Effects of personal air pollutant exposure on oxidative stress: Potential confounding by natural variation in melatonin levels.
Urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) are commonly used biomarkers of oxidative stress. However, their associations with air pollutant exposure have not been consistent across studies. We hypothesize that the inconsistency is partly due to confounding of circulating melatonin. We analyzed urinary 6-sulfatoxymelatonin (aMT6s), a surrogate of circulating melatonin, along with 8-OHdG and MDA, in 159 healthy adults who had not taken melatonin supplementation. Within the natural range of endogenously-generated aMT6s (0.3-93.5 ng/mg creatinine) measured in this study, increasing aMT6s levels were significantly associated with increasing concentrations of 8-OHdG and MDA. Measurements of PM2.5, ozone (O3), and nitrogen dioxide (NO2), coupled with time-activity data, were used to calculate time-averaged personal exposures 12 -hour (12h) and 24-hour (24h) prior to urine collection. Without controlling for aMT6s, the relationships between pollutant exposure and 8-OHdG or MDA were not clear. After controlling for aMT6s, an interquartile range (IQR) increase in 12h PM2.5 and 12h NO2 exposure was associated with 6.1% [95%CI: 1.6%-10.8%] and 8.6% [1.3%-16.5%] increase in MDA, respectively. An IQR increase in 12h O3 exposure was associated with a 5.7% [1.9%-9.7%] in 8-OHdG. The findings suggest the need for controlling for aMT6s as a confounder in using urinary 8-OHdG and MDA as biomarkers of oxidative stress related to short-term air pollution exposure.


The effect of plating density on granulosa cell growth and differentiation in vitro.
The extent of FSH-mediated LH/hCG receptor induction and of basal and FSH-stimulated progesterone production by porcine granulosa cells in vitro, in serum-containing medium, is directly related to the plating density. Relative to pre-culture levels, low- and high-density cultures of cells routinely exhibited 1-2- and 10-11-fold increases in [125I]iodo-hCG binding, respectively. Monolayer growth, i.e. cell division, as measured by increases in cell protein or DNA content, was inversely related to plating density. This density-directed inverse relationship between growth and differentiation did not appear to be coupled under the conditions utilized. Whereas monolayer growth was dependent upon the cell surface density, i.e. the number of cells per unit surface area, differentiation was dependent upon cell concentration, i.e. cells per unit volume of medium. Cells plated at low density in medium containing 10% serum exhibited 50% less [125I]iodo-hCG binding than cells in 5% serum (P less than 0.025). Conversely, cells plated at high density exhibited a 14% increase (P less than 0.025) in binding at the higher serum level. Thus, it appears that the extent of differentiation depends upon the capacity of cells to neutralize serum inhibition which in turn is dependent upon the cell concentration. Serum neutralization by granulosa cells is an FSH-dependent process. Conditioned medium derived from insulin-treated, high-density cultures did not facilitate optimum LH/hCG receptor induction in low-density cultures. Conditioned medium from cultures treated with insulin plus FSH, however, facilitated LH/hCG receptor induction in low-density cultures to the same extent as obtained in high-density cultures. The enhancement by FSH-conditioned medium cannot be attributed to residual FSH or to dilution of serum components during the preparation of the conditioned medium. The phenomena of serum-attenuated granulosa cell differentiation in vitro, and of a density-dependent reversal of this process, may have regulatory implications in vivo since follicular fluid contains many serum components and since the granulosa cell complement is an important determinant of follicle maturation.


Effect of methylphenidate on neurocognitive test battery: an evaluation according to the diagnostic and statistical manual of mental disorders, fourth edition, subtypes.
The aims of this study were to evaluate the neuropsychological characteristics of the restrictive (R) subtype according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition and the attention-deficit/hyperactivity disorder (ADHD) combined (CB) type and predominantly inattentive (PI) type subtypes and to evaluate whether methylphenidate (MPH) affects neurocognitive test battery scores according to these subtypes. This study included 360 children and adolescents (277 boys, 83 girls) between 7 and 15 years of age who had been diagnosed with ADHD and compared the neuropsychological characteristics and MPH treatment responses of patients with the R subtype-which has been suggested for inclusion among the ADHD subtypes in the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition-with those of patients with the PI and CB subtypes. They did not differ from the control subjects in the complex attention domain, which includes Continuous Performance Test, Stroop test, and Shifting Attention Test, which suggests that the R subtype displayed a lower level of deterioration in these domains compared with the PI and CB subtypes. The patients with the CB and PI subtypes did not differ from the control subjects in the Continuous Performance Test correct response domain, whereas those with the R subtype presented a poorer performance than the control subjects. The R subtype requires a more detailed evaluation because it presented similar results in the remaining neuropsychological evaluations and MPH responses.


Sensitivity of fibroblast growth factor 23 measurements in tumor-induced osteomalacia.
Tumor-induced osteomalacia (TIO) is a paraneoplastic syndrome of hypophosphatemia, decreased renal phosphate reabsorption, normal or low serum 1,25-dihydryxyvitamin-D concentration, myopathy, and osteomalacia. Fibroblast growth factor 23 (FGF23) is a phosphaturic protein overexpressed in tumors that cause TIO and is, at least partly, responsible for the manifestations of TIO. The objective of this study was to determine the sensitivity of FGF23 measurements in TIO. FGF23 concentrations were measured on stored samples with three ELISAs. This study was conducted at subspecialty referral centers. Twenty-two patients with suspected TIO, 13 with confirmed tumors, were studied. There were no interventions in this study. FGF23 concentration was the main outcome measure of this study. Elevated FGF23 concentrations were detected using the Immunotopics C-terminal assay in 16 of 22 TIO patients (for a sensitivity of 73%), the Immunotopics Intact assay in five of 22 patients (sensitivity, 23%), and the Kainos Intact assay in 19 of 22 patients (sensitivity, 86%). In the 13 patients with confirmed tumors, the sensitivity was higher with all assays: 92% for the Immunotopics C-terminal assay, 38% for the Immunotopics Intact assay, and 100% for the Kainos assay. The Kainos Intact assay was the most sensitive, followed by the Immunotopics C-terminal assay. The findings of normal FGF23 concentrations in some patients with TIO may indicate that FGF 23 is not responsible for the hypophosphatemia in these patients or that FGF23 secretion by some tumors is partially responsive to serum phosphate. Normal FGF23 concentrations should be interpreted in relation to the serum phosphate and 1,25-dihydryxyvitamin-D concentrations.


The RCK1 domain of the human BKCa channel transduces Ca2+ binding into structural rearrangements.
Large-conductance voltage- and Ca(2+)-activated K(+) (BK(Ca)) channels play a fundamental role in cellular function by integrating information from their voltage and Ca(2+) sensors to control membrane potential and Ca(2+) homeostasis. The molecular mechanism of Ca(2+)-dependent regulation of BK(Ca) channels is unknown, but likely relies on the operation of two cytosolic domains, regulator of K(+) conductance (RCK)1 and RCK2. Using solution-based investigations, we demonstrate that the purified BK(Ca) RCK1 domain adopts an alpha/beta fold, binds Ca(2+), and assembles into an octameric superstructure similar to prokaryotic RCK domains. Results from steady-state and time-resolved spectroscopy reveal Ca(2+)-induced conformational changes in physiologically relevant [Ca(2+)]. The neutralization of residues known to be involved in high-affinity Ca(2+) sensing (D362 and D367) prevented Ca(2+)-induced structural transitions in RCK1 but did not abolish Ca(2+) binding. We provide evidence that the RCK1 domain is a high-affinity Ca(2+) sensor that transduces Ca(2+) binding into structural rearrangements, likely representing elementary steps in the Ca(2+)-dependent activation of human BK(Ca) channels.


Difficulty-related changes in inter-regional neural synchrony are dissociated between target and non-target processing.
The major purpose of this study was to explore the changes in the local/global gamma-band neural synchronies during target/non-target processing due to task difficulty under an auditory three-stimulus oddball paradigm. Multichannel event-related potentials (ERPs) were recorded from fifteen healthy participants during the oddball task. In addition to the conventional ERP analysis, we investigated the modulations in gamma-band activity (GBA) and inter-regional gamma-band phase synchrony (GBPS) for infrequent target and non-target processing due to task difficulty. The most notable finding was that the difficulty-related changes in inter-regional GBPS (33-35 Hz) at P300 epoch (350-600 ms) completely differed for target and non-target processing. As task difficulty increased, the GBPS significantly reduced for target processing but increased for non-target processing. This result contrasts with the local neural synchrony in gamma-bands, which was not affected by task difficulty. Another major finding was that the spatial patterns of functional connectivity were dissociated for target and non-target processing with regard to the difficult task. The spatial pattern for target processing was compatible with the top-down attention network, whereas that for the non-target corresponded to the bottom-up attention network. Overall, we found that the inter-regional gamma-band neural synchronies during target/non-target processing change significantly with task difficulty and that this change is dissociated between target and non-target processing. Our results indicate that large-scale neural synchrony is more relevant for the difference in information processing between target and non-target stimuli.


New Insights into the TIFY Gene Family of Brassica napus and Its Involvement in the Regulation of Shoot Branching.
As plant-specific transcription factors, the TIFY family genes are involved in the responses to a series of biotic and abiotic stresses and the regulation of the development of multiple organs. To explore the potential roles of the TIFY gene family in shoot branching, which can shape plant architecture and finally determine seed yield, we conducted comprehensive genome-wide analyses of the TIFY gene family in Brassica napus. Here, HMMER search and BLASTp were used to identify the TIFY members. A total of 70 TIFY members were identified and divided into four subfamilies based on the conserved domains and motifs. These TIFY genes were distributed across 19 chromosomes. The predicted subcellular localizations revealed that most TIFY proteins were located in the nucleus. The tissue expression profile analyses indicated that TIFY genes were highly expressed in the stem, flower bud, and silique at the transcriptional level. High-proportioned activation of the dormant axillary buds on stems determined the branch numbers of rapeseed plants. Here, transcriptome analyses were conducted on axillary buds in four sequential developing stages, that is, dormant, temporarily dormant, being activated, and elongating (already activated). Surprisingly, the transcription of the majority of TIFY genes (65 of the 70) significantly decreased on the activation of buds. GO enrichment analysis and hormone treatments indicated that the transcription of TIFY family genes can be strongly induced by jasmonic acid, implying that the TIFY family genes may be involved in the regulation of jasmonic acid-mediated branch development. These results shed light on the roles of TIFY family genes in plant architecture.


