Transcriptional and replicational activation functions in the bovine papillomavirus type 1 E2 protein are encoded by different structural determinants.
A set of E2 proteins with mutations in the amino-terminal transactivation domain was made by a scheme called clustered charged-to-alanine scan. These mutant E2 proteins were tested for expression, stability, and compartmentalization in cells and for sequence-specific DNA binding, as well as in functional assays for transcriptional and replicational activation. We identified four groups of mutants. First, mutants K111A, K112A, and E176A were unable to activate replication and transcription because of oligomerization-induced retention of oligomers in the cytoplasm. Second, although fractions of the mutant proteins E74A and D143A/ R172C existed in the oligomeric form, they were localized in the nucleus. Certain fractions of these proteins existed as a dimer able to form a specific complex and activate replication; however, these proteins were inactive in transcriptional activation. Third, mutants R37A and D122A were localized in the nucleus, existed in the dimeric form, supported replication efficiently, and were severely crippled in transcriptional activation. The fourth group of mutants did not differ considerably from the wild-type protein. The activation of transcription by the wild type as well as mutant E2 proteins was dependent on the concentration of input E2 expression vector DNA and had a bell-like shape. We suggest that the reduction of transcriptional activation at higher E2 concentrations, the self-squelching activity, is caused by oligomerization of the E2 transactivator and is one of the mechanisms for the regulation of E2 activity. Our results also show that transcriptional and replicational activation activities are encoded by different determinants in the E2 protein.


A Methodological Quality Evaluation of Meta-Analyses on Nursing Home Research: Overview and Suggestions for Future Directions.
(1) Background: The nursing home (NH) research field lacks quality reporting about meta-analyses (MAs), and most gradings of MA evidence are biased on analyzing the effectiveness of independent variables in randomized control trials. (2) Objectives: This study aimed to perform a critical methodological review of MAs in the NH research field. (3) Methods: We searched the articles from four databases (PubMed, MEDLINE, CINAHL, and PsycINFO) until 15th January 2021. We reviewed a total of 41 published review articles in the NH research field. (4) Results: The studies primarily fell into the following categories: medicine (17/41), nursing (7/41), and psychiatry or psychology (6/41); 36.6% of the reviewed studies did not use any validated MA guidelines. The lowest correctly reported PRISMA 2000 guideline item was protocol and registration (14.6%), and more than 50% of articles did not report risk of bias. Moreover, 78.0% of studies did not describe missing reports of effect size formula. (5) Discussion: NH researchers must follow appropriate and updated guidelines for their MAs in order to provide validated reviews, as well as consider statistical issues such as the complexity of interventions, proper grouping, and scientific effect-size calculations to improve the quality of their study. Future quality review studies should investigate more diverse studies.


[CT enteroclysis for detection of small bowel tumors].
To assess the feasibility and the usefulness of CT enteroclysis (helical CT with enteroclysis) in detecting small bowel tumors. Fifty patients were referred for suspicion of small bowel tumor. CT enteroclysis is performed by injecting a large volume of water using a pomp through a nasojejunal tube followed by a thin section helical acquisition. Forty-eight helical CT enteroclysis were performed in order to detect 25 small bowel tumors. Among them 22 were confirmed by histological study. The mean size of tumors was 23 mm. In 12 of 17 cases, diagnosis was missed or incomplete at conventional barium study. Enteroscopy was performed in 12 of 22 cases, with discordant result in one case and incomplete result in 3 cases. In 8 cases, including 5 carcinoid tumors, patients had surgery after CT enteroclysis only, enteroscopy would probably have not made the diagnosis because the lesions were far from the duodenojejunal junction and ileocaecal valve. Helical CT enteroclysis is a new method for detecting small bowel tumors, easy to perform, well tolerated. It seems to be more sensitive than conventional barium studies and less invasive than enteroscopy. Tumor characterization and staging can be performed using a single examination. It seems to be justified to perform CT enteroclysis to detect small bowel tumors or in the evaluation of patients with polyposis.


