Immunohistochemical analysis of Metadherin in proliferative and cancerous breast tissue.
Metadherin (MTDH) has been reported to be associated with cancer progression in various types of human cancers including breast cancer. Whether MTDH contributes to carcinogenesis of breast cancer is still unknown. In the present study, we investigated the expression of MTDH in normal, UDH (usual ductal hyperplasia), ADH (atypical ductal hyperplasia), DCIS (ductal carcinoma in situ) and invasive cancer to explore the possible role of MTDH for breast cancer carcinogenesis. Immunohistochemistry was employed on paraffin sections of surgical removed breast samples. The immunohistochemical results showed almost no staining in normal tissue, moderate staining in ADH and UDH, intense MTDH stains in DCIS and cancer. Statistical analysis demonstrated significant different MTDH expression between proliferative and cancerous breast lesions (p < 0.001). MTDH was positively correlated with the histological differentiation of DCIS (p = 0.028). In breast cancer, statistical analysis revealed a significant correlation between MTDH expression with patients' age (p = 0.042), ER status (p = 0.018) and p53 status (p = 0.001). We also examined the effect of MTDH on cell proliferation in DCIS and cancer, and we found that MTDH overexpression was significantly correlated with high Ki67 index (p = 0.008 and p = 0.036, respectively). MTDH overexpression could be identified in proliferative breast lesions and may contribute to breast cancer progression.


Zinc, a novel structural element found in the family of bacterial adenylate kinases.
The adk gene from Bacillus stearothermophilus was cloned and overexpressed in Escherichia coli under the control of the lac promoter. The primary structure of B. stearothermophilus adenylate kinase exhibited 76% identity with the enzyme from Bacillus subtilis, 60% identity with the enzyme from Lactococcus lactis, and 42% identity with the enzyme from E. coli. The most striking property of the adenylate kinase from B. stearothermophilus is the presence of a structural zinc atom bound to four cysteines in a zinc finger-like fashion. The ability to coordinate zinc is predicted also for a number of other isoforms of bacterial adenylate kinases. Furthermore, the tightly bound metal ion contributes to the high thermodynamic stability of adenylate kinase from B. stearothermophilus.


A human lung xenograft mouse model of Nipah virus infection.
Nipah virus (NiV) is a member of the genus Henipavirus (family Paramyxoviridae) that causes severe and often lethal respiratory illness and encephalitis in humans with high mortality rates (up to 92%). NiV can cause Acute Lung Injury (ALI) in humans, and human-to-human transmission has been observed in recent outbreaks of NiV. While the exact route of transmission to humans is not known, we have previously shown that NiV can efficiently infect human respiratory epithelial cells. The molecular mechanisms of NiV-associated ALI in the human respiratory tract are unknown. Thus, there is an urgent need for models of henipavirus infection of the human respiratory tract to study the pathogenesis and understand the host responses. Here, we describe a novel human lung xenograft model in mice to study the pathogenesis of NiV. Following transplantation, human fetal lung xenografts rapidly graft and develop mature structures of adult lungs including cartilage, vascular vessels, ciliated pseudostratified columnar epithelium, and primitive "air" spaces filled with mucus and lined by cuboidal to flat epithelium. Following infection, NiV grows to high titers (10(7) TCID50/gram lung tissue) as early as 3 days post infection (pi). NiV targets both the endothelium as well as respiratory epithelium in the human lung tissues, and results in syncytia formation. NiV infection in the human lung results in the production of several cytokines and chemokines including IL-6, IP-10, eotaxin, G-CSF and GM-CSF on days 5 and 7 pi. In conclusion, this study demonstrates that NiV can replicate to high titers in a novel in vivo model of the human respiratory tract, resulting in a robust inflammatory response, which is known to be associated with ALI. This model will facilitate progress in the fundamental understanding of henipavirus pathogenesis and virus-host interactions; it will also provide biologically relevant models for other respiratory viruses.


Screening for anthelmintic resistance in equid strongyles (Nematoda) in Romania.
Resistance to anthelmintic medication of equid strongyles is a worldwide phenomenon and for this reason systematic investigations of resistant parasite populations are necessary. The purpose of the present study was to investigate the presence and distribution of equid strongyles resistant to the anthelmintics used in Romania, as well as the pre-treatment and post-treatment prevalence of species of strongylid nematodes. The Faecal Egg Count Reduction Test was performed between 2010 and 2013 on a total number of 588 horses and 23 donkeys from 26 locations (subgroups). Animals of the first group (I) consisting of subgroups no. 1-11 were treated with Albendazole (ABZ), those of the second group (II) consisting of subgroups no. 12-23 with Fenbendazole (FBZ), while Ivermectin (IVM) was used on animals of the third group (III) consisting of subgroups no. 24-26. Resistant strongyles have been found in 82% (average lower limit of the 95% confidence interval, LCL95%, was 65) of the total equids from the group treated with ABZ. In the group of horses treated with FBZ, resistant strongyles were identified in 75% of the subgroups (LCL95% = 44). No resistant strongyles have been identified in IVM-treated horse groups (LCL95% = 98). The pre-treatment prevalence of the species of the Strongylinae Müller, 1780 was 22%, whereas that with nematodes of the subfamily Cyathostominae Molin, 1861 78%. Post-treatment reduction of strongyline nematodes was observed (5%), which demonstrates the sensitivity of large strongyles to common anthelmintics. The post-treatment prevalence of cyathostomes was of 95%, which proves their resistance, especially to ABZ- and FBZ-based anthelmintics.


Large-scale identification of ubiquitination sites by mass spectrometry.
Ubiquitination is essential for the regulation of cellular protein homeostasis. It also has a central role in numerous signaling events. Recent advances in the production and availability of antibodies that recognize the Lys-ɛ-Gly-Gly (K-ɛ-GG) remnant produced by trypsin digestion of proteins having ubiquitinated lysine side chains have markedly improved the ability to enrich and detect endogenous ubiquitination sites by mass spectrometry (MS). The following protocol describes the steps required to complete a large-scale ubiquitin experiment for the detection of tens of thousands of distinct ubiquitination sites from cell lines or tissue samples. Specifically, we present detailed, step-by-step instructions for sample preparation, off-line fractionation by reversed-phase chromatography at pH 10, immobilization of an antibody specific to K-ɛ-GG to beads by chemical cross-linking, enrichment of ubiquitinated peptides using these antibodies and proteomic analysis of enriched samples by LC-tandem MS (MS/MS). Relative quantification can be achieved by performing stable isotope labeling by amino acids in cell culture (SILAC) labeling of cells. After cell or tissue samples have been prepared for lysis, the described protocol can be completed in ∼5 d.


Positive feedback between transcriptional and kinase suppression in nematodes with extraordinary longevity and stress resistance.
Insulin/IGF-1 signaling (IIS) regulates development and metabolism, and modulates aging, of Caenorhabditis elegans. In nematodes, as in mammals, IIS is understood to operate through a kinase-phosphorylation cascade that inactivates the DAF-16/FOXO transcription factor. Situated at the center of this pathway, phosphatidylinositol 3-kinase (PI3K) phosphorylates PIP(2) to form PIP(3), a phospholipid required for membrane tethering and activation of many signaling molecules. Nonsense mutants of age-1, the nematode gene encoding the class-I catalytic subunit of PI3K, produce only a truncated protein lacking the kinase domain, and yet confer 10-fold greater longevity on second-generation (F2) homozygotes, and comparable gains in stress resistance. Their F1 parents, like weaker age-1 mutants, are far less robust-implying that maternally contributed trace amounts of PI3K activity or of PIP(3) block the extreme age-1 phenotypes. We find that F2-mutant adults have <10% of wild-type kinase activity in vitro and <60% of normal phosphoprotein levels in vivo. Inactivation of PI3K not only disrupts PIP(3)-dependent kinase signaling, but surprisingly also attenuates transcripts of numerous IIS components, even upstream of PI3K, and those of signaling molecules that cross-talk with IIS. The age-1(mg44) nonsense mutation results, in F2 adults, in changes to kinase profiles and to expression levels of multiple transcripts that distinguish this mutant from F1 age-1 homozygotes, a weaker age-1 mutant, or wild-type adults. Most but not all of those changes are reversed by a second mutation to daf-16, implicating both DAF-16/ FOXO-dependent and -independent mechanisms. RNAi, silencing genes that are downregulated in long-lived worms, improves oxidative-stress resistance of wild-type adults. It is therefore plausible that attenuation of those genes in age-1(mg44)-F2 adults contributes to their exceptional survival. IIS in nematodes (and presumably in other species) thus involves transcriptional as well as kinase regulation in a positive-feedback circuit, favoring either survival or reproduction. Hyperlongevity of strong age-1(mg44) mutants may result from their inability to reset this molecular switch to the reproductive mode.


Clinical use of cabozantinib in the treatment of advanced kidney cancer: efficacy, safety, and patient selection.
Clear cell (cc) renal cell carcinoma (RCC) is the most common type of cancer found in the kidney accounting for ~90% of all kidney cancers. In 2012, there were ~337,000 new cases of RCC diagnosed worldwide with an estimated 143,000 deaths, with the highest incidence and mortality in Western countries. Despite improvements in cancer control achieved with VEGF- and mTOR-targeted therapy for RCC, progression remains virtually universal and additional therapies are needed. The pivotal results of the METEOR trial led to cabozantinib's designation as a breakthrough drug by the US Food and Drug Administration and its approval for treatment of advanced RCC in 2016. Subsequent data from the CABOSUN trial, where caboxantinib is compared with sunitinib, will provide information on the relative activity of cabozantinib as first-line therapy for ccRCC. We review the development of cabozantinib in advanced RCC and its role in the treatment landscape for advanced RCC.