Involving patients to improve service quality in primary care.
A sample of 92 UK patients volunteered to take part in focus groups to discuss what elements of local primary care provision were important to them. Issues raised were prioritised by the patients and then fashioned into 18 quality indicators which nine local practices were invited to assess themselves against. At the assessment meeting three months later over 40 changes in service provision were noted in the nine participating practices. A patient questionnaire carried out in each practice, however, indicated a tendency for practices to overestimate the services they felt they provided. Patients rated the experience of generating standards as very worthwhile and enjoyed being asked. Further research needs to be carried out to assess the effectiveness of this methodology in different settings.


Differences in the acidic sensitivity of transient receptor potential vanilloid 1 (TRPV1) between chickens and mice.
Chickens, one of the most important industrial animals, are a biological animal model. Here we focused on the transient receptor potential vanilloid 1 (TRPV1) to understand the pain system for acidic stimuli in chickens compared with mice. By using a whole-cell patch clamp system, we confirmed that acidic stimuli activate both chicken TRPV1 (cTRPV1) and mouse TRPV1 (mTRPV1), but the peak current of cTRPV1 is lower than that of mTRPV1, and it is difficult to desensitize cTRPV1 with an acidic stimulus compared to mTRPV1. Since the C-terminal of the calmodulin (CaM) binding site in TRPV1 was reported as one of the important structures for TRPV1 desensitization, we made chimeric cTRPV1 in which the CaM binding site of chicken is changed to that of mouse (cTRPV1-mCaM). We also compared the acidic responses of native chicken dorsal root ganglion (DRG) cells with that of mouse DRG cells. The TRPV1-mCaM results showed that the desensitization of mutant cTRPV1 was similar to that of mTRPV1, and that the basal activities of mutant cTRPV1 were significantly higher than those of cTRPV1. It was also difficult to desensitize the chicken DRG cells with an acidic stimulus, unlike the mouse DRG cells. These results suggest that there are differences in the pain transduction systems for acidic stimuli between chickens and mice that are caused by the dysfunction of the C-terminal CaM biding site of cTRPV1. These results imply that chickens repeatedly feel weak pain from an acidic stimulus, without desensitization.


Dendritic cell subtypes from lymph nodes and blood show contrasted gene expression programs upon Bluetongue virus infection.
Human and animal hemorrhagic viruses initially target dendritic cells (DCs). It has been proposed, but not documented, that both plasmacytoid DCs (pDCs) and conventional DCs (cDCs) may participate in the cytokine storm encountered in these infections. In order to evaluate the contribution of DCs in hemorrhagic virus pathogenesis, we performed a genome-wide expression analysis during infection by Bluetongue virus (BTV), a double-stranded RNA virus that induces hemorrhagic fever in sheep and initially infects cDCs. Both pDCs and cDCs accumulated in regional lymph nodes and spleen during BTV infection. The gene response profiles were performed at the onset of the disease and markedly differed with the DC subtypes and their lymphoid organ location. An integrative knowledge-based analysis revealed that blood pDCs displayed a gene signature related to activation of systemic inflammation and permeability of vasculature. In contrast, the gene profile of pDCs and cDCs in lymph nodes was oriented to inhibition of inflammation, whereas spleen cDCs did not show a clear functional orientation. These analyses indicate that tissue location and DC subtype affect the functional gene expression program induced by BTV and suggest the involvement of blood pDCs in the inflammation and plasma leakage/hemorrhage during BTV infection in the real natural host of the virus. These findings open the avenue to target DCs for therapeutic interventions in viral hemorrhagic diseases.


Endocrine differentiation of gastric adenocarcinoma. The prevalence as evaluated by immunoreactive chromogranin A and its biologic significance.
The prevalence of endocrine differentiation of conventional gastric adenocarcinoma was evaluated on the 212 cases (including 62 mucosal carcinomas) of consecutively resected stomach for adenocarcinoma in our hospital using anti-chromogranin A (CGA) antibodies. CGA-positive cells were found in 28 of 150 cases (18.7%) as an integral tumor component. In immunocytochemistry and electron microscopic examinations, we could classify these 28 cases into three groups according to the distribution patterns of CGA-positive cells. The first group consisted of 12 cases in which scattered CGA-positive cells were located in neoplastic glands. The second group consisted of six cases of scirrhous carcinoma in which CGA-positive cells were separated by fibrovascular tissue. The third group consisted of ten cases in which the positive cells were present in clusters. No definite correlation was recognized between the appearance of CGA cells and histologic types of predominance. In the analysis of the hormonal substances coexpressed by CGA-positive cells, immunoreactive serotonin (SER) was found most frequently, and somatostatin (SS), gastrin (GAS), glucagon/glicentin (GLU/GLI), and peptide-tyrosine-tyrosine (PYY) like immunoreactivities were found in a few tumor cells. CGA-positive cells occupied limited parts of the tumors in most cases, and they were noticeably more frequent in advanced stage cases. This might explain why endocrine differentiation reflects the dysexpression of the neoplastic stem cells. Furthermore, absence of mitotic figures in this type of cell and negativity of a single colony composed exclusively of CGA cells in metastatic foci suggested that these cells are in a dormant phase and are probably postmitotic.


An evidence-based review of the literature on the consequences of conservative versus aggressive discectomy for the treatment of primary disc herniation with radiculopathy.
It remains unknown whether aggressive disc removal with curettage versus conservative removal of a disc fragment with little disc invasion provides a better outcome for the treatment of lumbar disc herniation with radiculopathy. Determine the level of evidence within the clinical literature that supports the performance of a conservative versus aggressive technique for discectomy. Systematic evidence-based review of clinical literature. Patients with primary lumbar disc herniation with radiculopathy. Operative time, return to work status, recurrent disc herniation, self-reported, and functional measures assessed less than 2 years (short term) and greater than 2 years (long term) after surgery. Systematic Medline search was performed to identify all published studies relating to outcome after aggressive or conservative discectomy. Levels of evidence (I-V) were assessed for each study and grades of recommendation were generated (Good, Fair, Poor, Insufficient evidence) based on the NASS Clinical Guidelines' Levels of Evidence and Grades of Recommendation. There is fair evidence that conservative discectomy will result in shorter operative times and a quicker return to work despite similar lengths of hospital stay, similar pain levels at discharge, similar 6-month functional status, and a similar 2-year incidence of persistent/recurrent back and leg pain. There is poor quality evidence that conservative discectomy will result in a lower incidence of recurrent back pain beyond 2 years postoperatively. There is fair quality evidence that conservative discectomy will result in a higher incidence of recurrent disc herniation. There are no Level I studies to support conservative versus aggressive discectomy for the treatment of primary disc herniation. However, systematic review of the literature suggests that conservative discectomy may result in shorter operative time, quicker return to work, and a decreased incidence of long-term recurrent low back pain but with an increased incidence of recurrent disc herniation. Prospective randomized trails are needed to firmly assess this possible benefit.


Systematic review of cost-effectiveness analyses for combinations of prevention strategies against human papillomavirus (HPV) infection: a general trend.
Due to the arrival of multi-valent HPV vaccines, it is more and more important to have a better understanding of the relationship between vaccination and screening programmes. This review aimed to: (1) collect published evidence on the cost-effectiveness profile of different HPV prevention strategies and, in particular, those combining vaccination with changes in screening practices; (2) explore the cost-effectiveness of alternative preventive strategies based on screening and vaccination. A systematic literature review was conducted in order to identify the relevant studies regarding the cost-effectiveness of prevention strategies against HPV infection. Analysis comparing the modelling approaches between studies was made along with an assessment of the magnitude of impact of several factors on the cost-effectiveness of different screening strategies. A total of 18 papers were quantitatively summarised within the narrative. A high degree of heterogeneity was found in terms of how HPV prevention strategies have been assessed in terms of their economic and epidemiological impact, with variation in screening practice and valence of HPV vaccination found to have large implications in terms of cost-effectiveness. This review demonstrated synergies between screening and vaccination. New prevention strategies involving multi-valence vaccination, HPV DNA test screening, delayed commencement and frequency of screening could be implemented in the future. Strategies implemented in the future should be chosen with care, and informed knowledge of the potential impact of all possible prevention strategies. Highlighted in this review is the difficulty in assessing multiple strategies. Appropriate modelling techniques will need to be utilised to assess the most cost-effective strategies.


Biologic stability of plasma ion-implanted miniscrews.
To gain basic information regarding the biologic stability of plasma ion-implanted miniscrews and their potential clinical applications. Sixteen plasma ion-implanted and 16 sandblasted and acid-etched (SLA) miniscrews were bilaterally inserted in the mandibles of 4 beagles (2 miniscrews of each type per quadrant). Then, 250 - 300 gm of force from Ni-Ti coil springs was applied for 2 different periods: 12 weeks on one side and 3 weeks contralaterally. Thereafter, the animals were sacrificed and mandibular specimens including the miniscrews were collected. The insertion torque and mobility were compared between the groups. The bone-implant contact and bone volume ratio were calculated within 800 µm of the miniscrews and compared between the loading periods. The number of osteoblasts was also quantified. The measurements were expressed as percentages and analyzed by independent t-tests (p < 0.05). No significant differences in any of the analyzed parameters were noted between the groups. The preliminary findings indicate that plasma ion-implanted miniscrews have similar biologic characteristics to SLA miniscrews in terms of insertion torque, mobility, bone-implant contact rate, and bone volume rate.