First-Principles Vibrational Electron Energy Loss Spectroscopy of β-Guanine.
A general approach to model vibrational electron energy loss spectra obtained using an electron beam positioned away from the specimen is presented. The energy-loss probability of the fast electron is evaluated using first-principles quantum mechanical calculations (density functional theory) of the dielectric response of the specimen. The validity of the method is assessed using recently measured anhydrous β-guanine, an important molecular solid used by animals to produce structural colors. The good agreement between theory and experiments lays the basis for a quantitative interpretation of this spectroscopy in complex systems.


Distribution and role of lipopolysaccharide in the pathogenesis of acute renal proximal tubule injury.
Endotoxins (lipopolysaccharides; LPS) are known to cause multiple organ failure, including renal dysfunction. The present report elucidates LPS distribution and effect on renal proximal tubules in an attempt to gain a better understanding of the cellular mechanism underlying the pathogenesis of renal dysfunction in endotoxemia and sepsis. Rats were intravenously treated with biotin-linked or regular Escherichia coli (0111:B4) LPS (3 mg/kg) and sacrificed at different times. Kidneys were retrieved and examined for LPS localization, tubular permeability, ultracytochemical alterations, leukocyte sequestration, and ICAM-1 expression. The functional impact of endotoxemia was also assessed by monitoring the changes in urine levels of glucose in timed collections up to 6 h. LPS was localized on the plasma membranes of the apical microvilli, the labyrinth of the lateral intercellular spaces, in various organelles of epithelial cells, and in the endothelial cells of the peritubular capillaries. LPS caused structural damage and calcium accumulation in the mitochondria, leakage of tight junctions, widening of the basolateral intercellular spaces, intracellular and extracellular edema, leukocyte margination and accumulation, vascular expression of ICAM-1, and decrease of plasma membrane and mitochondrial Ca2(+)-ATPase. Physiological study showed that both urine volume and glucose were greatly increased after LPS infusion. The pathological alterations in the proximal tubules may directly contribute to the reduction in the reabsorption ability of the proximal tubules.


Unconstrained parameter estimation for assessment of dynamic cerebral autoregulation.
Measurement of dynamic cerebral autoregulation (CA), the transient response of cerebral blood flow (CBF) to changes in arterial blood pressure (ABP), has been performed with an index of autoregulation (ARI), related to the parameters of a second-order differential equation model, namely gain (K), damping factor (D) and time constant (T). Limitations of the ARI were addressed by increasing its numerical resolution and generalizing the parameter space. In 16 healthy subjects, recordings of ABP (Finapres) and CBF velocity (ultrasound Doppler) were performed at rest, before, during and after 5% CO(2) breathing, and for six repeated thigh cuff maneuvers. The unconstrained model produced lower predictive error (p < 0.001) than the original model. Unconstrained parameters (K'-D'-T') were significantly different from K-D-T but were still sensitive to different measurement conditions, such as the under-regulation induced by hypercapnia. The intra-subject variability of K' was significantly lower than that of the ARI and this parameter did not show the unexpected occurrences of zero values as observed with the ARI and the classical value of K. These results suggest that K' could be considered as a more stable and reliable index of dynamic autoregulation than ARI. Further studies are needed to validate this new index under different clinical conditions.


Lunge filter feeding biomechanics constrain rorqual foraging ecology across scale.
Fundamental scaling relationships influence the physiology of vital rates, which in turn shape the ecology and evolution of organisms. For diving mammals, benefits conferred by large body size include reduced transport costs and enhanced breath-holding capacity, thereby increasing overall foraging efficiency. Rorqual whales feed by engulfing a large mass of prey-laden water at high speed and filtering it through baleen plates. However, as engulfment capacity increases with body length (engulfment volume∝body length3.57), the surface area of the baleen filter does not increase proportionally (baleen area∝body length1.82), and thus the filtration time of larger rorquals predictably increases as the baleen surface area must filter a disproportionally large amount of water. We predicted that filtration time should scale with body length to the power of 1.75 (filter time∝body length1.75). We tested this hypothesis on four rorqual species using multi-sensor tags with corresponding unoccupied aircraft systems-based body length estimates. We found that filter time scales with body length to the power of 1.79 (95% CI: 1.61-1.97). This result highlights a scale-dependent trade-off between engulfment capacity and baleen area that creates a biomechanical constraint to foraging through increased filtration time. Consequently, larger whales must target high-density prey patches commensurate to the gulp size to meet their increased energetic demands. If these optimal patches are absent, larger rorquals may experience reduced foraging efficiency compared with smaller whales if they do not match their engulfment capacity to the size of targeted prey aggregations.


