Significance and implementation of RBE variations in proton beam therapy.
Key to radiation therapy is to apply a high tumor-destroying dose while protecting healthy tissue, especially near organs at risk. To optimize treatment for ion therapy not the dose but the dose multiplied by the relative biological effectiveness (RBE) is decisive. Proton therapy has been based on the use of a generic RBE, which is applied to all treatments independent of dose/fraction, position in the spread-out Bragg peak (SOBP), initial beam energy or the particular tissue. Dependencies of the RBE on various physical and biological properties are disregarded. The variability of RBE in clinical situations is believed to be within 10-20%. This is in the same range of effects that receive high attention these days, i.e., patient set-up uncertainties, organ motion effects, and dose calculation accuracy all affecting proton as well as conventional radiation therapy. Elevated RBE values can be expected near the edges of the target, thus probably near critical structures. This is because the edges show lower doses and, depending on the treatment plan, may be identical with the beam's distal edge, where dose is deposited in part by high-LET protons. We assess the rationale for the continued use of a generic RBE and whether the magnitude of RBE variation with treatment parameters is small relative to our abilities to determine RBE's. Two aspects have to be considered. Firstly, the available information from experimental studies and secondly, our ability to calculate RBE values for a given treatment plan based on parameters extracted from such experiments. We analyzed published RBE values for in vitro and in vivo endpoints. The values for cell survival in vitro indicate a substantial spread between the diverse cell lines. The average value at mid SOBP over all dose levels is approximately 1.2 in vitro and approximately 1.1 in vivo. Both in vitro and in vivo data indicate a statistically significant increase in RBE for lower doses per fraction, which is much smaller for in vivo systems. The experimental in vivo data indicate that continued employment of a generic RBE value of 1.1 is reasonable. At present, there seems to be too much uncertainty in the RBE value for any human tissue to propose RBE values specific for tissue, dose/fraction, etc. There is a clear need for prospective assessments of normal tissue reactions in proton irradiated patients and determinations of RBE values for several late responding tissues in animal systems, especially as a function of dose in the range of 1-4 Gy. However, there is a measurable increase in RBE over the terminal few mm of the SOBP, which results in an extension of the bio-effective range of the beam of a few mm. This needs to be considered in treatment planning, particularly for single field plans or for an end of range in or close to a critical structure. To assess our ability to calculate RBE values we studied two approaches, which are both based on the track structure theory of radiation action. RBE calculations are difficult since both the physical input parameters, i.e., LET distributions, and, even more so, the biological input parameters, i.e., local cellular response, have to be known with high accuracy. Track structure theory provides a basis for predicting dose-response curves for particle irradiation. However, designed for heavy ion applications the models show weaknesses in the prediction of proton radiation effects. We conclude that, at present, RBE modeling in treatment planning involves significant uncertainties. To incorporate RBE variations in treatment planning there has to be a reliable biological model to calculate RBE values based on the physical characteristics of the radiation field and based on well-known biological input parameters. In order to do detailed model calculations more experimental data, in particular for in vivo endpoints, are needed


Biochemical investigations of bowel inflammation in gastroschisis.
Neonates with gastroschisis have perivisceritis resulting from contact between the bowel and amniotic fluid. Here, we characterized the mediators involved in this inflammatory process in humans and ewes, to find a reliable marker of this process. We have diagnosed 41 cases of gastroschisis since 1995. Amniotic fluid sampled for karyotyping between 15 and 32 wk of gestation was also used to assay cytokines and inflammatory proteins. The findings were compared with those in 93 age-matched controls. Amniotic fluid cells were analyzed by means of cytology. Histologic examination of the bowel was performed when neonatal appendectomy was performed. The findings were compared with those obtained in a ewe model of gastroschisis. In gastroschisis, amniotic total protein, IL-6, IL-8, and ferritin levels were significantly higher than in controls. Gastroschisis was associated with significantly higher cell counts (mainly mononuclear cells) in amniotic fluid. At birth, macrophages were abundant in the fibrous peel covering the bowel. Similar results were obtained in the ewe model. Gastroschisis may be associated with a subchronic inflammatory process of variable intensity. This inflammation is restricted to the bowel wall and involves inflammatory cells such as macrophages, which may secrete ferritin, neopterin, and calprotectin.


Local Application of Growth Hormone to Enhance Osseointegration in Osteoporotic Bones: A Morphometric and Densitometric Study.
The aim of this study was to assess the effect of local application of growth hormone on osseointegration of dental implants inserted in osteoporotic bones. Twenty female New Zealand rabbits were used in this study. Ten were ovariectomized and fed a low-calcium diet for 6 weeks, and the others remained intact. A titanium implant was inserted into each tibia, in both groups. In half of the rabbits, 2 IU of growth hormone was placed into the ostectomy prior to the implant insertion. Two weeks after implant surgery, all animals were sacrificed. Tibiae were dissected from soft tissues, and included in methacrylate to be studied under light microscopy. Bone-to-implant contact (BIC) and bone mineral density (BMD) were measured by morphometric and densitometric analysis, respectively. Multifactorial analysis of variance (ANOVA) was used for statistical evaluation. P < .05 was considered to be significant. Ovariectomy induced less BIC and BMD in regions closer to the implant compared with the control group. Local application of growth hormone was able to increase the BIC in the ovariectomized group, with statistically significant differences with respect to the control group (P < .01). Regarding the BMD, no statistically significant differences were found. Within the limitations of this experimental study, local application of 2 IU of recombinant human growth hormone at the moment of titanium implant insertion in rabbit tibiae significantly enhanced the BIC around titanium implants 15 days after the implantation in this experimental osteoporotic animal model, without affecting the BMD.


Intracellular trafficking of sphingolipids: relationship to biosynthesis.
The intracellular routes of sphingolipid trafficking are related to the compartmentalized nature of sphingolipid metabolism, with synthesis beginning in the endoplasmic reticulum, continuing in the Golgi apparatus, and degradation occurring mainly in lysosomes. Whereas bulk sphingolipid transport between subcellular organelles occurs primarily via vesicle-mediated pathways, evidence is accumulating that sphingolipids are found in subcellular organelles that are not connected to each other by vesicular flow, implying additional trafficking routes. After discussing how sphingolipids are transported through the secretory pathway, I will review evidence for sphingolipid metabolism in organelles such as the mitochondria, and then discuss how this impacts upon our current understanding of the regulation of intracellular sphingolipid transport.


CREB promotes laryngeal cancer cell migration via MYCT1/NAT10 axis.
CREB, MYCY1 and NAT10 are involved in cancer cell migration. However, the relationship between these three proteins and their role in laryngeal cancer cell migration remains unknown. Transient gene transfection was performed in laryngeal cancer cells. Bioinformatics analysis was used to predict the binding of CREB to MYCT1 promoter. Binding of CREB to the promoter of MYCT1 was monitored by luciferase reporter assay and chromatin immuno-precipitation method in vitro and in vivo, respectively. Real-time RT-PCR and Western bolt were applied to detect gene transcription and translation levels, respectively. Laryngeal cancer cell migration was assayed by transwell chamber experiment. CREB protein expression was significantly up-regulated in laryngeal cancer tissues and associated with cancer differentiation, tumor stage, and lymphatic metastasis. CREB inhibits MYCT1 expression by direct binding to its promoter. Meanwhile, MYCT1 has a negative impact on the NAT10 gene expression. Furthermore, CREB promotes NAT10 expression via down-regulating the MYCT1 gene expression. In addition, contrary to MYCT1, CREB and NAT10 enhanced laryngeal cancer cell migration. MYCT1 and NAT10 significantly rescued the effects of CREB and MYCT1 on Hep2 cell migration, respectively. CREB promotes laryngeal cancer cell migration via MYCT1/NAT10 axis, suggesting that CREB might be a potential prognostic marker in laryngeal cancer.


Assembly of the aspartate transcarbamoylase holoenzyme from transcriptionally independent catalytic and regulatory cistrons.
The cistrons encoding the regulatory and catalytic polypeptides of aspartate transcarbamoylase (EC 188.8.131.52) from Escherichia coli K-12 have been cloned separately on plasmids from different incompatability groups. The catalytic cistron (pyrB) was carried by pACYC184 and expressed from its own promoter, whereas the regulatory cistron was expressed from the lac po of pBH20. The catalytic polypeptide chains assembled into enzymatically active trimers (c3) in vivo when expressed in the absence of regulatory subunits. Similarly, the regulatory polypeptide chains assembled into regulatory dimers (r2) in vivo in the absence of catalytic subunits. When cellular extracts containing regulatory dimers and catalytic trimers synthesized in separate cells were combined in vitro, partial spontaneous holoenzyme assembly occurred. When pyrB and pyrI were expressed from transcriptionally independent cistrons in the same cell, all detectable catalytic polypeptides were incorporated into the functional aspartate transcarbamoylase holoenzyme, 2(c3):3(r2). Thus, it is clear that the in vivo assembly of ATCase holoenzyme is a direct, spontaneous process involving the association of preformed regulatory subunits (r2) and catalytic subunits (c3). This procedure provides a general method for the construction of hybrid aspartate transcarbamoylase in vivo and may be applicable to other oligomeric enzymes constructed from different polypeptides.


Arrhythmias in the coronary care unit.
Symptomatic sustained cardiac arrhythmias are frequently observed in in the coronary care unit and often lead to hemodynamic compromise, especially in the presence of multisystem disease. The predominant arrhythmias noted in intensive care units are tachyarrhythmias, particularly atrial fibrillation and flutter, and ventricular tachycardia. Bradycardias, arguably less life-threatening than tachyarrhythmias, can arise from sinus node dysfunction or atrioventricular conduction block; transient vagally-mediated bradycardias are frequently encountered as well. Prompt diagnosis of the patient with tachycardia is critical as treatment depends on the accurate diagnosis of tachycardia mechanism. The electrocardiogram remains the most important diagnostic tool for the evaluation of both wide and narrow complex tachycardia. The electrocardiographic diagnosis of wide complex tachycardia is based on evaluation of atrioventricular relationship and QRS morphology while the diagnosis of narrow complex tachycardia is based on the location and morphology of P waves. It is important for critical care specialists to understand the principles of cardiac arrhythmia diagnosis and remain current with the recent advances in the pharmacologic and non-pharmacologic management of patients with arrhythmias.


