induced 70%, 70%, 40% and 60% histamine release from BMMC, respectively. Thrombin induced rapid degranulation, whereas the kinetics of histamine release by other stimuli reached a plateau after 5 min. ADP, AMP and GTP as well as ATP but not adenosine caused histamine release from BMMC, suggesting the presence of P2-purinoceptors on these cells.


Ball variance in a Harken mitral prosthesis. Echocardiographic and phonocardiographic features.
A case of ball variance in a caged-ball prosthetic valve is presented, illustrating the echocardiographic and phonocardiographic features. The swollen silicone ball was observed on echocardiographic studies to have diminished motion and to incompletely open within its cage. On phonocardiographic studies, the Q-S1 interval was prolonged, and no opening sound could be recorded. These noninvasive techniques may be helpful in predicting the need for replacement of prosthetic valves that have silicone rubber balls.


Requirements for heterologous production of a complex metalloenzyme: the membrane-bound [NiFe] hydrogenase.
By taking advantage of the tightly clustered genes for the membrane-bound [NiFe] hydrogenase of Ralstonia eutropha H16, broad-host-range recombinant plasmids were constructed carrying the entire membrane-bound hydrogenase (MBH) operon encompassing 21 genes. We demonstrate that the complex MBH biosynthetic apparatus is actively produced in hydrogenase-free hosts yielding fully assembled and functional MBH protein.


A comparison of interfraction setup error, patient comfort, and therapist acceptance for 2 different prostate radiation therapy immobilization devices.
Our purpose was to investigate interfraction setup error of the immobilization device required to implement transperineal ultrasound compared with the current, standard immobilization device. Patient comfort and radiation therapist (RT) satisfaction were also assessed. Cone beam computed tomography images were acquired before 4069 fractions from 111 patients (control group, n = 56; intervention group, n = 55) were analyzed. The intervention group was immobilized using the Clarity Immobilization System (CIS), comprising a knee rest with autoscan probe kit and transperineal ultrasound probe (n = 55), and control group using a leg immobilizer (LI) (n = 56). Interfraction setup errors were compared for both groups. Weekly questionnaires using a 10-point visual analog scale were administered to both patient groups to measure and compare patient comfort. RT acceptance for both devices was also compared using a survey. There was no significant difference in the magnitude of interfraction cone beam computed tomography-derived setup shifts in the lateral and anteroposterior direction between the LI and CIS (P = .878 and .690, respectively). However, a significant difference (P = .003) was observed in the superoinferior direction between the 2 groups of patients. Patient-reported level of comfort and stability demonstrated no significant difference between groups (P = .994 and .132). RT user acceptance measures for the LI and CIS were ease of handling (100% vs 53.7%), storage (100% vs 61.1%), and cleaning of the devices (100% vs 64.8%), respectively. The CIS demonstrated stability and reproducibility in prostate treatment setup comparable to LI. The CIS device had no impact on patient comfort; however, RTs indicated a preference for LI over the CIS mainly because of its weight and bulkiness.


Interaction of biomolecules with anatase, rutile and amorphous TiO2 surfaces: A molecular dynamics study.
The adhesion of biomolecules to dental and orthopedic implants is a fundamental step in the process of osseointegration. Short peptide motifs, such as RGD or KRSR, carried by extracellular matrix proteins or coated onto implant surfaces, accelerate cell adhesion and tissue formation. For this reason, understanding the binding mechanisms of adhesive peptides to oxidized surfaces of titanium implants is of paramount importance. We performed molecular dynamics simulations to compare the adhesion properties of 6 peptides, including the tripeptide RGD, its variants KGD and LGD, as well as the tetrapeptide KRSR, its variant LRSR and its truncated version RSR, on anatase, rutile, and amorphous titanium dioxide (TiO2) surfaces. The migration of these molecules from the water phase to the surface was simulated in an aqueous environment. Based on these simulations, we calculated the residence time of each peptide bound to the three different TiO2 structures. It was found that the presence of an N-terminal lysine or arginine amino acid residue resulted in more efficient surface binding. A pulling simulation was performed to detach the adhered molecules. The maximum pulling force and the binding energy were determined from the results of these simulations. The tri- and tetrapeptides had slightly greater adhesion affinity to the amorphous and anatase structure than to rutile in general, however specific surface and peptide binding characters could be detected. The binding energies obtained from our simulations allowed us to rank the adhesion strengths of the studied peptides.


Human immunodeficiency virus and type 2 diabetes.
The prevalence of diabetes is higher amongst individuals infected with HIV. The major contributor to hyperglycaemia is thought to be iatrogenic, with protease inhibitors being most commonly associated to insulin resistance. This article is to update general practitioners on the diagnosis and management of diabetes in HIV-infected patients. Specific considerations are highlighted including interactions of particular diabetic drugs with antiretroviral therapy (ART). We articulate why the use of Hemoglobin A1c (HbA1c) testing is not recommended as a diagnostic tool.


Pathological consequences of the vanishing twin.
Fetus papyraceus is a mummified, compressed fetus occurring in association with a viable twin. The death of the fetus usually occurs early in the second trimester. A co-twin dying earlier may be absorbed completely, whilst later fetal death usually results in macerated, but not compressed fetuses. This course of events can be well demonstrated by ultrasonography. The death of one fetuses may be associated with minor malformations of the surviving one. After termination of twin pregnancies the detailed check-up of the newborn and histopathological examination of the placenta is essential.


Photoprecursor approach as an effective means for preparing multilayer organic semiconducting thin films by solution processes.
The vertical composition profile of active layer has a major effect on the performance of organic photovoltaic devices (OPVs). While stepwise deposition of different materials is a conceptually straightforward method for controlled preparation of multi-component active layers, it is practically challenging for solution processes because of dissolution of the lower layer. Herein, we overcome this difficulty by employing the photoprecursor approach, in which a soluble photoprecursor is solution-deposited then photoconverted in situ to a poorly soluble organic semiconductor. This approach enables solution-processing of the p-i-n triple-layer architecture that has been suggested to be effective in obtaining efficient OPVs. We show that, when 2,6-dithienylanthracene and a fullerene derivative PC71BM are used as donor and acceptor, respectively, the best p-i-n OPV affords a higher photovoltaic efficiency than the corresponding p-n device by 24% and bulk-heterojunction device by 67%. The photoprecursor approach is also applied to preparation of three-component p-i-n films containing another donor 2,6-bis(5'-(2-ethylhexyl)-(2,2'-bithiophen)-5-yl)anthracene in the i-layer to provide a nearly doubled efficiency as compared to the original two-component p-i-n system. These results indicate that the present approach can serve as an effective means for controlled preparation of well-performing multi-component active layers in OPVs and related organic electronic devices.


Immunogenicity and protective efficacy of an RSV G S177Q central conserved domain nanoparticle vaccine.
Respiratory syncytial virus (RSV) can cause lower respiratory tract disease in infants and elderly populations. Despite decades of research, there remains no safe and approved RSV vaccine. Previously, we showed that an RSV G glycoprotein subunit vaccine candidate with a single point mutation within the central conserved domain (CCD), i.e. S177Q, considerably improved immunogenicity. Here, we examine the development of nanoparticle (NP) vaccines having either an RSV G protein CCD with wild-type sequence (NPWT) or an S177Q mutation (NP-S177Q). The NP vaccine immunogens were adjuvanted with monophosphoryl lipid A (MPLA), a TLR4 agonist to improve Th1- type responses. BALB/c mice were primed with 10 μg of NP-WT vaccine, NPS177Q, or vehicle, rested, and then boosted with a high (25 μg) or low (10 μg) dose of the NP-WT or NP-S177Q homologous candidate and subsequently challenged with RSV A2. The results showed that mice boosted with NP-S177Q developed superior immunogenicity and neutralizing antibodies compared to NP-WT boosting. IgG from either NP-S177Q or NP-WT vaccinated mice did not interfere with fractalkine (CX3CL1) binding to CX3CR1 and effectively blocked G protein CX3C-CX3CR1 binding. Both NP-WT and NP-S177Q vaccination induced similar neutralizing antibodies to RSV in challenged mice compared to vehicle control. NP-S177Q boosting improved correlates of protection including reduced BAL cell infiltration following RSV challenge. However, the NP vaccine platform will require improvement due to the poor solubility and the unexpectedly weaker Th1-type IgG2a response. The results from this study support further NP-S177Q vaccine candidate development.


Primary mediastinal seminoma.
The records and histopathology of 4 patients with primary mediastinal seminoma were reviewed. All of them were male and at presentation had one or more poor prognostic factors, namely superior vena cava obstruction, hilar lymphadenopathy, bulky tumor, and raised alpha-fetoprotein titers. Their treatment consisted partly or entirely of chemotherapy. Three of them survived free of disease 2-5 years off treatment, 1 of them died of pulmonary fibrosis with no tumor found at postmortem examination. The prognostic factors and treatment of this tumor and the risk of combined radiotherapy and chemotherapy containing bleomycin are discussed.