Glycerol hyperhydration in resting horses.
To determine whether administration of glycerol-containing solutions induces a state of transient hyperhydration in resting euhydrated horses, changes in plasma and urine constituents were measured in four horses for 1 h before and 5 h after nasogastric administration of each of four treatments (Experiment 1). Treatments were applied in a randomized fashion and included: (1) 1.0 g.kg(-)(1)glycerol in 8 L of water (G); (2) 8 L of water (W); (3) 8 L of 0.9% NaCl solution (S); and (4) 1.0 g.kg(-)(1)glycerol in 8 L of 0.9% NaCl solution (GS). In a subsequent study, voluntary water intake was measured hourly for 5 h after nasogastric administration of each treatment (Experiment 2). All treatments produced mild plasma volume expansion ranging from 3.2 to 5.8% in Experiment 1. Administration of glycerol containing solutions increased serum glycerol concentration approximately 100-fold and plasma osmolality (P(osm)) by approximately 10 mOsm/kg and resulted in a tendency towards increased renal water conservation despite increased osmole excretion. In contrast, W treatment decreased plasma and urine osmolality and was accompanied by increased urine production and decreased renal water conservation. Plasma and urine osmolality, as well as renal osmole and water excretion, were unchanged after S administration. In Experiment 2, horses treated with GS voluntarily drank an additional 5.2 +/- 0.9 L of water during the initial hour following nasogastric administration of 8 L of solution. Voluntary water intake with the other treatments was less than 1.0 L for the entire 5 h observation period. Collectively, the results of both experiments suggest that administration of glycerol in saline would produce transient hyperhydration in resting euhydrated horses by enhancing renal water conservation and stimulating voluntary water intake.


Evaluating Skill-Mix Models of Care: A Rapid Scoping Review of Measures and Outcomes.
To synthesize the literature on measures and outcomes for skill-mix models of care. To address the human health resource crisis, changes to skill mix within models of care are being implemented emphasizing the need to synthesize evaluation methods for skill-mix models in the future. A scoping review of the literature using a rigorous search strategy and selection process was completed to identify articles that examined skill-mix models in an effort to identify related concepts. Ten studies examined skill-mix models. Areas of measurement in assessing the impact of skill-mix models included patient outcomes, patient satisfaction, nurse satisfaction, cost, and nurse perceptions of role changes, model effectiveness, and quality of care. Studies examining nurse satisfaction, patient satisfaction, and/or cost generally reported improvements upon skill-mix model implementation. Studies examining patient outcomes related to skill mix were inconsistent. Factors for consideration upon implementation of a skill-mix change include education of role clarity, the number of unregulated staff who require supervision, and professional practice support.


A noncoherent optical analog image processor.
The description of a machine that performs a variety of image processing operations is given, together with a theoretical discussion of its operation. Spatial processing is performed by corrective convolution techniques. Density processing is achieved by means of an electrical transfer function generator included in the video circuit. Examples of images processed for removal of image motion blur, defocus, and atmospheric seeing blur are shown.


Does measurement of small intestinal diameter increase diagnostic accuracy of radiography in dogs with suspected intestinal obstruction?
The ratio between maximal small intestinal (SI) diameter and the height of the body of the fifth lumbar vertebra (L5) in radiographs has been reported as a diagnostic test in dogs with suspected intestinal obstruction. In order to assess the effect of the SI/L5 ratio on the accuracy of radiographic diagnosis of intestinal obstruction, lateral abdominal radiographs of 37 dogs with small intestinal obstruction and 48 nonobstructed dogs were mixed and examined independently by six observers who were unaware of the final diagnosis and who represented a range of experience. Observers first examined radiographs subjectively and stated the likelihood of obstruction (definitely not, probably not, equivocal, probably, definitely). Observers subsequently reexamined the radiographs, determined the SI/L5 ratio, and again stated the likelihood of obstruction. The most frequent cause of obstruction was foreign body (29/37, 78%). Dogs with SI obstruction had a significantly larger median SI/L5 ratio than nonobstructed dogs (P = 0.0002). Using an SI/L5 ratio of 1.7 for diagnosis of intestinal obstruction, sensitivity and specificity were 66%. Use of the SI/L5 ratio was not associated with increased accuracy of diagnosis for any observer, regardless of experience, hence this test may have no diagnostic impact.