Reversal of nonlocal vortex motion in the regime of strong nonequilibrium.
We investigate nonlocal vortex motion in weakly pinning a-NbGe nanostructures, which is driven by a transport current I and remotely detected as a nonlocal voltage V{nl}. At a high I of a given polarity, V{nl} changes sign dramatically. This is followed by V{nl} becoming even in I, with the opposite sign at low and high temperatures T. These findings can be explained by a Nernst-like effect resulting from local electron overheating (low T), and a magnetization enhancement due to a nonequilibrium quasiparticle distribution that leads to a gap enhancement near the vortex core (high T).


Long-Term Clinical Outcomes of Patients with Colorectal Cancer with Metastatic Epidural Spinal Cord Compression Treated with Hybrid Therapy (Surgery Followed by Stereotactic Body Radiation Therapy).
Hybrid therapy, consisting of separation surgery followed by stereotactic body radiation therapy, has become the mainstay treatment for radioresistant spinal metastases. Histology-specific outcomes for hybrid therapy are scarce. In clinical practice, colorectal cancer (CRC) is particularly thought to have poor outcomes regarding spinal metastases. The goal of this study was to evaluate clinical outcomes for patients treated with hybrid therapy for spinal metastases from CRC. This retrospective study was performed at a tertiary cancer center. Adult patients with CRC spinal metastasis who were treated with hybrid therapy for high-grade epidural spinal cord or nerve root compression from 2005 to 2020 were included. Outcome variables evaluated included patient demographics, overall survival and progression-free survival, surgical and radiation complications, and clinical-genomic correlations. Inclusion criteria were met by 50 patients. Progression of disease occurred in 7 (14%) patients at the index level, requiring reoperation and/or reirradiation at a mean of 400 days after surgery. Postoperative complications occurred in 16% of patients, with 3 (6%) requiring intervention. APC exon 14 and 16 mutations were found in 15 of 17 patients tested and in all 3 of 7 local failures tested. Twenty patients (40%) underwent further radiation due to disease progression at other spinal levels. Hybrid therapy in patients with CRC resulted in 86.7% local control at 2 years after surgery, with limited complications. APC mutations are commonly present in CRC patients with spine metastases and may suggest worse prognosis. Patients with CRC spinal metastases commonly progress outside the index treatment level.


The miR-145-MMP1 axis is a critical regulator for imiquimod-induced cancer stemness and chemoresistance.
Cancer stemness, chemoresistance, and metastasis are related biological events. However, whether they have common molecular mechanisms remains to be determined. Here, we report that imiquimod (IMQ) facilitates the acquisition of stem-cell-like properties and chemoresistance via the upregulation of matrix metalloproteinase 1 (MMP1) and downregulation of microRNA-145 (miR-145). MiR-145-5p was found to suppress MMP1 expression through direct binding, and miR-145-mediated downregulation of MMP1 reversed the effects of IMQ. In addition, IMQ downregulated miR-145 by promoting DNA methylation at its promoter. DNA methyltransferase inhibitors limited IMQ-induced MMP1 expression, stemness, and chemoresistance. Collectively, our results highlight the miR-145-MMP1 axis as a potential coordinator of cancer stemness and chemoresistance. Given the role of MMP1 in the initiation of metastasis, the miR-145-MMP1 axis serves as a promising therapeutic target for improved cancer treatment.


An evolutionary perspective on anti-tumor immunity.
The challenges associated with demonstrating a durable response using molecular-targeted therapies in cancer has sparked a renewed interest in viewing cancer from an evolutionary perspective. Evolutionary processes have three common traits: heterogeneity, dynamics, and a selective fitness landscape. Mutagens randomly alter the genome of host cells creating a population of cells that contain different somatic mutations. This genomic rearrangement perturbs cellular homeostasis through changing how cells interact with their tissue microenvironment. To counterbalance the ability of mutated cells to outcompete for limited resources, control structures are encoded within the cell and within the organ system, such as innate and adaptive immunity, to restore cellular homeostasis. These control structures shape the selective fitness landscape and determine whether a cell that harbors particular somatic mutations is retained or eliminated from a cell population. While next-generation sequencing has revealed the complexity and heterogeneity of oncogenic transformation, understanding the dynamics of oncogenesis and how cancer cells alter the selective fitness landscape remain unclear. In this technology review, we will summarize how recent advances in technology have impacted our understanding of these three attributes of cancer as an evolutionary process. In particular, we will focus on how advances in genome sequencing have enabled quantifying cellular heterogeneity, advances in computational power have enabled explicit testing of postulated intra- and intercellular control structures against the available data using simulation, and advances in proteomics have enabled identifying novel mechanisms of cellular cross-talk that cancer cells use to alter the fitness landscape.


Transverse + posterior wall fractures of the acetabulum: epidemiology, operative management and long-term results.
PURPOUSE OF THE STUDY: Associated transverse and posterior wall fractures account for approximately 20% of all acetabular fractures. To asses the risk of these concommitant bone injuries on early joint failure despite a high rate of postoperative congruency. The analysis of 104 surgically treated patients with associated transverse and posterior wall fractures showed that more than half of these patients had associated injuries. The mean age was 35 years, and > 75% of these patients were male. A high energy trauma was the trauma mechanism in 94.2%. The mean ISS was 26.3 points. The majority of patients showed a juxta- or transtectal fracture line. The mean articular fracture displacement was 13.5 mm. 87.5% of the patients showed a femoral head dislocation. An acetabular roof comminution was present in 16.3%. 20.2% of patients received a fracture related preoperative nerve injury to the sciatic nerve. Osteosynthesis was performed 9.9 days after trauma. The Kocher-Langenbeck approach was used in > 90% for stabilization with a combination of plate and screw fixation in 71.1%. The mean operative time was 190 minutes with a blood loss of 855 ml. Postoperatively the hip joint was congruent in 90.3% with anatomical or near-anatomical joint reconstruction in > 90%. Iatrogenic nerve injury occurred in 12 patients (8.9%). 67 patients (67.7%) could be followed after a mean of 42.7 months. The average subjective Visual Analog Scale pain score was 42.7. Mild or no pain was seen in 58.2%. The mean Merle d'Aubigné score was 15.4 with 56.7% of patients having a functionally perfect or good result. 52.2% had no post-traumatic osteoarthritic changes of their hip joint. A joint failure was diagnosed in 32.8% of the patients. Analyzing only patients with anatomically reconstructed hip joints, patients showed comparable results with 61.3% having no or mild pain and 59.2% a good or excellent functional result. Posttraumatic arthrotic changes occur in only 26.5% of these patients. A joint failure was present in 32.7%. In this group, a joint failure was significantly more likely to be present with an additional acetabular comminution zone. Associated transverse and posterior wall fractures have a significant risk of early joint failure despite a high rate of postoperative congruency.


Modification of a Novel Umami Octapeptide with Trypsin Hydrolysis Sites via Homology Modeling and Molecular Docking.
Increasing the copy number of peptides is an effective method to genetically engineer recombinant expression and obtain umami peptides in large quantities. However, the umami taste value of multicopy number umami peptides is lower than the single ones, thus limiting the industrial application of recombinantly expressed umami peptides. With aims to solve this problem, modification of an umami beefy meaty peptide (BMP) with trypsin hydrolysis sites was carried out via homology modeling and molecular docking in this study. A total of 1286 modified peptide sequences were created and molecularly simulated for docking with the homology modeling-constructed umami receptor (T1R1/T1R3), and 837 peptides were found to be better docked than the BMP. Afterward, the MLSEDEGK peptide with the highest docking score was synthesized. And umami taste evaluation results demonstrated that this modified peptide was close to that of monosodium glutamate (MSG) and BMP, as confirmed by electronic tongue and sensory evaluation (umami value: 8.1 ± 0.2 for BMP; 8.2 ± 0.3 for MLSEDEGK peptide). Meanwhile, mock trypsin digestion of eight copies of MLSEDEGK peptide results showed that the introduced digestion sites were effective. Therefore, the novel modified BMP in this study has the potential for large-scale production by genetic engineering.


Precepting: a win-win for students and pharmacists.
The education of pharmacy students includes hundreds of hours of experiential education under the supervision of a licensed pharmacist called a preceptor. While this real-world education is instrumental in preparing students for their pharmacy career, many experienced pharmacists are hesitant to commit to becoming a preceptor. By addressing some of their fears about precepting and becoming knowledgeable about the available resources, pharmacists can embrace the opportunity to become preceptors as a mutual learning experience between pharmacist and student. Experienced pharmacists can learn about innovations in practice and new drugs on the market, and, with some training, students can help with basic tasks in everyday practice.


Three gonadotropin-releasing hormone neuronal groups with special reference to teleosts.
Gonadotropin-releasing hormone (GnRH) is a decapeptide, which has been isolated from the hypothalamus as a releasing hormone of gonadotropins from the pituitary. However, subsequent morphological studies have demonstrated the presence of multiple GnRH neuronal groups outside the hypothalamus and preoptic area. In most vertebrate lineages studied to date, GnRH neuronal groups are present along the terminal nerve and in the midbrain tegmentum, in addition to a population in the preoptico-hypothalamic areas. The presence of GnRH fibers in extrahypothalamic areas has also been demonstrated, indicating a significance for GnRH neurons in functions other than those that are purely hypophysiotropic. Among vertebrate lineages, GnRH neurons have been most extensively studied in teleost fish through morphological, electrophysiological, behavioral and molecular approaches. To date, studies on differential roles of GnRH neuronal groups have been mostly restricted to teleosts. In the present review, the anatomy and functions of each GnRH neuronal group are reconsidered, based mainly on knowledge from teleosts. Recent findings in teleosts indicate that the preoptico-hypothalamic GnRH neurons are hypophysiotropic and that GnRH neurons of the terminal nerve and midbrain tegmentum regulate neural activities in various regions, including extrahypothalamic areas. The latter populations presumably serve as neuromodulatory systems to control aspects of neural functions such as reproductive behavior. Similar functional differentiation may be generalized to other vertebrate lineages as well.