Who is Publishing in Facial Cosmetic Surgery? A Citation Analysis Across Specialties Over Five Decades.
Competition among our sister cosmetic specialties continues to increase. Once a field dominated by plastic surgeons, there is a clear trend toward increased competition from core and non-core disciplines. While these marketplace trends are obvious, how such competition has affected academia or peer reviewed publications is less clear. We analyzed the most cited peer reviewed facial aesthetic literature over the past five decades to see if marketplace trends are echoed in a similar manner across the academic disciplines of plastic surgery, otolaryngology, dermatology, and ophthalmology. The top 50 cited articles for each decade from the 1970s to the 2010s were identified for the topics of facelift, rhinoplasty, browlift, and blepharoplasty using the Thomson/Reuters Web of Knowledge. Data collected were: the number of citations/article, first authors' specialty affiliation, and journal specialty affiliation. Data were plotted graphically and trends were analyzed. With regards to first authorship, plastic surgery had the highest percentage across all surgeries at every time point, except for rhinoplasty from 2010-present, when otolaryngology had a higher percentage (48% vs 40%). Observed trends demonstrated: (1) increasing contributions from otolaryngology in rhinoplasty, facelift, and browlift; and (2) increasing contributions from ophthalmology in blepharoplasty. Plastic surgery journals are the most common platform for publication across all four surgeries. Plastic surgeons, and plastic surgery journals, still remain a strong force in academic facial cosmetic surgery. However, it appears that the competition from non-plastic surgeons observed in clinical practice is being mirrored in the area of journal publications. We as a specialty need to continue to strive for high quality academic productivity.


Obligatory coupling of colicin release and lysis in mitomycin-treated Col+ Escherichia coli.
Previous work has shown that Escherichia coli K12 strains carrying the small, high copy number ColE2-P9 plasmid produce large amounts of colicin and then lyse and release colicin when grown in broth culture containing mitomycin C. Strains carrying the larger, low copy number ColIa-CA53 plasmid produced much less colicin and did not lyse or discharge more than 15% of their colicin when grown under the same conditions. Naturally-occurring Col+ strains and E. coli K12 derivatives carrying different Col plasmids could be classified either as ColE2+-like or ColIa+-like according to whether or not they produced large amounts of colicin and lysed and discharged colicin when grown in the presence of mitomycin, and also by the size and presumed copy number of the Col plasmid they carried. Strains carrying multiple copies of the cloned colicin Ia structural gene produced large amounts of colicin but did not lyse or release colicin when grown in the presence of mitomycin. This result rules out the possibility that high level accumulation of colicin is sufficient to cause lysis. Conditions were sought under which colicin Ia could be released from the producing cells. It was found that mitomycin-treated cultures of strains carrying both ColE2 and ColIa plasmids released both colicins when they lysed, although colicin Ia release occurred later than colicin E2 release. It was also noted that colicin Ia-laden cells released their colicin when diluted into fresh culture medium.


3-aryl-4-(3,4,5-trimethoxyphenyl)pyridines inhibit tubulin polymerisation and act as anticancer agents.
A series of cis-restricted 3-aryl-4-(3,4,5-trimethoxyphenyl)pyridines as novel tubulin polymerisation inhibitors was designed based on molecular docking. Compound 9p, exhibited potent antiproliferative activity against HeLa, MCF-7, and A549 cell lines. Mechanism studies indicated that 9p potently inhibited tubulin polymerisation and disrupted the microtubule dynamics of tubulin in HeLa cells. Moreover, 9p could cause G2/M phase cell cycle arrest and apoptosis in HeLa cells. In addition, the prediction of physicochemical properties disclosed that 9p conformed well to the Lipinski's rule of five. The initial results suggest that the 3-aryl-4-(3,4,5-trimethoxyphenyl)pyridines could serve as a promising scaffold for the development of novel anticancer drugs.


Familial Atypical Hemolytic Uremic Syndrome with Positive p.S1191L (c.3572C>T) Mutation on the CFH Gene: A Single-center Experience.
The atypical hemolytic uremic syndrome (aHUS) is characterized by thrombocytopenia, microangiopathic hemolytic anemia and acute kidney injury (AKI), which can exhibit a poor prognosis. Complement factor H (CFH) gene mutations play a key role in this disease, which may be sporadic or familial. We studied 13 people from the same family, investigated for gene mutations of the familial aHUS after a family member presented to our emergency clinic with the aHUS and reported a family history of chronic renal failure. The p.S1191L mutation on the CFH gene was heterozygous in six people from the patient's family with the aHUS. One of these family members is our patient with acute kidney injury, and the other two are followed at the Nephrology Clinic, Medeniyat University, Goztepe Training and Research Hospital, Istanbul, Turkey, due to chronic renal failure. The other three family members showed no evidence of renal failure. The index case had a history of six sibling deaths; three died of chronic renal failure. Plasmapheresis and fresh frozen plasma treatment were administered to our patient. When the patient showed no response to this treatment, eculizumab (ECZ) therapy was started. The study demonstrated that thorough family history should be taken in patients with the aHUS. These patients may have the familial type of the disease, and they should be screened genetically. Eculizumab should be the first choice in the treatment with plasmapheresis. It should be kept in mind that the use of ECZ as prophylaxis in posttransplant therapy is extremely important for preventing rejection.


The student experience of applied equivalence-based instruction for neuroanatomy teaching.
Esoteric jargon and technical language are potential barriers to the teaching of science and medicine. Effective teaching strategies which address these barriers are desirable. Here, we created and evaluated the effectiveness of stand-alone 'equivalence-based instruction' (EBI) learning resources wherein the teaching of a small number of direct relationships between stimuli (e.g., anatomical regions, their function, and pathology) results in the learning of higher numbers of untaught relationships. We used a pre and post test design to assess students' learning of the relations. Resources were evaluated by students for perceived usefulness and confidence in the topic. Three versions of the resources were designed, to explore learning parameters such as the number of stimulus classes and the number of relationships within these classes. We show that use of EBI resulted in demonstrable learning of material that had not been directly taught. The resources were well received by students, even when the quantity of material to be learned was high. There was a strong desire for more EBI-based teaching. The findings are discussed in the context of an ongoing debate surrounding 'rote' vs. 'deep' learning, and the need to balance this debate with considerations of cognitive load and esoteric jargon routinely encountered during the study of medicine. These standalone EBI resources were an effective, efficient and well-received method for teaching neuroanatomy to medical students. The approach may be of benefit to other subjects with abundant technical jargon, science and other areas of medicine.


Transduction of cultured fish cells with recombinant baculoviruses.
Five fish cell lines were tested for their ability to be transduced by Ac-CAlacZ, a recombinant baculovirus that is capable of expressing a beta-galactosidase reporter gene from the CAG promoter (consisting of a cytomegalovirus enhancer element, a chicken actin promoter and rabbit beta-globin termination sequences). TO (Tilapia ovary), EPC (carp), CHH-1 (Chum salmon heart fibroblast) and CHSE-214 (chinook salmon embryo) cells were transducible, as demonstrated by an in situ beta-galactosidase assay, whereas RTG-2 (rainbow trout gonad) cells were not. The EPC cell line was used for more detailed studies on baculovirus transduction. The transduction frequency was found to be higher at 28 degrees C than at 21 degrees C. Addition of the histone deacetylase inhibitor sodium butyrate increased the number of blue cells detected 5- to 7-fold. The m.o.i. was positively correlated with transduction frequency, although the relationship did not appear to be strictly linear, as has been observed with mammalian cells. The temperature at which baculoviruses were adsorbed to EPC cells did not affect levels of beta-galactosidase expression. We also examined expression levels of beta-galactosidase in EPC cells after infection with a baculovirus construct that overexpresses the vesicular stomatitis virus G protein and displays it on the virion surface. Expression levels with this virus were approximately 15-fold higher than were observed with Ac-CAlacZ.


Dissolution and redistribution of trace elements and nutrients during dredging of iron monosulfide enriched sediments.
The increased use of estuarine waters for commercial and recreational activities is one consequence of urbanisation. Western Australia's Peel-Harvey Estuary highlights the impacts of urbanisation, with a rapidly developing boating industry and periodic dredging activity. The aim of this research is to evaluate the potential mobility of nutrients and trace elements during dredging, and the influence of flocculation on iron and sulfur partitioning in iron monosulfide enriched sediments. Our findings indicate a short-term increase in nitrate, phosphate and ammonium, during dredging through the resuspension of sediments. However, no increase in metal mobilisation during dredging was observed except copper (Cu) and zinc (Zn). Flocculant addition increased the release of nutrients, zinc (Zn) and arsenic (As) from sediments, had no effect on acid volatile sulfides and pyritic sulfur, but corresponded with an initial sharp rise in elemental sulfur concentrations. The run-off water from geofabric bags should be treated to decrease the concentrations of Zn and As to their background levels before releases into the estuary. Long-term impact of dredging on organic matter mineralisation and its subsequent effect on nutrients and trace elements dynamics needs further investigation.


Subepithelial fibrosis after myopic epikeratoplasty. Report of a case.
A 29-year-old man with a -22.00-diopter myopia in the right eye underwent a planar, nonfreeze myopic epikeratoplasty and postoperatively developed a central subepithelial opacity. The opacity recurred after two superficial keratectomies; finally, another epikeratoplasty was performed. Light microscopic examination of the original button showed that the epithelium was thickened, collagenous subepithelial fibrocellular tissue was present, and Bowman's layer was focally disrupted. The stroma was edematous and peripherally scarred with scattered keratocytes. Examination by transmission electron microscopy revealed keratocytes lying anterior and posterior to Bowman's layer. The subepithelial fibrocellular tissue corresponded to an area of a break in Bowman's layer and probably originated from the donor stroma. The break in Bowman's layer may have been caused by the microkeratome cuts, but its origin is not certain.