Robust markers associated with floral traits in roses are suitable for marker-assisted selection across gene pools.
We investigated the potential of markers associated with floral traits for parental selection in a cut rose breeding program. We analysed six Kompetitive Allele Specific PCR (KASP) markers for three important floral traits, petal length, petal number and scent, derived from experiments in a garden rose population. The six markers were applied to genotype a collection of 384 parental genotypes used for commercial cut rose breeding. We phenotyped a selection of progeny derived from pairs of parents having either high or low dosages of (contrasting) marker alleles associated with these traits. Significant differences were found between the contrasting progeny groups for each of the traits, although parents with the optimal allele dosage combinations could not always be used for the crosses. This not only supports the robustness of these marker‒trait associations but also demonstrates their potential for commercial rose breeding. It also demonstrates the use of marker information generated in garden rose populations for cut rose breeding. The online version contains supplementary material available at 10.1007/s11032-023-01438-5.


Connective tissue growth factor inhibits adipocyte differentiation.
Adipocyte differentiation is a key process implicated in the pathogenesis of obesity and insulin resistance. Its regulation is triggered by a cascade of transcription factors, including the CCAAT/enhancer binding proteins (C/EBPs) and peroxisome proliferator-activated receptor-gamma (PPARgamma). Growth factors such as transforming growth factor-beta1 (TGF-beta1) are known to inhibit adipocyte differentiation in vitro, via the C/EBP pathway, and in vivo, but whether a downstream mediator of TGF-beta1, connective tissue growth factor (CTGF), also known as CCN2, has a similar role is unknown. Mouse 3T3-L1 cells were differentiated into adipocytes by using standard methods, and effects and regulation of CTGF were studied. Intervention with recombinant human CTGF during differing stages of differentiation caused an inhibition in the development of the adipocyte phenotype, according to the gene expression of the differentiation markers adiponectin and PPARgamma, as well as suppression of lipid accumulation and expression of the lipogenic enzyme glycerol-3-phosphate dehydrogenase. Whereas CTGF gene expression promptly fell by 90% as 3T3-L1 preadipocytes differentiated into mature adipocytes, CTGF mRNA expression was induced by added TGF-beta1. CTGF applied to cells early in the course of differentiation inhibited total cell protein levels and nuclear localization of the beta-isoform of C/EBP (C/EBP-beta) and, subsequently, total cell C/EBP-alpha levels. CTGF also inhibited the adipocyte differentiation program in primary cultures of mouse preadipocytes. Expression of CTGF mRNA was twofold higher in the central fat depots of mice compared with subcutaneous fat, suggesting a potential role for CTGF in vivo. In summary, these data show that CTGF inhibits the adipocyte differentiation program.


Implantation failure of peritoneal dialysis catheters: a scanning electron microscopical study.
Patients with end stage renal failure may be treated by continuous ambulatory peritoneal dialysis. The transcutaneous portion of the catheters used in this treatment is covered with porous expanded polytetrafluoroethylene (PTFE) to provide a surface suitable for tissue infiltration. Following some instances where catheters failed to become fixed in the abdominal wall, a scanning electron microscopical study was carried out to compare the infiltration of catheters having successful or unsuccessful implantation. The porous layer of a well-fixed catheter, removed after successful renal transplantation, was infiltrated with collagen fibrils and overlaid by layered connective tissue composed of fibroblasts and collagen fibre bundles, sometimes linking to surrounding muscle fibres. The examination of four unsuccessful catheters revealed no evidence for infection being the cause of implantation failure. However the porous surface of these catheters was filled with blood components and products, sometimes apparently laid down in layers, suggesting that frequent bleeding resulting from repeated trauma may be responsible for the failure of catheter fixation. These findings led to two changes in clinical practice with apparent patient benefit. The implantation site has been relocated to reduce chafing by clothing and the post-operative wound dressing technique has been altered to minimise catheter movement.