Clinical, radiological and ultrasonographic findings related to knee pain in osteoarthritis.
Pain is the predominant symptom of knee osteoarthritis (OA) and the main reason of disability. Ultrasound is now one of the new imaging modality in Musculoskeletal medicine and its role in assessing the pain severity in the knee osteoarthritis is evaluated in this study. (1) To study the correlation between ultrasonographic (US) findings and pain score and (2) whether ultrasonographic findings show a better association of pain level than conventional X-rays in patients suffering from primary knee osteoarthritis. In this multi-center study, 193 patients with primary knee OA were asked to score their average knee pain using the Western Ontario and McMaster Universities Arthritis (WOMAC) questionnaire;patients would then go for a radiological and an US evaluation of their painful knee. Findings from both imaging modalities will be studied with the associated pain score. Ultrasound showed that knee effusion has positive correlation with pain score upon walking (r = 0.217) and stair climbing (r = 0.194). Presence of suprapatellar synovitis had higher pain score on sitting (Spearman's Rank correlation = 0.355). The medial(r = 0.170) and lateral meniscus protrusion (r = 0.201) were associated with pain score upon stair climbing. Our study found that both imaging modalities shown some significant association with the aspect of pain; neither one is clearly better but rather complementary to each other. A trend is found in both modalities: walking pain is related to pathologies of the either the lateral or medial tibiofemoral joint(TFJ)while stair climbing pain is related to both tibiofemoral joint pathologies and also to the patellofemoral joint (PFJ) pathology. This suggested that biomechanical derangement is an important aspect in OA knee pain.


Accuracy of Cup Positioning With the Computed Tomography-Based Two-dimensional to Three-Dimensional Matched Navigation System: A Prospective, Randomized Controlled Study.
The accuracy of various navigation systems used for total hip arthroplasty has been described, but no publications reported the accuracy of cup orientation in computed tomography (CT)-based 2D-3D (two-dimensional to three-dimensional) matched navigation. In a prospective, randomized controlled study, 80 hips including 44 with developmental dysplasia of the hips were divided into a CT-based 2D-3D matched navigation group (2D-3D group) and a paired-point matched navigation group (PPM group). The accuracy of cup orientation (absolute difference between the intraoperative record and the postoperative measurement) was compared between groups. Additionally, multiple logistic regression analysis was performed to evaluate patient factors affecting the accuracy of cup orientation in each navigation. The accuracy of cup inclination was 2.5° ± 2.2° in the 2D-3D group and 4.6° ± 3.3° in the PPM group (P = .0016). The accuracy of cup anteversion was 2.3° ± 1.7° in the 2D-3D group and 4.4° ± 3.3° in the PPM group (P = .0009). In the PPM group, the presence of roof osteophytes decreased the accuracy of cup inclination (odds ratio 8.27, P = .0140) and the absolute value of pelvic tilt had a negative influence on the accuracy of cup anteversion (odds ratio 1.27, P = .0222). In the 2D-3D group, patient factors had no effect on the accuracy of cup orientation. The accuracy of cup positioning in CT-based 2D-3D matched navigation was better than in paired-point matched navigation, and was not affected by patient factors. It is a useful system for even severely deformed pelvises such as developmental dysplasia of the hips.


Low yield of early postoperative imaging after anastomotic urethroplasty.
To evaluate the necessity and clinical effect of posturethroplasty imaging. We reviewed our database of all urethroplasties performed by a single surgeon at our referral center during a 2-year period. The patients underwent voiding cystourethrography at a mean of 24 days postoperatively. The data analyzed included patient history and demographics, operative details, imaging results, and clinical outcomes. From 2007 to 2009, 210 patients underwent urethral reconstruction at our center. The patients undergoing meatoplasty or staged repairs were excluded, leaving 156 patients with postoperative imaging studies for analysis. Of 110 anterior urethroplasties, 59 (54%) consisted of excision and primary anastomosis, 28 (25%) an augmented anastomotic procedure, and 23 (21%) a pure ventral onlay with a flap or graft. All 46 posterior urethroplasties were performed with scar excision and primary anastomosis. Of the 156 patients, only 4 (3%) had extravasation on postoperative voiding cystourethrography (2 after posterior urethroplasty, 1 after augmented anastomosis, and 1 after ventral onlay)--all were successfully managed with catheter replacement and removal at a mean of 8 days afterward. None of the 59 men undergoing excision and primary anastomosis demonstrated extravasation. Extravasation on posturethroplasty voiding cystourethrography is rare after approximately 3 weeks of catheter drainage. Imaging can be omitted after uncomplicated excision and primary anastomosis urethroplasty.


Structural and mechanistic insights into Hsp104 function revealed by synchrotron X-ray footprinting.
Hsp104 is a hexameric AAA+ ring translocase, which drives protein disaggregation in nonmetazoan eukaryotes. Cryo-EM structures of Hsp104 have suggested potential mechanisms of substrate translocation, but precisely how Hsp104 hexamers disaggregate proteins remains incompletely understood. Here, we employed synchrotron X-ray footprinting to probe the solution-state structures of Hsp104 monomers in the absence of nucleotide and Hsp104 hexamers in the presence of ADP or ATPγS (adenosine 5'-O-(thiotriphosphate)). Comparing side-chain solvent accessibilities between these three states illuminated aspects of Hsp104 structure and guided design of Hsp104 variants to probe the disaggregase mechanism in vitro and in vivo We established that Hsp104 hexamers switch from a more-solvated state in ADP to a less-solvated state in ATPγS, consistent with switching from an open spiral to a closed ring visualized by cryo-EM. We pinpointed critical N-terminal domain (NTD), NTD-nucleotide-binding domain 1 (NBD1) linker, NBD1, and middle domain (MD) residues that enable intrinsic disaggregase activity and Hsp70 collaboration. We uncovered NTD residues in the loop between helices A1 and A2 that can be substituted to enhance disaggregase activity. We elucidated a novel potentiated Hsp104 MD variant, Hsp104-RYD, which suppresses α-synuclein, fused in sarcoma (FUS), and TDP-43 toxicity. We disambiguated a secondary pore-loop in NBD1, which collaborates with the NTD and NBD1 tyrosine-bearing pore-loop to drive protein disaggregation. Finally, we defined Leu-601 in NBD2 as crucial for Hsp104 hexamerization. Collectively, our findings unveil new facets of Hsp104 structure and mechanism. They also connect regions undergoing large changes in solvation to functionality, which could have profound implications for protein engineering.


Air caloric testing in otitis media. (preliminary studies).
On certain occasions it becomes important to evaluate vestibular function in a patient with otitis media. The potential application of the air caloric test in evaluating such patients was examined. Patients with unilateral otitis media before and after surgery were studied to answer certain clinical questions. More questions were raised than answers provided. The preliminary conclusions from this study are: 1. Patients with tympanostomy tubes or small perforation of one ear may show a caloric response in the perforated ear equal to that of the intact ear. 2. Patients with a large tympanic membrane perforation on one side may show hyperactive caloric responses on the perforated side. 3. Patients with a moist ear may show inverted horizontal nystagmus to warm air caloric testing. This applies to patients with a large perforation or mastoidectomy cavity. 4. Patients with a dry open mastoid or fenestration cavity are likely to show a hyperactive caloric response on the side of previous surgery, accompanied by vegetative symptoms. This size of the cavity appears to be less important than the presence of the cavity per se. 5. Patients may be safely tested in the early postoperative period.


Physiology, behaviour and welfare of fish during recreational fishing and after release.
This review investigates how recreational fishing affects the physiology, behaviour, and welfare of fish. Sentience and the capacity of fish to experience pain, suffering and fear are discussed, and practical recommendations for improving the treatment of fish during recreational fishing are provided. Handling procedures used in recreational fishing should match the environment where the fish is caught and the size and strength of the fish. Minimising the number of hooks on lures and baits, and using barbless hooks and circle hooks generally reduce rates of injury and the severity of tissue trauma. Capture time, handling time, and exposure to air play significant roles in the stress responses of fish and should be minimised by anglers. Keep-nets, gaffs, landing-nets, live-wells and other restraining devices should only be used when necessary because each device can prolong or intensify the negative influences of catching fish by hook-and-line. The use of fish as bait is discouraged unless they have been euthanised. Euthanasia of fish used as bait or fish captured recreationally should include a stunning blow to the head, followed by bleeding-out, or pithing the brain and spinal cord. Fish should be euthanised if they are bleeding, injured, deeply-hooked, foul-hooked, or severely exhausted, but local fisheries' regulations must be obeyed. Fish that are released after being caught may be subjected to additional factors that influence their welfare, such as elevated stress, barotrauma, suppressed feeding and growth, impaired reproductive function, increased potential for disease, infection, and delayed mortality.


Determinants of glucose toxicity and its reversibility in the pancreatic islet beta-cell line, HIT-T15.
HIT-T15 cells, a clonal beta-cell line, were cultured and passaged weekly for 6 mo in RPMI 1640 media containing various concentrations of glucose. Insulin content decreased in the intermediate- and late-passage cells as a continuous rather than a threshold glucose concentration effect. In a second series of experiments, cells were grown in media containing either 0.8 or 16.0 mM glucose from passages 76 through 105. Subcultures of passages 86, 92, and 99 that had been grown in media containing 16.0 mM glucose were switched to media containing 0.8 mM glucose and also carried forward to passage 105. Dramatic increases in insulin content and secretion and insulin gene expression were observed when the switches were made at passages 86 and 92 but not when the switch was made at passage 99. These findings suggest that glucose toxicity of insulin-secreting cells is a continuous rather than a threshold function of glucose concentration and that the shorter the period of antecedent glucose toxicity, the more likely that full recovery of cell function will occur.


Wartime traumatic aneurysms: acute presentation, diagnosis, and multimodal treatment of 64 craniocervical arterial injuries.
Operation Iraqi Freedom has resulted in a significant number of closed and penetrating head injuries, and a consequence of both has been the accompanying neurovascular injuries. Here we review the largest reported population of patients with traumatic neurovascular disease and offer our experience with both endovascular and surgical management. A retrospective analysis of all military casualties returning to the Walter Reed Army Medical Center and the National Naval Medical Center, Bethesda, Maryland, from April 2003 until April 2008 was performed. All patients undergoing diagnostic cerebral angiography during their inpatient stay were included in the study. A total of 513 war trauma-related consults were performed from April 2003 to April 2008, resulting in the evaluation of 408 patients with closed and penetrating head injuries. In this population, 279 angiographic studies were performed in 187 patients (25 closed craniocervical injuries, 162 penetrating craniocervical injuries), resulting in the detection of 64 vascular injuries in 48 patients (26.2% of those studied, 34% prevalence). Vascular injuries were characterized by traumatic intracranial aneurysms (TICAs) (n = 31), traumatic extracalvarial aneurysms (TECAs) (n = 19), arterial dissections (n = 11), and arteriovenous fistulae (n = 3). The average TICA size on admission was 4.1 mm, with an observed increase in aneurysm size in 11 cases. In the TICA/TECA group, 24 aneurysms in 23 patients were treated endovascularly with either coiling or stent-assisted coiling, resulting in preservation of the parent artery in 12 of 24 vessels (50%). The injuries in 3 patients in this group progressed despite endovascular treatment and required definitive clip exclusion. Thirteen additional aneurysms in 8 patients were treated surgically, resulting in parent artery preservation in 4 cases (30.8%). Eleven of the 13 remaining TICAs/TECAs resolved spontaneously without treatment. A total of 6 aneurysm ruptures (average size, 8.25 mm) occurred, resulting in 3 deaths. Four of 6 ruptures occurred in TICAs in which the interval size increase was noted angiographically. The management of traumatic vascular injury has evolved with technological advancement and the willingness of the neurosurgeon to intervene. Although open surgical intervention remains a viable solution, endovascular options are available and safe and can effectively temporize a patient while acute sequelae of serious head injury resolve.