Magic-angle spinning solid-state NMR spectroscopy of the beta1 immunoglobulin binding domain of protein G (GB1): 15N and 13C chemical shift assignments and conformational analysis.
Magic-angle spinning solid-state NMR (SSNMR) studies of the beta1 immunoglobulin binding domain of protein G (GB1) are presented. Chemical shift correlation spectra at 11.7 T (500 MHz 1H frequency) were employed to identify signals specific to each amino acid residue type and to establish backbone connectivities. High sensitivity and resolution facilitated the detection and assignment of every 15N and 13C site, including the N-terminal (M1) 15NH3, the C-terminal (E56) 13C', and side-chain resonances from residues exhibiting fast-limit conformational exchange near room temperature. The assigned spectra lend novel insight into the structure and dynamics of microcrystalline GB1. Secondary isotropic chemical shifts report on conformation, enabling a detailed comparison of the microcrystalline state with the conformation of single crystals and the protein in solution; the consistency of backbone conformation in these three preparations is the best among proteins studied so far. Signal intensities and line widths vary as a function of amino acid position and temperature. High-resolution spectra are observed near room temperature (280 K) and at <180 K, whereas resolution and sensitivity greatly degrade substantially near 210 K; the magnitude of this effect is greatest among the side chains of residues at the intermolecular interface of the microcrystal lattice, which we attribute to intermediate-rate translational diffusion of solvent molecules near the glass transition. These features of GB1 will enable its use as an excellent model protein not only for SSNMR methods development but also for fundamental studies of protein thermodynamics in the solid state.


[Ionotropic receptors of acetylcholine and ATP: the mechanisms of inhibition].
In this review two kinds of receptors connected with ionic channels are analyzed. First kind is the receptor whose channel can be open with acetylcholine; they a named ionotropic cholinoreceptors (AChRn). The second kind is the receptor whose channel can be open with extracellular ATP. They a named ionotropic purinoreceptors and are designated P2X. Mathematical modelling of the kinetics of interactions with agonists and inhibitors of cholinergic and purine receptors has shown that each of the investigated mechanisms of inhibition of postsynaptic currents (competitive block, channel block, allosteric modulation and acceleration of desensitization) has different influence on the basic characteristics of postsynaptic currents. For unambiguous classification of inhibitory substances according to the molecular mechanisms of their action it is necessary to consider: a trend and size of change under inhibitory action of a decay time constant of the agonist-induced currents; dependence of development of changes and of speed of washing out of substance's effect from duration of its action, from number of activations of receptors and its frequency.


UV-induced mutations in epidermal cells of mice defective in DNA polymerase η and/or ι.
Xeroderma pigmentosum variant (XP-V) is a human rare inherited recessive disease, predisposed to sunlight-induced skin cancer, which is caused by deficiency in DNA polymerase η (Polη). Polη catalyzes accurate translesion synthesis (TLS) past pyrimidine dimers, the most prominent UV-induced lesions. DNA polymerase ι (Polι) is a paralog of Polη that has been suggested to participate in TLS past UV-induced lesions, but its function in vivo remains uncertain. We have previously reported that Polη-deficient and Polη/Polι double-deficient mice showed increased susceptibility to UV-induced carcinogenesis. Here, we investigated UV-induced mutation frequencies and spectra in the epidermal cells of Polη- and/or Polι-deficient mice. While Polη-deficient mice showed significantly higher UV-induced mutation frequencies than wild-type mice, Polι deficiency did not influence the frequencies in the presence of Polη. Interestingly, the frequencies in Polη/Polι double-deficient mice were statistically lower than those in Polη-deficient mice, although they were still higher than those of wild-type mice. Sequence analysis revealed that most of the UV-induced mutations in Polη-deficient and Polη/Polι double-deficient mice were base substitutions at dipyrimidine sites. An increase in UV-induced mutations at both G:C and A:T pairs associated with Polη deficiency suggests that Polη contributes to accurate TLS past both thymine- and cytosine-containing dimers in vivo. A significant decrease in G:C to A:T transition in Polη/Polι double-deficient mice when compared with Polη-deficient mice suggests that Polι is involved in error-prone TLS past cytosine-containing dimers when Polη is inactivated.


[A case of lupus cystitis in a 74-year-old woman].
The patient was a 74-year-old woman. As the history of the present illness, Raynaud's phenomenon appeared in 1998, antinuclear antibody positivity was detected in 2002, and she visited our department for the first time. Leukopenia and positivity for anti-DNA and anti-RNP antibodies were present, but active lesions were not, and thus, course observation was selected. Pollakiuria and a sensation of residual urine appeared in February 2005, diarrhea and nausea developed in November, and she was admitted to our hospital. Abdominal CT detected bilateral hydronephrosis, marked hydroureter, and hypertrophy of the urinary bladder wall, cystoscopy detected trabeculation, and features of interstitial cystitis were noted on biopsy. Edematous colon mucosa was noted on lower endoscopy, submucosal inflammatory cell infiltration on biopsy, and IgG deposition in the small vascular wall on immunostaining. Systemic lupus erythematosus (SLE) that developed as lupus cystitis was diagnosed. The clinical findings were improved by 50 mg of prednisolone. Although she developed lupus cystitis at an elderly age of 74 years, IgG deposition in the small vascular wall was detected by immunostaining of the intestinal mucosa. It is a valuable case proved that causative disease of a digestive tract symptom was enterocolitis through an immune complex as autoimmune reaction by SLE immunohistologically. There are 46 cases of lupus cystitis in Japan by 2007 since Kato reported lupus cystitis in 1985. We summarize clinical features of 46 cases and discuss difference with this case.


Drug use, homelessness and health: responding to the opioid overdose crisis with housing and harm reduction services.
Canada is in the midst of an opioid overdose crisis and Alberta has one of the highest opioid use rates across the country. Populations made vulnerable through structural inequities who also use opioids, such as those who are unstably housed, are at an increased risk of experiencing harms associated with opioid use. The main purpose of this study was to explore if there was an association between unstable housing and hospital use for people who use opioids. Analysis utilized self-reported data from the Alberta Health and Drug Use Survey which surveyed 813 Albertans in three cities. Hospital use was modeled using a logistic regression with our primary variable of interest being housing unstable status. Chi square tests were conducted between hospital use and variables associated with demographics, characteristics of drug use, health characteristics, and experiences of receiving services to establish model inclusion. Results revealed a significant association between housing instability and hospital use with unstably housed individuals twice as likely torequire hospital care. Results highlight the importance of concurrently addressing housing instability alongside the provision of harm reduction services such as safe supply and supervised consumption sites. These findings have significant implications for policy and policymakers during the opioid overdose epidemic, and provide a foundation for future areas of research.


Toward a Quantitative Colorimeter for Point-of-Care Nitrite Detection.
This paper reports on a low-cost, quantitative, point-of-care solution for the early detection of nitrite, a common biomarker for urinary tract infections (UTIs). In a healthy individual, nitrite is not found in the urine. However, a subject with a suspected UTI will produce nitrite in their urine since the bacteria present will convert nitrate into nitrite. Traditionally, nitrite is monitored by urinary dipsticks that are either read by eye or using a reflectance spectrophotometer. Both methods provide a semiquantitative positive or negative result at best. In this paper, we described a novel, affordable, portable transmission-based colorimeter for the quantitative measurement of nitrite. A unique permutation of the Griess reaction was optimized for the clinical detection of nitrite in urine and is reported. By using nitrite spiked in a salt buffer, artificial, and human urine samples, the performance of the colorimeter was evaluated against dipsticks read using two commercial dipstick analyzers, Urisys 1100 (Roche Diagnostics) and Clinitek Status+ (Siemens Medical Solutions). The colorimeter was able to detect the clinically relevant range of nitrite from 0.78 to 200 μM in a salt buffer. The detection limit in artificial urine was determined as 1.6 μM, which is ∼16× more sensitive than commercial dipstick reflectance analyzers, enabling the possibility for earlier detection of urinary infections. The colorimeter is assembled using off-the-shelf components (<$80) and controlled by a smartphone application via low-energy bluetooth. It has a built-in color correction algorithm and is designed to enable for a turbidity correction in samples containing bacteria or other cellular debris as well. The mobile application can display the nitrite concentration for a single sample or display the results over a period of time. Tracking urinalysis results longitudinally can help identify trends such as increases in nitrite concentrations over an individual's baseline and identify possible infections earlier. While the detection of nitrite was showcased here, this portable analyzer can be expanded to other colorimetric-based chemistries to detect a panel of biomarkers, which can improve the overall sensitivity and specificity of the desired assay.


1CPN: A coarse-grained multi-scale model of chromatin.
A central question in epigenetics is how histone modifications influence the 3D structure of eukaryotic genomes and, ultimately, how this 3D structure is manifested in gene expression. The wide range of length scales that influence the 3D genome structure presents important challenges; epigenetic modifications to histones occur on scales of angstroms, yet the resulting effects of these modifications on genome structure can span micrometers. There is a scarcity of computational tools capable of providing a mechanistic picture of how molecular information from individual histones is propagated up to large regions of the genome. In this work, a new molecular model of chromatin is presented that provides such a picture. This new model, referred to as 1CPN, is structured around a rigorous multiscale approach, whereby free energies from an established and extensively validated model of the nucleosome are mapped onto a reduced coarse-grained topology. As such, 1CPN incorporates detailed physics from the nucleosome, such as histone modifications and DNA sequence, while maintaining the computational efficiency that is required to permit kilobase-scale simulations of genomic DNA. The 1CPN model reproduces the free energies and dynamics of both single nucleosomes and short chromatin fibers, and it is shown to be compatible with recently developed models of the linker histone. It is applied here to examine the effects of the linker DNA on the free energies of chromatin assembly and to demonstrate that these free energies are strongly dependent on the linker DNA length, pitch, and even DNA sequence. The 1CPN model is implemented in the LAMMPS simulation package and is distributed freely for public use.