Analysis of inteins in the Candida parapsilosis complex for simple and accurate species identification.
Inteins are coding sequences that are transcribed and translated with flanking sequences and then are excised by an autocatalytic process. There are two types of inteins in fungi, mini-inteins and full-length inteins, both of which present a splicing domain containing well-conserved amino acid sequences. Full-length inteins also present a homing endonuclease domain that makes the intein a mobile genetic element. These parasitic genetic elements are located in highly conserved genes and may allow for the differentiation of closely related species of the Candida parapsilosis (psilosis) complex. The correct identification of the three psilosis complex species C. parapsilosis, Candida metapsilosis, and Candida orthopsilosis is very important in the clinical setting for improving antifungal therapy and patient care. In this work, we analyzed inteins that are present in the vacuolar ATPase gene VMA and in the threonyl-tRNA synthetase gene ThrRS in 85 strains of the Candida psilosis complex (46 C. parapsilosis, 17 C. metapsilosis, and 22 C. orthopsilosis). Here, we describe an accessible and accurate technique based on a single PCR that is able to differentiate the psilosis complex based on the VMA intein. Although the ThrRS intein does not distinguish the three species of the psilosis complex by PCR product size, it can differentiate them by sequencing and phylogenetic analysis. Furthermore, this intein is unusually present as both mini- and full-length forms in C. orthopsilosis. Additional population studies should be performed to address whether this represents a common intraspecific variability or the presence of subspecies within C. orthopsilosis.


CancerPPD: a database of anticancer peptides and proteins.
CancerPPD (http://crdd.osdd.net/raghava/cancerppd/) is a repository of experimentally verified anticancer peptides (ACPs) and anticancer proteins. Data were manually collected from published research articles, patents and from other databases. The current release of CancerPPD consists of 3491 ACP and 121 anticancer protein entries. Each entry provides comprehensive information related to a peptide like its source of origin, nature of the peptide, anticancer activity, N- and C-terminal modifications, conformation, etc. Additionally, CancerPPD provides the information of around 249 types of cancer cell lines and 16 different assays used for testing the ACPs. In addition to natural peptides, CancerPPD contains peptides having non-natural, chemically modified residues and D-amino acids. Besides this primary information, CancerPPD stores predicted tertiary structures as well as peptide sequences in SMILES format. Tertiary structures of peptides were predicted using the state-of-art method, PEPstr and secondary structural states were assigned using DSSP. In order to assist users, a number of web-based tools have been integrated, these include keyword search, data browsing, sequence and structural similarity search. We believe that CancerPPD will be very useful in designing peptide-based anticancer therapeutics.


Rapid turnover of phosphatidylinositol-4,5-bisphosphate in insulin-secreting cells mediated by Ca2+ and the ATP-to-ADP ratio.
Phosphatidylinositol-4,5-bisphosphate (PIP(2)) is important for a variety of cellular processes as a precursor for second messengers and by regulating ion channels, the cytoskeleton, and vesicle traffic in many types of cells, including insulin-secreting beta-cells. Here, we applied evanescent wave microscopy and the PIP(2)-binding pleckstrin homology domain from phospholipase C (PLC)-delta fused to the green fluorescent protein to characterize the regulation of plasma membrane PIP(2) in individual insulin-secreting MIN6 beta-cells. Elevation of the glucose concentration from 3 to 11 mmol/l evoked antisynchronous oscillations of [PIP(2)] and cytoplasmic Ca(2+)concentration, consistent with PLC being periodically activated by the voltage-dependent Ca(2+) influx. The effect of adenine nucleotides on [PIP(2)] was studied in cells permeabilized with alpha-toxin. ATP dose- dependently stimulated PIP(2) synthesis with half-maximal effect at 300 mumol/l. Omission of the nucleotide resulted in rapid loss of PIP(2) with t(1/2) < 40 s. ADP also stimulated PIP(2) formation, but this effect reflected local ATP formation and was prevented by the adenylate kinase inhibitor diadenosine-pentaphosphate. The ATP-induced PIP(2) synthesis was counteracted by the ADP analog adenosine-5'-O-2-thiodiphosphate. We conclude that plasma membrane PIP(2) is dynamically regulated by intracellular Ca(2+) and the ATP-to-ADP ratio in insulin-secreting cells. The rapid turnover allows maintenance of PIP(2) levels while generating second messengers of critical importance for insulin secretion.