Large-scale separation of antipsychotic alkaloids from Rauwolfia tetraphylla L. by pH-zone-refining fast centrifugal partition chromatography.
pH-zone-refining centrifugal partition chromatography was successively applied in the large-scale separation of close R(f) antipsychotic indole alkaloids directly from CHCl(3) fraction of Rauwolfia tetraphylla leaves. Two experiments with increasing mass from 500 mg to 3 g of crude alkaloid extracts (1C) of R. tetraphylla were carried out in normal-displacement mode using a two-phase solvent system composed of methyl tert-butyl ether/ACN/water (4:1:5, v/v/v) where HCl (12 mM) was added to the lower aqueous stationary phase as a retainer and triethylamine (5 mM) to the organic mobile phase as an eluter. The two centrifugal partition chromatography separations afforded a total of 162.6 mg of 10-methoxytetrahydroalstonine (1) and 296.5 mg of isoreserpiline (2) in 97% and 95.5% purity, respectively, along with a 400.9 mg mixture of α-yohimbine and reserpiline (3 and 4). Further, this mixture was resolved over medium pressure LC using TLC grade silica gel H (average particle size 10 μm), which afforded 160.4 mg of α-yohimbine (3) and 150.2 mg of reserpiline (4) in >95% purities. The purity of the isolated antipsychotic alkaloids was analyzed by high-performance LC and their structures were characterized on the basis of their 1D, 2D NMR and electrospray ionization-mass spectroscopic data.


[Psychosomatic correlations of sympathetic reflex dystrophy (Sudeck's disease). Review of the literature and initial clinical results].
The present state of the art in psychosomatic research on reflex sympathetic dystrophy or Sudeck's atrophy (also known as Sudeck's disease) is reviewed. The survey confirms the impression prevailing with surgeons and orthopaedists that patients suffering from this disease are "psychically peculiar", i.e. they appear strange or odd. Psychometric examinations show that they suffer from enhanced anxiety, are emotionally rather unstable and display a tendency to depressiveness associated with a marked self-esteem rating problem complex. Basing upon these criteria the occurrence of reflex sympathetic dystrophy in the postoperative course of patients with Dupuytren's contracture could be correctly predicted in 43 of 47 patients, the remaining 4 patients being forecast with a false-positive prediction (Zachariae 1964). Studies on Children with reflex sympathetic dystrophy likewise confirmed these characteristic features. In a study of our own we examined in 12 patients their biographical development as well as life events that had happened 6 months before onset of the disease. We obtained a surprisingly uniform result: In all patients we found that at least one or in most cases several severely stressful life events had occurred; all patients had been suffering from chronic headache and/or pain in the lumbar vertebral column for many years; and all of them had shown signs in the course of their development in childhood and adolescence that have been described by Engel as "pain proneness". Over and above this, the anamnesis of most patients revealed other psychogenic and psychosomatic diseases or signs and symptoms. The case reports of all the 12 patients served as basis for initial approaches to a psychosomatic disease concept of reflex sympathetic dystrophy.


Oral immunotherapy of recurrent urinary tract infections: a double-blind placebo-controlled multicenter study.
We treated 166 patients suffering from recurrent urinary tract infections under double-blind conditions for 3 months with 1 capsule daily of either the immunostimulating bacterial extract (85) or a placebo (81), followed by a 3-month observation period without the test drugs. The bacterial extract exerted a significant beneficial curative action and long-term consolidative effect on the frequency of recurrent urinary tract infections with marked improvements in the characteristic signs and symptoms. It was significantly superior to placebo for the majority of the assessed parameters: number of recurrent urinary tract infections, bacteriuria, leukocyturia, erythrocyturia, nitrituria, albuminuria and casts in urine. Consumption of antibiotics, chemotherapeutics, urinary antiseptics or disinfectants was significantly less under active drug therapy compared to placebo. Tolerance was good with only 2 side effects reported in 2 patients (2%) in the active group compared to 11 among 5 (6%) in the placebo group. Therefore, the bacterial extract can be considered an efficient and well tolerated drug for the treatment of urinary tract infections, and their accompanying signs and symptoms, as well as for decreasing the risk of recurrences and the need for antibiotics and other antibacterial drugs.


Ag-doped multiwalled carbon nanotube/polymer composite electrodes.
Two types of carbon nanotube (CNT) based films were fabricated by adsorbing two different types of multiwalled carbon nanotubes (MWCNTs) on bacterial cellulose membrane templates. A bacterial cellulose membrane consists of ribbon-shaped nanofibrils that are arranged to form a porous 3D network. These characteristics lead to the uniform deposition of MWCNTs on the membrane structure and the final CNT based films with high surface areas. Two types of MWCNTs were used: (1) MWCNTs that had been purified by an acid treatment and (2) Ag-doped MWCNTs, which consisted of MWCNTs with Ag nanoparticles attached to their surfaces. The morphologies of the Ag-doped MWCNTs and CNT based films were examined using transmission electron microscopy. In addition, the electrical conductivity and electrochemical properties of the CNT based films were measured using a four-point probe and a cyclic voltammeter, respectively. The cyclic voltammetry data showed that the Ag particles attached to the MWCNT surfaces influenced the electrochemical properties of the CNT based films.


Influence of IS256 on Genome Variability and Formation of Small-Colony Variants in Staphylococcus aureus.
Staphylococcus aureus has acquired resistance to nearly all antibiotics used in clinical practice. Whereas some resistance mechanisms are conferred by uptake of resistance genes, others evolve by mutation. In this study, IS256 has been shown to play a role, e.g., in S. aureus strains displaying intermediate resistance to vancomycin (VISA). To characterize the IS256 insertion sites in the genomes of two closely related sequence type 247 (ST247) VISA strains, all insertions were mapped in both VISA and a susceptible control strain. The results showed that the three ST247 strains contained the highest number so far of IS256 insertions for all sequenced S. aureus strains. Furthermore, in contrast to the case with the other IS elements in these genomes, the IS256 insertion sites were not identical in the closely related strains, indicating a high transposition frequency of IS256 When IS256 was introduced into a laboratory strain which was then cultured in the presence of antibiotics, it was possible to isolate small-colony variants (SCVs) that possessed IS256 insertions in guaA and hemY that displayed increased resistance to vancomycin and aminoglycosides, respectively. For these clones, a very rapid reversion to the wild type that resembled the fast reversion of clinical SCVs was observed. The reversion was caused by excision of IS256 in a small number of fast-growing clones that quickly outcompeted the SCVs in broth cultures. In conclusion, the presence of IS256 confers a strong genomic plasticity that is useful for adaptation to antibiotic stress.


CDK9 and SPT5 proteins are specifically required for expression of herpes simplex virus 1 replication-dependent late genes.
DNA replication greatly enhances expression of the herpes simplex virus 1 (HSV-1) γ2 late genes by still unknown mechanisms. Here, we demonstrate that 5,6-dichloro-1-β-d-ribofuranosylbenzimidazole (DRB), an inhibitor of CDK9, suppresses expression of γ2 late genes with an IC50 of 5 μm, which is at least 10 times lower than the IC50 value required for inhibition of expression of early genes. The effect of DRB could not be explained by inhibition of DNA replication per se or loading of RNA polymerase II to late promoters and subsequent reduction of transcription. Instead, DRB reduces accumulation of γ2 late mRNA in the cytoplasm. In addition, we show that siRNA-mediated knockdown of the transcription factor SPT5, but not NELF-E, also gives rise to a specific inhibition of HSV-1 late gene expression. Finally, addition of DRB reduces co-immunoprecipitation of ICP27 using an anti-SPT5 antibody. Our results suggest that efficient expression of replication-dependent γ2 late genes is, at least in part, regulated by CDK9 dependent co- and/or post-transcriptional events involving SPT5 and ICP27.


High density polyethylene prosthetic femoral head replacemnt in the dog.
A high density polyethylene (RCH 1000) femoral head prosthesis was inserted in 5 large dogs. Periodic clinical and roentgenographic evaluations, and histopathological examination of the replaced joints as well as biomechanical evaluation of the prosthetic joints were done at intervals from 3 weeks to 15 months. HDP is NOT suitable for movable weight-bearing prosthesis articulating with cartilage or bone. Wear of the cartilage and the polyethylene occurred within a few weeks of function and became extensive proportionally to the length of use. The accumulating wear particles of HDP caused a significant foreign body granulomatous reaction which resulted in erosion of bone adjacent to the acetabulum in one hip.


The revenue engine that could: "think you can" by refining the revenue cycle with the right people, processes, and tools.
Many physicians continue to wrestle with an economy-in-recovery and declining reimbursements. In this business climate, practices can't afford reimbursement process mistakes and inefficiencies; they're simply too expensive. Just a few denied surgical claims can cost a practice thousands of dollars. That's the cost of the annual electronic health records license or the T1 line. Uncovering any and all opportunities to improve the speed and efficiency of getting paid can positively contribute to the bottom line. This article reiterates the basics and "best" practices for efficient revenue cycle operations. The goal is to have the right tasks performed by the right number of people at the right time and with the right tools to optimize revenue.


Preliminary experiences with CO2-laser and ultrasonic aspirator in brain surgery.
The basic problems and potential of CO2-laser and ultrasound aspirator in brain surgery are presented on the basis of experience with the Sharplan 733A CO2-laser and the Sonotec ultrasound aspirator. Complete removal of meningiomas and gliotic scar and partial removal of malignant astrocytomas by CO2-laser and ultrasound aspirator was performed. There were no complications related to the surgical procedure. The main advantage of the CO2-laser was precise cutting of neural tissue and rapid evaporation for debulkment of tumor. The ultrasound aspirator was especially useful for rapid internal debulkment by fragmentation and aspiration of soft tumors. On the basis of these experiences the surgical theater should be equipped with these and other instruments in such a way that they can be readily and interchangeably used in any neurosurgical procedure.


HBV DNA and other hepatitis B virus markers in sera from long-term hemodialysis and kidney transplant patients.
Sera from 191 long-term hemodialysis patients and from 115 renal transplant patients were studied for the presence of HBsAg and other HBV markers. In 19.1% of the hemodialysis patients and 28.7% of the transplant patients, the sera were positive for HBeAg.HBV DNA in sera was detected by molecular hybridization. HBV DNA was present in the sera of 15 out of 16 hemodialysis patients positive for HBeAg, and in one hemodialysis patient positive for anti-HBe. All renal transplant patients positive for HBeAg were also positive for HBV DNA. Twelve transplant patients were positive for HBV DNA, but negative for both HBeAg and anti-HBe. Determination of HBV DNA was the most sensitive marker of infectivity in patients with end-stage renal disease, and in renal transplant patients.