Transfected Plasmodium knowlesi produces bioactive host gamma interferon: a new perspective for modulating immune responses to malaria parasites.
Transgenic pathogenic microorganisms expressing host cytokines such as gamma interferon (IFN-gamma) have been shown to manipulate host-pathogen interaction, leading to immunomodulation and enhanced protection. Expression of host cytokines in malaria parasites offers the opportunity to investigate the potential of an immunomodulatory approach by generating immunopotentiated parasites. Using the primate malaria parasite Plasmodium knowlesi, we explored the conditions for expressing host cytokines in malaria parasites. P. knowlesi parasites transfected with DNA constructs for expressing rhesus monkey (Macaca mulatta) IFN-gamma under the control of the heterologous P. berghei apical membrane antigen 1 promoter, produced bioactive IFN-gamma in a developmentally regulated manner. IFN-gamma expression had no marked effect on in vitro parasite development. Bioactivity of the parasite-produced IFN-gamma was shown through inhibition of virus cytopathic effect and confirmed by using M. mulatta peripheral blood cells in vitro. These data indicate for the first time that it is feasible to generate malaria parasites expressing bioactive host immunomodulatory cytokines. Furthermore, cytokine-expressing malaria parasites offer the opportunity to analyze cytokine-mediated modulation of malaria during the blood and liver stages of the infection.


Impact of implantoplasty on strength of the implant-abutment complex.
Implantoplasty, a procedure that is done to smooth contaminated implant surfaces, has been used in the treatment of peri-implantitis. It reduces the implant diameter, which might compromise the implant's strength. This in vitro study was designed to evaluate the effect of implantoplasty on implant strength. Thirty-two tapered implants were used; half were 3.75 mm in diameter (narrow) and the other half were 4.7 mm in diameter (wide). All implants were connected to 20-degree angled abutments. The apical half of each implant was embedded in acrylic resin. Eight 3.75-mm- and eight 4.7-mm-diameter implants were randomly assigned to receive implantoplasty. The remaining implants did not receive implantoplasty (control group). Implantoplasty was performed with a series of diamond and polishing burs. The specimens were then loaded 30 degrees off-axis in a universal testing machine until fracture failure occurred. Bending and fracture strength values were recorded and analyzed statistically (α = .05). The fractured surfaces were evaluated under a scanning electron microscope. All narrow implants failed by fracture at the implant platform. The mean bending strength of narrow implants was statistically significantly reduced by implantoplasty (511.4 ± 55.9 N versus 613.9 ± 42.8 N). Implantoplasty did not affect the strength of wide implants; fracture failures occurred at the abutment screw. The fracture mode was ductile and the crack growth was oblique in direction, indicating complex stress distribution and concentration under loading. Within the limits of this study, implantoplasty appeared to weaken the strength of narrower implants. Therefore, this procedure should be performed with caution on narrower, freestanding implants that are subject to greater occlusal force (eg, posterior regions). Validation of these results is needed for different implant systems.


Plane wave versus focused transmissions for contrast enhanced ultrasound imaging: the role of parameter settings and the effects of flow rate on contrast measurements.
Contrast enhanced ultrasound (CEUS) and dynamic contrast enhanced ultrasound (DCE-US) can be used to provide information about the vasculature aiding diagnosis and monitoring of a number of pathologies including cancer. In the development of a CEUS imaging system, there are many choices to be made, such as whether to use plane wave (PW) or focused imaging (FI), and the values for parameters such as transmit frequency, F-number, mechanical index, and number of compounding angles (for PW imaging). CEUS image contrast may also be dependent on subject characteristics, e.g. flow speed and vessel orientation. We evaluated the effect of such choices on vessel contrast for PW and FI in vitro, using 2D ultrasound imaging. CEUS images were obtained using a VantageTM (Verasonics Inc.) and a pulse-inversion (PI) algorithm on a flow phantom. Contrast (C) and contrast reduction (CR) were calculated, where C was the initial ratio of signal in vessel to signal in background and CR was its reduction after 200 frames (acquired in 20 s). Two transducer orientations were used: parallel and perpendicular to the vessel direction. Similar C and CR was achievable for PW and FI by choosing optimal parameter values. PW imaging suffered from high frequency grating lobe artefacts, which may lead to degraded image quality and misinterpretation of data. Flow rate influenced the contrast based on: (1) false contrast increase due to the bubble motion between the PI positive and negative pulses (for both PW and FI), and (2) contrast reduction due to the incoherency caused by bubble motion between the compounding angles (for PW only). The effects were less pronounced for perpendicular transducer orientation compared to a parallel one. Although both effects are undesirable, it may be more straight forward to account for artefacts in FI as it only suffers from the former effect. In conclusion, if higher frame rate imaging is not required (a benefit of PW), FI appears to be a better choice of imaging mode for CEUS, providing greater image quality over PW for similar rates of contrast reduction.


Sulfasalazine could modulate the CD44v9-xCT system and enhance cisplatin-induced cytotoxic effects in metastatic bladder cancer.
The prognostic role of CD44v9, a variant isoform of CD44 and a new cell surface marker of cancer stem cells, remains unclear in bladder cancer (BC) patients. Furthermore, limited information is available on the functional role of sulfasalazine (SSZ), which could modulate the CD44v9-xCT system in order to enhance cisplatin (CDDP)-induced cytotoxicity and inhibit the metastatic potential of BC. CD44v9 protein expression was examined immunohistochemically in 63 muscle invasive BC (MIBC) patients who underwent radical cystectomy. CD44v9 expression was independently associated with disease recurrence and cancer-specific death in MIBC. Cytotoxic effects, glutathione levels, and reactive oxygen species production by SSZ and CD44v9 and phospho-p38MAPK protein expression by SSZ with or without CDDP were assessed in MBT-2V cells with highly metastatic potential. Sulfasalazine exerted cytotoxic effects against MBT-2V cells by inhibiting glutathione levels and inducing the production of reactive oxygen species. Sulfasalazine in combination with CDDP appeared to exert strong cytotoxic effects against MBT-2V cells by inhibiting CD44v9 expression and upregulating phospho-p38MAPK expression. The inhibitory effects of SSZ with or without CDDP were also investigated using an MBT-2V lung metastatic model. In the murine lung metastatic BC model, SSZ significantly prolonged animal survival. Furthermore, the combination of SSZ with CDDP exerted stronger inhibitory effects on the establishment of lung tumor nodules than SSZ or CDDP alone. CD44v9 expression could be a clinical biomarker for predicting poor outcomes in MIBC patients. Sulfasalazine in combination with CDDP has potential as a novel therapy against metastatic BC.


Reproducible extracellular vesicle size and concentration determination with tunable resistive pulse sensing.
The size of extracellular vesicles (EVs) can be determined with a tunable resistive pulse sensor (TRPS). Because the sensing pore diameter varies from pore to pore, the minimum detectable diameter also varies. The aim of this study is to determine and improve the reproducibility of TRPS measurements. Experiments were performed with the qNano system (Izon) using beads and a standard urine vesicle sample. With a combination of voltage and stretch that yields a high blockade height, we investigate whether the minimum detected diameter is more reproducible when we configure the instrument targeting (a) fixed stretch and voltage, or (b) fixed blockade height. Daily measurements with a fixed stretch and voltage (n=102) on a standard urine sample show a minimum detected vesicle diameter of 128±19 nm [mean±standard deviation; coefficient of variation (CV) 14.8%]. The vesicle concentration was 2.4·109±3.8·109 vesicles/mL (range 1.4·108-1.8·1010). When we compared setting a fixed stretch and voltage to setting a fixed blockade height on 3 different pores, we found a minimum detected vesicle diameter of 118 nm (CV 15.5%, stretch), and 123 nm (CV 4.5%, blockade height). The detected vesicle concentration was 3.2-8.2·108 vesicles/mL with fixed stretch and 6.4-7.8·108 vesicles/mL with fixed blockade height. Summary/conclusion: Pore-to-pore variability is the cause of the variation in minimum detected size when setting a fixed stretch and voltage. The reproducibility of the minimum detectable diameter is much improved by setting a fixed blockade height.


B-cell tumor development in Tet2-deficient mice.
The TET2 gene encodes an α-ketoglutarate-dependent dioxygenase able to oxidize 5-methylcytosine into 5-hydroxymethylcytosine, which is a step toward active DNA demethylation. TET2 is frequently mutated in myeloid malignancies but also in B- and T-cell malignancies. TET2 somatic mutations are also identified in healthy elderly individuals with clonal hematopoiesis. Tet2-deficient mouse models showed widespread hematological differentiation abnormalities, including myeloid, T-cell, and B-cell malignancies. We show here that, similar to what is observed with constitutive Tet2-deficient mice, B-cell-specific Tet2 knockout leads to abnormalities in the B1-cell subset and a development of B-cell malignancies after long latency. Aging Tet2-deficient mice accumulate clonal CD19+ B220low immunoglobulin M+ B-cell populations with transplantable ability showing similarities to human chronic lymphocytic leukemia, including CD5 expression and sensitivity to ibrutinib-mediated B-cell receptor (BCR) signaling inhibition. Exome sequencing of Tet2-/- malignant B cells reveals C-to-T and G-to-A mutations that lie within single-stranded DNA-specific activation-induced deaminase (AID)/APOBEC (apolipoprotein B messenger RNA editing enzyme, catalytic polypeptide-like) cytidine deaminases targeted motif, as confirmed by the lack of a B-cell tumor in compound Tet2-Aicda-deficient mice. Finally, we show that Tet2 deficiency accelerates and exacerbates T-cell leukemia/lymphoma 1A-induced leukemogenesis. Together, our data establish that Tet2 deficiency predisposes to mature B-cell malignancies, which development might be attributed in part to AID-mediated accumulating mutations and BCR-mediated signaling.