Hydrophobic, Superabsorbing Aerogels from Choline Chloride-Based Deep Eutectic Solvent Pretreated and Silylated Cellulose Nanofibrils for Selective Oil Removal.
Superabsorbents are highly appealing materials for use in cleaning up oil and chemical spills. However, the development of a low-cost, highly efficient superabsorbent remains a major challenge. This paper demonstrates a straightforward method of producing a cellulose nanofibril aerogel that is low-cost, ultralight, highly porous, hydrophobic, and reusable superabsorbing cellulose nanofibril aerogel from recycled waste fibers using a simple, environmentally friendly nanofibrillation treatment involving deep eutectic solvent and freeze-drying. Nanofibrillation and hydrophobic modification (silylation) of waste cellulose fibers resulted in nanofibril sponges with ultralow density (0.0029 g/cm3) and high porosity (up to 99.81%) after freeze-drying. These sponges exhibited excellent absorption performances for various oils and organic solvents and were reusable. In particular, the nanofibril aerogels showed selectivity in absorbing marine diesel oil from an oil-water mixture and possessed ultrahigh absorption capacities of up to 142.9 g/g, much higher than those of the commercial absorbent materials (i.e., polypropylene-based material) (8.1-24.6 g/g) that were used as references. The absorbed oil could easily be recovered by means of simple mechanical squeezing. In addition, the nanofibril sponges exhibited excellent reusability, maintaining a high capacity to absorb diesel oil for at least 30 cycles at 71.4-81.0% of capacity compared to a fresh absorbent. The above-mentioned advantages make cellulose nanofibril superabsorbents created from recycled waste cellulose fibers promising material for cleaning oil and chemical spills.


[Rho-kinase signaling pathway participates in endothelin-1-induced human airway smooth muscle cell migration and cytoskeletal reorganization].
To investigate the role of Rho-kinase signaling pathway in human airway smooth muscle cell (ASMCs) migration and cytoskeletal reorganization induced by endothelin-1 (ET-1). Primary cultured human ASMCs obtained by tracheal explant culture method were examined for cell migration in response to ET-1 treatment using modified Boyden chambers. The changes in actin cytoskeletal reorganization were observed under confocal laser scanning microscope, and the phosphorylation of myosin-phosphatase target 1 (p-MYPT1) was examined using Western blot analysis. At the concentration of 0.1, 1, 10, and 100 nmol/L, ET-1 induced migration of the ASMCs, and 10 nmol/L ET-1 produced the most obvious effect (P<0.01). Rho-kinase inhibitor Y-27632 showed a dose-dependent inhibitory effect on ET-1-induced ASMC migration, and in cells exposed to 10 nmol/L ET-1, Y-27632 at 10 micromol/L significantly blocked ASMC migration (P<0.01). ET-1 (10 nmol/L) exposure resulted in reorganization of actin cytoskeleton and formation of stress fibers in the ASMCs, which were obviously inhibited by Y-27632. Compared with the control group, the AMSCs showed significant enhancement of p-MYPT1 protein expression after ET-1 exposure for 15 and 30 min (P<0.01), but prolonged exposure failed to result in the expression enhancement (P>0.05). Rho-kinase signaling pathway may play an important role in ET-1-induced ASMC migration and reorganization of actin cytoskeleton.


Role of PCNA and ependymal cells in ascidian neural development.
The central nervous system (CNS) of the ascidian tadpole larva consists of only 370 cells, yet it develops similarly to the elaborate vertebrate CNS. There are two phases in ascidian CNS development: an early phase, in which cells of mixed neuronal and non-neuronal (ependymal) fates are formed, and a late phase, in which only additional ependymal cells are produced. Proliferating cell nuclear antigen (PCNA), the auxiliary factor for DNA polymerase , is expressed zygotically in the developing CNS. Here we employ antisense oligonucleotides directed against PCNA mRNA to determine the role of late dividing ependymal cells in embryos of the ascidian Styela clava. An antisense oligonucleotide beginning at the translation start site in PCNA mRNA was effective in inhibiting zygotic PCNA accumulation. In normal tadpoles, the head is located in the same anteroposterior plane as the tail, which promotes larval swimming and dispersal. Embryos treated with PCNA antisense oligonucleotides developed into boomerang-shaped tadpoles with their heads positioned at an acute angle with respect to their tails, causing them to swim in circles rather than in the usual forward direction. Further studies showed that PCNA inhibition arrested DNA synthesis and induced nuclear DNA fragmentation typical of programmed cell death in the developing CNS. No other defects were apparent in these abnormal tadpoles, and they were eventually able to metamorphose into juveniles. The results suggest that PCNA and late dividing ependymal cells are required for normal CNS development and larval morphogenesis in ascidians.


Effects of selective inhibition of cytochrome P-450 omega-hydroxylases and ischemic preconditioning in myocardial protection.
Cytochrome P-450 (CYP) omega-hydroxylases and their arachidonic acid (AA) metabolite, 20-hydroxyeicosatetraenoic acid (20-HETE), produce a detrimental effect on ischemia-reperfusion injury in canine hearts, and the inhibition of CYP omega-hydroxylases markedly reduces myocardial infarct size expressed as a percentage of the area at risk (IS/AAR, %). In this study, we demonstrated that a specific CYP omega-hydroxylase inhibitor, N-methylsulfonyl-12,12-dibromododec-11-enamide (DDMS), markedly reduced 20-HETE production during ischemia-reperfusion and reduced myocardial infarct size compared with control [19.5 +/- 1.0% (control), 9.6 +/- 1.5% (0.40 mg/kg DDMS), 4.0 +/- 2.0% (0.81 mg/kg DDMS), P < 0.01]. In addition, 20-hydroxyeicosa-6(Z),15(Z)-dienoic acid (20-HEDE, a putative 20-HETE antagonist) significantly reduced myocardial infarct size from control [10.3 +/- 1.3% (0.032 mg/kg 20-HEDE) and 5.9 +/- 1.9% (0.064 mg/kg 20-HEDE), P < 0.05]. We further demonstrated that one 5-min period of ischemic preconditioning (IPC) reduced infarct size to a similar extent as that observed with the high doses of DDMS and 20-HEDE, and the higher dose of DDMS given simultaneously with IPC augmented the infarct size reduction [9.9 +/- 2.8% (IPC) to 2.5 +/- 1.4% (0.81 mg/kg DDMS), P < 0.05] to a greater degree than that observed with either treatment alone. These results suggest an important negative role for endogenous CYP omega-hydroxylases and their product, 20-HETE, to exacerbate myocardial injury in canine myocardium. Furthermore, for the first time, this study demonstrates that the effect of IPC and the inhibition of CYP omega-hydroxylase synthesis (DDMS) or its actions (20-HEDE) may have additive effects in protecting the canine heart from ischemia-reperfusion injury.


Immunohistochemical detection of p53 expression in patients with preoperative chemoradiation for rectal cancer: association with prognosis.
The expression of p53 in patients with rectal cancer who underwent preoperative chemoradiation and and its potential prognostic significance were evaluated. p53 expression was examined using immunohistochemistry in pathologic specimens from 210 rectal cancer patients with preoperative chemoradiotherapy and radical surgery. All patients were classified into two groups according to the p53 expression: low p53 (<50% nuclear staining) and high p53 (≥50%) groups. p53 expression was significantly associated with tumor location from the anal verge (p=0.036). In univariate analysis, p53 expression was not associated with disease-free survival (p=0.118) or local recurrence-free survival (p=0.089). Multivariate analysis showed that tumor distance from the anal verge (p=0.006), ypN category (p=0.011), and perineural invasion (p=0.048) were independent predictors of disease-free survival; tumor distance from the anal verge was the only independent predictor of local recurrence-free survival. When the p53 groups were subdivided according to ypTNM category, disease-free survival differed significantly in patients with ypN+ disease (p=0.027) only. Expression of p53 in pathologic specimens as measured by immunohistochemical methods may have a significant prognostic impact on survival in patients with ypN+ rectal cancer with preoperative chemoradiotherapy. However, it was not an independent predictor of recurrence or survival.


Anxiety and depression among infertile women: a cross-sectional survey from Hungary.
Infertility is often associated with a chronic state of stress which may manifest itself in anxiety-related and depressive symptoms. The aim of our study is to assess the psychological state of women with and without fertility problems, and to investigate the background factors of anxiety-related and depressive symptoms in women struggling with infertility. Our study was conducted with the participation of 225 (134 primary infertile and 91 fertile) women, recruited in a clinical setting and online. We used the following questionnaires: Spielberger Trait Anxiety Inventory (STAI-T), Shortened Beck Depression Inventory (BDI) and Fertility Problem Inventory (FPI). We also interviewed our subjects on the presence of other sources of stress (the quality of the relationship with their mother, financial and illness-related stress), and we described sociodemographic and fertility-specific characteristics. We tested our hypotheses using independent-samples t-tests (M ± SD) and multiple linear regression modelling (ß). Infertile women were younger (33.30 ± 4.85 vs. 35.74 ± 5.73, p = .001), but had significantly worse psychological well-being (BDI = 14.94 ± 12.90 vs. 8.95 ± 10.49, p < .0001; STAI-T = 48.76 ± 10.96 vs. 41.18 ± 11.26, p < .0001) than fertile subjects. Depressive symptoms and anxiety in infertile women were associated with age, social concern, sexual concern and maternal relationship stress. Trait anxiety was also associated with financial stress. Our model was able to account for 58% of the variance of depressive symptoms and 62% of the variance of trait anxiety. Depressive and anxiety-related symptoms of infertile women are more prominent than those of fertile females. The measurement of these indicators and the mitigation of underlying distress by adequate psychosocial interventions should be encouraged.


The pathogenic roles of the p.R130S prestin variant in DFNB61 hearing loss.
DFNB61 is a recessively inherited nonsyndromic hearing loss caused by mutations in SLC26A5 , the gene that encodes the voltage-driven motor protein, prestin. Prestin is abundantly expressed in the auditory outer hair cells that mediate cochlear amplification. Two DFNB61-associated SLC26A5 variants, p.W70X and p.R130S, were identified in patients who are compound heterozygous for these nonsense and missense changes ( SLC26A5 W70X/R130S ). Our recent study showed that mice homozygous for p.R130S ( Slc26a5 R130S/R130S ) suffer from hearing loss that is ascribed to significantly reduced motor kinetics of prestin. Given that W70X-prestin is nonfunctional, compound heterozygous Slc26a5 R130S/- mice were used as a model for human SLC26A5 W70X/R130S . By examining the pathophysiological consequences of p.R130S prestin when it is the sole allele for prestin protein production, we determined that this missense change results in progressive outer hair cell loss in addition to its effects on prestin's motor action. Thus, this study fully defines the pathogenic roles for the p.R130S prestin, which points to the presence of a limited time window for potential clinical intervention.