Effect on growth in pemoline-treated children with attention deficit disorder.
The growth of 22 children with attention deficit disorder (ADD) was monitored longitudinally for up to four years. Each child received at least one year of continuous, successful pemoline (Cylert) therapy, after which drug "vacations" were allowed. The effective dosage of pemoline ranged from 56 to 150 mg/day during the first year of treatment. Stature and weight measurements at six-month intervals were matched to those of paired "normal" children from the Fels Longitudinal Study. Significant deficits were observed for mean weight change at six and 12 months after baseline, and for mean stature change at 12 and 18 months after baseline. However, all subsequent six-month results up to four years did not differ significantly between the two groups. These results show a temporary retardation in the rate of growth in weight and stature with later catch-up growth in children treated wih pemoline.


Potentiometric analysis of sialic acid with a flexible carbon cloth based on boronate affinity and molecularly imprinted polymers.
An electrochemical platform was proposed for highly sensitive and selective analysis of sialic acid based on molecularly imprinted polymers (MIPs), which were electropolymerized with the monomer molecules of 3-aminophenylboronic acid (ABA) on a carbon cloth (CC) electrode in the presence of template molecules. The fabricated sensor, named the PABA/CC-based MIP electrode, could be used for the detection of sialic acid because the reversible and covalent boronic acid-diol binding was sensitive to the electrochemical potential of the prepared sensor. The utilization of a CC film as the substrate could improve the sensitivity due to its good electrical conductivity and large surface area. Under the optimized conditions, a good relationship between the change in potential and the concentration of sialic acid was obtained in the range from 40 μM to 440 μM with a detection limit of 0.5 μM. The resulting MIP sensor also displayed good selectivity, reproducibility and stability. Moreover, this sensor was successfully applied for the evaluation of the sialic acid level in infant formulas.


Indirect Co-Culture of Testicular Cells with Bone Marrow Mesenchymal Stem Cells Leads to Male Germ Cell-Specific Gene Expressions.
Non-obstructive azoospermia is mostly irreversible. Efforts to cure this type of infertility have led to the application of stem cells in the reproduction field. In the present study, testicular cell-mediated differentiation of male germ-like cells from bone marrow-derived mesenchymal stem cells (BM-MSCs) in an in vitro indirect co-culture system is investigated. In this experimental study, mouse BM-MSCs were isolated and cultured up to passage three. Identification of the cells was evaluated using specific surface markers by flow-cytometry technique. Four experimental groups were investigated: control, treatment with retinoic acid (RA), indirect co-culture with testicular cells, and combination of RA and indirect co-culture with testicular cells. Finally, following differentiation, the quantitative expression of germ cell-specific markers including Dazl, Piwil2 and Stra8 were evaluated by real-time polymerase chain reaction (PCR). Molecular analysis revealed a significant increase in Dazl expression in the indirect co-culture with testicular cells group in comparison to the control group. Quantitative expression level of Piwil2 was not significantly changed in comparison to the control group. Stra8 expression was significantly higher in RA group in comparison to other groups. Indirect co-culture of BM-MSCs in the presence of testicular cells leads to expression of male germ cell-specific gene, Dazl, in the induced cells. Combination of co-culture with testicular cells and RA did not show any positive effect on the specific gene expressions.