Rotation lines of hydrogen chloride for wavenumber calibration of far infrared spectrometers.
Contours are calculated for the pairs of H(35)Cl and H(37)Cl rotation lines for a range of spectral slitwidths between 0.10 cm(-1) and 3.0 cm(-1). The peak positions are compared with the frequencies of the H(35)Cl lines with a view to predicting the displacement of each peak due to overlap of the H(37)Cl lines. Experimental measurements of the J = 12 and 13 lines show that the predictions are correct. It is suggested that the calculated peaks be used as calibration data for instruments, although it is pointed out that if the wavenumber error of the instrument exceeds one-twentieth of the spectral slitwidth, the displacements can be ignored.


Segmentation of diesel spray images with log-likelihood ratio test algorithm for non-Gaussian distributions.
A methodology for processing images of diesel sprays under different experimental situations is presented. The new approach has been developed for cases where the background does not follow a Gaussian distribution but a positive bias appears. In such cases, the lognormal and the gamma probability density functions have been considered for the background digital level distributions. Two different algorithms have been compared with the standard log-likelihood ratio test (LRT): a threshold defined from the cumulative probability density function of the background shows a sensitive improvement, but the best results are obtained with modified versions of the LRT algorithm adapted to non-Gaussian cases.


Drugs associated with hepatotoxicity and their reporting frequency of liver adverse events in VigiBase: unified list based on international collaborative work.
Challenges exist in the clinical diagnosis of drug-induced liver injury (DILI) and in obtaining information on hepatotoxicity in humans. (i) To develop a unified list that combines drugs incriminated in well vetted or adjudicated DILI cases from many recognized sources and drugs that have been subjected to serious regulatory actions due to hepatotoxicity; and (ii) to supplement the drug list with data on reporting frequencies of liver events in the WHO individual case safety report database (VigiBase). (i) Drugs identified as causes of DILI at three major DILI registries; (ii) drugs identified as causes of drug-induced acute liver failure (ALF) in six different data sources, including major ALF registries and previously published ALF studies; and (iii) drugs identified as being subjected to serious governmental regulatory actions due to their hepatotoxicity in Europe or the US were collected. The reporting frequency of adverse events was determined using VigiBase, computed as Empirical Bayes Geometric Mean (EBGM) with 90% confidence interval for two customized terms, 'overall liver injury' and 'ALF'. EBGM of >or=2 was considered a disproportional increase in reporting frequency. The identified drugs were then characterized in terms of regional divergence, published case reports, serious regulatory actions, and reporting frequency of 'overall liver injury' and 'ALF' calculated from VigiBase. After excluding herbs, supplements and alternative medicines, a total of 385 individual drugs were identified; 319 drugs were identified in the three DILI registries, 107 from the six ALF registries (or studies) and 47 drugs that were subjected to suspension or withdrawal in the US or Europe due to their hepatotoxicity. The identified drugs varied significantly between Spain, the US and Sweden. Of the 319 drugs identified in the DILI registries of adjudicated cases, 93.4% were found in published case reports, 1.9% were suspended or withdrawn due to hepatotoxicity and 25.7% were also identified in the ALF registries/studies. In VigiBase, 30.4% of the 319 drugs were associated with disproportionally higher reporting frequency of 'overall liver injury' and 83.1% were associated with at least one reported case of ALF. This newly developed list of drugs associated with hepatotoxicity and the multifaceted analysis on hepatotoxicity will aid in causality assessment and clinical diagnosis of DILI and will provide a basis for further characterization of hepatotoxicity.


H-NS and RNA polymerase: a love-hate relationship?
Histone-like nucleoid structuring (H-NS) protein is a component of bacterial chromatin and influences gene expression both locally and on a global scale. Although H-NS is broadly considered a silencer of transcription, the mechanisms by which H-NS inhibits gene expression remain poorly understood. Here we discuss recent advances in the context of a 'love-hate' relationship between H-NS and RNA polymerase, in which these factors recognise similar DNA sequences but interfere with each other's activity. Understanding the complex relationship between H-NS and RNA polymerase may unite the multiple models that have been proposed to describe gene silencing by H-NS.


Suppressor function of peripheral blood mononuclear cells in normal individuals and in patients with systemic lupus erythematosus.
Normal peripheral blood mononuclear cells demonstrated increased DNA synthesis and secretion of newly synthesized protein when suboptimal concentrations of Concanavalin A (Con A) were added to the cultures after 24-h incubation in vitro. Cells stimulated by Con A, 1 mug/ml, after 24-h incubation demonstrated 3.0 times more tritiated thymidine incorporation, and 4.4 times more 14C-amino acid incorporation into newly synthesized secreted protein, than cells stimulated at 0 h (P less than 0.001). The acquisition of increased responsiveness was not abrogated by washing and resuspending the cells in fresh medium. Since the increased responsiveness could be inhibited by the addition to the cultures of small numbers of cells previously activated by Con A it is suggested that the enhanced reactivity acquired in culture represents the loss of a subpopulation of suppressor cells that modulate the T-lymphocyte response. Cells from nine patients with active, untreated systemic lupus erythematosus demonstrated normal responses to optimal concentrations of Con A added at 0 h, but an impaired response to Con A, 1 mug/ml. When these cells were incubated for 24 h, a significant increased response to Con A was not observed. This observation suggests that patients with active SLE lack circulating suppressor cells. When seven SLE patients were again studied after corticosteroid therapy had led to clinical improvement, the response to Con A, 1 mug/ml, added after 24-h incubation was similar to that observed in normal controls, suggesting that suppressor function in SLE returns as disease activity declines.


Lactate acquisition promotes successful colonization of the murine genital tract by Neisseria gonorrhoeae.
Previous studies on Neisseria gonorrhoeae have demonstrated that metabolism of lactate in the presence of glucose increases the growth rate of the bacterium and enhances its resistance to complement-mediated killing. Although these findings in vitro suggest that the acquisition of lactate promotes gonococcal colonization, the significance of this carbon source to the survival of the gonococcus in vivo remains unknown. To investigate the importance of lactate utilization during Neisseria gonorrhoeae genital tract infection, we identified the gene lctP, which encodes the gonococcal lactate permease. A mutant that lacks a functional copy of lctP was unable to take up exogenous lactate and did not grow in defined medium with lactate as the sole carbon source, in contrast to the wild-type and complemented strains; the mutant strain exhibited no growth defect in defined medium containing glucose. In defined medium containing physiological concentrations of lactate and glucose, the lctP mutant demonstrated reduced early growth and increased sensitivity to complement-mediated killing compared with the wild-type strain; the enhanced susceptibility to complement was associated with a reduction in lipopolysaccharide sialylation of the lctP mutant. The importance of lactate utilization during colonization was evaluated in the murine model of lower genital tract infection. The lctP mutant was significantly attenuated in its ability to colonize and survive in the genital tract, while the complemented mutant exhibited no defect for colonization. Lactate is a micronutrient in the genital tract that contributes to the survival of the gonococcus.


Suicide and other causes of mortality after post-partum psychiatric admission.
The risk of suicide in postnatal women is low and those suicides that occur appear to be associated with severe psychiatric illness. No previous study has specifically studied the risk of suicide following post-partum psychiatric disorder. We calculated standardised mortality ratios (SMRs) for suicide, unnatural deaths and deaths from natural causes for women admitted to psychiatric hospital in the first year after childbirth, using computerised cross-linkages between the Danish Psychiatric Case Register and the Danish registers of birth and causes of death for 1973-1993. During the study period 1567 women were admitted to psychiatric hospital of whom 107 (6.8%) died. The SMRs (compared with 100) were 1719 (95% CI1284-2254) for suicide, 1329 (95% CI1038-1676) for all unnatural causes and 238 (95% CI167-329) for natural causes. Suicides and deaths from all unnatural causes were most likely to occur in the first year after childbirth, the SMR for suicide within one year being 7216 (95% CI 3945-12 108). Although postnatal women as a whole appear to have a low rate of suicide, severe post-partum psychiatric disorder is associated with a high rate of deaths from natural and unnatural causes, particularly suicide. The risk is especially high in the first postnatal year, when the suicide risk is increased 70-fold. Close clinical supervision at this time is indicated.


Community-Based Policies and Support for Free Drinking Water Access in Outdoor Areas and Building Standards in U.S. Municipalities.
We examined community-level characteristics associated with free drinking water access policies in U.S. municipalities using data from a nationally representative survey of city managers/officials from 2,029 local governments in 2014. Outcomes were 4 free drinking water access policies. Explanatory measures were population size, rural/urban status, census region, poverty prevalence, education, and racial/ethnic composition. We used multivariable logistic regression to test differences and presented only significant findings. Many (56.3%) local governments had at least one community plan with a written objective to provide free drinking water in outdoor areas; municipalities in the Northeast and South regions and municipalities with ≤ 50% of non-Hispanic whites were less likely and municipalities with larger population size were more likely to have a plan. About 59% had polices/budget provisions for free drinking water in parks/outdoor recreation areas; municipalities in the Northeast and South regions were less likely and municipalities with larger population size were more likely to have it. Only 9.3% provided development incentives for placing drinking fountains in outdoor, publicly accessible areas; municipalities with larger population size were more likely to have it. Only 7.7% had a municipal plumbing code with a drinking fountain standard that differed from the statewide plumbing code; municipalities with a lower proportion of non-Hispanic whites were more likely to have it. In conclusion, over half of municipalities had written plans or a provision for providing free drinking water in parks, but providing development incentives or having a local plumbing code provision were rare.


Incidence of hypoglycemia following insulin-based acute stabilization of hyperkalemia treatment.
The aim of this study was to assess the incidence of hypoglycemia in hospitalized patients following acute treatment of hyperkalemia with insulin. A characterization of the affected patients and the administered insulin/dextrose regimens was also performed. A retrospective search of the electronic records of a large university-based tertiary care hospital was conducted, from June 1, 2009 to December 1, 2009, to identify patients who developed hypoglycemia following acute stabilization of hyperkalemia treatment with regular insulin. Of 219 hyperkalemic patients who met the criteria of the study, 19 patients (8.7%) were identified ashypoglycemic (blood glucose <70 mg/dl), and 5 of these patients (2.3% of study patients) were classified as severely hypoglycemic (blood glucose <40 mg/dl). Fifteen (79%) of the hypoglycemic patients had acute kidney injury or were end-stage renal disease patients on hemodialysis at the time of treatment. Fifty-eight percent of the hypoglycemic events occurred following the commonly employed 10 units of regular insulin and 25 gm of dextrose 50% treatment regimen. Iatrogenic hypoglycemia, as a result of treatment for hyperkalemia, is a common occurrence. Hyperkalemia occurs disproportionately in patients with acute kidney injury or end-stage renal disease, and these patients are predisposed to an increased risk of hypoglycemia. The risk of severe hypoglycemia escalates inpatients with lower body weight and creatinine clearance.Hypoglycemia risk can be minimized by providing sufficient dextrose in the treatment regimen, however, patient variability in treatment response dictates careful blood glucose monitoring before and after treatment.