Underlying Mechanisms and Related Diseases Behind the Complex Regulatory Role of NOD-Like Receptor X1.
Among nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs), NOD-like receptor X1 (NLRX1) is the only known NLR family member that is targeted to the mitochondria, which contains a C-terminal leucine-rich repeat domain, a central conserved nucleotide-binding domain, and an unconventional N-terminal effector domain. It is unique due to several atypical features, such as mitochondrial localization, noninflammasome forming, and relatively undefined N-terminal domain. NLRX1 has multiple functions, including negative regulation of type-I interferon signaling, attenuation of proinflammatory nuclear factor kappa B (NF-κB) signaling, autophagy induction, modulation of reactive oxygen species production, cell death regulation, and participating in cellular senescence. In addition, due to its diverse functions, NLRX1 has been associated with various human diseases, including respiratory, circulatory, motor, urinary, nervous, and digestive systems, to name but a few. However, the exact regulatory mechanisms of NLRX1 are still unclear in many related diseases since conflicting and controversial topics on NLRX1 in the previous studies remain. In this review, we review recent research advances on the underlying mechanisms and related disorders behind the complex regulatory role of NLRX1, which may provide a promising target to prevent and/or treat the corresponding diseases.


Targeting mTOR in the Context of Diet and Whole-body Metabolism.
The mechanistic target of the rapamycin (mTOR) signaling pathway is the central regulator of cell growth and proliferation by integrating growth factor and nutrient availability. Under healthy physiological conditions, this process is tightly coordinated and essential to maintain whole-body homeostasis. Not surprisingly, dysregulated mTOR signaling underpins several diseases with increasing incidence worldwide, including obesity, diabetes, and cancer. Consequently, there is significant clinical interest in developing therapeutic strategies that effectively target this pathway. The transition of mTOR inhibitors from the bench to bedside, however, has largely been marked with challenges and shortcomings, such as the development of therapy resistance and adverse side effects in patients. In this review, we discuss the current status of first-, second-, and third-generation mTOR inhibitors as a cancer therapy in both preclinical and clinical settings, with a particular emphasis on the mechanisms of drug resistance. We focus especially on the emerging role of diet as an important environmental determinant of therapy response, and posit a conceptual framework that links nutrient availability and whole-body metabolic states such as obesity with many of the previously defined processes that drive resistance to mTOR-targeted therapies. Given the role of mTOR as a central integrator of cell metabolism and function, we propose that modulating nutrient inputs through dietary interventions may influence the signaling dynamics of this pathway and compensatory nodes. In doing so, new opportunities for exploiting diet/drug synergies are highlighted that may unlock the therapeutic potential of mTOR inhibitors as a cancer treatment.


[Alternations of bronchial smooth muscle and effect of therapy on remodeling in asthma].
Alternations in airway wall architecture, particularly increased smooth muscle mass are associated with pathogenesis of asthma. Muscle fiber hyperplasia and hypertrophy is a major contributor to the increase in smooth muscle mass. Airway smooth muscle was traditionally considered to have only contractile and proliferative functions and has little attention with regard to its ability to express and release inflammatory mediators. Airway smooth muscle cells have been shown to release cytokines such as: GM-CSF, IL-11, IL-6, IL-1, IL-5, IL-8, PGs and NO. Airway remodeling has been shown to respond to some degree anti-inflammatory therapy. Several study results indicate that steroid can positively influence progressive airflow limitation. Combined use of a beta2-agonist and steroid can reduced the remodeling progression.


Preference for a fixed combination of brinzolamide/timolol versus dorzolamide/timolol among patients with open-angle glaucoma or ocular hypertension.
To assess ocular discomfort upon instillation and patient preference for brinzolamide/timolol relative to dorzolamide/timolol, in patients with open-angle glaucoma or ocular hypertension. This was a multicenter, prospective, patient-masked, randomized, crossover study. On day 0, patients received one drop of brinzolamide/timolol in one eye and one drop of dorzolamide/timolol in the contralateral eye. On day 1, patients were randomly assigned to receive one drop of either brinzolamide/timolol or dorzolamide/timolol in both eyes; on day 2, patients received one drop of the alternate treatment in both eyes. Measures included a patient preference question on day 2 (primary) and mean ocular discomfort scale scores on days 1 and 2 (secondary). Safety assessments included adverse events, visual acuity, and slit-lamp examinations. Of 120 patients who enrolled, 115 completed the study. Of these, 112 patients instilled both medications and expressed a study medication preference on day 2. A significantly greater percentage preferred brinzolamide/timolol to dorzolamide/timolol (67.0% versus 30.4%; P < 0.001). The ocular discomfort (expressed as mean [standard deviation]) with brinzolamide/timolol was significantly lower than with dorzolamide/timolol (day 2:1.9 [2.3] versus 3.7 [2.8], respectively [P = 0.0003]; both days combined: 2.1 [2.5] versus 3.5 [2.9], respectively [P = 0.00014]). On day 1, five patients receiving brinzolamide/timolol reported five nonserious adverse events (AEs): flu (n = 1), bitter taste (n = 2), and headache (n = 2). Four events, bitter taste (two events) and headache (two events), were considered related to brinzolamide/timolol. Events were mild in intensity, except bitter taste of moderate intensity reported by one patient. No AEs were reported at day 2. All AEs resolved without additional treatment. No clinically relevant changes from baseline were observed in best-corrected visual acuity or slit-lamp examinations of ocular signs. Patients had less discomfort with brinzolamide/timolol than with dorzolamide/timolol, and more expressed a preference for brinzolamide/timolol. Both treatments were generally safe and well tolerated.


Energetic Constraints Produce Self-sustained Oscillatory Dynamics in Neuronal Networks.
Overview: We model energy constraints in a network of spiking neurons, while exploring general questions of resource limitation on network function abstractly. Background: Metabolic states like dietary ketosis or hypoglycemia have a large impact on brain function and disease outcomes. Glia provide metabolic support for neurons, among other functions. Yet, in computational models of glia-neuron cooperation, there have been no previous attempts to explore the effects of direct realistic energy costs on network activity in spiking neurons. Currently, biologically realistic spiking neural networks assume that membrane potential is the main driving factor for neural spiking, and do not take into consideration energetic costs. Methods: We define local energy pools to constrain a neuron model, termed Spiking Neuron Energy Pool (SNEP), which explicitly incorporates energy limitations. Each neuron requires energy to spike, and resources in the pool regenerate over time. Our simulation displays an easy-to-use GUI, which can be run locally in a web browser, and is freely available. Results: Energy dependence drastically changes behavior of these neural networks, causing emergent oscillations similar to those in networks of biological neurons. We analyze the system via Lotka-Volterra equations, producing several observations: (1) energy can drive self-sustained oscillations, (2) the energetic cost of spiking modulates the degree and type of oscillations, (3) harmonics emerge with frequencies determined by energy parameters, and (4) varying energetic costs have non-linear effects on energy consumption and firing rates. Conclusions: Models of neuron function which attempt biological realism may benefit from including energy constraints. Further, we assert that observed oscillatory effects of energy limitations exist in networks of many kinds, and that these findings generalize to abstract graphs and technological applications.


Negative dispersion: a backward wave or fast light? Nanoplasmonic examples.
Propagation of light in a negative dispersion regime (antiparallel phase and group velocities) may be attributed to either fast light or a backward wave. We show that by applying causality, only one of these is valid for each scenario. A nanoplasmonic structure is shown to support both types of solution depending on the parameters.


[Analysis of a small quatity of polypeptides by two-dimensional electrophoresis: application to characterization of proteins].
A new two-dimensional electrophoresis is described. The first electrophoresis is performed in a 10 p. cent gel (1 X 110 mm) in the presence of 7 M urea; the migration in the second dimension proceeds on a 20 p. cent polyacrylamide sheet (160 X 110 X 1 mm) in the presence of sodium dodecyl sulphate. This system resolves most of the polypeptides obtained by treating a very small amount (about 100 mug) of proteins with cyanogene bromide. Macromolecules can thus be quickly and easily characterized.


Effects of amburoside A and isokaempferide, polyphenols from Amburana cearensis, on rodent inflammatory processes and myeloperoxidase activity in human neutrophils.
The present study evaluated the anti-inflammatory activity of amburoside A (a phenol glucoside) and isokaempferide (a flavonol) isolated from the trunk bark of Amburana cearensis, a medicinal plant used in northeast Brazil for the treatment of asthma. Animals (male Wistar rats or Swiss mice) pre-treated with amburoside A (25 and 50 mg/kg) or isokaempferide (12.5, 25 and 50 mg/kg), orally or intraperitoneally, showed a significant inhibition of the paw oedema induced by carrageenan (1%), prostaglandin E(2) (30 nmol/paw), histamine (200 microg/paw) or serotonin (200 microg/paw). Histological and morphometric evaluations of the rat paw oedema induced by carrageenan showed that amburoside A and isokaempferide also inhibited the accumulation of inflammatory cells. Amburoside A reduced significantly the paw oedema and the increase in vascular permeability induced by dextran, as related to the control group. Similar results were observed with the isokaempferide pre-treatment. Furthermore, amburoside A or isokaempferide inhibited both leucocyte and neutrophil migrations, in mouse peritoneal cavity, after the carrageenan injection. The polyphenols were not cytotoxic and blocked N-formyl-methyl-leucyl-phenylalanine-induced myeloperoxidase release and activity in human neutrophils. In addition, amburoside A and isokaempferide at 50 and 100 microg/ml concentrations reduced significantly the lipopolysaccharide-mediated increase in tumour necrosis factor-alpha (TNF-alpha) levels. These results provide, for the first time, evidence to support the anti-inflammatory activity of amburoside A and isokaempferide that seems to be related to an inhibition of inflammatory mediators, such as TNF-alpha, as well as histamine, serotonin and prostaglandin E(2), besides leucocyte infiltration in a dose- or concentration-dependent manner. These anti-inflammatory effects can be explained, at least in part, by the ability of these compounds to reduce neutrophil degranulation, myeloperoxidase activity, mediators as well as TNF-alpha secretion.


Cost analysis of cordotomy and intrathecal pain pump placement for refractory cancer pain.
Cancer pain can be debilitating and 10-20% of patients will have refractory pain despite optimal medical management. Here, we present a cost comparison of treating terminal cancer patients with intravenous (IV) narcotics, anterolateral cordotomy, or intrathecal pain pump (ITPP) placement. We evaluated and treated 2 patients with metastatic breast cancer and expected survivals of <1 year. The first patient, a 53-year-old female, had tumor invasion of the right chest wall and had failed oral pain regimens; she was admitted to receive IV Dilaudid as patient-controlled analgesia (PCA). After 7 days of treatment without improvement, she underwent a left-sided C1-2 cordotomy. For her, the cost of the cordotomy was $18,462 and the expenses for 7 days hospital stay with PCA was $89,884; the total was $108,346. The second patient, a 60-year-old female, had severe somatic pain due to invasion by tumor of the left knee cap. She, too, has failed oral therapy and was receiving in-hospital IV Dilaudid PCA. Following 2 days of failed treatment, a morphine ITPP was placed and effectively treated her pain. In patient 2, the cost of the ITPP was $80,603 and the expenses for 8 days of the hospital stay with PCA came to $84,785; the total was $165,389. The treatment of refractory pain in cancer patients is challenging. It requires invasive procedures such as cordotomy or ITPP. Although procedures may yield comparable pain control, there was a significant cost savings for cordotomy versus ITPP ($57,043 saved).