Solubilization and partial purification of the thiazide diuretic receptor from rabbit renal cortex.
This study was designed to solubilize, characterize and begin to purify the thiazide-sensitive Na/Cl transporter from mammalian kidney. Metolazone, a thiazide-like diuretic drug, binds to receptors in rat renal cortex closely related to the thiazide-sensitive Na/Cl transport pathway of the renal distal tubule. In the current study, [3H]metolazone bound to receptors in rabbit renal cortical microsomes. The portion of [3H]metolazone binding that was inhibited by hydrochlorothiazide reflected binding to a high-affinity class of receptor. The affinity (Kd 2.0 +/- 0.1 nM) and number (Bmax = 0.9 +/- 0.4 pmol/mg protein) of high-affinity receptors in rabbit renal cortical membranes were similar to values reported previously for rat. When proximal and distal tubule fragments were separated by Percoll gradient centrifugation, receptors were restricted to the fraction that contained distal tubules. When compared with cortical homogenates, receptor density was enriched 12-fold by magnesium precipitation and differential centrifugation. The zwitterionic detergent CHAPS solubilized 25-35% of the receptors (at 6 mM). Chloride inhibited and Na stimulated binding of [3H]metolazone to solubilized high-affinity receptors. The receptors could be purified significantly by hydroxyapatite chromatography and size exclusion high performance liquid chromatography (HPLC). The combination of magnesium precipitation and differential centrifugation, hydroxyapatite chromatography, and size exclusion HPLC resulted in a 213-fold enrichment of receptors, compared to renal cortical homogenate. The current results indicate that thiazide receptors from rabbit kidney share characteristics with receptors from rat, and that rabbit receptors can be solubilized in CHAPS and purified significantly by hydroxyapatite chromatography and size exclusion HPLC.


Dynamic modeling of genes controlling cancer stem cell proliferation.
The growing evidence that cancer originates from stem cells (SC) holds a great promise to eliminate this disease by designing specific drug therapies for removing cancer SC. Translation of this knowledge into predictive tests for the clinic is hampered due to the lack of methods to discriminate cancer SC from non-cancer SC. Here, we address this issue by describing a conceptual strategy for identifying the genetic origins of cancer SC. The strategy incorporates a high-dimensional group of differential equations that characterizes the proliferation, differentiation, and reprogramming of cancer SC in a dynamic cellular and molecular system. The deployment of robust mathematical models will help uncover and explain many still unknown aspects of cell behavior, tissue function, and network organization related to the formation and division of cancer SC. The statistical method developed allows biologically meaningful hypotheses about the genetic control mechanisms of carcinogenesis and metastasis to be tested in a quantitative manner.


Degeneration of hippocampal fibers and spatial memory deficit in the aged rat.
Old and young Fisher 344 rats were compared for their ability to learn a delayed alternation task. The old animals displayed significant impairment of alternation learning, and were slower than the young animals. The brains of these animals were examined using a silver degeneration stain, and among old rats there was conspicuous degeneration. The greatest density of degenerating fibers was seen in the hippocampus and in anatomically related tracts, but there was substantial fiber staining in the corpus callosum, anterior commissure, and internal capsule. Examination of the young brains revealed only an occasional fiber. There were no signs of cortical atrophy in the old animals. The histopathology of the aged animals' hippocampus and fiber tracts supports the possibility that the delayed alternation impairment shown by these animals was a result of age related degenerative changes.


[Serotype profile and antibiotyping of Streptococcus pneumoniae strains isolated from nasal carriage in pediatric patients].
The aim of this study was to determine the serotype profile and antibiotyping of Streptococcus pneumoniae from nasal isolates in children. Nasopharyngeal swab samples obtained from 125 children were cultivated and screened for the presence of S. pneumoniae. Strains were identified according to standard microbiological methods. The isolates were serotyped by the Quellung reaction and resistance patterns were determined by the microdilution method according to NCCLS guidelines. Results indicate an overall pneumococcal carriage rate of 24% (n = 30). The most commonly isolated serotypes were 23F (20%), 6B (20%) and 14 (13%). 73% of isolates were resistant at least to one of the tested antibiotics. 47% of the strains were consistently resistant to penicillin and the serotypes 6B and 23F were frequently associated with this marker. 60% of the strains were resistant to doxycycline, 37% to trimethoprim/sulfamethoxazole and clindamycin, 30% to erythromycin, 23% to cloramphenicol, 7% to ceftriaxone and 3% to cefepime. All strains were sensible to ofloxacin, rifampin and vancomycin. The most common combined resistance patterns were PNC-ERI-TMP/SMX-DOX-CLO-CLI (16.59%) and PNC-DOX (13.27%). The results obtained in this study will allow to orient the empiric therapy for pneumococcal infections and a rational use of antibiotics in clinical practice, as well as the application of an appropriate vaccination program specially adapted to the serotypes more frequently found in children.


A novel primary antibiotic cement-coated locking plate as a temporary fixation for the treatment of open tibial fracture.
Complex lower extremity trauma reconstruction remains a challenge. This study used an internal fixation composite structure of antibiotic cement plates as a temporary fixation to treat lower extremity Grade III open fractures; thus, reducing the treatment period and complications of external fixation. We aimed to assess the safety and efficacy of this technique in the initial surgery stage. Between January 2018 and March 2021, 20 patients with Gustilo grade IIIB/C open fractures received an antibiotic cement-coated locking plate as a temporary internal fixator during initial surgery. Thorough debridement and temporary internal fixation were performed with a 3.5-mm system antibiotic cement-coated locking plate. Ten patients required free bone fragment removal, followed by bone cement packing. The final stage involved internal fixation and wound repair with a free anterolateral thigh flap. Clinical and imaging results were retrospectively analysed. The repair time ranged 1-7 days. All flaps survived. Two patients experienced wound infection, and one developed severe bone infection 3 months after three-stage bone graft surgery. Autologous cancellous bone grafting was performed on 10 patients with bone defects 6 weeks after surgery. Bone union was universally achieved after 1 year. This method proved safe and effective, successfully repairing Grade III open fractures of the lower extremity 1-7 days post-treatment.


A systematic analysis of misleading evidence in unsafe rulings in England and Wales.
Evidence has the potential to be misleading if its value when expressing beliefs in hypotheses is not fully understood or presented. Although the knowledge base to understand uncertainties is growing, a challenge remains to prioritise research and to continuously assess the magnitude and consequences of misleading evidence in criminal cases. This study used a systematic content analysis to identify misleading evidence, drawing information from case transcripts of rulings argued unsafe by the Court of Appeal of England and Wales. In the 7-year study period, 218 applications were successful on appeal, containing 235 cases of misleading evidence. The majority (76%) of successful appeals were based upon the same materials available in the original trial, rather than the presentation of new relevant information. Witness (39%), forensic (32%), and character evidence (19%) were the most commonly observed evidence types, with the validity of witnesses (26%), probative value of forensic evidence (12%), and relevance of character evidence (10%) being the most prevalent combinations of identified issues. Additionally, the majority (66%) of misleading evidence types relate to their interpretation at activity level. The findings suggest that many of these misleading aspects could have been prevented by providing more transparency in the relationship between evidence and hypotheses. Generally, the results contribute to gaining a more complete picture of the role of misleading evidence in the criminal justice system.


Inferences about mental states.
Human social cognition relies on an ability to predict what others will think, feel or do in novel situations. Research in social neuroscience has consistently observed several brain regions that contribute ubiquitously to these abilities, including medial prefrontal cortex and aspects of lateral and medial parietal cortex. Interestingly, parallel work has suggested that this same network of regions subserves several seemingly distinct phenomena-notably, the abilities to remember the past, imagine the future and visualize spatial layouts-suggesting the existence of a common set of cognitive processes devoted to projecting oneself into worlds that differ mentally, temporally or physically from one's current experience. This use of self-projection to understand others' minds requires perceivers to solve three distinct cognitive challenges: (i) generating a simulated facsimile of one's own hypothetical mental states in a given situation, (ii) suppressing one's own current mental states, and (iii) deciding on the appropriateness of simulated states for understanding a particular other person. The present paper reviews recent psychology and neuroscience research aimed at understanding the underlying mechanisms that allow humans to solve each of these cognitive challenges to use self-projection to predict and understand the mental states of others.


Initial conditions influence the characteristics of ballistic contractions in the ankle dorsiflexors.
This study investigated the influence of different initial conditions on a subsequent fast (ballistic) isometric contraction of the ankle dorsiflexor muscles. Surface electromyograms (EMGs) of dorsiflexor and plantarflexor muscles were recorded during ballistic contractions performed without any pre-activation (BAL) and in ballistic contractions preceded by a sustained submaximal contraction (20% MVC) that was followed either by a rapid voluntary relaxation of the agonist muscle (VRBAL) or by a rapid antagonist (reversal) contraction (ARBAL). In the latter condition, three different antagonist torque levels were compared (25, 50 and 75% MVC). The results showed that the mean average rate of torque development was significantly (P < 0.001) greater for the ARBAL condition (968.5 ± 183.9% MVC/s) compared with the VRBAL (509.3 ± 78.7% MVC/s) and BAL (461.8 ± 79.9% MVC/s) conditions. Furthermore, the mean value recorded for VRBAL was significantly (P < 0.05) greater than for BAL condition. The faster increases in torque during the VRBAL and ARBAL conditions were associated with a greater agonist EMG activity. Compared with VRBAL, performance during the ARBAL condition was improved by a greater level of antagonist coactivation and, in some trials, by the presence of a silent EMG period between the end of the antagonist activation and the onset of the agonist ballistic contraction. Together, these results indicate that the initial conditions can have a substantial influence on the rate of torque development during ballistic contractions performed in isometric conditions.


[Travellers in medical practice].
In order to competently advise people who are planning to travel abroad the doctor needs information both about the planned journey (the mode of travel, the itinerary, the time of the trip) and about possible preexisting health problems. The area where the traveller intends to stay determines the specific risk of endemic diseases such as malaria, dengue-fever, yellow-fever etc. The recommendations for prophylaxis of malaria and for the different vaccinations are constantly modified and can be attained from the internet. Pre-existing health hazards such as cardiovascular diseases, diabetes mellitus, HIV-infection need to be assessed. Special consideration should be taken for pregnant women. The traveller returning from an endemic malarious area who feels ill or is febrile is a medical emergency and must be assessed for the possibility of having malaria since nontreated malaria in non-immune people has a very high mortality rate. Diarrhea is a common problem but is rarely dangerous. Long standing or bloody diarrhea, however, calls for a thorough search for the responsible pathogen. Healthy return travellers do not need a check-up examination because there are virtually no consequences for treatment.