Extravasation of Noncytotoxic Drugs: A Review of the Literature.
Extravasation is a potential complication associated with intravenous therapy administration. Inadvertent leakage of medications with vesicant properties can cause severe tissue necrosis, which can lead to devastating long-term consequences. Recognizing potential agents is an essential step in mitigating the risk of extravasation. A literature search was carried out using PubMed with the following key words: extravasation, soft tissue injury, phlebitis, and infiltration, from January 1961 through January 2014. The publications were screened manually and reviewed to identify reports for medications that included synonyms of the International Nonproprietary Name, while excluding antineoplastic agents, radiographic contrast material, investigational or nonmarketed drugs, and animal data, to yield 70 articles. Furthermore, reference citations from publications were also reviewed for relevance and yielded 4 articles. We discovered 232 cases of extravasation involving 37 agents (in order of frequency): phenytoin, parenteral nutrition, calcium gluconate, potassium chloride, calcium chloride, dopamine, dextrose solutions, epinephrine, sodium bicarbonate, nafcillin, propofol, norepinephrine, mannitol, arginine, promethazine, vancomycin, tetracycline, dobutamine, vasopressin, sodium thiopental, acyclovir, amphotericin, ampicillin, cloxacillin, gentamicin, metronidazole, oxacillin, penicillin, amiodarone, albumin, furosemide, lipids, lorazepam, immunoglobulin, morphine, and sodium valproate. Potential properties contributing to extravasation include the following: pH, osmolarity, diluent, vasoactive properties, and inactive ingredients. Antidotes and supportive care agents used in the management of these cases of extravasation include hyaluronidase, phentolamine, terbutaline, topical anesthetics (such as lidocaine and prilocaine cream), topical antimicrobials (such as silver sulfadiazine and chlorhexidine), topical debridement agents (collagenase ointment), topical steroids, and topical vasodilators (nitroglycerin). Data on the management of noncytotoxic extravasations is sparse, consisting primarily of case reports and anecdotal evidence. Fortunately, this adverse outcome is preventable and identification of vesicant agents plays a pivotal role. The intent of this review is to provide a reference identifying noncytotoxic vesicants and the management of extravasations associated with specific agents.


The experiences of making infant feeding choices by African, Caribbean and Black HIV-positive mothers in Ontario, Canada.
Mothers in HIV-endemic countries are advised to exclusively breastfeed their babies until six months because of lack of resources and better chances for child survival, while in developed countries, replacement feeding is advised. What are the experiences of HIV-positive women who migrate from HIV-endemic countries to developed countries, when making infant feeding choices? In-depth interviews and focus group discussions with a total of 25 women living with HIV in Toronto and Hamilton, Ontario. Free infant formula alleviates the practical constraints in making infant feeding choices. However, cultural beliefs and social expectations constrain HIV-positive mothers' decision not to breastfeed. This is further complicated by the different policies. Service providers should understand the psychological and emotional experiences of the mothers in order to provide the appropriate support. Peers could be potential sources of support. The differences in policies are issues of global justice that need to be addressed.


The Implementation of an Interpersonal Violence Screening Program in Primary Care Settings: Lessons Learned.
Interpersonal violence (IPV) is a public health issue that disproportionately affects women. IPV screening improves likelihood of survivor disclosure and access to additional support. To enhance primary care IPV screening, Technology Enhanced Screening and Supportive Assistance (TESSA) uses integrated technological systems to deliver bidirectional, evidence-informed health navigation, health management, and safety interventions. This study evaluates TESSA implementation in primary care clinics using the Consolidated Framework for Implementation Research (CFIR). CFIR is a metatheoretical framework used for evaluating clinical intervention implementation. Salient constructs within CFIR's five domains (intervention characteristics, outer setting, inner setting, characteristics of individuals, and process) were identified (23 constructs), and pertinent implementation details were examined. Key lessons learned included intervention characteristic constructs like intervention source (e.g., selecting tablets that can screen for items integral to the program's aims) and adaptability (e.g., ensuring tablets worked with electronic medical records for each clinic), process constructs like engaging champions (e.g., garnering buy-in from key clinic stakeholders and staff), outer setting constructs like patient needs and resources (e.g., addressing pertinent patient resource needs) and external policies and incentives (e.g., incentivizing clinics by addressing clinic needs), and inner setting constructs like leadership engagement (e.g., ensuring buy-in from organizational leaders as leadership changed frequently). CFIR identifies important implementation factors for programs like TESSA that screen for high-risk populations and implement in primary care settings. The TESSA program implementation permits increased IPV screening among primary health care populations, thus promoting access to resources for otherwise hard-to-reach populations.


Image registration under illumination variations using region-based confidence weighted M-estimators.
We present an image registration model for image sets with arbitrarily shaped local illumination variations between images. Any nongeometric variations tend to degrade the geometric registration precision and impact subsequent processing. Traditional image registration approaches do not typically account for changes and movement of light sources, which result in interimage illumination differences with arbitrary shape. In addition, these approaches typically use a least-square estimator that is sensitive to outliers, where interimage illumination variations are often large enough to act as outliers. In this paper, we propose an image registration approach that compensates for arbitrarily shaped interimage illumination variations, which are processed using robust M -estimators tuned to that region. Each M-estimator for each illumination region has a distinct cost function by which small and large interimage residuals are unevenly penalized. Since the segmentation of the interimage illumination variations may not be perfect, a segmentation confidence weighting is also imposed to reduce the negative effect of mis-segmentation around illumination region boundaries. The proposed approach is cast in an iterative coarse-to-fine framework, which allows a convergence rate similar to competing intensity-based image registration approaches. The overall proposed approach is presented in a general framework, but experimental results use the bisquare M-estimator with region segmentation confidence weighting. A nearly tenfold improvement in subpixel registration precision is seen with the proposed technique when convergence is attained, as compared with competing techniques using both simulated and real data sets with interimage illumination variations.


Mitochondrial tRNA methylation in Alzheimer's disease and progressive supranuclear palsy.
Methylation of mitochondrial tRNAs (mt-tRNA) at the 9th position ("p9 site") is known to impact translational efficiency and downstream mitochondrial function; however, direct assessment of mt-RNA methylation is challenging. Recent RNA sequence-based methods have been developed to reliably identify post-transcriptional methylation. Though p9 methylation has been studied in healthy human populations and in the context of cancer, it has not yet been analyzed in neurodegenerative disease, where mitochondrial dysfunction is a prominent and early hallmark of disease progression. Mitochondrial p9 methylation was inferred from multi-allelic calls in RNA-seq data. Gene-based association studies were performed in FUMA. Correlations between nuclear gene expression and p9 methylation were tested using Spearman's rho. Fisher's Exact test was used in PANTHER and IPA to test for overrepresentation and enrichment of biological processes and pathways in the top nuclear genes correlated with p9 methylation. Variable methylation was observed at 11 p9 sites in post-mortem cerebellar tissue of elderly subjects who were either healthy or diagnosed with Alzheimer's disease (AD), progressive supranuclear palsy (PSP) or pathological aging (PA). Similarities in degree of methylation were observed between AD and PSP. Certain nuclear encoded genes were identified as significantly associated with p9 methylation. Expression of 5300 nuclear encoded genes was significantly correlated with p9 methylation, with AD and PSP subjects exhibiting similar expression profiles. Overrepresentation and enrichment testing using the top transcripts revealed enrichment for a number of molecular processes, terms and pathways including many of which that were mitochondrial-related. With mitochondrial dysfunction being an established hallmark of neurodegenerative disease pathophysiology, this work sheds light on the potential molecular underpinnings of this dysfunction. Here we show overlap in cerebellar pathophysiology between common tauopathies such as Alzheimer's disease and progressive supranuclear palsy. Whether p9 hypermethylation is a cause or consequence of pathology remains an area of focus.


A picture speaks a thousand words: evaluation of a pictorial post-vaccination care resource in Australia.
Evaluating the 'Common Reactions to Vaccination' post-vaccination care resource was seen as an opportunity to contribute to the limited literature base in this important area, learn from the strengths and weaknesses of the resource and gain insight into post-vaccination care practices. Semi-structured in-depth interviews were conducted with 12 general practitioners and 29 practice nurses in New South Wales and Australian Capital Territory, Australia. Structured interview guides were used and data was analysed thematically. A self-administered survey was also distributed to parents or guardians during routine childhood vaccination visits. When compared with previous resources, participants felt the new resource was more appropriate as it had a simple layout; it was colourful, incorporated pictures and had basic and practical information. Information about post-vaccination care and common reactions to vaccination must be provided in written form accompanied by a verbal reinforcement so that patients can revisit the information at a later stage if required. The 'Common Reactions to Vaccination' post-vaccination care resource provides comprehensive information in an easy-to-understand pictorial way and was appreciated by both vaccination providers and patients.


In vivo imaging of autophagy in a mouse stroke model.
Recent studies have suggested that autophagy is involved in a neural death pathway following cerebral ischemia. In vivo detection of autophagy could be important for evaluating ischemic neural cell damage for human stroke patients. Using novel green fluorescent protein (GFP)-fused microtubule-associated protein 1 light chain 3 (LC3) transgenic (Tg) mice, in vivo imaging of autophagy was performed at 1, 3 and 6 d after 60 min transient middle cerebral artery occlusion (tMCAO). Ex vivo imaging of autophagy, testing of the autophagy inhibitor 3-methyladenine (3-MA), estern blot analysis, immunohistochemistry, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) and fluorescent analyses were performed on brain sections following tMCAO. In vivo fluorescent signals were detected above the ischemic hemisphere through the skull bone at 1, 3 and 6 d after tMCAO, with a peak at 1 d. Similar results were obtained with ex vivo fluorescence imaging. western blot analysis revealed maximum LC3-I and LC3-II expression at 1 d after tMCAO and fluorescence immunohistochemistry demonstrated that GFP-LC3-positive cells were primarily neuronal, not astroglial or microglial, cells. The number of GFP-LC3/TUNEL double-positive cells was greater in the periischemic area than in the core. These results provided evidence of in vivo autophagy detection, with a peak at 1 d, in a live animal model following cerebral ischemia. This novel technique could be valuable for monitoring autophagic processes in vivo in live stroke patients, as well as for clarifying the detailed role of autophagy in the ischemic brain, as well as in other neurological diseases.