IRF7 as an Immune-Related Prognostic Biomarker and Associated with Tumor Microenvironment in Low-Grade Glioma.
Tumor microenvironment (TME) affects the occurrence and progression of low-grade glioma (LGG). The aim of this study is to identify TME-related genes that influence prognosis in LGG patients and to explore their function and role in tumor immunity. The TME components of LGG samples in the Cancer Genome Atlas (TCGA) database were identified by the ESTIMATE method, and differentially expressed genes (DEGs) with significant differences in immune scores and stromal scores were screened out. The core genes of DEGs were screened out by protein-protein interaction (PPI) network. Furthermore, immune-related target genes significantly correlated with prognosis were identified. Survival analysis and correlation analysis showed the correlation between target genes and clinical features and prognosis. The expression differences of target genes were verified by external database Chinese Glioma Genome Atlas (CGGA). CIBERSORT software identified the proportion of tumor-infiltrating immune cells (TICs) that were significantly related to target genes. Gene set enrichment analysis (GSEA) could enrich the main functions related to high and low expression of target genes. A total of 1567 DEGs were screened out from 529 LGG samples in the TCGA database, and 146 immune-related genes affecting prognosis were found. A total of 403 core genes were obtained from PPI network. The target gene interferon regulatory factor 7 (IRF7) was significantly associated with prognosis and clinical features of the tumor. The CGGA database verified the relationship between high and low expression groups of IRF7 and prognosis. GSEA indicated that IRF7 was mainly enriched in immune-related activities, significantly correlated with T cells CD8, macrophages M1, macrophages M2 and monocytes. The IRF7 is involved in immune responses in TME of LGG, which in turn influenced tumor occurrence and progression. IRF7 can act as a potential biomarker for prognosis in patients with LGG and provide a target for tumor immunotherapy.


Competition studies with repressors and activators of viral enhancer function in F9 mouse embryonal carcinoma cells.
DNA competition studies have been used to investigate the presence of a repressor of viral enhancer function in F9 mouse embryonal carcinoma cells. The complete polyoma virus enhancer region, cotransfected into F9 cells with the SV40 promoter/enhancer attached to a chloramphenicol acetyl transferase marker gene, induced a small increase in pSV2CAT expression. This can be explained by preferential but weak binding by polyoma sequences of a molecule repressing pSV2CAT transcription. Repressor activity substantially disappeared when the cells were induced to differentiate by retinoic acid. Repressor binding was localised to one half of the polyoma enhancer, but was lost on further fragmentation of this region. It appears that multiple sequence elements may be required for repressor binding and that these are at least partially separable from the complement of elements binding enhancer activating molecules.


Changes in mineral elements and starch quality of grains during the improvement of japonica rice cultivars.
The improvement of rice cultivars plays an important role in yield increase. However, little is known about the changes in starch quality and mineral elements during the improvement of rice cultivars. This study was conducted to investigate the changes in starch quality and mineral elements in japonica rice cultivars. Twelve typical rice cultivars, applied in the production in Jiangsu province during the last 60 years, were grown in the paddy fields. These cultivars were classified into six types according to their application times, plant types and genotypes. The nitrogen (N), phosphorus (P) and, and potassium (K) were mainly distributed in endosperm, bran and bran, respectively. Secondary and micromineral nutrients were distributed throughout grains. With the improvement of cultivars, total N contents gradually decreased, while total P, K and magnesium contents increased in grains. Total copper and zinc contents in type 80'S in grains were highest. The improvement of cultivars enhanced palatability (better gelatinisation enthalpy and amylose content), taste (better protein content) and protein quality (better protein components and essential amino acids). Correlation analysis indicated the close relationship between mineral elements and starch quality. The mineral elements and starch quality of grains during the improvement of japonica rice cultivars are improved. © 2017 Society of Chemical Industry.


Raman spectroscopy of dorsal root ganglia from streptozotocin-induced diabetic neuropathic rats submitted to photobiomodulation therapy.
In this study, we used Raman spectroscopy as a new tool to investigate pathological conditions at the level of chemical bond alterations in biological tissues. Currently, there have been no reports on the spectroscopic alterations caused by diabetic neuropathy in the dorsal root ganglia (DRG). DRG are a target for the treatment of neuropathic pain, and the need for more effective therapies is increasing. Photobiomodulation therapy (PBMT) through infrared low-level laser irradiation (904 nm) has shown analgesic effects on the treatment of neuropathy. Thus, the aim of this study was to use Raman spectroscopy to characterize the spectral DRG identities of streptozotocin (STZ)-induced diabetic neuropathic (hyperalgesic) rats and to study the influence of PBMT over such spectra. Characteristic DRG peaks were identified at 2704, 2850, 2885, 2940, 3061 and 3160 cm-1 , whose assignments are CH2 /CH3 symmetric/asymmetric stretches, and C─H vibrations of lipids and proteins. DRG from hyperalgesic rats showed an increased normalized intensity of 2704, 2850, 2885 and 3160 cm-1 . These same peaks had their normalized intensity reduced after PBMT treatment, accompanied by an anti-hyperalgesic effect. Raman spectroscopy was able to diagnose spectral alterations in DRG of hyperalgesic rats and the PBMT reduced the intensity of hyperalgesia and the altered Raman spectra.


Novel alternative splicing by exon skipping in KIT associated with whole-body roan in an intercrossed population of Landrace and Korean Native pigs.
The KIT locus has been suggested to be a strong candidate region linked with whole-body roan in the F(2) population produced by intercrosses between Landrace and Korean Native pigs. In this manuscript, we report the finding of a novel alternative splicing event in the porcine KIT gene that results in the skipping of exon 5 in the I(Rn) allele. KIT mRNAs that lack exon 5 were identified in the large intestine and skin, suggesting that the mechanism responsible for the skipping of exon 5 may be tissue specific. A U(26) repeat in intron 5 showed complete linkage (LOD = 11.8) with the roan phenotype and absolute association with the black phenotype of the Korean Native pig (KNP) population samples, inferring that the repeat pattern may alter the complementary base-pairing-mediated looping-out of introns 4 and 5, which may mediate the exon 5-skipping event. Although the sample size in our study was relatively small, we speculate that the R3 allele containing the U(26) repeat is a causative element for the roan phenotype via alternative control of the exon skipping in our roan pedigree.


Application of linear discriminant analysis to the differentiation of pure milk from different species and mixtures.
Discriminant analysis is used to identify different milk samples on the basis of the gas chromatographic data for fatty acids in 20 samples each of milk fat from cows, sheep, and goats. The method can differentiate mixtures and pure milks with a high degree of correct classifications. A good discrimination can also be obtained by using a reduced set of variables. The method is useful for the interpretation of gas chromatographic data and should allow a higher proportion of correct classifications than is possible by visual inspection of the chromatograms.


Aggregation and charge behavior of metallic and nonmetallic nanoparticles in the presence of competing similarly-charged inorganic ions.
The influence of competing, similarly charged, inorganic ions on the size and charge behavior of suspended titanium-dioxide (nTiO(2)), silver (nAg) and fullerene (nC(60)) nanoparticles (NPs) was investigated. Under pH and ionic conditions similar to natural water bodies, Ca(2+) induced aggregation of nTiO(2) and nAg NPs more strongly than K(+) and Na(+). Although K(+) and Na(+) had a similar effect on aggregation, K(+) provided better screening of the particle surface charge presumably because of its small hydrated radius. These effects were decidedly more prominent for TiO(2) than Ag. Anions (co-ions), SO(4)(2-) and Cl(-), affected the surface charge behavior of nTiO(2) but not of nAg NPs. The zeta potential (ZP) of nTiO(2) NPs was more negative at higher SO(4)(2-)/Cl(-) ratios than lower. When Mg(2+) was the counterion, charge inversion and rapid aggregation of nC(60) NPs occurred under alkaline conditions, with a more pronounced effect for Cl(-) than SO(4)(2-). Response dissimilarities suggest fundamental differences in the interfacial-interaction characteristics of these NPs in the aquatic environment with corresponding differences in transport of these particles. Our study also shows the important role played by the iso-electric point pH (pH(iep)) of the NPs in determining their aggregation kinetics in the environment.


Food consumption, physical activity and socio-economic status related to BMI, waist circumference and waist-to-height ratio in adolescents.
To examine the association between obesity and food group intakes, physical activity and socio-economic status in adolescents. A cross-sectional study was carried out in 2008. Cole's cut-off points were used to categorize BMI. Abdominal obesity was defined by a waist circumference at or above the 90th percentile, as well as a waist-to-height ratio at or above 0·500. Diet was evaluated using an FFQ, and the food group consumption was categorized using sex-specific tertiles of each food group amount. Physical activity was assessed via a self-report questionnaire. Socio-economic status was assessed referring to parental education and employment status. Data were analysed separately for girls and boys and the associations among food consumption, physical activity, socio-economic status and BMI, waist circumference and waist-to-height ratio were evaluated using logistic regression analysis, adjusting the results for potential confounders. Public schools in the Azorean Archipelago, Portugal. Adolescents (n 1209) aged 15-18 years. After adjustment, in boys, higher intake of ready-to-eat cereals was a negative predictor while vegetables were a positive predictor of overweight/obesity and abdominal obesity. Active boys had lower odds of abdominal obesity compared with inactive boys. Boys whose mother showed a low education level had higher odds of abdominal obesity compared with boys whose mother presented a high education level. Concerning girls, higher intake of sweets and pastries was a negative predictor of overweight/obesity and abdominal obesity. Girls in tertile 2 of milk intake had lower odds of abdominal obesity than those in tertile 1. Girls whose father had no relationship with employment displayed higher odds of abdominal obesity compared with girls whose father had high employment status. We have found that different measures of obesity have distinct associations with food group intakes, physical activity and socio-economic status.