Virulence of Emerging Arthrotropic Avian Reoviruses Correlates With Their Ability to Activate and Traffic Interferon-γ Producing Cytotoxic CD8+ T Cells Into Gastrocnemius Tendon.
Newly emerging arthrotropic avian reoviruses (ARVs) are genetically divergent, antigenically heterogeneous, and economically costly. Nevertheless, the mechanism of emerging ARV-induced disease pathogenesis and potential differences in virulence between virus genotypes have not been adequately addressed. In this study, the life cycle of ARV, including the formation of cytoplasmic ARV neo-organelles, paracrystalline structures, and virus release mechanisms, were characterized in the infected host cell by transmission electron microscopy (TEM). In addition, progressive changes in the structure of infected cells were investigated by time-lapse and field emission scanning electron (FE-SE) microscopy. ARVs from the four genotypic cluster groups included in the study caused gross and microscopic lesions in the infected birds. Marked infiltration of γδT cells, CD4+ and CD8+ T lymphocytes were observed in ARV infected tendon tissues starting day 3 post-infection. The ARV variant from genotype cluster-2 triggered significantly high trafficking of IFN-γ producing CD8+ T lymphocytes in tendon tissues and concomitantly showed high morbidity and severe disease manifestations. In contrast, the ARV variant from genotype cluster-4 was less virulent, caused milder disease, and accompanied less infiltration of IFN-γ producing CD8+ T cells. Interestingly, when we blunted antiviral immune responses using clodronate liposomes (which depletes antigen-presenting cells) or cyclosporin (which inhibits cytokine production that regulates T-cell proliferation), significantly lower IFN-γ producing CD8+ T cells infiltrated into tendon tissues, resulting in reduced tendon tissues apoptosis and milder disease manifestations. In summary, these data suggest that the degree of ARV virulence and tenosynovitis/arthritis are potentially directly associated with the ability of the virus to traffic massive infiltration of cytotoxic CD8+ T cells into the infected tissues. Moreover, the ability to traffic cytotoxic CD8+ T cells into infected tendon tissues and the severity of tenosynovitis differ between variants from different ARV genotype cluster groups. However, more than one virus isolate per genotype group needs to be tested to further confirm the association of pathogenicity with genotype. These findings can be used to further examine the interaction of viral and cellular pathways which are essential for the pathogenesis of the disease at the molecular level and to develop effective disease control strategies.


The surface hydrophobicity and avirulent character of an encapsulated strain of Klebsiella pneumoniae.
In order to elucidate how virulence is controlled in encapsulated bacteria, some surface properties of an encapsulated but avirulent strain of Klebsiella pneumoniae, strain 277, were examined. Although strain 277 was heavily fimbriated, the fimbriae did not demonstrate an avirulent character and were not responsible for the surface hydrophobicity of this strain. The surface hydrophobicity was well correlated with the capacity of the bacteria to associate with polymorphonuclear cells. More bacteria with hydrophobic surfaces associated with the PMN than nonhydrophobic bacteria. The hydrophobic surface character of this strain was not affected by either trypsin treatment or extraction with salt solution. We assume that the capsule of strain 277 has more hydrophobic polysaccharides than that of the virulent strain. Some chemical modifications might therefore exist in the capsular polysaccharides of the avirulent strain.


Exercise And Heart Failure: Advancing Knowledge And Improving Care.
Exercise limitation is the hallmark of heart failure, and an increasing degree of intolerance is associated with poor prognosis. Objective evaluation of functional class (e.g., cardiopulmonary exercise testing) is essential for adequate prognostication in patients with advanced heart failure and for implementing an appropriate exercise training program. A graded exercise program has been shown to be beneficial in patients with heart failure and has become an essential component of comprehensive cardiac rehabilitation in these patients. An exercise program tailored to the patient's preferences, possibilities, and physiologic reserve has the greatest chance of being successful. Despite being safe, effective, and a guideline-recommended treatment to improve quality of life, exercise training remains grossly underutilized. Patient, physician, insurance and practice barriers need to be addressed to improve this quality gap.


The Prediction of Survival in Hepatocellular Carcinoma Based on A Four Long Non-coding RNAs Expression Signature.
Prognostic stratification in hepatocellular carcinoma (HCC) patients is still challenging. Long non-coding RNAs (lncRNAs) have been proven to play a crucial role in tumorigenesis and progression of cancers. The aim of this study is to develop a useful prognostic index based on lncRNA signature to identify patients at high risk of disease progression. We obtained lncRNA expression profiles from three publicly available datasets from Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). By the risk scoring method, we built an individualized four-lncRNA signature (HCCLnc-4) to predict survival of HCC patients in the discovery set (ROC curve, AUC: 0.83, 95% CI: 0.65-1.00, P < 0.05, Kaplan-Meier analysis and log-rank test, P < 0.01). Similar prognostic value of HCCLnc-4 has been further verified in two other independent sets. Stratified analysis and multivariate Cox regression analysis suggested the independence of HCCLnc-4 for prediction of HCC patient survival from traditional clinicopathological factors. Area under curve (AUC) analysis suggested that HCCLnc-4 could compete sufficiently with, or might be even better than classical pathological staging systems to predict HCC patient prognosis in the same data sets. Functional analysis and network analysis suggested the potential implication of lncRNA biomarkers. Our study developed and validated the lncRNA prognostic index of HCC patients, warranting further clinical evaluation and preventive interventions.


Contributions of heterosis and epistasis to hybrid fitness.
Early-generation hybrid fitness is difficult to interpret because heterosis can obscure the effects of hybrid breakdown. We used controlled reciprocal crosses and common garden experiments to distinguish between effects of heterosis and nuclear and cytonuclear epistasis among morphotypes and advanced-generation hybrid derivative populations in the Piriqueta caroliniana (Turneraceae) plant complex. Seed germination, growth, and sexual reproduction of first-generation hybrids, inbred parental lines, and outbred parental lines were compared under field conditions. Average vegetative performance was greater for hybrids than for inbred lines, and first-season growth was similar for hybrids and outbred parental lines. Hybrid survival surpassed that of inbred lines and was equal to or greater than outbred lines' survival, and more F(1) than parental plants reproduced. Reductions in hybrid fitness due to Dobzhansky-Muller incompatibilities (epistasis among divergent genetic elements) were expressed as differences in vegetative growth, survival, and reproduction between plants from reciprocal crosses for both F(1) and backcross hybrid generations. Comparing performance of hybrids against parental genotypes from intra- and interpopulation crosses allowed a more robust prediction of F(1) hybrids' success and more accurate interpretations of the genetic architecture of F(1) hybrid vigor.


Metachronous rectal metastasis from primary transverse colon cancer: a case report.
Colorectal metastases from primary colorectal cancers are very rare, and little is known about their epidemiological aspects or the best diagnostic and therapeutic strategies. Herein, we report a case of a 65-year-old woman with suspected metachronous metastasis to the rectum from primary transverse colon cancer. The patient underwent a laparoscopic extended right hemicolectomy for primary transverse colon cancer. Histopathological examination showed moderately differentiated adenocarcinoma, and the tumor was diagnosed as stage IIA (T3, N0, M0). Fifteen months after her colectomy, a computed tomography scan demonstrated a rectal tumor and a right ovarian tumor. Colonoscopy revealed a superficial elevated lesion in the middle rectum, and histological analysis showed moderately differentiated adenocarcinoma. Laparoscopic low anterior resection preserving the left colic artery and bilateral adnexectomy were performed. Histological examination of the rectal tumor showed that adenocarcinoma was mainly present in the submucosa and muscularis propria, while the carcinoma-involved region of the mucosal layer had mucosal colonization representing the spread of metastatic tumor cells along the basement membrane of preexisting crypts and/or villi. The right ovarian tumor proved to be moderately differentiated adenocarcinoma that was positive for cytokeratin 20 and negative for cytokeratin 7 staining, indicating metastasis from the colorectal cancer. The rectal and ovarian tumors were similar to transverse colon cancer in architectural and cytological atypia. Both adenocarcinomas of the transverse colon and rectum were negative for p53 in immunohistochemical staining and RAS wild type in genetic assessment. These findings support a possible diagnosis of rectal and ovarian metastasis from the primary transverse colon cancer. The patient recovered well after surgery, and neither relapse nor metastasis was observed 18 months after surgery. Distinguishing primary from metastatic colorectal cancer can be challenging, but a comprehensive evaluation of histological features, clinical history, and tumor distribution can enable making a correct diagnosis and implementing optimal treatment.


Selection for reduced translation costs at the intronic 5' end in fungi.
It is generally believed that introns are not translated; therefore, the potential intronic features that may be related to the translation step (occurring after splicing) have yet to be thoroughly studied. Here, focusing on four fungi, we performed for the first time a comprehensive study aimed at characterizing how translation efficiency is encoded in introns and affects their evolution. By analysing their intronome we provide evidence of selection for STOP codons close to the intronic 5' end, and show that the beginning of introns are selected for significantly high translation, presumably to reduce translation and metabolic costs in cases of non-spliced introns. Ribosomal profiling data analysis in Saccharomyces cerevisiae supports the conjecture that in this organism intron retention frequently occurs, introns are partially translated, and their translation efficiency affects organismal fitness. We show that the reported results are more significant in highly translated and highly spliced genes, but are not associated only with genes with a specific function. We also discuss the potential relation of the reported signals to efficient nonsense-mediated decay due to splicing errors. These new discoveries are supported by population-genetics considerations. In addition, they are contributory steps towards a broader understanding of intron evolution and the effect of silent mutations on gene expression and organismal fitness.


Field migration rates of tidal meanders recapitulate fluvial morphodynamics.
The majority of tidal channels display marked meandering features. Despite their importance in oil-reservoir formation and tidal landscape morphology, questions remain on whether tidal-meander dynamics could be understood in terms of fluvial processes and theory. Key differences suggest otherwise, like the periodic reversal of landscape-forming tidal flows and the widely accepted empirical notion that tidal meanders are stable landscape features, in stark contrast with their migrating fluvial counterparts. On the contrary, here we show that, once properly normalized, observed migration rates of tidal and fluvial meanders are remarkably similar. Key to normalization is the role of tidal channel width that responds to the strong spatial gradients of landscape-forming flow rates and tidal prisms. We find that migration dynamics of tidal meanders agree with nonlinear theories for river meander evolution. Our results challenge the conventional view of tidal channels as stable landscape features and suggest that meandering tidal channels recapitulate many fluvial counterparts owing to large gradients of tidal prisms across meander wavelengths.