Raloxifene and stroke risks in Japanese postmenopausal women with osteoporosis on postmarketing surveillance.
The aim of this study was to assess whether the risks of stroke and stroke death in Japanese postmenopausal women with osteoporosis exposed to raloxifene increased in comparison with those of the general Japanese female population. Safety data associated with use of raloxifene were collected in observational settings from 6,970 women with a median follow-up period of 366 days on postmarketing surveillance conducted in Japan. The stroke incidences in raloxifene-treated women were compared with population stroke rates derived from epidemiology studies conducted at three distinct locations in Japan, yielding standardized morbidity ratios for stroke for the purpose of stroke risk assessment. Exposure to raloxifene totaled 7,474 patient-years and 23 stroke cases were reported, including 4 fatal cases. Multiple stroke risk factors were present in three of four women with fatal outcome. The standardized stroke morbidity ratios in raloxifene-treated women versus women from the three reference regions, including Akita Prefecture, Takashima town in Shiga Prefecture, and Okinawa Prefecture, were 0.68 (95% CI, 0.45-1.02), 0.54 (95% CI, 0.35-0.83), and 0.82 (95% CI, 0.54-1.24), respectively. In this initial interim analysis, there seems to be no significant increased risk of stroke among Japanese women with 1 year of raloxifene use in comparison to Japanese epidemiological data. An increased risk of fatal stroke in the treatment population was indeterminable because of the small number of observed fatal cases. However, ongoing safety monitoring of stroke risk among raloxifene women will continue.


Effect of polymorphic colour vision for fruit detection in the spider monkey Ateles geoffroyi, and its implications for the maintenance of polymorphic colour vision in platyrrhine monkeys.
Most platyrrhine monkeys have an X-linked tri-allelic polymorphism for medium and long wavelength (M/L) sensitive cone photopigments. These pigments' sensitivity maxima (lambdamax) range from 535 to 562 nm. All animals also have an autosomally coded short-wavelength-sensitive (S) cone pigment. In populations with three M/L alleles there are six different colour vision phenotypes. Heterozygous females have trichromatic colour vision, while males and homozygous females are dichromats. The selective basis for this polymorphism is not understood, but is probably affected by the costs and benefits of trichromatic compared to dichromatic colour vision. For example, it has been suggested that trichromats are better equipped than dichromats to detect fruit against a leaf background. To investigate this possibility, we modeled fruit detection by various colour vision phenotypes present in the frugivorous spider monkey, Ateles geoffroyi. Our study population is thought to have three M/L alleles with cone pigment lambdamax values close to 535, 550 and 562 nm. The model predicted that all trichromat phenotypes had an advantage over dichromats, and the 535/562 nm phenotype was best; however, the model predicted that dichromats could detect all of the fruit species consumed by spider monkeys. We conclude that the heterozygote advantage experienced by females may be the most plausible explanation for the maintenance of this polymorphism in A. geoffroyi. Nevertheless, more studies need to evaluate social foraging behaviour and the performance of different phenotypes of other New World monkeys to determine if this is a global explanation for this phenomena or more specific to A. geofforyi.


Complete genome sequence of the phenanthrene-degrading soil bacterium Delftia acidovorans Cs1-4.
Polycyclic aromatic hydrocarbons (PAH) are ubiquitous environmental pollutants and microbial biodegradation is an important means of remediation of PAH-contaminated soil. Delftia acidovorans Cs1-4 (formerly Delftia sp. Cs1-4) was isolated by using phenanthrene as the sole carbon source from PAH contaminated soil in Wisconsin. Its full genome sequence was determined to gain insights into a mechanisms underlying biodegradation of PAH. Three genomic libraries were constructed and sequenced: an Illumina GAii shotgun library (916,416,493 reads), a 454 Titanium standard library (770,171 reads) and one paired-end 454 library (average insert size of 8 kb, 508,092 reads). The initial assembly contained 40 contigs in two scaffolds. The 454 Titanium standard data and the 454 paired end data were assembled together and the consensus sequences were computationally shredded into 2 kb overlapping shreds. Illumina sequencing data was assembled, and the consensus sequence was computationally shredded into 1.5 kb overlapping shreds. Gaps between contigs were closed by editing in Consed, by PCR and by Bubble PCR primer walks. A total of 182 additional reactions were needed to close gaps and to raise the quality of the finished sequence. The final assembly is based on 253.3 Mb of 454 draft data (averaging 38.4 X coverage) and 590.2 Mb of Illumina draft data (averaging 89.4 X coverage). The genome of strain Cs1-4 consists of a single circular chromosome of 6,685,842 bp (66.7 %G+C) containing 6,028 predicted genes; 5,931 of these genes were protein-encoding and 4,425 gene products were assigned to a putative function. Genes encoding phenanthrene degradation were localized to a 232 kb genomic island (termed the phn island), which contained near its 3' end a bacteriophage P4-like integrase, an enzyme often associated with chromosomal integration of mobile genetic elements. Other biodegradation pathways reconstructed from the genome sequence included: benzoate (by the acetyl-CoA pathway), styrene, nicotinic acid (by the maleamate pathway) and the pesticides Dicamba and Fenitrothion. Determination of the complete genome sequence of D. acidovorans Cs1-4 has provided new insights the microbial mechanisms of PAH biodegradation that may shape the process in the environment.


Arginine usage in mycobacteria-infected macrophages depends on autocrine-paracrine cytokine signaling.
Nitric oxide (NO) produced by macrophages is toxic to host tissues and invading pathogens, and its regulation is essential to suppress host cytotoxicity. Macrophage arginase 1 (Arg1) competes with NO synthases for arginine, a substrate common to both types of enzymes, to inhibit NO production. Two signal transduction pathways control the production of Arg1 in macrophages: One pathway dependent on the Toll-like receptor adaptor protein myeloid differentiation marker 88 (MyD88) induces the expression of Arg1 during intracellular infections, whereas another pathway, which depends on signal transducer and activator of transcription 6 (STAT6), is required for Arg1 expression in alternatively activated macrophages. We found that mycobacteria-infected macrophages produced soluble factors, including interleukin-6 (IL-6), IL-10, and granulocyte colony-stimulating factor (G-CSF), that induced expression of Arg1 in an autocrine-paracrine manner. Arg1 expression was controlled by the MyD88-dependent production of these cytokines rather than by cell-intrinsic MyD88 signaling to Arg1. Our study revealed that the MyD88-dependent pathway that induced the expression of Arg1 after infection by mycobacteria required STAT3 activation and that this pathway may cause the development of an immunosuppressive niche in granulomas because of the induced production of Arg1 in surrounding uninfected macrophages.


Evolution and maintenance of microbe-mediated protection under occasional pathogen infection.
Every host is colonized by a variety of microbes, some of which can protect their hosts from pathogen infection. However, pathogen presence naturally varies over time in nature, such as in the case of seasonal epidemics. We experimentally coevolved populations of Caenorhabditis elegans worm hosts with bacteria possessing protective traits (Enterococcus faecalis), in treatments varying the infection frequency with pathogenic Staphylococcus aureus every host generation, alternating host generations, every fifth host generation, or never. We additionally investigated the effect of initial pathogen presence at the formation of the defensive symbiosis. Our results show that enhanced microbe-mediated protection evolved during host-protective microbe coevolution when faced with rare infections by a pathogen. Initial pathogen presence had no effect on the evolutionary outcome of microbe-mediated protection. We also found that protection was only effective at preventing mortality during the time of pathogen infection. Overall, our results suggest that resident microbes can be a form of transgenerational immunity against rare pathogen infection.


RNA interference-mediated inhibition of hepatocyte nuclear factor 1alpha identifies target genes.
Hepatocyte nuclear factor 1alpha (HNF1alpha) is a homeodomain transcription factor that is central to co-ordinated differentiation of a number of cell lineages, including hepatocytes in the liver and islet cells in the pancreas. HNF1alpha interacts directly with other transcription factors and co-factors and is involved in chromatin modification to alter gene expression. To further investigate the pivotal role of HNF1alpha in transcriptional control pathways we utilized RNA interference. An siRNA oligonucleotide specific for HNF1alpha reduced HNF1alpha protein levels by up to 70% in transient transfections of Caco2 cells. The same sequence incorporated into an shRNAi reduced protein levels by up to 90% in stable transfections. Microarray analysis of RNA from cell lines with stable RNAi-mediated down-regulation of HNF1alpha, identified genes known to be regulated by this transcription factor and also novel genes.


Lysine 63-linked polyubiquitin chain may serve as a targeting signal for the 26S proteasome.
Recruitment of substrates to the 26S proteasome usually requires covalent attachment of the Lys48-linked polyubiquitin chain. In contrast, modifications with the Lys63-linked polyubiquitin chain and/or monomeric ubiquitin are generally thought to function in proteasome-independent cellular processes. Nevertheless, the ubiquitin chain-type specificity for the proteasomal targeting is still poorly understood, especially in vivo. Using mass spectrometry, we found that Rsp5, a ubiquitin-ligase in budding yeast, catalyzes the formation of Lys63-linked ubiquitin chains in vitro. Interestingly, the 26S proteasome degraded well the Lys63-linked ubiquitinated substrate in vitro. To examine whether Lys63-linked ubiquitination serves in degradation in vivo, we investigated the ubiquitination of Mga2-p120, a substrate of Rsp5. The polyubiquitinated p120 contained relatively high levels of Lys63-linkages, and the Lys63-linked chains were sufficient for the proteasome-binding and subsequent p120-processing. In addition, Lys63-linked chains as well as Lys48-linked chains were detected in the 26S proteasome-bound polyubiquitinated proteins. These results raise the possibility that Lys63-linked ubiquitin chain also serves as a targeting signal for the 26S proteaseome in vivo.