Maxi-anion channel and pannexin 1 hemichannel constitute separate pathways for swelling-induced ATP release in murine L929 fibrosarcoma cells.
The maxi-anion channel plays a classically recognized role in controlling the membrane potential through the chloride conductance. It also has novel functions as a regulated pathway for the release of the anionic signaling molecules ATP and excitatory amino acids from cells subjected to osmotic perturbation, ischemia, or hypoxia. Because hemichannels formed by pannexins and connexins have been reported to mediate ATP release from a number of cell types, these hemichannels may represent the molecular correlate of the maxi-anion channel. Here, we found that L929 fibrosarcoma cells express functional maxi-anion channels which mediate a major portion of swelling-induced ATP release, and that ATP released via maxi-anion channels facilitates the regulatory volume decrease after osmotic swelling. Also, it was found that the cells express the mRNA for pannexin 1, pannexin 2, and connexin 43. Hypotonicity-induced ATP release was partially suppressed not only by known blockers of the maxi-anion channel but also by several blockers of pannexins including the pannexin 1-specific blocking peptide (10)Panx1 and small interfering (si)RNA against pannexin 1 but not pannexin 2. The inhibitory effects of maxi-anion channel blockers and pannexin 1 antagonists were additive. In contrast, maxi-anion channel activity was not affected by pannexin 1 antagonists and siRNAs against pannexins 1 and 2. Although a connexin 43-specific blocking peptide, Gap27, slightly suppressed hypotonicity-induced ATP release, maxi-anion channel activity was not affected by Gap27 or connexin 43-specific siRNA. Thus, it is concluded that the maxi-anion channel is a molecular entity distinct from pannexin 1, pannexin 2, and connexin 43, and that the maxi-anion channel and the hemichannels constitute separate pathways for swelling-induced ATP release in L929 cells.


Leukotriene receptor blockade in experimental heart failure.
The pathophysiological role of endogenous leukotrienes in cardiovascular control and the regulation of renal function in congestive heart failure is not known. Therefore, in six conscious dogs with or without heart failure induced by right ventricular pacing (270/min, 10 days) we studied the effects of the leukotriene receptor antagonist FPL55712 on hemodynamics, plasma hormones and renal function. In healthy dogs, FPL55712 (1 mg kg-1 + 0.01 mg kg-1 min-1 i.v.) had little effect on hemodynamics, only reducing heart rate by 11% and insignificantly increasing systemic vascular resistance. Plasma levels of norepinephrine (-57%), renin (-30%) and aldosterone (-24%) were significantly decreased. Renal function parameters were not changed. In dogs with heart failure, FPL55712 significantly increased systemic vascular resistance (+16%) and decreased cardiac output (-15%). Plasma hormone levels were not changed, but renal plasma flow was decreased (-13%) and glomerular filtration rate (+12%), renal vascular resistance (+13%) and filtration fraction (+23%) were increased. It is concluded that there is no evidence for a contribution of endogenous leukotrienes to the systemic vasoconstriction in experimental heart failure. Whether the increase in systemic and renal vascular resistance induced by the leukotriene antagonist in dogs with heart failure reflects a role for endogenous leukotrienes with vasodilator action is still unclear and deserves further investigation.


A survival benefit of combination antibiotic therapy for serious infections associated with sepsis and septic shock is contingent only on the risk of death: a meta-analytic/meta-regression study.
To assess whether a potential benefit with combination antibiotic therapy is restricted to the most critically ill subset of patients, particularly those with septic shock. OVID MEDLINE (1950-October 2009), EMBASE (1980-October 2009), the Cochrane Central Register of Controlled Trials (to third quarter 2009), the ClinicalTrial.gov database, and the SCOPUS database. Randomized or observational studies of antimicrobial therapy of serious bacterial infections potentially associated with sepsis or septic shock. Fifty studies met entry criteria. Study design, mortality/clinical response, and other variables were extracted independently by two reviewers. When possible, study datasets were split into mutually exclusive groups with and without shock or critical illness. Although a pooled odds ratio indicated no overall mortality/clinical response benefit with combination therapy (odds ratio, 0.856; 95% confidence interval, 0.71-1.03; p = .0943; I = 45.1%), stratification of datasets by monotherapy mortality risk demonstrated substantial benefit in the most severely ill subset (monotherapy risk of death >25%; odds ratio of death, 0.51; 95% confidence interval, 0.41-0.64; I = 8.6%). Of those datasets that could be stratified by the presence of shock/critical illness, the more severely ill group consistently demonstrated increased efficacy of a combination therapy strategy (odds ratio, 0.49; 95% confidence interval, 0.35-0.70; p < .0001; I = 0%). An increased risk of death was found in low-risk patients (risk of death <or=15% in the monotherapy arm) exposed to combination therapy (odds ratio, 1.53; 95% confidence interval, 1.16-2.03; p = .003; I = 8.2%). Meta-regression indicated that efficacy of combination therapy was dependent only on the risk of death in the monotherapy group. Combination antibiotic therapy improves survival and clinical response of high-risk, life-threatening infections, particularly those associated with septic shock but may be detrimental to low-risk patients.


Involvement of tumor necrosis factor alpha in hippocampal development and function.
Tumor necrosis factor alpha (TNFalpha) is a cytokine produced mainly by cells of the immune system. It is also expressed by brain neurons and glia. The physiological role of TNFalpha in the brain is not yet fully clear. Using TNFalpha-deficient mice, we have examined its role in hippocampal development and function. We report here that TNFalpha is involved in the regulation of morphological development in the hippocampus. TNFalpha-deficient mice exhibited an accelerated maturation of the dentate gyrus region and smaller dendritic trees in CA1 and CA3 regions in young mouse. In addition to its involvement in hippocampal morphogenesis, TNFalpha deficiency specifically improved performance of affected mice in behavioral tasks related to spatial memory. Moreover, lack of TNFalpha increased the expression of nerve growth factor (NGF), but not brain-derived neurotrophic factor (BDNF), following performance of the learning task. Our results suggest that TNFalpha actively influences hippocampal development and function. In adult mice, TNFalpha may interfere with memory consolidation, perhaps by regulating NGF levels.


[Selective IgA deficiency].
Selective IgA deficiency may be defined as an inborn state characterized by a decrease of serum IgA levels below 8 IU/1 (approximately 5 mg/dl) which may be associated with clinical symptoms of disease. The frequency of this condition in the general population varies between 1 : 400 and 1 : 3000 in different countries. Patients with defects of chromosome 18, ataxia teleangiectatica and with connatal rubella syndrome have a high incidence of IgA deficiency. Inspite of the decrease in circulating IgA there are B-lymphocytes containing IgA molecules in the peripheral blood. Thus it has been concluded that transformation of B-lymphocytes into IgA bearing plasmacells is stunted by another mechanism. While small amounts of IgA may be released by transformed plasmacells the capacity of B-lymphocytes to mature into fully functioning plasmacells releasing normal amounts of IgA is defective. T-cells acting as suppressor cells for IgA differentiation have been demonstrated in peripheral blood and are a possible explanation for this phenomenon. The majority of individuals with IgA deficiency are healthy. Evaluations of increased susceptibility for infections have to consider the fact that 6 respiratory tract infections per year are the average for any preschool child. However a number of children with IgA deficiency suffer from recurrent bacterial infections such as sinusitis, bronchitis and pneumonia, usually responding well to antibiotic treatment. IgA deficiency has an established correlation with atopic disease. There is an 40 fold increase in incidence of allergies and autoimmune diseases such as rheumatoid arthritis, lupus erythematodes and thyroiditis in individuals with IgA deficiency.(ABSTRACT TRUNCATED AT 250 WORDS)


Functional diversity among seed dispersal kernels generated by carnivorous mammals.
1. Knowledge of the spatial scale of the dispersal service provided by important seed dispersers (i.e. common and/or keystone species) is essential to our understanding of their role on plant ecology, ecosystem functioning and, ultimately, biodiversity conservation. 2. Carnivores are the main mammalian frugivores and seed dispersers in temperate climate regions. However, information on the seed dispersal distances they generate is still very limited. We focused on two common temperate carnivores differing in body size and spatial ecology - red fox (Vulpes vulpes) and European pine marten (Martes martes) - for evaluating possible functional diversity in their seed dispersal kernels. 3. We measured dispersal distances using colour-coded seed mimics embedded in experimental fruits that were offered to the carnivores in feeding stations (simulating source trees). The exclusive colour code of each simulated tree allowed us to assign the exact origin of seed mimics found later in carnivore faeces. We further designed an explicit sampling strategy aiming to detect the longest dispersal events; as far we know, the most robust sampling scheme followed for tracking carnivore-dispersed seeds. 4. We found a marked functional heterogeneity among both species in their seed dispersal kernels according to their home range size: multimodality and long-distance dispersal in the case of the fox and unimodality and short-distance dispersal in the case of the marten (maximum distances = 2846 and 1233 m, respectively). As a consequence, emergent kernels at the guild level (overall and in two different years) were highly dependent on the relative contribution of each carnivore species. 5. Our results provide the first empirical evidence of functional diversity among seed dispersal kernels generated by carnivorous mammals. Moreover, they illustrate for the first time how seed dispersal kernels strongly depend on the relative contribution of different disperser species, thus on the composition of local disperser assemblages. These findings provide a key starting point for understanding and modelling plant population processes that include mammal-mediated seed dispersal, such as connectivity, range expansion and colonization.


Association of sedentary time with risk of cardiovascular diseases and cardiovascular mortality: A systematic review and meta-analysis of prospective cohort studies.
Given the high prevalence of cardiovascular disease (CVD), we meta-analysed CVD relative risk (RR) in relation to high vs. low categories of self-reported and objectively assessed sedentary behaviours from cohort studies; in a sub-sample (n = 4 studies), the theoretical substitution of one hour spent sedentary with the same amount of time spent in light-intense physical activity was evaluated. Based on 19 studies (60,526 fatal and non-fatal CVD, 1,473,354 individuals and 13,559,139 persons-year) we estimated a 30% increased CVD risk for high vs. low categories of sedentary behaviour (RR = 1.29, confidence interval (CI) = 1.22;1.37). Every hour spent sedentary corresponds to a 5% increased fatal and non-fatal CVD risk (RR = 1.05, CI = 1.02;1.07). Dose-response meta-analysis revealed that sedentary behaviour is statistically significantly associated to fatal and non-fatal CVD risk following a J-shaped relation. Substituting one hour spent sedentary with physical activity of light intensity reduced the risk of fatal and non-fatal CVD events by one-fifth (RR =0.84, CI = 0.73;0.97). In meta-regression analysis, potential influential factors such as age, sex, and medical condition did not essentially alter the results.