Superantigen-based immunotherapy: a phase I trial of PNU-214565, a monoclonal antibody-staphylococcal enterotoxin A recombinant fusion protein, in advanced pancreatic and colorectal cancer.
To establish the maximum-tolerated dose (MTD) and define the toxicities of a single-dose infusion of PNU-214565, a recombinant Escherichia coli-derived fusion protein of Staphylococcal enterotoxin A (SEA) and the Fab-fragment of the C242 monoclonal antibody in patients with advanced colorectal and pancreatic carcinomas. To investigate the capability of PNU-214565 to induce a superantigen (SAg) response resulting in cytokine production and tumor regression. Twenty-one patients (age range, 39 to 76 years; median, 64; 12 men, nine women; 18 colorectal, three pancreatic cancers) were treated with a single 3-hour infusion of PNU-214565, with doses ranging from 0.01 to 1.5 ng/kg. All patients had prior chemotherapy and a good performance status Eastern Cooperative Oncology Group [ECOG] performance status [PS] = 0 [n = 10]; PS = 1 [n = 11]), 10 had prior radiation, and 18 had prior surgery. Fever and hypotension were the most common toxicities. Fever of any grade occurred in 16 of 21 patients (76%): four of 21 (19%) with grade 2 and two of 21 (9.5%) with grade 3. Hypotension of any grade occurred in 13 of 21 (62%): four of 21 with grade 2 and one of 21 (5%) with grade 3. Interleukin-2 (IL-2) and tumor necrosis factor alpha (TNF alpha) induction correlated with toxicity. In the two patients with grade 3 fever, peak IL-2 and TNF alpha levels were 2.9 IU/mL and 165 pg/mL, and 8.3 IU/mL and 245 pg/mL, respectively. Transient, > or = 50% decreases in circulating monocytes were observed in 17 of 21 patients as early as 0.5 hours (median time, 2 hours) from the start of infusion. Decreases (mean 33%) in circulating lymphocytes were observed in seven of 21 patients. All three patients with grade 3 toxicity were treated at the 0.5-ng/kg dose. The significance of baseline anti-SEA, human antimouse antibody (HAMA), CA242-soluble antigen levels, and T-cell receptor variable beta region (TCR V beta) subsets and histocompatibility leukocyte antigen-DR (HLA-DR) genotypes was assessed as possible predictors of toxicity. All toxicities were transient and easily managed. No grade 3 toxicity occurred at the higher dose levels. PNU-214565, a SAg-based tumor targeted therapy, is safe when given as a single 3-hour infusion at doses up to 1.5 ng/kg. The MTD for a single dose was not determined. The safety of a repeated dose schedule is currently under investigation, beginning with doses determined to be safe in this trial.


Polyolefin Recyclates for Rigid Packaging Applications: The Influence of Input Stream Composition on Recyclate Quality.
In order to shift to a circular plastics economy, high quality recyclates are required to effectively substitute virgin materials. Current approaches to empirically quantify the substitutability for recyclates are mainly limited by the abundance of virgin material grades along with a lack of adequate application-specific property profiles. In contrast, this work aims for a holistic analysis of the substitution potential of polyolefin recyclates intended for rigid packaging applications. This approach is fundamentally based on the classification of virgin polyolefins into different application-specific sub-groups with defined property windows derived from supplier data sheets, which allows for a generalization within one polymer type without neglecting the various available material grades. Moreover, the findings should provide valuable information for improvements of quality-defining process steps along the value chain of mechanical recycling. Therefore, it is of great importance to correlate the input stream composition of the investigated recyclates with the obtained qualities. The investigation of the substitution potential for selected recyclates clearly highlights the necessity of functional recycling for enhanced quality levels, which especially affects the sorting step in the recycling value chain. This work illustrates that a homogeneous waste stream directly correlates with a high substitution potential. Thus, the development of economically viable sorting strategies which take the functionality of plastic waste products into account must be targeted in future research. Furthermore, the development of detailed application-specific property windows in a joint effort with manufacturers should be pursued, as it allows for a meaningful empirical quantification of the substitutability for recyclates obtained from mechanical recycling.


Bile duct injury repair: when? what? who?
Laparoscopic cholecystectomy is associated with a two-to-four times higher risk of bile duct injury (BDI) than open cholecystectomy. BDI can lead to significant morbidity and even mortality. The first priority in BDI is to control peritoneal and biliary sepsis and to convert an acute BDI to a controlled external biliary fistula (EBF) - this can be achieved by endoscopic and/ or radiological intervention in most cases. This should be followed by assessment of the extent of injury - both biliary and vascular. Immediate management of BDI recognized during cholecystectomy depends on the type of injury, the condition of the patient, and the experience of the surgeon. For BDI recognized after cholecystectomy, early repair is not recommended, as the results are poor. The EBF may evolve into a benign biliary stricture (BBS), which should be electively repaired by a Roux-en-Y hepatico-jejunostomy. The use of an endoscopic stent as definitive management of BDI is not recommended. Long-term follow-up is essential after the repair of a BBS, as recurrence can occur several years after repair. Recurrent BBS is best treated with endoscopic balloon dilatation. Excellent early and long-term results can be obtained in specialized units at tertiary care referral centers.


Ionic strength and transition metals control PrPSc protease resistance and conversion-inducing activity.
The essential component of infectious prions is a misfolded protein termed PrPSc, which is produced by conformational change of a normal host protein, PrPC. It is currently unknown whether PrPSc molecules exist in a unique conformation or whether they are able to undergo additional conformational changes. Under commonly used experimental conditions, PrPSc molecules are characteristically protease-resistant and capable of inducing the conversion of PrPC molecules into new PrPSc molecules. We describe the effects of ionic strength, copper, and zinc on the conformation-dependent protease resistance and conversion-inducing activity of PrPSc molecules in scrapie-infected hamster brains. In the absence of divalent cations, PrPSc molecules were > 20-fold more sensitive to proteinase K digestion in low ionic strength buffers than in high ionic strength buffers. Addition of micromolar concentrations of copper or zinc ions restored the protease resistance of PrPSc molecules under conditions of low ionic strength. These transition metals also controlled the conformation of purified truncated PrP-(27-30) molecules at low ionic strength, confirming that the N-terminal octapeptide repeat region of PrPSc is not required for binding to copper or zinc ions. The protease-sensitive and protease-resistant conformations of PrPSc were reversibly interchangeable, and only the protease-resistant conformation of PrPSc induced by high ionic strength was able to induce the formation of new protease-resistant PrP (PrPres) molecules in vitro. These findings show that PrPSc molecules are structurally interconvertible and that only a subset of PrPSc conformations are able to induce the conversion of other PrP molecules.


Structured writing and processing major stressful events: a controlled trial.
Traumatic events may result in a variety of physical and psychological health problems. Self-confrontation with traumatic memories, by putting painful emotions and thoughts into (verbal) words, is associated with psychophysiological benefits. Self-confrontation may be invoked during sessions and enhanced by structural assignments, which the client carries out between the sessions. In this context, writing assignments could be a useful tool in reprocessing traumatic events. The effects of writing assignments have been demonstrated in several case studies and in a number of experimental studies. However, the experimental studies have several limitations, for example the effects of writing on the impact of trauma are not examined. Furthermore, the psychological mechanisms that mediate the effects of trauma on health are less clear. These two issues are the main issues of the current study. A group of 26 participants were instructed to write about their negative events during five 45-min sessions over a period of 2 weeks. They were compared at pre-treatment, post-treatment and at 6-week follow-up to a waiting-list control group (n = 22). The trauma-writing groups experienced fewer intrusions and showed less avoidance behavior from pre-treatment to follow-up, whereas the waiting-list control group did not change significantly. Similar results were found on depressive symptoms. No effects on mood measures were found. Implications for future research and clinical practice are discussed.


Effects of diet before endurance exercise on hepcidin response in young untrained females.
We examined the effects of diet before endurance exercise on hepcidin response in young untrained females. Ten young untrained females [age: 20.6 ± 0.8 y, height: 157.5 ± 1.0 cm, weight: 54.4 ± 1.5 kg, and maximal oxygen uptake (VO2max): 35.9 ± 1.1 mL/kg/min] were involved in two experimental conditions with a crossover design. The two conditions were separated by approximately 1 month, and each condition was performed during the follicular phase. Subjects completed 60 min of pedaling at 65% of VO2max after consuming a meal (FED) or not consuming a meal (CON). Blood samples were collected before, immediately after, and 3 h after exercise. Serum ferritin levels before exercise did not differ between the two conditions (P > 0.05). Blood glucose and lactate levels were significantly elevated immediately after exercise only under the FED condition (P < 0.05). Serum iron levels were significantly elevated after exercise under both conditions. However, the plasma interleukin-6 and serum hepcidin levels were not significantly different 3 h after exercise under either condition (P > 0.05). Consuming a meal before endurance exercise at moderate intensity did not affect exercise-induced hepcidin elevation in young untrained females.