Perspectives of key stakeholders and smokers on a very low nicotine content cigarette-only policy: qualitative study.
To investigate views of New Zealand key stakeholders (stakeholders) and smokers on very low nicotine content (VLNC) cigarettes, and a policy mandating that only VLNC cigarettes are available for sale. Using a semi-structured interview schedule, we interviewed 17 stakeholders and held focus groups with 21 smokers. Questions were asked about VLNC cigarettes and a VLNC cigarette-only policy. Smokers were given approximately 15 VLNC cigarettes to take home and smoke. One week after the focus groups, 17 smokers were interviewed. Data were analysed using a general inductive approach. Stakeholders and smokers were largely unconvinced of the value of a mandated reduction in nicotine in cigarettes. After smoking VLNC cigarettes, smokers had less interest in them but would support them being sold alongside high nicotine content (HNC) cigarettes at a much cheaper price. The government is not likely to mandate nicotine reduction in cigarettes if there is a perceived lack of support from stakeholders or smokers. However, they could make VLNC cigarettes available as an option for smokers utilising a differential tax favouring VLNC cigarettes. If this were combined with better access to nicotine containing e-cigarettes, smokers may shift away from HNC cigarettes.


Witnessing the evolution of transcription in mitochondria: the mitochondrial genome of the primitive brown alga Pylaiella littoralis (L.) Kjellm. Encodes a T7-like RNA polymerase.
A region of the mitochondrial genome of the primitive brown alga Pylaiella littoralis containing a plasmid-like insert which contains a transcribed T7-phage-type RNA polymerase gene is described. This is a first report of a phage-type RNA polymerase gene integrated in a mitochondrial genome. As the mitochondrial genome of this alga also contains sigma-70 proteobacterial promoter regions, i.e. traces of the ancestral alpha2betabeta'sigma-70 proteobacterial RNA polymerase, this genome witnesses two types of RNA polymerases. As such the mitochondrial genome of P. littoralis represents a unique stage in the evolution of transcription in mitochondria, which contrasts with that of the primitive protist Reclinomonas americana, which still retains the ancestral alpha2betabeta'sigma-70 proteobacterial RNA polymerase genes, and with animals, land plants and fungi, which use phage-type polymerases.


Roles of pH, cation valence, and ionic strength in the stability and aggregation behavior of zinc oxide nanoparticles.
The effects of pH, cation valence, and ionic strength (IS) on the stability and aggregation behavior of zinc oxide nanoparticles (ZnO NPs) were investigated in this study. Results showed that ZnO NPs were most prone to aggregation at the isoelectric point (pH = 8.7), with an aggregation rate (ΔD/Δt) of 30.1. ZnO NPs showed a greater propensity for dissolution at lower pH (pH < 7), and Zn2+ was more rapidly released into the aqueous phase in acidic solutions than neutral or alkaline conditions. The C/C0 of ZnO NPs was about 21.56% and remained stable in acidic solution of pH 4.0. Additionally, slow sedimentation with a C/C0 ratio of 95.0% was observed due to an increase in repulsive interactions between nanoparticles under pH = 10. The effect of cations on the ΔD/Δt of ZnO NPs decreased in strength as follows: Ca2+ > Mg2+ > K+ > Na+. High-valence metal cations (Ca2+, Mg2+) were more competitively adsorbed onto the surface of ZnO NPs with a hydrogen atom due to Coulomb's law, increasing the zeta potential and stabilizing the suspension of ZnO NPs at IS < 10 mM. Furthermore, compression of the electric double layer (EDL) became stronger than electrostatic adsorption with increasing IS, reaching a maximum ΔD/Δt of 23.3 (Ca2+, pH = 7, IS = 1 M). The C/C0 ratio of ZnO NPs decreased from 100% to 56.5% (Na+), 52.2% (K+), 45.2% (Mg2+), and 40.1% (Ca2+) at pH = 7 and an IS of 0.5 M. In addition to the cation valence, the hydration forces and ionic radii of the metal cations might be other factors that affected the interactions of metal cations with ZnO NPs. Finally, the total interaction energy between ZnO NPs was calculated using the Derjaguin-Landau-Verwey-Overbeek (DLVO) theoretical formula, and the calculated results were in agreement with the experimental outcomes under various aquatic environmental conditions.


Overuse Injuries Among Female Combat Warriors in the Israeli Defense Forces: A Cross-sectional Study.
Integration of females in combat units poses a unique challenge for army commanders around the world. The purpose of this study is to provide a detailed up-to-date situation report regarding overuse injuries among combat female warriors in the IDF (Israeli Defense Forces) in order to enable evidence-based decision-making, prevention policy, and further research of this highly significant military public health issue. A cross-sectional descriptive study was conducted including 2,519 females recruited to combat duties during the year of 2013. The main data source was the IDF's computerized medical consultation records package (CPR). Descriptive statistics was performed and some results were compared with males using data from other reports and studies of the IDF. The overall injury rate was 28.3%. Of all injuries, 86% were in the ankle and calf (41%), the lower back (23%), and the knee (22%) regions. The average lost training days was 11 d for females as compared with 8 d for males. The overall rates of stress fractures and the rates of femur and femoral neck stress fractures were significantly higher among females as compared with males (11.2% vs. 2.5%, p = 0.0032, and 7.8% vs. 1.6% p = 0.00001, respectively). The overuse injury rates among females in the IDF are high and may be considered a significant military public health problem. In order to reduce the numbers of overuse injuries, which is expected to significantly increase after the IDF's policy change regarding to combat duties open for females, we recommend planning and implementation of policies and intervention programs and further research regarding to overuse injuries among female combat warriors with special focus on the calf and ankle, lower back, and knee regions and femoral stress fractures.


Citation indices for social media articles in urology.
To evaluate the impact of publications on urological participation in social media (SoMe) by virtue of citations in the urological and non-urological literature. On 15 March 2016, a PubMed search was undertaken using the names of the major SoMe platforms in current use and associated with the field of urology. The search term 'urolog*' was used to specifically capture articles that could be associated with 'urology', 'urologist' or 'urological'. Exclusion criteria for analysis included non-English language articles, articles published for the first time online in any form after 1 March 2015, articles irrelevant to the topic of SoMe, and letters of correspondence. Included articles were then searched in Google Scholar and citations analysed to determine if citations were from the urological literature or non-urological literature. Citations from non-urological journals were considered to be as such even if authored by urologists and on the subject of urology and SoMe. Prior to exclusions as defined in the methods, our PubMed search yielded 232 articles of which 17 were non-English language and 66 had been published after 1 March 2015. Allowing for 12 months after the most recent articles were published, we found that the mean number of total citations in any journal was 20.8. There were more citations in journals not specific to urology, with 8.3 citations in urological journals, compared to 12.6 citations in non-urological journals. Urological SoMe journal articles are highly cited, particularly in the non-urological literature. It is likely that the magnitude of citations has positively contributed to the impact factors of the almost all journals publishing these manuscripts.


2-Amino-4-(piperidin-1-yl)-11H-pyrimido[4,5-b][1,5]benzodiazepin-6-ium chloride monohydrate and 2-amino-4-[methyl(2-methylphenyl)amino]-11H-pyrimido[4,5-b][1,5]benzodiazepin-6-ium chloride-benzene-1,2-diamine (1/1): complex sheets generated by multiple hydrogen bonds.
In each of 2-amino-4-(piperidin-1-yl)-11H-pyrimido[4,5-b][1,5]benzodiazepin-6-ium chloride monohydrate, C(16)H(19)N(6)(+).Cl(-).H(2)O, (I), and 2-amino-4-[methyl(2-methylphenyl)amino]-11H-pyrimido[4,5-b][1,5]benzodiazepin-6-ium chloride-benzene-1,2-diamine (1/1), C(19)H(19)N(6)(+).Cl(-).C(6)H(8)N(2), (II), the seven-membered ring in the cation adopts a boat conformation. The pyrimidine ring in (II) adopts a twist-boat conformation, but the corresponding ring in (I) is essentially planar. The amino groups of the benzene-1,2-diamine component of (II) are both pyramidal. The independent components of (I) are linked into complex sheets by a combination of N-H...O, N-H...N, N-H...Cl and O-H...Cl hydrogen bonds. In the crystal structure of (II), one N-H...N hydrogen bond and six independent N-H...Cl hydrogen bonds combine to link the components into complex sheets.


Interventions for improving outcomes for pregnant women who have experienced genital cutting.
Female genital cutting (FGC) refers to all procedures that involve the partial or total removal of the external female genitalia, or other injury to the female genital organs for cultural or other non-therapeutic reasons. There are no known medical benefits to FGC, and it can be potentially dangerous for the health and psychological well-being of women and girls who are subjected to the practice resulting in short- and long-term complications. Health problems of significance associated with FGC faced by most women are maternal and neonatal mortality and morbidity, the need for assisted delivery and psychological distress. Under good clinical guidelines for caring for women who have undergone genital cutting, interventions could provide holistic care that is culturally sensitive and non-judgemental to improve outcomes and overall quality of life of women. This review focuses on key interventions carried out to improve outcome and overall quality of life in pregnant women who have undergone FGC. To evaluate the impact of interventions to improve all outcomes in pregnant women or women planning a pregnancy who have undergone genital cutting. The comparison group consisted of those who have undergone FGC but have not received any intervention. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (31 December 2012) and organisations engaged in projects regarding FGC. Randomised controlled trials (RCTs), cluster-randomised trials or quasi-RCTs with reported data comparing intervention outcomes among pregnant women or women planning a pregnancy who have undergone genital cutting compared with those who did not receive any intervention. We did not identify any RCTs, cluster-randomised trials or quasi-RCTs. There are no included studies. FGC research has focused mainly on observational studies to describe the social and cultural context of the practice, and we found no intervention trials conducted to improve outcomes for pregnant women presenting with complications of FGC. While RCTs will provide the most reliable evidence on the effectiveness of interventions, there remains the issue of what is considered ethically appropriate and the willingness of women to undergo randomisation on an issue that is enmeshed in cultural traditions and beliefs. Consequently, conducting such a study might be difficult.