Long-term culture of mouse embryonic stem cell-derived adherent neurospheres and functional neurons.
Innumerous protocols, using the mouse embryonic stem (ES) cells as model for in vitro study of neurons functional properties and features, have been developed. Most of these protocols are short lasting, which, therefore, does not allow a careful analysis of the neurons maturation, aging, and death processes. We describe here a novel and efficient long-lasting protocol for in vitro ES cells differentiation into neuronal cells. It consists of obtaining embryoid bodies, followed by induction of neuronal differentiation with retinoic acid of nonadherent embryoid bodies (three-dimensional model), which further allows their adherence and formation of adherent neurospheres (AN, bi-dimensional model). The AN can be maintained for at least 12 weeks in culture under repetitive mechanical splitting, providing a constant microenvironment (in vitro niche) for the neuronal progenitor cells avoiding mechanical dissociation of AN. The expression of neuron-specific proteins, such as nestin, sox1, beta III-tubulin, microtubule-associated protein 2, neurofilament medium protein, Tau, neuronal nuclei marker, gamma-aminobutyric acid, and 5-hydroxytryptamine, were confirmed in these cells maintained during 3 months under several splitting. Additionally, expression pattern of microtubule-associated proteins, such as lissencephaly (Lis1) and nuclear distribution element-like (Ndel1), which were shown to be essential for differentiation and migration of neurons during embryogenesis, was also studied. As expected, both proteins were expressed in undifferentiated ES cells, AN, and nonrosette neurons, although presenting different spatial distribution in AN. In contrast to previous studies, using cultured neuronal cells derived from embryonic and adult tissues, only Ndel1 expression was observed in the centrosome region of early neuroblasts from AN. Mature neurons, obtained from ES cells in this work, display ionic channels and oscillations of membrane electrical potential typical of electrically excitable cells, which is a characteristic feature of the functional central nervous system (CNS) neurons. Taken together, our study demonstrated that AN are a long-term culture of neuronal cells that can be used to analyze the process of neuronal differentiation dynamics. Thus, the protocol described here provides a new experimental model for studying neurological diseases associated with neuronal differentiation during early development, as well as it represents a novel source of functional cells that can be used as tools for testing the effects of toxins and/or drugs on neuronal cells.


Indications for Non-Invasive Ventilation in Respiratory Failure.
Non-invasive ventilation (NIV) is increasingly being used to treat episodes of acute respiratory failure not only in critical care and respiratory wards, but also in emergency departments. Aim of this review is to summarize the current indications for the management of NIV for respiratory failure. Current literature about the topic was reviewed and critically reported to describe the rationale and physiologic advantages of NIV in various situations of respiratory failure. Early NIV use is commonly associated with the significant decrease in endotracheal intubation rate, the incidence of infective complications (especially ventilatory associated pneumonia), Intensive Care Units and the length of hospital stay and, in selected conditions, also in mortality rates. Severe acute exacerbation of chronic obstructive pulmonary disease (pH<7.35 and relative hypercarbia) and acute cardiogenic pulmonary oedema are the most common NIV indications; in these conditions NIV advantages are clearly documented. Not so evident are the NIV benefits in hypoxaemic respiratory failure occurring without prior chronic respiratory disease (De novo respiratory failure). One recent randomized control trial reported in hypoxaemic respiratory failure a survival benefit of high-flow nasal cannulae over standard oxygen therapy and bilevel NIV. Evidence suggests the advantages of NIV also in respiratory failure in immunocompromised patients or chest trauma patients. Use during a pandemic event has been assessed in several observational studies but remains controversial; there also is not sufficient evidence to support the use of NIV treatment in acute asthma exacerbation. NIV eliminates morbidity related to the endotracheal tube (loss of airway defense mechanism with increased risk of pneumonia) and in selected conditions (COPD exacerbation, acute cardiogenic pulmonary edema, immunosuppressed patients with pulmonary infiltrates and hypoxia) is clearly associated with a better outcome in comparison to conventional invasive ventilation. However, NIV is associated with complications, especially minor complications related to interface. Major complications like aspiration pneumonia, barotrauma and hypotension are infrequent.


Response to therapy in myasthenia gravis with anti-MuSK antibodies.
Myasthenia gravis (MG) with antibodies against the muscle-specific tyrosine kinase (MuSK abs) is often a severe disease requiring aggressive treatment. Various immunosuppressive (IS) regimens have been employed; the efficacy of plasma exchange is unanimously recognized, while the indication for thymectomy is controversial. We evaluated the response to therapy in 57 MuSK-positive patients (12 M/45 F) comparing our experience with other authors' results. Disease severity and response to treatment were graded according to MG Foundation of America; follow-up ranged from 0.5-29 years. Owing to both MG severity and the unsatisfactory response to cholinesterase inhibitors, most patients (54/57) needed IS treatment, and 35 received one or more courses of plasma exchange and intravenous immunoglobulin. At the end of follow-up, the rate of complete remission was 8.8%, and IS treatment had been withdrawn in only 10/54 patients. The extent of therapeutic response varied considerably. With conventional IS therapy (prednisone alone or in combination with azathioprine or cyclosporine), most patients achieved good control of their disease, but 30% of them were left with permanent facial and bulbar weakness. In patients with refractory disease, the use of mycophenolate mofetil and rituximab proved very effective, as also reported by other authors. In our and others' experience, MuSK-positive MG markedly improves with IS therapy, although, in comparison with the AChR-positive disease, it is characterized by a lower remission rate, as a higher proportion of patients remain dependent on treatment. Thymectomy is mostly considered scarcely effective; however, at present, no firm conclusions can be drawn on its role in the treatment of this form of MG.


Parasphingopyxis lamellibrachiae gen. nov., sp. nov., isolated from a marine annelid worm.
A Gram-stain-negative, aerobic, motile, orange-pigmented, slightly halophilic, rod-shaped bacterium, designated strain JAMH 0132(T), was isolated from the trophosome of a tubeworm in Kagoshima Bay, Japan, and its taxonomic position was investigated using a polyphasic approach. The novel strain grew optimally at 28-30 °C and with about 2.0 % (w/v) NaCl. Chemotaxonomic analysis showed that Q-10 was the predominant respiratory quinone and that C(18 : 1)ω7c, C(16 : 0) 2-OH and C(16 : 0) were the major fatty acids. Sphingoglycolipid, phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylcholine were the major polar lipids. The genomic DNA G+C content was 60.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain JAMH 0132(T) belonged to the family Sphingomonadaceae, within the class Alphaproteobacteria. The novel strain appeared most closely related to Sphingopyxis baekryungensis SW-150(T) (95.1 % 16S rRNA gene sequence similarity) and showed less sequence similarity with representatives of the genera Blastomonas, Sphingomonas, Sphingosinicella and Novosphingobium (<94.8 %). In having no detectable polyamine, strain JAMH 0132(T) differed from members of all genera currently in the family Sphingomonadaceae. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain JAMH 0132(T) represents a novel species of a new genus in the family Sphingomonadaceae for which the name Parasphingopyxis lamellibrachiae gen. nov., sp. nov. is proposed. The type strain of Parasphingopyxis lamellibrachiae gen. nov., sp. nov. is JAMH 0132(T) (= JCM 15549(T) = NCIMB 14486(T)).


Prolonged intrathecal baclofen withdrawal syndrome. Case report and discussion of current therapeutic management.
The authors describe a patient who experienced a prolonged course of intrathecal baclofen withdrawal syndrome after removal of an implantable baclofen pump for treatment of pump infection and meningitis. The current literature outlines management options for the acute management of this syndrome. In this report the authors discuss the long-term presentation of this syndrome and suggest a treatment strategy for management of the syndrome. A 37-year-old man who presented with a baclofen pump infection and meningitis experienced acute onset of intrathecal baclofen withdrawal syndrome 12 hours after the pump had been surgically removed. The patient's symptoms evolved into a severe, treatment-refractory withdrawal syndrome lasting longer than 1 month. Oral baclofen replacement with adjunctive administration of parenteral gamma-aminobutyric acid agonists only served to stabilize the patient's critical condition throughout his hospital course. Replacement of the baclofen pump and restoration of intrathecal delivery of the medication was necessary to trigger the patient's dramatic recovery and complete reversal of the withdrawal syndrome within approximately 48 hours. These findings indicate that a more direct method of treating infected baclofen pumps than immediate surgical removal is necessary to prevent the onset of intrathecal baclofen withdrawal syndrome. Various options for preventing the onset of the syndrome while simultaneously treating the infection are discussed.


A complex of poly(4-vinylpyridine) and tolane based hemi-phasmid benzoic acid: towards luminescent supramolecular side-chain liquid crystalline polymers.
A supramolecular side-chain liquid crystalline (LC) polymer P4VP(1)x was prepared using poly(4-vinylpyridine) and a tolane based hemi-phasmid benzoic acid. P4VP(1)x exhibits good processability, forming smectic and hexagonal columnar LC phases at different compositions. Its photoluminescence properties depend on the LC structures.


Vitamin D status and response to treatment in post-menopausal osteoporosis.
Treatment with anti-resorptive agents over 13 months was associated with for three to fivefold lower bone mineral density changes and 1.5-fold increased risk of incidence fracture in vitamin D insufficient as compared to vitamin D repleted postmenopausal osteoporotic women. Several drugs were registered for the treatment of osteoporosis on the basis of clinical trials in which vitamin D repletion was a pre-requisite inclusion criteria and vitamin D supplements were used as adjunctive therapy. However, in routine clinical practice these supplements are not consistently recommended. We studied 1515 women with postmenopausal osteoporosis under treatment with anti-resorbing agents (alendronate, risedronate, raloxifene) for 13.1 months with an adherence > 75%. The patients were classified as vitamin D deficient (N = 514) or vitamin D repleted (N = 1001) according to risk factors (N = 1062) or the level of 25(OH) vitamin D [25(OH)D] above or below 50 nmol/l (N = 453). Vitamin D deficient and vitamin D repleted subjects differed significantly for annualized spine and hip bone mineral density (BMD) changes adjusted for all available confounding factors (type of treatment, age, global calcium intake, baseline BMD values). One hundred fifty one patients suffered from a new incident clinical fracture. The adjusted odds ratio for incident fractures in vitamin D deficient as compared to vitamin D repleted women was 1.77 (1.20 - 2.59, 95% CI; p = 0.004). Optimal vitamin D repletion seems to be necessary to maximize the response to anti-resorbers in terms of both BMD changes and anti-fracture efficacy.