Gamma-aminobutyric acid-mediated neurotransmission in the pontine reticular formation modulates hypnosis, immobility, and breathing during isoflurane anesthesia.
Many general anesthetics are thought to produce a loss of wakefulness, in part, by enhancing gamma-aminobutyric acid (GABA) neurotransmission. However, GABAergic neurotransmission in the pontine reticular formation promotes wakefulness. This study tested the hypotheses that (1) relative to wakefulness, isoflurane decreases GABA levels in the pontine reticular formation; and (2) pontine reticular formation administration of drugs that increase or decrease GABA levels increases or decreases, respectively, isoflurane induction time. To test hypothesis 1, cats (n = 5) received a craniotomy and permanent electrodes for recording the electroencephalogram and electromyogram. Dialysis samples were collected from the pontine reticular formation during isoflurane anesthesia and wakefulness. GABA levels were quantified using high-performance liquid chromatography. For hypothesis 2, rats (n = 10) were implanted with a guide cannula aimed for the pontine reticular formation. Each rat received microinjections of Ringer's (vehicle control), the GABA uptake inhibitor nipecotic acid, and the GABA synthesis inhibitor 3-mercaptopropionic acid. Rats were then anesthetized with isoflurane, and induction time was quantified as loss of righting reflex. Breathing rate was also measured. Relative to wakefulness, GABA levels were significantly decreased by isoflurane. Increased power in the electroencephalogram and decreased activity in the electromyogram caused by isoflurane covaried with pontine reticular formation GABA levels. Nipecotic acid and 3-mercaptopropionic acid significantly increased and decreased, respectively, isoflurane induction time. Nipecotic acid also increased breathing rate. Decreasing pontine reticular formation GABA levels comprises one mechanism by which isoflurane causes loss of consciousness, altered cortical excitability, muscular hypotonia, and decreased respiratory rate.


Adaptive and maladaptive actions of angiotensin II in patients with severe congestive heart failure.
The renin-angiotensin system appears to have evolved millions of years ago as a primary attempt to preserve circulatory homeostasis at a time when the principal cause of a low cardiac output was intravascular volume depletion. Angiotensin II supported systemic BP by direct systemic vasoconstriction, by facilitating the central and peripheral effects of the sympathetic nervous system, by promoting renal sodium retention by the production of aldosterone, and by increasing total body water by enhancing thirst and the synthesis of vasopressin. In addition, angiotensin II evolved as an important mechanism to preserve the glomerular filtration rate in low-flow states. These actions of angiotensin II were beneficial when the system first evolved, but its activation in patients with heart failure not only fails to reverse the low-output state but further exacerbates loading conditions in the left ventricle, thereby leading to worsening heart failure. Moreover, increased levels of angiotensin II cause heightened sympathetic nervous activity, potassium depletion, and hyponatremia, each of which can lead to further clinical deterioration. Therefore, activation of the renin-angiotensin system in heart failure might appear (at first) to be a maladaptive response. Recent evidence, however, suggests that this hormonal system continues (even in heart failure) to carry out the primary functions for which it was designed. Angiotensin II plays an important role in preserving systemic BP and in preserving the glomerular filtration rate as renal artery pressure and renal blood flow decline; in addition, by stimulating the synthesis of aldosterone, the renin-angiotensin system provides an important role for potassium disposal.(ABSTRACT TRUNCATED AT 250 WORDS)


Melatonin ameliorates hepatic steatosis by inhibiting NLRP3 inflammasome in db/db mice.
Type 2 diabetes mellitus (T2DM) is commonly accompanied by obesity and non-alcoholic fatty liver disease (NAFLD), yet the mechanism underlying diabetes-related NAFLD is not fully understood. It has been reported that melatonin can regulate glucose and lipid metabolism. This study aims to investigate the actions and mechanisms of melatonin toward the development of diabetes-related NAFLD. Melatonin (bid, 30 mg/kg/day, i.p.) was administrated to db/db mice for 8 weeks, while saline was administrated to db/m mice. The metabolic parameters of mice were measured using an automatic biochemistry analyzer. The oxidative stress indexes and mitochondrial membrane potential (MMP) were determined with kits. Pathological assessment in liver tissues was used to analyze the effects of melatonin on hepatic steatosis. The levels of IL-1β and IL-18 were detected with ELISA kits. The mRNA levels of NLRP3 inflammasome were detected using quantitative real-time PCR assay, and protein expressions were estimated using Western blotting assay. Immunofluorescence staining was used to evaluate the caspase-1 expression in the liver. Melatonin treatment significantly reduced blood glucose, serum insulin, body weight, related liver weight, serum lipids, and hepatic enzymes in db/db mice. Melatonin markedly corrected the NAFLD phenotypes, including lipid accumulation, steatohepatitis, fibrosis, and oxidative stress levels. Melatonin significantly improved the MMP level and decreased the serum IL-1β and IL-18 concentrations. The mRNA levels of the NLRP3 inflammasome could also be remarkably reversed by melatonin in the liver tissues. The activation of the NLRP3 inflammasome was also suppressed, evidenced by the downregulated proteins of NLRP3, caspase-1, IL-1β, and IL-18. The enhanced fluorescence intensity of caspase-1 in the liver tissues was also obviously weakened by the melatonin treatment. Our study concluded that melatonin could safeguard against NAFLD by improving hepatic steatosis in db/db mice, and this action could be associated with the regulation of the NLRP3 inflammasome activation.


[Olfactory esthesioneurocytoma: ultrastructural study of a case (author's transl)].
One case of malignant tumour of the left nasal cavity is reported in a woman 56 year old, affected by the disease 24 years. Numerous recurrences appeared and various histological diagnoses were performed. At the last surgery, the tumour invaded the ethmoid and was a typical olfactory esthesioneurocytoma. By electron microscopy, mature ganglion cells with dense cored vesicles (neurosecretory granules) were densely packed. Neuritic processes with microtubules were rarely normal in size and their content was most often abnormal; furthermore dystrophic axons were noted in great number.


Distinct mechanisms control the accumulation of the Rb-related p107 and p130 proteins during cell growth.
A variety of studies have demonstrated the critical role of the Rb/E2F pathway in the control of cell growth and have highlighted a complexity in the accumulation of both the E2F family proteins and the Rb family of proteins. Whereas the Rb protein is found in both growing and quiescent cells, the accumulation of p130 and p107 is tightly regulated with respect to the growth state of the cell. The p130 protein is found in quiescent cells but not in growing cells, whereas the inverse is true for the p107 protein. Control of p130 accumulation is posttranscriptional, because p130 RNA is relatively constant in growing and quiescent cells. The disappearance of the p130 protein after stimulation of cell growth coincides with cyclin-dependent kinase-mediated phosphorylation and is blocked by inhibitors of the 26S proteasome. In contrast, the cell growth-dependent regulation of p107 expression reflects the transcriptional regulation of the p107 gene. Similar to several other growth-regulated genes, the control of p107 expression is largely the result of E2F-dependent repression in quiescent cells. These experiments thus demonstrate a control of Rb family member expression mediated through distinct mechanisms of both transcriptional and posttranslational control and also suggest an intimate relationship in which p130 controls the expression of p107.


[Beneficial effects of continuous hemodiafiltration after cardiac surgery in patients with chronic renal failure].
From December 1993 to June 1996, fourteen patients with chronic renal failure (CRF) were underwent cardiac surgery. Thirteen of them had coronary artery bypass grafting (CABG) and one patient had aortic valve replacement. We employed continuous hemodiafiltration (CHDF) with or without peritoneal dialysis in eleven cases in the early postoperative course of open heart surgery. In these cases, the usefulness of CHDF were examined. CHDF was started after 16.9 +/- 13.7 hours of surgery, and continued for 31.9 +/- 18.0 hours. We adjusted blood flow as 80 ml/min, perfusate flow of hemofilter as 500 ml/hr, and the activated clotting time (ACT) as 150-200 seconds with heparin or nafamostat mesilate. No hemorrhagic tendency was observed. Mean serum creatinine level before and after 24 hours of CHDF was 5.6 +/- 2.2 and 5.6 +/- 2.1 mg/dl respectively, and it was not elevated. The average removal volume was 2,223 +/- 1,256 ml for the first 24 hours, and pulmonary wedge pressure was decreased from 9.0 +/- 2.3 to 6.4 +/- 3.0 mmHg (p < 0.05). However, there was no significant change at cardiac output before and after CHDF. Total dose of dopamine and dobutamine were unchanged. No significant change were noted at systemic vascular resistance and pulmonary vascular resistance. We conclude that CHDF allows an adequate removal of fluid and waste products without deleterious effects on hemodynamics even in patients after cardiac surgery. CHDF is a safe and effective alternative in patients with CRF after cardiac surgery.


A Tight Spot: How Personality Moderates the Impact of Social Norms on Sojourner Adaptation.
How do you navigate the norms of your new culture when living abroad? Taking an interactionist perspective, we examined how contextual factors and personality traits jointly affect sojourners' adaptation to the host-country culture. We hypothesized that tightness (strong, rigidly imposed norms) of the host culture would be associated with lower levels of adaptation and that tightness of the home culture would be associated with higher levels of adaptation. Further, we proposed that the impact of tightness should be dependent on personality traits associated with navigating social norms (agreeableness, conscientiousness, and honesty-humility). We analyzed longitudinal data from intercultural exchange students ( N = 889) traveling from and to 23 different countries. Multilevel modeling showed that sojourners living in a tighter culture had poorer adaptation than those in a looser culture. In contrast, sojourners originating from a tighter culture showed better adaptation. The negative effect of cultural tightness was moderated by agreeableness and honesty-humility but not conscientiousness.