NFATc1 signaling drives chronic ER stress responses to promote NAFLD progression.
Non-alcoholic fatty liver disease (NAFLD) can persist in the stage of simple hepatic steatosis or progress to steatohepatitis (NASH) with an increased risk for cirrhosis and cancer. We examined the mechanisms controlling the progression to severe NASH in order to develop future treatment strategies for this disease. NFATc1 activation and regulation was examined in livers from patients with NAFLD, cultured and primary hepatocytes and in transgenic mice with differential hepatocyte-specific expression of the transcription factor (Alb-cre, NFATc1c.a . and NFATc1Δ/Δ ). Animals were fed with high-fat western diet (WD) alone or in combination with tauroursodeoxycholic acid (TUDCA), a candidate drug for NAFLD treatment. NFATc1-dependent ER stress-responses, NLRP3 inflammasome activation and disease progression were assessed both in vitro and in vivo. NFATc1 expression was weak in healthy livers but strongly induced in advanced NAFLD stages, where it correlates with liver enzyme values as well as hepatic inflammation and fibrosis. Moreover, high-fat WD increased NFATc1 expression, nuclear localisation and activation to promote NAFLD progression, whereas hepatocyte-specific depletion of the transcription factor can prevent mice from disease acceleration. Mechanistically, NFATc1 drives liver cell damage and inflammation through ER stress sensing and activation of the PERK-CHOP unfolded protein response (UPR). Finally, NFATc1-induced disease progression towards NASH can be blocked by TUDCA administration. NFATc1 stimulates NAFLD progression through chronic ER stress sensing and subsequent activation of terminal UPR signalling in hepatocytes. Interfering with ER stress-responses, for example, by TUDCA, protects fatty livers from progression towards manifest NASH.


[Porcelain veneers: 4 application technics].
The present paper describes the different techniques currently employed for the use of porcelain laminate veneer, from the simplest stock teeth to the most complex casting system. Advantages and disadvantages of each are described herein.


DNA under Force: Mechanics, Electrostatics, and Hydration.
Quantifying the basic intra- and inter-molecular forces of DNA has helped us to better understand and further predict the behavior of DNA. Single molecule technique elucidates the mechanics of DNA under applied external forces, sometimes under extreme forces. On the other hand, ensemble studies of DNA molecular force allow us to extend our understanding of DNA molecules under other forces such as electrostatic and hydration forces. Using a variety of techniques, we can have a comprehensive understanding of DNA molecular forces, which is crucial in unraveling the complex DNA functions in living cells as well as in designing a system that utilizes the unique properties of DNA in nanotechnology.


The DIACOM study (effect of DosIng frequency of oral Antidiabetic agents on the COMpliance and biochemical control of type 2 diabetes).
Sulphonylureas are widely used in the management of type 2 diabetes. The effectiveness of treatment with oral antidiabetic drugs depends largely on patient compliance. The objective of the DIACOM (effect of DosIng frequency of oral Antidiabetic agents on the COMpliance and biochemical control of type 2 diabetes) study was to compare the compliance of patients treated with once-daily (od) or twice-daily (bid) sulphonylureas. One hundred and five patients, previously treated with glibenclamide, were randomized to receive gliclazide in modified-release formulation (MR) once daily or glibenclamide twice daily for 16 weeks, using an electronic monitoring system (MEMS). A significant difference in compliance was observed between the two groups. The overall compliance was 93.5+/-14.0% in the once-daily gliclazide MR group and 87.2+/-21.1% in the twice-daily glibenclamide group (p<0.05), and the correct number of doses was taken on 86.3+/-15.4 and 66.9+/-29.0% of treatment days respectively (p<0.0001). The percentage of missed doses was 9.3+/-12.5% in the once-daily group and 17.5+/-18.0% in the twice-daily group (p<0.01). The percentage of doses taken in the correct time window and correct inter-dose interval was higher in the once-daily group, as was therapeutic coverage. Patients in the gliclazide MR group also achieved significantly better glycaemic control [fasting plasma glucose and glycated haemoglobin (HbA(1c))] than those treated with glibenclamide (p<0.0001). The study demonstrates that patient compliance with once-daily gliclazide MR is significantly better than with twice-daily glibenclamide. Consistently better efficacy was observed for short-term (fasting glucose) and long-term glycaemic control (HbA(1c)) in the once-daily group. These results demonstrate the possible therapeutic advantages of once-daily agents over twice-daily agents in the treatment of type 2 diabetes.


Diazepam dependence prevented by glutamate antagonists.
Long-term treatment leads to tolerance to and dependence on benzodiazepines. Abrupt termination of benzodiazepine administration triggers the expression of signs of dependence. Mice withdrawn from chronic treatment with diazepam showed a time-related evolution of anxiety, muscle rigidity, and seizures between days 4 and 21 after treatment discontinuation. A period between withdrawal days 1 and 3 was symptom-free. Surprisingly, during this "silent phase" the susceptibility of mice to alpha-amino-3-hydroxy-5-tert-butyl-4-isoxazolepropionate (ATPA) and kainate seizures and the magnitude of monosynaptic reflexes mediated by non-N-methyl-D-aspartate (NMDA) mechanisms were enhanced. In apparent contrast, the "active phase", between withdrawal days 4 and 21, was characterized by increased susceptibility to NMDA seizures and enhanced magnitude of polysynaptic reflexes, which are NMDA dependent. Treatment of mice with alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) antagonists 1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine (GYKI 52466) or 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo(f)quinoxaline but not with the NMDA antagonist 3-[(+/-)-2-carboxypiperazin-4-yl]-propyl-1-phosphonate (CPP) during the silent phase prevented signs of dependence. In contrast, treatment with CPP but not with GYKI 52466 during the active phase prevented the symptoms. The development of tolerance to and dependence on diazepam was prevented by concurrent treatment of mice with CPP but was not prevented by GYKI 52466. These data indicate that NMDA-dependent mechanisms contribute to the development of tolerance to diazepam and to the expression of signs of dependence in mice after termination of long-term treatment with diazepam. Nevertheless, the non-NMDA-mediated silent phase is essential for triggering the symptoms. Therefore, AMPA antagonists may offer a therapeutic approach for preventing dependence on benzodiazepines that is an alternative to NMDA antagonism.


An indirect comparison of HbA1c treatment effect with albiglutide and exenatide 2.0 mg QW using the Bucher method.
No head-to-head comparisons exist between once-weekly (QW) glucagon-like peptide-1 receptor agonists; accordingly, this indirect comparison was conducted to evaluate the comparative efficacy of QW albiglutide vs QW exenatide. Following a systematic literature search, it was determined that HARMONY 7 and DURATION 6, Phase III trials for albiglutide and exenatide, respectively, were similar in study design and baseline characteristics and included a common comparator arm, making them suitable for an indirect comparison using the Bucher method. The primary endpoint of change from baseline in glycated hemoglobin (HbA1c) with albiglutide 50 mg QW and exenatide 2.0 mg QW was compared and tested for noninferiority. The indirect comparison showed a treatment difference of 0.0% (95% confidence interval: -0.189% to 0.189%) in mean change in HbA1c from baseline, and albiglutide 50 mg was noninferior to exenatide 2.0 mg QW at the noninferiority margin of 0.3%. In the absence of a head-to-head trial, these results can be used in pharmacoeconomic analysis and to inform health technology assessment and clinical decision making.


Rainbow trout glucocorticoid receptor overexpression in Escherichia coli: production of antibodies for western blotting and immunohistochemistry.
Fragments of cDNA that encode the N-terminal and DNA-binding domains (DBD) of the rainbow trout glucocorticoid receptor (rtGR) were expressed in Escherichia coli as fusion proteins with glutathione-S-transferase (GST). The fusion proteins induced by IPTG could readily be detected as 45- and 40-kDa bands, respectively, in crude extracts, as well as in proteins purified on glutathione-agarose. These purified hybrid proteins were used to immunize rabbits. The antisera produced were tested for specificity by Western blot analysis using extracts from COS-1 cells transfected with an rtGR expression vector and from trout liver cells. The antisera raised against the DBD domain did not detect any bands on Western blots, even at low antiserum dilution. However, the purified DBD fusion protein specifically bound GRE-containing DNA fragments in gel-shift assays, and the retarded complexes were supershifted by these antibodies. The antisera raised against the N-terminal domain consistently detected two protein bands at 104 and 100 kDa in the two cell extracts and allowed specific immunohistochemical staining in fish brain and pituitary. For the first time in fish, these antibodies will allow analysis of GR expression in different cortisol target tissues.


Phylogenetic analysis of 17271 Indian SARS-CoV-2 genomes to identify temporal and spatial hotspot mutations.
The second wave of SARS-CoV-2 has hit India hard and though the vaccination drive has started, moderate number of COVID affected patients is still present in the country, thereby leading to the analysis of the evolving virus strains. In this regard, multiple sequence alignment of 17271 Indian SARS-CoV-2 sequences is performed using MAFFT followed by their phylogenetic analysis using Nextstrain. Subsequently, mutation points as SNPs are identified by Nextstrain. Thereafter, from the aligned sequences temporal and spatial analysis are carried out to identify top 10 hotspot mutations in the coding regions based on entropy. Finally, to judge the functional characteristics of all the non-synonymous hotspot mutations, their changes in proteins are evaluated as biological functions considering the sequences by using PolyPhen-2 while I-Mutant 2.0 evaluates their structural stability. For both temporal and spatial analysis, there are 21 non-synonymous hotspot mutations which are unstable and damaging.


[Case of pulmonary aspergilloma in the apices of both lungs which was associated with welder's lung].
A 48-year-old man, who had worked as a welder for 30 years, was admitted to our hospital with bloody sputum. His chest CT scan showed diffuse centrilobular micronodules and fungus balls within cavities in the apices of both lungs. We diagnosed pulmonary aspergilloma associated with welder's lung in the apices of both lungs, by bronchoscopy. He was treated with oral antifungal drugs for 1 month, but he had massive hemoptysis. Bronchoscopy showed that the hemoptysis originated from the right upper lobe. We performed right upper lobectomy after right bronchial and intercostal arterial embolization. About 2 months after surgery, he had bloody sputum again. We then performed left bronchial and intercostal arterial embolization because bronchoscopy showed that the bloody sputum originated from the left upper lobe. His bloody sputum disappeared after the last embolization treatment. We report a rare case of pulmonary aspergilloma associated with welder's lung, with a discussion based on a review of the literature.