Delimiting species without nuclear monophyly in Madagascar's mouse lemurs.
Speciation begins when populations become genetically separated through a substantial reduction in gene flow, and it is at this point that a genetically cohesive set of populations attain the sole property of species: the independent evolution of a population-level lineage. The comprehensive delimitation of species within biodiversity hotspots, regardless of their level of divergence, is important for understanding the factors that drive the diversification of biota and for identifying them as targets for conservation. However, delimiting recently diverged species is challenging due to insufficient time for the differential evolution of characters--including morphological differences, reproductive isolation, and gene tree monophyly--that are typically used as evidence for separately evolving lineages. In this study, we assembled multiple lines of evidence from the analysis of mtDNA and nDNA sequence data for the delimitation of a high diversity of cryptically diverged population-level mouse lemur lineages across the island of Madagascar. Our study uses a multi-faceted approach that applies phylogenetic, population genetic, and genealogical analysis for recognizing lineage diversity and presents the most thoroughly sampled species delimitation of mouse lemur ever performed. The resolution of a large number of geographically defined clades in the mtDNA gene tree provides strong initial evidence for recognizing a high diversity of population-level lineages in mouse lemurs. We find additional support for lineage recognition in the striking concordance between mtDNA clades and patterns of nuclear population structure. Lineages identified using these two sources of evidence also exhibit patterns of population divergence according to genealogical exclusivity estimates. Mouse lemur lineage diversity is reflected in both a geographically fine-scaled pattern of population divergence within established and geographically widespread taxa, as well as newly resolved patterns of micro-endemism revealed through expanded field sampling into previously poorly and well-sampled regions.


Top-down or bottom-up regulation of intra-host blood-stage malaria: do malaria parasites most resemble the dynamics of prey or predator?
Knowledge of the factors that limit parasite numbers offers hope of improved intervention strategies as well as exposing the selective forces that have shaped parasite life-history strategies. We develop a theoretical framework with which to consider the intra-host regulation of malaria parasite density. We analyse a general model that relates timing and magnitude of peak parasite density to initial dose under three different regulatory processes. The dynamics can be regulated either by top-down processes (upgradable immune regulation), bottom-up processes (fixed immune response and red blood cell (RBC) limitation) or a mixture of the two. We define and estimate the following key parameters: (i) the rate of RBC replenishment; (ii) the rate of destruction of uninfected RBCs; and (iii) the maximum parasite growth rate. Comparing predictions of this model with experimental results for rodent malaria in laboratory mice allowed us to reject functional forms of immune upregulation and/or effects of RBC limitation that were inconsistent with the data. Bottom-up regulation alone was insufficient to account for observed patterns without invoking either localized depletion of RBC density or merozoite interference. By contrast, an immune function upregulated in proportion to either merozoite or infected RBC density was consistent with observed dynamics. An immune response directed solely at merozoites required twice the level of activation of one directed at infected RBCs.


The moderating effects of children's fear and activity level on relations between parenting practices and childhood symptomatology.
Parenting practices have been previously linked to childhood symptomatology. However, little consideration has been given to the potential effect of individual differences within the child on this relation. The current study assessed the moderating effects of children's activity level and fear on relations between parenting practices and childhood aggression and depressive symptoms using a sample of 64 fourth-, and fifth-grade boys. The findings showed that poorly monitored active boys and fearful boys who were exposed to harsh discipline exhibited high levels of aggression. Boys characterized by high fear who were exposed to harsh discipline or whose parents were extremely overinvolved showed elevated levels of depressive symptoms. These findings suggest that integrating children's individual differences with parenting models enhances our understanding of the etiology of childhood symptomatology. The intervention implications of such an integration are discussed.


American Thyroid Association Statement on Postoperative Hypoparathyroidism: Diagnosis, Prevention, and Management in Adults.
Hypoparathyroidism (hypoPT) is the most common complication following bilateral thyroid operations. Thyroid surgeons must employ strategies for minimizing and preventing post-thyroidectomy hypoPT. The objective of this American Thyroid Association Surgical Affairs Committee Statement is to provide an overview of its diagnosis, prevention, and treatment. HypoPT occurs when a low intact parathyroid hormone (PTH) level is accompanied by hypocalcemia. Risk factors for post-thyroidectomy hypoPT include bilateral thyroid operations, autoimmune thyroid disease, central neck dissection, substernal goiter, surgeon inexperience, and malabsorptive conditions. Medical and surgical strategies to minimize perioperative hypoPT include optimizing vitamin D levels, preserving parathyroid blood supply, and autotransplanting ischemic parathyroid glands. Measurement of intraoperative or early postoperative intact PTH levels following thyroidectomy can help guide patient management. In general, a postoperative PTH level <15 pg/mL indicates increased risk for acute hypoPT. Effective management of mild to moderate potential or actual postoperative hypoPT can be achieved by administering either empiric/prophylactic oral calcium and vitamin D, selective oral calcium, and vitamin D based on rapid postoperative PTH level(s), or serial serum calcium levels as a guide. Monitoring for rebound hypercalcemia is necessary to avoid metabolic and renal complications. For more severe hypocalcemia, inpatient management may be necessary. Permanent hypoPT has long-term consequences for both objective and subjective well-being, and should be prevented whenever possible.


The Microbiota of Breast Tissue and Its Association with Breast Cancer.
In the United States, 1 in 8 women will be diagnosed with breast cancer in her lifetime. Along with genetics, the environment contributes to disease development, but what these exact environmental factors are remains unknown. We have previously shown that breast tissue is not sterile but contains a diverse population of bacteria. We thus believe that the host's local microbiome could be modulating the risk of breast cancer development. Using 16S rRNA amplicon sequencing, we show that bacterial profiles differ between normal adjacent tissue from women with breast cancer and tissue from healthy controls. Women with breast cancer had higher relative abundances of Bacillus, Enterobacteriaceae and Staphylococcus Escherichia coli (a member of the Enterobacteriaceae family) and Staphylococcus epidermidis, isolated from breast cancer patients, were shown to induce DNA double-stranded breaks in HeLa cells using the histone-2AX (H2AX) phosphorylation (γ-H2AX) assay. We also found that microbial profiles are similar between normal adjacent tissue and tissue sampled directly from the tumor. This study raises important questions as to what role the breast microbiome plays in disease development or progression and how we can manipulate this for possible therapeutics or prevention. This study shows that different bacterial profiles in breast tissue exist between healthy women and those with breast cancer. Higher relative abundances of bacteria that had the ability to cause DNA damage in vitro were detected in breast cancer patients, as was a decrease in some lactic acid bacteria, known for their beneficial health effects, including anticarcinogenic properties. This study raises important questions as to the role of the mammary microbiome in modulating the risk of breast cancer development.


Development of a switched integrator amplifier for high-accuracy optical measurements.
In the field of low flux optical measurements, the development and use of large area silicon detectors is becoming more frequent. The current/voltage conversion of their photocurrent presents a set of problems for traditional transimpedance amplifiers. The switched integration principle overcomes these limitations. We describe the development of a fully characterized current-voltage amplifier using the switched integrator technique. Two distinct systems have been developed in parallel at the United Kingdom's National Physical Laboratory (NPL) and Czech Metrology Institute (CMI) laboratories. We present the circuit theory and best practice in the design and construction of switched integrators. In conclusion the results achieved and future developments are discussed.


Tenascin expression in cancer cells and stroma of human breast cancer and its prognostic significance.
Sections of formalin-fixed, paraffin-embedded tissues from 210 human breast cancers were immunohistochemically examined using the mAb against human tenascin (TN) RCB1. Immunoreactive TN was detected in the breast cancer stroma in 77 (36.7%) cases, whereas the remaining 133 (63.3%) were negative. Of the 77, 12 (5.7%) cases also showed positive staining in the carcinoma cell cytoplasm. The positive cells were often observed in the margin of the cancer nests at the site adjacent to the stroma. According to the staining pattern of TN, the breast cancer cases were classified into the three groups of cancer cell TN(+)/stromal TN(+), cancer cell(-)/stromal TN(+), and cancer cell(-)/stromal TN(-). Analysis of the relationship of these TN patterns with various clinicopathological characteristics of the tumors and the patient outcome revealed that, in comparison to the cancer cell(-)/stromal TN(-) group, the cancer cell TN(+)/stromal TN(+) group exhibited increased frequency of lymph node metastasis and exceptionally poor outcome, and the cancer cell(-)/stromal TN(+) group also showed more frequent metastasis and poorer outcome. Most of the cancer cell TN(+)/stromal TN(+) cases were c-erbB-2 positive and estrogen receptor negative. Furthermore, in situ hybridization of freshly obtained breast cancer tissues demonstrated that both cancer cells and stromal cells express TN mRNA. These results indicate that the TN in breast cancer is produced by cancer epithelial cells as well as by stromal mesenchymal cells, and that cancer cell TN might be involved in cancer spreading, resulting in unfavorable patient prognosis.


Identifying educational priorities for occupational therapy students to prepare for mental health practice in Australia and New Zealand: Opinions of practising occupational therapists.
The effective preparation of occupational therapy students for mental health practice is critical to facilitate positive consumer outcomes, underpin optimal practice and support new graduates' professional identity. This project was established to determine a set of 'educational priorities' for occupational therapy students to prepare them for current (and future) entry-level practice in mental health, from the perspective of mental health occupational therapists in Australia and New Zealand. The study included two phases. In Phase One, participants identified what they considered to be important educational priorities for occupational therapy students to prepare them for practice in mental health. For Phase Two, an 'expert panel' was assembled to review and rank these using a Policy Delphi approach. Eighty-five participants provided educational priorities in Phase One. These were grouped into a total of 149 educational themes. In Phase Two, the expert panel (consisting of 37 occupational therapists from diverse locations and practice settings) prioritised these themes across three Delphi rounds. A final priority list was generated dividing educational themes into three prioritised categories: 29 'Essential', 25 'Important' and 44 'Optional' priorities. Highest-ranked priorities were: clinical reasoning, client-centred practice, therapeutic use of self, functional implications of mental illness, therapeutic use of occupation and mental health fieldwork experience. The priority list developed as part of this project provides additional information to support the review of occupational therapy curricula across Australia and New Zealand to ensure that new graduates are optimally prepared for mental health practice.


Management of seizures in children with falciparum malaria.
Falciparum malaria is the most common cause of convulsions in children admitted to hospital in malaria endemic areas. Detection of convulsions requires close monitoring and one should be on the look out for signs of subtle convulsions. About a third of acute seizures in children with cerebral malaria, do not manifest as convulsions, but as changes in eye deviation, salivation and/or eye deviation. It is important to terminate convulsions lasting more than 5 min, since prolonged convulsions are associated with neurological deficits in survivors of children with severe malaria. Initial management should include putting the child into the left lateral position, checking the blood glucose and administering oxygen if hypoxic. The benzodiazepines, particularly diazepam, are used as the initial anticonvulsants. Phenobarbital and phenytoin are used as second-line treatments. Prompt and effective management of falciparum malaria associated convulsions may contribute to a better outcome in children with severe malaria.