Co-delivery of siRNA and an anticancer drug for treatment of multidrug-resistant cancer.
To develop a novel nanomedicine approach for the treatment of multidrug-resistant (MDR) cancer by combining an anticancer drug and suppressors of cellular resistance within one multifunctional nanocarrier-based delivery system (NDS). The NDS consisted of cationic liposomes (carrier, 100-140 nm), doxorubicin (DOX, anticancer drug), siRNA targeted to MRP1 and BCL2 mRNA (suppressors of pump and nonpump cellular-resistance, respectively). The resulting approximately 500 nm complex has a zeta potential of +4 mV. The NDS provides an effective co-delivery of DOX and siRNA as well as cell-death induction and suppression of cellular resistance in MDR lung cancer cells. We demonstrate NDS-enhanced efficiency of chemotherapy to a level that cannot be achieved by applying its components separately.


A benchmarking method to evaluate the accuracy of a commercial proton monte carlo pencil beam scanning treatment planning system.
AcurosPT is a Monte Carlo algorithm in the Eclipse 13.7 treatment planning system, which is designed to provide rapid and accurate dose calculations for proton therapy. Computational run-time in minimized by simplifying or eliminating less significant physics processes. In this article, the accuracy of AcurosPT was benchmarked against both measurement and an independent MC calculation, TOPAS. Such a method can be applied to any new MC calculation for the detection of potential inaccuracies. To validate multiple Coulomb scattering (MCS) which affects primary beam broadening, single spot profiles in a Solidwater® phantom were compared for beams of five selected proton energies between AcurosPT, measurement and TOPAS. The spot Gaussian sigma in AcurosPT was found to increase faster with depth than both measurement and TOPAS, suggesting that the MCS algorithm in AcurosPT overestimates the scattering effect. To validate AcurosPT modeling of the halo component beyond primary beam broadening, field size factors (FSF) were compared for multi-spot profiles measured in a water phantom. The FSF for small field sizes were found to disagree with measurement, with the disagreement increasing with depth. Conversely, TOPAS simulations of the same FSF consistently agreed with measurement to within 1.5%. The disagreement in absolute dose between AcurosPT and measurement was smaller than 2% at the mid-range depth of multi-energy beams. While AcurosPT calculates acceptable dose distributions for typical clinical beams, users are cautioned of potentially larger errors at distal depths due to overestimated MCS and halo implementation.


Acetylcholinesterase function in apoptotic retina pigment epithelial cells induced by H2O2.
To investigate the acetylcholinesterase (AChE) expression involved in retina pigment epithelial (RPE) apoptosis induced by higher concentrations H2O2. The human retinal pigment epithelium cell line ARPE-19 was from ATCC (Rockville, MD). Cultured ARPE-19 cells were treated with H2O2 at 0, 250, 500, 1 000, 2 000µmol/L and cell viability was measured with MTT assay. AChE expression and DNA fragments were analyzed by immunocytochemistry, TUNEL and PARP-1 Western blotting. Immunofluorescence detected AChE exist in the normal human retinal tissue. When H2O2 >500µmol/L, AChE expression showed an increase after 2h, and this concentration was selected for the present study. RPE cell was induced with 1 000µmol/L H2O2 for 2h, compared to the control group, cell activity decline detected by MTT, AChE and PARP-1 protein expression was significantly increased detected by Western blotting. AChE immunofluorescence staining was positive in RPE cell after H2O2 incubate 2h. In addition, pretreatment with 100µmol/L epigallocatechin gallate (EGCG), cell viability increased from 31.20%±3.90% to 70.23%±12.96%. AChE is weakly expressed in normal human RPE cells. Stimulation with H2O2 caused the stable increase of AChE expression in RPE cells, which may indicate that AChE may be an important role in AMD.


Molecular cloning and characterization of ARS elements from the mud loach (Misgurnus mizolepis).
Autonomously replicating sequences (ARSs) are thought to occur within, or adjacent to, the matrix attachment regions (MARs). To identify fish ARSs, MARs of the mud loach fish were obtained from nuclear matrices using a modified LIS method. These DNA fragments were screened for their ability to act as ARSs by being cloned into the ARS cloning vector, pURY19, and transformed into Saccharomyces cerevisiae. Sixteen ARSs were isolated, most of which were more efficient in transformation than the positive control vector, pURY19-2 microm, which contained the 2 microm circle origin of yeast. In particular, one clone, pURY19-ARS223, was 18 times more efficient in back-transforming E. coli than the positive control vector. Therefore, ARS223, which has strong ARS activity in yeast, could be a good candidate for inclusion in expression vehicles that are used to transfect fish cell lines or embryos. A DNA sequence analysis showed that the essential ARS elements contain potential ARS consensus sequences, and are predicted to have hairpin loop structures, or curved or kinked DNA. In addition, the MAR-Finder program suggested that ARSs also contain MAR motifs. These include AT tracts, ORI patterns, kinked DNA, ATC tracts, and Topoisomerase II consensus sequences. The in vitro matrix binding assay confirmed that all of the cloned ARSs could associate with the nuclear matrix. This indicates that ARSs elements may be located in or near the MARs. This is the first study that has identified and characterized ARSs in fish.


Detection of carious lesions utilizing depolarization imaging by polarization sensitive optical coherence tomography.
As dental caries is one of the most common diseases, the early and noninvasive detection of carious lesions plays an important role in public health care. Optical coherence tomography (OCT) with its ability of depth-resolved, high-resolution, noninvasive, fast imaging has been previously recognized as a promising tool in dentistry. Additionally, polarization sensitive imaging provides quantitative measures on the birefringent tissue properties and can be utilized for imaging dental tissue, especially enamel and dentin. By imaging three exemplary tooth samples ex vivo with proximal white spot, brown spot, and cavity, we show that the combination of polarization sensitive OCT and the degree of polarization uniformity (DOPU) algorithm is a promising approach for the detection of proximal carious lesions due to the depolarization contrast of demineralized tissue. Furthermore, we investigate different sizes of the DOPU evaluation kernel on the resulting contrast and conclude a suitable value for this application. We propose that DOPU provides an easy to interpret image representation and appropriate contrast for possible future screening applications in early caries diagnostics.


Metabolism of 5-fluorouracil in various human normal and tumor tissues.
The metabolism of 5-fluorouracil (5-FU) in tumors and normal tissues of humans was investigated in vitro. Phosphorylation of 5-FU was faster in tumor tissues than in normal tissues. Phosphorylating activity with 2-deoxy-alpha-D-ribose 1-phosphate (dRiblP) and ATP as cofactors was more active than that with alpha D-ribose 1-phosphate (RiblP) and ATP, or 5-phospho-alpha-D-ribosyldiphosphate (PPRibP) as cofactors. Phosphorylating activity in squamous cell carcinoma of the lung was similar to that in adenocarcinomas. Degradation of 5-FU was much faster in the liver than in other tissues including tumor tissues.


Relationships among airway reactivity, pupillary alpha-adrenergic and cholinergic responsiveness, and age.
Healthy adult volunteers (n = 122), who denied personal history of lung disease or family history of cystic fibrosis or asthma, took no interfering medications, and had forced expiratory volume in 1 s greater than or equal to 80% predicted, underwent methacholine challenge and pupillary reactivity testing. Pupil diameter measured in dark and light test conditions declined with age (Pearson's r = -0.54 and -0.36). Pupillary alpha-adrenergic responsiveness (expressed as the concentration of phenylephrine required to dilate the pupil 1 mm) was significantly correlated with age. Older subjects required lower concentrations for dilation and therefore were more sensitive to phenylephrine. Pupillary cholinergic responsiveness (the concentration of carbachol required for 1-mm constriction) was not significantly correlated with age. Therefore the significantly smaller baseline pupil size in the elderly cannot be explained by failure of alpha-adrenergic receptor responses or by increased pupillary cholinergic responsiveness. We found no significant correlation of methacholine bronchial reactivity with age. In addition, there was no relation between airway reactivity and pupillary alpha-adrenergic or cholinergic responsiveness in this sample of healthy adults. These findings, taken with others in the literature, suggest that the contribution of alpha-adrenergic and cholinergic responsiveness to nonspecific airway reactivity in healthy persons is small, if it exists at all, and that there is no significant change in airway reactivity with age in healthy adults.


Malignant tumors of the small intestine.
Malignant tumors of the small bowel are unusual and account for only 1% to 5% of all gastrointestinal tract malignancies. Thirteen cases of malignant tumors of the small bowel identified at the VAMC in Puerto Rico from January 1999 to September 2001 and a review of the literature are presented. The mean age of our cases was 67 (range: 45-78). Vague abdominal pain, nausea, vomiting, and melena were the most frequently reported symptoms. The average time from first symptoms to diagnosis was 3.2 months. A positive test for occult blood or hypochromic microcytic anemia was invariably present. Hyperbilirubinemia and increased alkaline phosphatase were warning signs that allowed earlier diagnosis in patients with duodenal tumors. Fifty-four percent of the lesions were detected by endoscopic examination while 46% relied on radiographic studies. Eleven had carcinomas and two malignant gastrointestinal stromal tumors. Neither carcinoids nor lymphomas were identified. Our report of thirteen cases of malignant small bowel tumors is unusual and exhibits differences with the previously reported data. The cases were identified in a limited Hispanic population in a short period of time. Aggressive evaluation and a high suspicion of these malignancies should be entertained whenever subtle symptoms and unexplained gastrointestinal blood loss are assessed.