Chemokine RANTES is upregulated in monocytes from patients with hyperhomocysteinemia.
To investigate the changes in plasma level of the chemokine RANTES (regulated upon activation, normal T cells expressed and secreted) and the responsiveness of lipopolysaccharide (LPS)-induced RANTES secretion from monocytes in patients with hyperhomocysteinemia (HHcy). The plasma levels of homocysteine (Hcy), folate, and RANTES were measured in 38 control patients with normal Hcy levels and 40 patients with HHcy and the mRNA synthesis of RANTES in isolated human monocytes was determined by RNase protection assays. The plasma level of RANTES was elevated in HHcy patients compared with controls (median 5.3 vs 3.5 ng/mL, P<0.01). LPS-induced RANTES production from monocytes of HHcy patients was also increased significantly. In addition, Hcy directly increased the mRNA level of RANTES in isolated normal human monocytes in a time- and dose-dependent manner. Upregulated RANTES from monocytes in HHcy patients may be involved in the atherogenesis of HHcy-induced atherosclerosis.


The association between cryptococcal variety and immunocompetent and immunocompromised hosts.
The varieties of Cryptococcus neoformans serotypes are neoformans and gattii. Data suggest an association between cryptococcal variety and host immune status. In addition, the two varieties differ in their epidemiology and pathogenicity. Furthermore, symptoms, outcome, and response of cryptococcosis to antifungal therapy may vary. The two varieties also differ in immune-modulating effects. Sparse clinical data suggest var. gattii is more virulent and may be more recalcitrant to antifungal therapy. Also, its infections produce more sequelae. A better understanding of how cryptococcal variety influences the clinical course and response to the treatment of cryptococcosis is needed. Clinicians should be aware of the association, especially in patients with refractory disease. It may be useful to type the isolate to the variety level and administer prolonged antifungal therapy.


Phosphodiesterase type 5 inhibition attenuates cyclosporine A induced nephrotoxicity in mice.
We investigated the renal protective effects of phophodiesterase type 5 (PDE5) inhibitors in mice with cyclosporine A (CyA; a calcineurin phosphatase inhibitor) induced nephrotoxicity. Fifty male mice were divided into five groups of 10. Group 1 received no treatment, group 2 received only saline orally, group 3 received 30 mg/kg/day CyA by subcutaneous injection, group 4 received only 30 mg/kg/day vardenafil orally, and group 5 received 30 mg/kg/day CyA by subcutaneous injection and 30 mg/kg/day vardenafil orally. At 28 days, platelet-derived growth factor A (PDGF-A) and C (PDGF-C), transforming growth factor-beta 1 (TGF-β1), cyclo-oxygenase 1 and 2 (COX-1 and COX-2), and P glycoprotein (Pgp) expression levels were measured in the renal tissues. In addition, expressions of COX-1 and COX-2 genes were determined using real-time PCR. PDE5 inhibitor administration ameliorated decreased PDGF-A and C, TGF-β1, COX-1 and -2, and Pgp expression levels by modulation of cyclic guanosine monophosphate (cGMP) activity in kidneys. The relative expressions of COX-1 and COX-2 genes to GAPDH revealed that the maximum increase was obtained in the group treated with CyA and vardenafil for both COX-1 and COX-2 genes. Our study revealed that long term oral treatment with vardenafil protects kidneys from CyA induced nephrotoxicity. We showed that long term oral treatment with PDE5 prevents pathological kidney changes caused by CyA induced nephrotoxicity.


Synthesis and Characterization of Dimeric Artesunate Glycerol Monocaprylate Conjugate and Formulation of Nanoemulsion Preconcentrate.
Malaria is one of the major life-threatening health problems worldwide. Artesunate is the most potent antimalarial drug to combat severe malaria. However, development of drug resistance, short plasma half-life, and poor bioavailability limit the efficacy of this drug. Here, we applied the dimerization concept to synthesize dimeric artesunate glycerol monocaprylate conjugate (D-AS-GC) by conjugating artesunate (AS) with glycerol monocaprylate (GC) via esterification reaction. D-AS-GC conjugate, AS, and GC were well characterized by 1H NMR, attached proton test (APT) 13C NMR and 2D NMR spectroscopy. D-AS-GC conjugate was further analyzed by ESI-TOF MS. Finally, a series of nanoemulsion preconcentrate (F1-F6) of D-AS-GC was prepared by mixing different ratios of oil and surfactant/cosurfactant and evaluated after dilution with an aqueous phase. The optimized formulation (F6) exhibits a clear nanoemulsion and the hydrodynamic diameter of the dispersed phase was determined by DLS and DOSY NMR spectroscopy. The morphology of the nanoemulsion droplets of F6 was investigated by AFM, which revealed the formation of tiny nanoemulsion droplets on a hydrophilic mica substrate. Moreover, using a less polar silicon wafer led to the formation of larger droplets with a spherical core shell-like structure. Overall, the rational design of the dimeric artesunate-based nanoemulsion preconcentrate could potentially be used in more efficient drug delivery systems.


Interfacial Polyelectrolyte Complex Spinning of Cellulose Nanofibrils for Advanced Bicomponent Fibers.
Fiber spinning of anionic TEMPO-oxidized cellulose (TOCN) nanofibrils with polycations by interfacial polyelectrolyte complexation is demonstrated. The formed fibers were mostly composed of cellulose nanofibrils and the polycations were a minor constituent, leading to yield and ultimate strengths of ca. 100 MPa and ca. 200 MPa, and Young's modulus of ca. 15 GPa. Stretching of the as-formed wet filaments of TOCN/polycation by 20% increased the Young's modulus, yield strength, and ultimate tensile strength by approximately 45, 36, and 26%, respectively. Importantly, feasibility of compartmentalized wound bicomponent fibers by simultaneous spinning of two fibers of different compositions and entwining them together was shown. This possibility was further exploited to demonstrate reversible shape change of a bicomponent fiber directly by humidity change, and indirectly by temperature changes based on thermally dependent humidity absorption. The demonstrated route for TOCN-based fiber preparation is expected to open up new avenues in the application of nanocelluloses in advanced fibrous materials, crimping, and responsive smart textiles.


Experimental and computational study of pulsatile flow characteristics in Romanesque and gothic aortic arch models.
Romanesque and Gothic are two types of deformed aortic arch geometries after surgical repair of coarctation of the aorta. The abnormal arch geometry and hemodynamics are associated with late systemic hypertension, aortic aneurysms, and other cardiovascular complications. To understand the fluid dynamic signatures of such flow, a combined experimental and computational fluid dynamic (CFD) study has been conducted to quantitatively compare the main (axial) and secondary flow characteristics. In the experiments, a pulsatile flow simulator was used to generate the pulsatile flow conditions. Phase-locked planar and tomographic particle image velocimetry techniques were employed to quantitatively study the flow fields. Three-dimensional CFD simulations were also performed and compared with the experimental data. The results show that in the Romanesque arch, the flow first accelerates along the inner wall and then becomes more uniform in the cross-section after the peak systole. A pair of wall vortices and Dean-type vortices develop during the systolic phase. The coherent structures are continuously extended into the descending aorta and persist throughout the cycle. In comparison, the Gothic arch exhibits a highly skewed velocity distribution with high velocity around the arch apex. The sharp curvature causes flow separation, jet impingement, and stagnant flow near the top. The coherent structures in the Gothic arch are less continuous in the descending aorta, which also differ from those observed in the Romanesque model. The distinct flow characteristics of the Gothic arch lead to more temporal and spatial variations of wall shear stress in the descending aorta, implying hemodynamic risks for aortic complications.


The ER luminal binding protein (BiP) mediates an increase in drought tolerance in soybean and delays drought-induced leaf senescence in soybean and tobacco.
The ER-resident molecular chaperone BiP (binding protein) was overexpressed in soybean. When plants growing in soil were exposed to drought (by reducing or completely withholding watering) the wild-type lines showed a large decrease in leaf water potential and leaf wilting, but the leaves in the transgenic lines did not wilt and exhibited only a small decrease in water potential. During exposure to drought the stomata of the transgenic lines did not close as much as in the wild type, and the rates of photosynthesis and transpiration became less inhibited than in the wild type. These parameters of drought resistance in the BiP overexpressing lines were not associated with a higher level of the osmolytes proline, sucrose, and glucose. It was also not associated with the typical drought-induced increase in root dry weight. Rather, at the end of the drought period, the BiP overexpressing lines had a lower level of the osmolytes and root weight than the wild type. The mRNA abundance of several typical drought-induced genes [NAC2, a seed maturation protein (SMP), a glutathione-S-transferase (GST), antiquitin, and protein disulphide isomerase 3 (PDI-3)] increased in the drought-stressed wild-type plants. Compared with the wild type, the increase in mRNA abundance of these genes was less (in some genes much less) in the BiP overexpressing lines that were exposed to drought. The effect of drought on leaf senescence was investigated in soybean and tobacco. It had previously been reported that tobacco BiP overexpression or repression reduced or accentuated the effects of drought. BiP overexpressing tobacco and soybean showed delayed leaf senescence during drought. BiP antisense tobacco plants, conversely, showed advanced leaf senescence. It is concluded that BiP overexpression confers resistance to drought, through an as yet unknown mechanism that is related to ER functioning. The delay in leaf senescence by BiP overexpression might relate to the absence of the response to drought.


Continuous Movement Tracking Performance for Predictable and Unpredictable Tasks with Vibrotactile Feedback.
The purpose of this paper was to determine human movement tracking performance in response to vibrotactile feedback tracking for predictable and unpredictable continuous movement tasks. Thirteen subjects performed elbow flexion/extension and knee flexion/extension continuous movement tracking tasks while receiving tactile stimulation proportional to limb joint position error. Subjects followed 0.2-2.0 Hz desired movements for predictable tasks (single sinusoid) and unpredictable tasks (combination of three sinusoids). Tactile stimulation reaction times at the forearm to induce elbow flexion/extension and at the shank to induce knee flexion/extension were also recorded. Results of frequency tracking showed that 100 percent of participants correctly tracked unpredictable tasks at all frequencies, but only 60-80 percent of participants correctly tracked predictable tasks at frequencies less than 1 Hz and only 20-60 percent of participants correctly tracked predictable tasks at frequencies greater than 1 Hz. Subjects had less phase lag for predictable tasks than for unpredictable tasks. Reaction times at the forearm were 379 ms and at the shank 437 ms. These findings suggest that continuous vibrotactile feedback based on position errors may not be the most effective means of training higher frequency human movements and serve to inform future vibrotactile feedback design related to training human limb movements for predictable and unpredictable tasks.