DEC1 (STRA13) protein expression relates to hypoxia- inducible factor 1-alpha and carbonic anhydrase-9 overexpression in non-small cell lung cancer.
Differentiated embryo-chondrocyte expressed gene 1 (DEC1) is involved in cell differentiation, proliferation, and apoptosis, and was recently shown to be regulated by hypoxia. The present immunohistochemical study demonstrates extensive nuclear expression of the protein in 38% of a series of 115 non-small cell lung carcinomas using a polyclonal antibody (Ab) recognizing DEC1 protein. Such expression was directly related to the expression of two hypoxia-regulated proteins, namely the hypoxia-inducible factor (HIF) 1alpha and carbonic anhydrase-9. Although DEC1 was not related to angiogenesis or to the expression of VEGF and thymidine phosphorylase, a direct association with up-regulated bFGF receptors was noted. DEC1 was persistently expressed in the nuclei of normal bronchial and alveolar tissue. It is suggested that loss of DEC1 expression is an early event in the development of lung cancer, while DEC1 gene expression occurs in a subset of tumours and parallels the overexpression of other hypoxia-regulated proteins.


How green is the grass on the other side? Frontopolar cortex and the evidence in favor of alternative courses of action.
Behavioral flexibility is the hallmark of goal-directed behavior. Whereas a great deal is known about the neural substrates of behavioral adjustment when it is explicitly cued by features of the external environment, little is known about how we adapt our behavior when such changes are made on the basis of uncertain evidence. Using a Bayesian reinforcement-learning model and fMRI, we show that frontopolar cortex (FPC) tracks the relative advantage in favor of switching to a foregone alternative when choices are made voluntarily. Changes in FPC functional connectivity occur when subjects finally decide to switch to the alternative behavior. Moreover, interindividual variation in the FPC signal predicts interindividual differences in effectively adapting behavior. By contrast, ventromedial prefrontal cortex (vmPFC) encodes the relative value of the current decision. Collectively, these findings reveal complementary prefrontal computations essential for promoting short- and long-term behavioral flexibility.


Reliability and validity of a Chinese version of the Multidimensional Fatigue Symptom Inventory-Short Form (MFSI-SF-C).
To examine the psychometric properties of the Chinese version of the Multidimensional Fatigue Symptom Inventory-Short Form (MFSI-SF-C) for use in Chinese-speaking countries. The assessment of fatigue is a challenging task for most researchers because culture may influence perceptions of meaning of fatigue. The lack of examination of the psychometric properties of the fatigue measures across studies limits the scientific rigour for generating additional research on the concept of 'fatigue.' A cross-sectional study. The study recruited 107 cancer inpatients from two medical centres in Taiwan. The MFSI-SF-C was examined using a two step process: (1) Translation and back-translation of the instrument; and (2) Examination of internal consistency reliability, test-retest reliability, content validity and construct validity. The results showed that the Cronbach's α of MFSI-SF-C total scale and subscales ranged between 0·83-0·92. The content validity index was 0·93. The difference between the fatigue of cancer patients and the comparison group of healthy people in the community was significant. The results demonstrated good convergent validity when comparing fatigue with depression and quality of life. Factor analysis confirmed the four dimensions of fatigue: physical, emotional, mental and vigour. It showed moderate intercorrelation between subscales and high factor loadings also helped to clarify the psychometric meaning. The reliability and validity information presented in this article support the use of the Chinese version of the MFSI-SF as a research instrument for measuring fatigue in Chinese populations. This study also provides evidence that the MFSI-SF possesses robust psychometric properties. The MFSI-SF-C is an effective and comprehensive tool for measuring fatigue in Chinese patients with cancer.


Evidence that the folate-dependent proteins YgfZ and MnmEG have opposing effects on growth and on activity of the iron-sulfur enzyme MiaB.
The folate-dependent protein YgfZ of Escherichia coli participates in the synthesis and repair of iron-sulfur (Fe-S) clusters; it belongs to a family of enzymes that use folate to capture formaldehyde units. Ablation of ygfZ is known to reduce growth, to increase sensitivity to oxidative stress, and to lower the activities of MiaB and other Fe-S enzymes. It has been reported that the growth phenotype can be suppressed by disrupting the tRNA modification gene mnmE. We first confirmed the latter observation using deletions in a simpler, more defined genetic background. We then showed that deleting mnmE substantially restores MiaB activity in ygfZ deletant cells and that overexpressing MnmE with its partner MnmG exacerbates the growth and MiaB activity phenotypes of the ygfZ deletant. MnmE, with MnmG, normally mediates a folate-dependent transfer of a formaldehyde unit to tRNA, and the MnmEG-mediated effects on the phenotypes of the ΔygfZ mutant apparently require folate, as evidenced by the effect of eliminating all folates by deleting folE. The expression of YgfZ was unaffected by deleting mnmE or overexpressing MnmEG or by folate status. Since formaldehyde transfer is a potential link between MnmEG and YgfZ, we inactivated formaldehyde detoxification by deleting frmA. This deletion had little effect on growth or MiaB activity in the ΔygfZ strain in the presence of formaldehyde, making it unlikely that formaldehyde alone connects the actions of MnmEG and YgfZ. A more plausible explanation is that MnmEG erroneously transfers a folate-bound formaldehyde unit to MiaB and that YgfZ reverses this.


Faking giants: the evolution of high prey clearance rates in jellyfishes.
Jellyfishes have functionally replaced several overexploited commercial stocks of planktivorous fishes. This is paradoxical, because they use a primitive prey capture mechanism requiring direct contact with the prey, whereas fishes use more efficient visual detection. We have compiled published data to show that, in spite of their primitive life-style, jellyfishes exhibit similar instantaneous prey clearance and respiration rates as their fish competitors and similar potential for growth and reproduction. To achieve this production, they have evolved large, water-laden bodies that increase prey contact rates. Although larger bodies are less efficient for swimming, optimization analysis reveals that large collectors are advantageous if they move through the water sufficiently slowly.


Natural zwitterionic organosulfurs as surface ligands for antifouling and responsive properties.
Natural sulfur-containing zwitterionic compounds, l-cysteine (Cys), l-methionine, and glutathionine (GSH), have been employed as surface ligands to prevent protein nonspecific adsorption on planar substrates. These organosulfur compounds form self-assembled monolayers (SAMs) on gold substrates by gold-sulfur interaction. The chemical elements of SAMs were confirmed using x-ray photoelectron spectroscopy. The surface wetting tests for SAMs show that films prepared from Cys and GSH exhibited super-hydrophilicity (contact angles of θ = ~5°) due to their high coverage and strong hydration via ionic solvation and formation of hydrogen bonding. Quartz crystal microbalance with dissipation sensor was used to quantitatively and qualitatively monitor the adsorption of bovine serum albumin (BSA) from buffer onto these SAMs. It was found that the GSH film enables the resistance of BSA adsorption to the best extent at a physiological pH. Moreover, the surface charges of modified substrates were modulated by varying the pH value to control BSA adsorption. The effect of electrostatic repulsion on the antifouling behavior becomes prominent at a pH where the protein and the surface carry same charges. Consolidating the BSA adsorption measurements at different pH values, the antifouling properties of GSH-modified Au should be attributed to prevention of entropy gain and enthalpy loss, making BSA adsorption energetically unfavorable. It is believed that the surface modification with natural organosulfur ligands holds great potential in improving the biocompatibility of medical devices and in offering intelligent biointerfaces in response to environmental stimuli.


Evaluation of Salmonella typhimurium mutants in a model of experimental gastroenteritis.
Salmonella typhimurium strains harboring independent, defined mutations in aroA, invA, ssrA, or msbB were assessed for their ability to induce fluid accumulation, tissue damage, and local inflammation in rabbit ileal loops. Three wild-type strains of S. typhimurium, TML, HWSH, and SL1344, and two mutant strains, S. typhimurium SL1344 ssrA and S. typhimurium SL1344 msbB, consistently induced fluid accumulation in the lumen of loops and inflammation of loop-associated tissues. In contrast, three different S. typhimurium aroA strains and an invA mutant of SL1344 did not induce significant fluid accumulation in the rabbit ileal loops. However, the S. typhimurium aroA strains did induce an inflammatory infiltrate and some local villus-associated damage, but the invA mutant did not. Histologically, wild-type S. typhimurium, S. typhimurium SL1344 ssrA, and S. typhimurium SL1344 msbB demonstrated more severe effects on villus architecture than S. typhimurium aroA strains, whereas S. typhimurium invA-infected loops showed no detectable damage. This suggests that villus damage most likely contributes to fluid accumulation within the loop.


Phase II study of R115777, a farnesyl transferase inhibitor, in myelodysplastic syndrome.
To perform a phase II study of the farnesyl transferase inhibitor R115777 (Zarnestra; Johnson and Johnson Pharmaceutical Research and Development, Raritan, NJ) in patients with myelodysplastic syndrome (MDS), using doses recommended in a phase I study in relapsed/refractory leukemia. Patients with MDS were treated with R115777 at doses of 600 mg orally (PO) bid in cycles of 4 weeks of therapy followed by a 2-week rest period. Dose reduction rules for toxicity were applied. Twenty-seven of the 28 patients treated were assessable. Three patients responded (complete remission, n = 2; partial remission, n = 1). Responders included two patients with refractory anemia with excess blasts and one patient with refractory anemia with excess blasts in transformation. Two of the responders had a diploid karyotype and one had multiple cytogenetic abnormalities including monosomy 5 and 7. The starting dose of 600 mg PO bid resulted in side effects (myelosuppression, fatigue, neurotoxicity, rash, or leg pain) necessitating dose reduction (n = 4) or discontinuation of therapy (n = 7) in 11 (41%) of 27 patients during the induction period (12 weeks). Lower doses of 300 mg PO bid were well tolerated. All responses occurred in patients who had been reduced to this dose level during the initial two cycles. This study suggests that R115777 has modest activity in MDS patients, but that, in this patient population, 4 weeks of daily doses of 600 mg PO bid is not tolerated. Further exploration of the optimal dose/schedule and correlation with biologic end points are warranted.