[Study of the mechanisms of levomycetin inactivation by the palgue causative agent and Escherichia coli with episomal and chromosomal resistance. The enzymatic acetylation of levomycetin].
One of the mechanisms of levomycetin inactivation, i.e. enzymatic acetylation of the plague causative agent and Coli bacteria with chromosomic and episomic type of resistance was studied. It was shown that resistance to levomycetin in the recombinants of the plague microbe and Coli bacteria stipulated by one and the same R-factor was associated with their capacity for the antibiotic inactivation mainly with the help of levomycetinacetyltransferase. At the same time the enzyme activity in the mutants was very low and in the cells of Y. pestis and E. coli sensitive to the antibiotic it was absent. Levomycetinacetyltransferase is to somee extent soluble, still a singificant amount of it is connected with the structures of the bacterial cells and in particular with the cytoplasmic membranes. Comparison of the activity of levomycetinacetyltransferase as dependent on the incubation time, substrate concentration, pH and thermal treatment provided determination of quantitative differences in the properties of the enzyme of Y. petis and E. coli with episomic multiple drug resistance. However, under the experimental conditions transfer of R-factor from one host to the other induced no qualitative changes in the mechanism of levomycetin inactivation.


Recent advances in pancreaticobiliary maljunction.
The purpose of this review is to evaluate our current knowledge of the embryologic etiology of pancreaticobiliary maljunction (PBM), its diagnosis, clinical aspects, and treatment, and to clarify the mechanisms of PBM involvement in carcinogenesis. Although the embryologic etiology of PBM still awaits clarification, an arrest of the migration of the common duct of the biliary and pancreatic ducts inwards in the duodenal wall has hitherto been speculated to result in a long common channel in PBM. However, we propose the hypothesis that the etiology of PBM is caused by a disturbance in the embryonic connections (misarrangement) of the choledochopancreatic duct system in the extremely early embryo. That is, PBM is an anomaly caused by a misarrangement whereby the terminal bile duct joins with a branch of the ventral pancreatic duct system, including the main pancreatic duct. PBM is frequently associated with congenital bile duct cyst (CCBD). However, these two anomalies are thought to have different embryonic etiologies. The diagnostic criteria for PBM are the radiological and anatomical detection of the extramural location of the junction of the pancreatic and biliary ducts in the duodenal wall. However, in PBM patients with a short common duct (less than 1 cm in length), detection of the extramural location is difficult. The clinical features of PBM are intermittent abdominal pain, with or without elevation of pancreatic enzyme levels; and obstructive jaundice, with or without acute pancreatitis, while the clinical features of PBM patients with CCBD are primary bile duct stone and acute cholangitis. The optimum approach for the treatment of PBM is the prevention of the reciprocal reflux of bile and pancreatic juice in the pancreas and the bile duct system. To achieve these aims, the surgical approach is most effective, and complete biliary diversion procedures with bile duct resection (for example, choledochoduodenostomy or choledochojejunostomy of the Roux-en-Y type) are most useful. Recently, it has been recognized that the development of biliary ductal carcinoma is associated with PBM. That is, the development of gallbladder cancer occurs frequently in PBM patients without CCBD, and bile duct cancer originating from the cyst wall also occurs in PBM patients with CCBD. It is speculated that the pathogenesis of the bile duct or gallbladder cancer in PBM patients involves the reciprocal reflux of bile and pancreatic juice. Investigations of epithelial cell proliferation in the gallbladder of PBM patients, and of K- ras mutations and p53 suppressor gene mutations, loss of heterozygosity of p53, and overexpression of the p53 gene product in gallbladder cancer and noncancerous lesions in PBM patients have been carried out in various laboratories around the world. The results support the conclusion that PBM is a high risk factor for the development of bile duct carcinoma.


Improving the quality of occupational health care in Washington State: new approaches to designing community-based health care systems.
The need for concerted action to improve quality was stressed in the recent Institute of Medicine report, Crossing the Quality Chasm: A New Health Care System for the 21st Century. This article describes an innovative community-based delivery system initiative designed to improve quality and health outcomes for occupational health conditions. Known as the Occupational Health Services (OHS) project, this Washington State initiative focuses on three targeted conditions: low back sprain, carpal tunnel syndrome, and fractures. To fulfill its purpose, which is to provide clinical resources and training opportunities and foster quality improvement activities on a community-wide basis, the OHS incorporates several key delivery system components, including systems to track patient and employer satisfaction and health outcomes; formal physician agreements that will provide incentives for using best-practices; and community-based Centers of Occupational Health and Education (COHEs) that will function as a resource for providers, patients, and employers. Our experience in developing the OHS quality improvement initiative should have relevance for health care clinicians, administrators, policy makers, and researchers engaged in similar pursuits outside the field of occupational health.


An optogenetic toolbox for unbiased discovery of functionally connected cells in neural circuits.
Optical imaging approaches have revolutionized our ability to monitor neural network dynamics, but by themselves are unable to link a neuron's activity to its functional connectivity. We present a versatile genetic toolbox, termed 'Optobow', for all-optical discovery of excitatory connections in vivo. By combining the Gal4-UAS system with Cre/lox recombination, we target the optogenetic actuator ChrimsonR and the sensor GCaMP6 to stochastically labeled, nonoverlapping and sparse subsets of neurons. Photostimulation of single cells using two-photon computer-generated holography evokes calcium responses in downstream neurons. Morphological reconstruction of neurite arbors, response latencies and localization of presynaptic markers suggest that some neuron pairs recorded here are directly connected, while others are two or more synapses apart from each other. With this toolbox, we discover wiring principles between specific cell types in the larval zebrafish tectum. Optobow should be useful for identification and manipulation of networks of interconnected neurons, even in dense neural tissues.Mechanisms of neural processing can only be understood by revealing patterns of connectivity among the cellular components of the circuit. Here the authors report a new genetic toolbox, 'Optobow', which enables simultaneous optogenetic activation of single neurons in zebrafish and measuring the activity of downstream neurons in the network.


Müller glia and phagocytosis of cell debris in retinal tissue.
Müller cells are the predominant glial cell type in the retina of vertebrates. They play a wide variety of roles in both the developing and the mature retina that have been widely reported in the literature. However, less attention has been paid to their role in phagocytosis of cell debris under physiological, pathological or experimental conditions. Müller glia have been shown to phagocytose apoptotic cell bodies originated during development of the visual system. They also engulf foreign molecules that are injected into the eye, cone outer segments and injured photoreceptors. Phagocytosis of photoreceptor cell debris in the light-damaged teleost retina is primarily carried out by Müller cells. Once the microglial cells become activated and migrate to the photoreceptor cell layer, the phagocytic activity of Müller cells progressively decreases, suggesting a possible mechanism of communication between Müller cells and neighbouring microglia and photoreceptors. Additionally, it has been shown that phagocytic Müller cells acquire proliferating activity in the damaged teleost retina, suggesting that engulfment of apoptotic photoreceptor debris might stimulate the Müller glia to proliferate during the regenerative response. These findings highlight Müller glia phagocytosis as an underlying mechanism contributing to degeneration and regeneration under pathological conditions.


Diabetic amyotrophy: reassessment of the clinical spectrum.
Diabetic amyotrophy is a syndrome whose recognition may be difficult or delayed. We reviewed thirteen patients with this disorder, all of whom had significant proximal lower limb wasting and weakness as the predominant feature and eleven of whom had pain in the affected limbs. Significant weight loss was common. In nine patients the deficits were largely or totally reversible. Important variations from the classical features were observed. Only five patients displayed asymmetric proximal lower limb wasting, weakness and pain, motor deficits in the remainder being either unilateral or bilateral and symmetrical. The shoulder girdle and arms were also involved in two patients. Proximal limb pain was not invariable, a distal sensory peripheral neuropathy was common, and diabetic control at diagnosis was likely to be good. No prognostic factors were identified. Thus, not all patients with diabetic amyotrophy exhibit the classically-described features. Other than careful clinical examination, a thorough bilateral electromyographic and nerve conduction study remains the most helpful diagnostic test. Appreciation of the clinical spectrum and context of diabetic amyotrophy should facilitate its differentiation from other disorders, including other forms of diabetic neuropathy.


Effect of ionotropic and metabotropic glutamate agonists and D-aspartate on prolactin release from anterior pituitary cells.
Although the presence of ionotropic (iGluRs) and metabotropic (mGluRs) glutamate receptors has been demonstrated in the anterior pituitary, recent reports on the direct effect of glutamate on prolactin (PRL) secretion by anterior pituitary cells have presented contradictory results. Hence, the aim of this study was to determine the effect of ionotropic (iGluRs) and metabotropic (mGluRs) glutamate receptor agonists on prolactin (PRL) release. In addition, since D-Aspartate (D-Asp) is found in the pituitary and is involved in neuroendocrine regulation, we also studied the direct action of D-Asp on PRL secretion. Finally, since the posterior pituitary participates in the regulation of PRL secretion, we examined the influence of the posterior pituitary on the effects of NMDA and D-Asp on PRL release. Glutamate (1000 microM) increased PRL secretion from cultured anterior pituitary cells. Both NMDA (100 microM) and kainate (100 microM) increased PRL secretion and these effects were blocked by a specific NMDA receptor antagonist. AMPA did not modify PRL release in these cultures. The group I and II mGluR agonist, trans-ACPD (1000 microM), and a specific group II mGluR agonist, L-CCG-I (100-1000 microM), inhibited whereas specific group I and III mGluR agonists, 3-HPG and L-AP4 respectively, had no effect on PRL release. Finally, D-Asp (100-1000 microM) stimulated PRL secretion and this effect was reduced by a NMDA receptor antagonist. When anterior pituitary cells were cultured in the presence of posterior pituitary cells, NMDA did not modify PRL or GABA release, while D-Asp increased PRL secretion and decreased GABA release in these cocultures. In conclusion, our results show that L-glutamate has a differential direct effect on PRL release: it exerts a stimulatory action via iGluRs and an inhibitory effect via mGluRs. D-Asp could directly stimulate PRL release through NMDA receptors. D-Asp may also stimulate PRL release by decreasing GABA release from the posterior pituitary.