When "Crack walnuts" lies in different brain regions: evidence from a voxel-based lesion-symptom mapping study.
Theories of lexical processing differ as to how multimorphemic words, such as compounds, are mentally processed. The most recent findings seem to support the dual route hypothesis, which assumes that complex words can be stored and retrieved either whole or by decomposition into their constituents. Despite great efforts to investigate the cognitive mechanisms involved in processing complex words, very little is known about how compounds are represented in the brain. The present study was designed to address this issue in a group of 20 left-hemispheric stroke patients who were submitted to four picture-naming tasks involving nouns, verbs, noun-noun (NN) and verb-noun (VN) compounds. To determine the brain lesions implicated in these tasks, we analyzed patients' performances together with their lesions using Voxel-based Lesion Symptom Mapping (VLSM). Results showed that while NN involved the same temporal areas as nouns, VN (although they belong to the noun category) involved different fronto-temporal regions. This latter finding is discussed within the view that distinct mechanisms process the different constituents of complex words.


Stability of a giant connected component in a complex network.
We analyze the stability of the network's giant connected component under impact of adverse events, which we model through the link percolation. Specifically, we quantify the extent to which the largest connected component of a network consists of the same nodes, regardless of the specific set of deactivated links. Our results are intuitive in the case of single-layered systems: the presence of large degree nodes in a single-layered network ensures both its robustness and stability. In contrast, we find that interdependent networks that are robust to adverse events have unstable connected components. Our results bring novel insights to the design of resilient network topologies and the reinforcement of existing networked systems.


Infections associated with locking reconstruction plates: a retrospective review.
In this study, we examined the incidence of infection with the use of a locking reconstruction bone plate/system. All patients, treated with a locking reconstruction bone plate/screw system for mandible fractures in the Oral and Maxillofacial Surgery service over a 28-month period at a level I trauma center, were evaluated through a retrospective chart review by independent examiners. The use of a locking reconstruction plate was determined by the attending staff involved in the patient's care. Patient population included single, bilateral, and comminuted fractures. Patient characteristics were noted and include dentate versus edentulous, smoking history, and history of previous infection. Fifty-six locking bone plates were placed in 42 patients. Eight (19%) of the patients were infected before treatment. A persistent infection remained in 3 of these 8 patients (37.5%). Two patients (5.8%), with 3 fracture sites (6.4%) developed postoperative infection that required further intervention. All 5 of the patients who were infected after surgery were heavy smokers. History of preoperative infection and smoking appear to be significant factors in the etiology of postoperative infection. All postoperative infections resolved successfully with local measures and with no loss of fixation. The use of locking reconstruction plates can facilitate the management of complicated fractures; however, it does not eliminate complications. Postoperative infections are related to numerous factors, including preoperative incidence of infection, smoking, and proper use of the plates.


Tissue fractions of cadmium in two hyperaccumulating Jerusalem artichoke genotypes.
In order to investigate the mechanisms in two Jerusalem artichoke (Helianthus tuberosus L.) genotypes that hyperaccumulate Cd, a sand-culture experiment was carried out to characterize fractionation of Cd in tissue of Cd-hyperaccumulating genotypes NY2 and NY5. The sequential extractants were: 80% v/v ethanol (FE), deionized water (FW), 1 M NaCl (FNaCl), 2% v/v acetic acid (FAcet), and 0.6 M HCl (FHCl). After 20 days of treatments, NY5 had greater plant biomass and greater Cd accumulation in tissues than NY2. In both genotypes the FNaCl fraction was the highest in roots and stems, whereas the FAcet and FHCl fractions were the highest in leaves. With an increase in Cd concentration in the culture solution, the content of every Cd fraction also increased. The FW and FNaCl ratios in roots were lower in NY5 than in NY2, while the amount of other Cd forms was higher. It implied that, in high accumulator, namely, NY5, the complex of insoluble phosphate tends to be shaped more easily which was much better for Cd accumulation. Besides, translocation from plasma to vacuole after combination with protein may be one of the main mechanisms in Cd-accumulator Jerusalem artichoke genotypes.


European Association of Urology Guidelines on Muscle-invasive and Metastatic Bladder Cancer: Summary of the 2023 Guidelines.
We present an overview of the updated 2023 European Association of Urology (EAU) guidelines for muscle-invasive and metastatic bladder cancer (MMIBC). To provide practical evidence-based recommendations and consensus statements on the clinical management of MMIBC with a focus on diagnosis and treatment. A broad and comprehensive scoping exercise covering all areas of the MMIBC guidelines has been performed annually since 2017. Searches cover the Medline, EMBASE, and Cochrane Libraries databases for yearly guideline updates. A level of evidence and strength of recommendation are assigned. The evidence cutoff date for the 2023 MIBC guidelines was May 4, 2022. Patients should be counselled regarding risk factors for bladder cancer. Pathologists should describe tumour and lymph nodes in detail, including the presence of histological subtypes. The importance of the presence or absence of urothelial carcinoma (UC) in the prostatic urethra is emphasised. Magnetic resonance imaging (MRI) of the bladder is superior to computed tomography (CT) for disease staging, specifically in differentiating T1 from T2 disease, and may lead to a change in treatment approach in patients at high risk of an invasive tumour. Imaging of the upper urinary tract, lymph nodes, and distant metastasis is performed with CT or MRI; the additional value of flurodeoxyglucose positron emission tomography/CT still needs to be determined. Frail and comorbid patients should be evaluated by a multidisciplinary team. Postoperative histology remains the most important prognostic variable, while circulating tumour DNA appears to be an interesting predictive marker. Neoadjuvant systemic therapy remains cisplatin-based. In motivated and selected women and men, sexual organ-preserving cystectomy results in better functional outcomes without compromising oncological outcomes. Robotic and open cystectomy have comparable outcomes and should be combined with (extended) lymph node dissection. The diversion type is an individual choice after taking patient and tumour characteristics into account. Radical cystectomy remains a highly complex procedure with considerable morbidity and risk of mortality, although lower rates are observed for higher hospital volumes (>20 cases/yr). With proper patient selection, trimodal therapy (chemoradiation) has comparable outcomes to radical cystectomy. Adjuvant chemotherapy after surgery improves disease-specific survival and overall survival (OS) in patients with high-risk disease who did not receive neoadjuvant treatment, and is strongly recommended. There is a weak recommendation for adjuvant nivolumab, as OS data are not yet available. Health-related quality of life should be assessed using validated questionnaires at baseline and after treatment. Surveillance is needed to monitor for recurrent cancer and functional outcomes. Recurrences detected on follow-up seem to have better prognosis than symptomatic recurrences. This summary of the 2023 EAU guidelines provides updated information on the diagnosis and treatment of MMIBC for incorporation into clinical practice. The European Association of Urology guidelines panel on muscle-invasive and metastatic bladder cancer has released an updated version of the guideline containing information on diagnosis and treatment of this disease. Recommendations are based on studies published up to May 4, 2022. Surgical removal of the bladder and bladder preservation are discussed, as well as updates on the use of chemotherapy and immunotherapy in localised and metastatic disease.


Correlation of injury occurrence data with estimated maximal aerobic capacity and body composition in a high-frequency manual materials handling task.
This article evaluates the correlation between injury occurrence, step test estimated maximal aerobic capacity (VO2max), and body composition in a high-frequency manual materials handling task. The study used 212 highly trained male manual material handlers working for a major materials handling company. Three locations across the United States (western, midwestern, and southeastern) were chosen based on similarity of size and function. An estimated maximal aerobic capacity was obtained for each participant using a submaximal bench step protocol. Also, a percentage body fat estimation was randomly obtained for approximately 25% of the participants. The correlation between injury occurrences, absolute VO2max, relative VO2max, and percentage body fat were analyzed. Also, the relationship between both VO2max estimations and percentage body fat was analyzed. Finally, the correlation between location VO2max and percentage body fat was studied. Results indicated no significant difference between absolute VO2max, injury, or percentage body fat. Relative VO2max suggested a significant relationship with injury occurrences and body composition. Body composition also indicated a significant correlation with injury occurrences. Finally, location played a significant factor in injury occurrence, step test estimated VO2max, and estimated body composition. This investigation demonstrates significant evidence of the predictability of employee injury occurrence and the fitness estimation methods used. In a high-frequency manual materials handling task, high occurrences of injury were significantly correlated with low estimated relative maximal aerobic capacity and high estimated percentage body fat.


Smart polymer brush nanostructures guide the self-assembly of pore-spanning lipid bilayers with integrated membrane proteins.
Nanopores in arrays on silicon chips are functionalized with pH-responsive poly(methacrylic acid) (PMAA) brushes and used as supports for pore-spanning lipid bilayers with integrated membrane proteins. Robust platforms are created by the covalent grafting of polymer brushes using surface-initiated atom transfer radical polymerization (ATRP), resulting in sensor chips that can be successfully reused over several assays. His-tagged proteins are selectively and reversibly bound to the nitrilotriacetic acid (NTA) functionalization of the PMAA brush, and consequently lipid bilayer membranes are formed. The enhanced membrane resistance as determined by electrochemical impedance spectroscopy and free diffusion of dyed lipids observed as fluorescence recovery after photobleaching confirmed the presence of lipid bilayers. Immobilization of the His-tagged membrane proteins on the NTA-modified PMAA brush near the pore edges is characterized by fluorescence microscopy. This system allows us to adjust the protein density in free-standing bilayers, which are stabilized by the polymer brush underneath. The potential application of the integrated platform for ion channel protein assays is demonstrated.