Age changes of jaws and soft tissue profile.
Age-related changes of jaws and soft tissue profile are important both for orthodontists and general dentists. Mouth profile is the area which is manipulated during dental treatment. These changes should be planned in accordance with other components of facial profile to achieve ultimate aim of structural balance, functional efficacy, and esthetic harmony. Through this paper, the authors wish to discuss age changes of the hard and soft tissues of human face which would help not only the orthodontists but also oral surgeons, prosthodontists, pedodontists, and general dentists.


Age-related changes in human tendo calcaneus collagen fibrils.
The ruptures of tendo calcaneus often occur between the age of 30-45 years as described by several textbooks. It is also described that some diseases and drugs are said to be responsible in the etiology; however, there are no studies related with the detailed histological structure of collagen fibrils found in the tendon in the age groups of humans. In view thereof, this study was aimed to obtain further information on the etiology and to find an answer regarding the frequency of the ruptures occurring between the age of 30-45 years in humans. In the study, the biopsy specimens taken from 28 patients (ages 1-68 year) who had undergone surgery due to tendo calcaneus ruptures or achilloplasty operation were examined by transmission electron microscope. All the specimens were prepared according to routine electron microscopic tissue preparation technique. The patients were divided into 7 age groups (1-9, 10-19, 20-29, 30-39, 40-49, 50-59, >60 years) and there were 4 patients in each group. The transverse diameters of collagen fibers were measured from the ultra thin sections and statistical analysis of the results were performed. The study was carried out in the electron microscopy laboratory of the Anatomy Department of Hacettepe University, Ankara, Turkey between January 2004 and September 2004. The diameters of the collagen fibers were higher in the 20-29 year-old group compared to other groups and it showed a statistically significant difference. In patients who were in the 30-39 year-old group or older, the diameters of the collagen fibers were lesser than the 20-29 year-old group. However, an increase was observed in the collagen fibril concentration of these groups. In examination of the specimens of patients who were under 20-year-old, the diameter of the collagen fibers were less than the 20-29 year-old group. The electron microscopic appearance of the tissue sample of a one-year-old patient had a specific organization and in this patient, both the diameters and concentration of collagen fibers were less. We believe that the decrease in the diameters of collagen fibers of 30-45 year-old patients who are in the active period of their life, can play a role in the etiology of the frequency of tendo calcaneus ruptures similar to other etiologic factors.


Prevalence and abundance of lactic acid bacteria in raw milk associated with forage types in dairy cow feeding.
Lactic acid bacteria (LAB) found in milk can be responsible for organoleptic defects in cheese. To identify sources of LAB that could potentially develop during cheese making, we evaluated their prevalence and abundance in milk according to the type of forage used in dairy cow feeding. Forages and bulk tank milk were sampled 3 times on 24 farms using either hay alone (control), or grass or legume silage supplemented with corn silage or not. Both types of silage were either non-inoculated or inoculated with commercial preparations containing at least a Lactobacillus buchneri strain along with Lactobacillus casei, Lactobacillus plantarum, Enterococcus faecium, or Pediococcus pentosaceus. Our results indicate that LAB viable counts in milk samples (2.56 log cfu/mL) did not differ according to the type of forage used. A total of 1,239 LAB were isolated and identified by partial 16S rRNA gene sequencing. Although inoculation increased lactobacilli abundance in grass silage by 35%, we did not observe an effect on the LAB profile of milk. Indeed, we found no significant difference in milk LAB prevalence and abundance according to the type of forage (P > 0.05). Moreover, isolates belonging to the L. buchneri group were rarely found in bulk tank milk (3 out of 481 isolates). Random amplified polymorphic DNA typing of 406 LAB isolates revealed the plausible transfer of some strains from silage to milk (~6%). Thus, forage is only a minor contributor to LAB contamination of milk.


Adenophora remotiflora protects human skin keratinocytes against UVB-induced photo-damage by regulating antioxidative activity and MMP-1 expression.
Chronic ultraviolet (UV) exposure-induced reactive oxygen species (ROS) are commonly involved in the pathogenesis of skin damage by activating the metalloproteinases (MMP) that break down type I collagen. Adenophora remotiflora (AR) is a perennial wild plant that inhabits Korea, China, and Japan. The present study investigated the protective effects of AR against UVB-induced photo-damage in keratinocytes. An in vitro cell-free system was used to examine the scavenging activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical and nitric oxide (NO). The effect of AR on ROS formation, antioxidant enzymes, elastase, MMP-1 level, and mRNA expression of MMP-1 were determined in UVB-irradiated human keratinocyte HaCaT cells. AR demonstrated strong DPPH free radical and NO scavenging activity in a cell-free system exhibiting IC50 values of 1.88 mg/mL and 6.77 mg/mL, respectively. AR pretreatment dose-dependently attenuated the production of UVB-induced intracellular ROS, and antioxidant enzymes (catalase and superoxide dismutase) were enhanced in HaCaT cells. Furthermore, pretreatment of AR prevented UVB-induced elastase and collagen degradation by inhibiting the MMP-1 protein level and mRNA expression. Accordingly, AR treatment elevated collagen content in UVB-irradiated HaCaT cells. The present study provides the first evidence of AR inhibiting UVB-induced ROS production and induction of MMP-1 as a result of augmentation of antioxidative activity in HaCaT human keratinocytes. These results suggest that AR might act as an effective inhibitor of UVB-modulated signaling pathways and might serve as a photo-protective agent.


Role of divalent cations in plasmid DNA adsorption to natural organic matter-coated silica surface.
The adsorption kinetics of supercoiled and linear plasmid DNA onto a natural organic matter (NOM)-coated silica surface are acquired using a quartz crystal microbalance with dissipation monitoring (QCM-D) in the presence of common divalent electrolytes CaCl2 and MgCl2. The adsorption kinetics of both DNA are noticeably higher in the presence of CaCl2 compared to MgCl2. We hypothesize that specific bridging between the DNA phosphate groups and NOM carboxyl functional groups in the presence of Ca2+ cations may lead to more efficient attachmentthan in the presence of Mg2+ cations, which are only likely to allow for charge neutralization. The influence of background Na+ cations on the adsorption kinetics in the presence of CaCl2 is found to be insignificant, while the presence of Na+ leads to slower attachment kinetics in MgCl2. Rinsing the DNA layer adsorbed in the presence of CaCl2 with a solution of low NaCl concentration followed by deionized water does not result in observable detachment, indicating irreversibility of DNA adsorption. Instead, softening of the DNA layer adsorbed in the presence of CaCl2 with background Na+ occurs with the rinses due to the increase in electrostatic repulsion between the phosphate functional groups along the DNA backbone. In the case of the DNA layer adsorbed in the presence of CaCl2 without background Na+, softening of the layer does not occur with the rinses.


Radiation induced pulmonary fibrosis as a model of progressive fibrosis: Contributions of DNA damage, inflammatory response and cellular senescence genes.
Purpose/Aim of Study: Studies of pulmonary fibrosis (PF) have resulted in DNA damage, inflammatory response, and cellular senescence being widely hypothesized to play a role in the progression of the disease. Utilizing these aforementioned terms, genomics databases were interrogated along with the term, "pulmonary fibrosis," to identify genes common among all 4 search terms. Findings were compared to data derived from a model of radiation-induced progressive pulmonary fibrosis (RIPF) to verify that these genes are similarly expressed, supporting the use of radiation as a model for diseases involving PF, such as human idiopathic pulmonary fibrosis (IPF). In an established model of RIPF, C57BL/6J mice were exposed to 12.5 Gy thorax irradiation and sacrificed at 24 hours, 1, 4, 12, and 32 weeks following exposure, and lung tissue was compared to age-matched controls by RNA sequencing. Of 176 PF associated gene transcripts identified by database interrogation, 146 (>82%) were present in our experimental model, throughout the progression of RIPF. Analysis revealed that nearly 85% of PF gene transcripts were associated with at least 1 other search term. Furthermore, of 22 genes common to all four terms, 16 were present experimentally in RIPF. This illustrates the validity of RIPF as a model of progressive PF/IPF based on the numbers of transcripts reported in both literature and observed experimentally. Well characterized genes and proteins are implicated in this model, supporting the hypotheses that DNA damage, inflammatory response and cellular senescence are associated with the pathogenesis of PF.


The U-shaped association between achieved blood pressure and risk of cardiovascular events and mortality in elderly and younger patients.
The main objective is to assess the appropriate level of achieved SBP and DBP to prevent cardiovascular events. We used the National Sample Cohort from the National Health Insurance Service in Korea and analyzed data of 44 462 hypertensive patients aged 20--84 years. Achieved SBP and DBP were categorized according to average achieved SBP (<120, 120-129, 130-139, 140-149, and ≥150 mmHg) and DBP (<70, 70-79, 80-89, 90-99, and ≥100 mmHg). We examined the association between achieved BP and composite outcome including cardiovascular death, admission of stroke, myocardial infarction, or heart failure, and all-caused death in elderly aged more than 65 years and in younger patients. After a median follow-up of 6.8 years, achieved SBP less than 120 mmHg and at least 150 mmHg in elderly and younger patients, respectively, were significantly associated with a higher risk of composite outcome than achieved SBP of 120-129 mmHg. Cox's proportional hazard analysis showed that the association between achieved SBP and risk of composite outcome and all-cause death had U-shaped relationships and identified a nadir of SBP of 135.6 and 128.9 mmHg, respectively, for composite outcome and 135.1 and 131.4 mmHg, respectively, for all-cause death in elderly and younger patients. Compared with SBP of 120-129 mmHg, not only low achieved SBP of less than 120 mmHg but also high BP are associated with risk of adverse cardiovascular event and all-cause death in both elderly and younger patients with a distinct U-shaped relationship.