Renal hematoma as a complication of extracorporeal shock wave lithotripsy.
To evaluate risk factors, clinical presentation, therapeutic management, and treatment of residual stones and subsequent development of renal hematoma following Extracorporeal Shock Wave Lithotripsy (ESWL). A retrospective review was made of 31 post-ESWL renal hematoma cases diagnosed between May 1987 and June 1996. Lithotripsy treatments were outpatient procedures without anaesthesia. Our center has three electromagnetic sources, two with biplane X-ray centering and one ultrasound-guided (SIEMENS Lithostar II, SIEMENS Lithostar System C and SIEMENS Lithostar Ultra, respectively). We analysed findings from patient history, physical examination, blood analysis, and renovesical sonographs. Follow-up involved periodical checks, blood analyses and renovesical sonographs, scheduled first at three-month intervals and later at six-month intervals. Our center performed 21 699 lithotripsies on a total of 10 953 patients in this period. Thirty-one renal hematomas were diagnosed, giving an incidence rate of 0.28%. Twenty-four patients presented clinical onsets and the commonest symptom for consultation was low back pain (74%). Eleven patients of this group (46%) were hypertensive. All patients received conservative treatment. With a mean follow-up time of 18 months, ultrasound showed persistent hematoma in 11 patients (36%). There were residual stones in 71% of patients; further lithotripsy was performed on seven patients with no clinical or ultrasonographic signs of change in the hematoma. Renal hematoma post-ESWL is a rare complication. Main risk factors are hypertension, clotting disorders and previous ESWL sessions. Flank pain is the main symptom at presentation. Elective management is conservative. Presence of hematoma is not a contraindication for further treatments of residual stones.


Iranian nurses' knowledge, attitude and behaviour on skin care, prevention and management of pressure injury: A descriptive cross-sectional study.
Pressure injuries as an indicator measuring the quality of nursing care and patient safety is a major health care problem worldwide. The aim of this study was to assess the knowledge, attitude and behaviour of nurses in preventing pressure injuries. Descriptive cross-sectional study. This descriptive cross-sectional study enrolled 214 registered nurses in Iran. Patient satisfaction was assessed using pieker pressure ulcer knowledge test, attitude towards pressure ulcer tool and behaviour of pressure ulcer questionnaire. Data analysed by SPSSv.24 applying descriptive and inferential statistics. The mean scores of knowledge, attitude and behaviour of nurses on the prevention of pressure injury were 27.24 (SD 5.23), 38.55 (SD 6.43) and 51.24 (SD 7.54), respectively. There was a correlation between knowledge, attitude and behaviour with the history of pressure injury training. Also, there was a significant relationship between knowledge with educational level and attitude with work experience. Knowledge, attitude and behaviour of nurses were in moderate level. Necessary measures to overcome problems such as the availability of pressure reducing equipments, motivating the nurses, eliminating the shortage of nurses and empowering nurses by holding practical workshops are important in providing patients safety.


[Recommendations for the diagnosis of candidemia in Latin America. Grupo Proyecto Épico].
Candidemia is one of the most frequent opportunistic mycoses worldwide. Limited epidemiological studies in Latin America indicate that incidence rates are higher in this region than in the Northern Hemisphere. Diagnosis is often made late in the infection, affecting the initiation of antifungal therapy. A more scientific approach, based on specific parameters, for diagnosis and management of candidemia in Latin America is warranted. 'Recommendations for the diagnosis and management of candidemia' are a series of manuscripts that have been developed by members of the Latin America Invasive Mycosis Network. They aim to provide a set of best-evidence recommendations for the diagnosis and management of candidemia. This publication, 'Recommendations for the diagnosis of candidemia in Latin America', was written to provide guidance to healthcare professionals on the diagnosis of candidemia, as well as on the usefulness and application of susceptibility testing in patients who have a confirmed diagnosis of candidemia. Computerized searches of existing literature were performed by PubMed. The data were extensively reviewed and analyzed by members of the group. The group also met on two occasions to pose questions, discuss conflicting views, and deliberate on a series of management recommendations. 'Recommendations for the diagnosis of candidemia in Latin America' includes diagnostic methods used to detect candidemia, Candida species identification, and susceptibility testing. The availability of methods, their costs and treatment settings are considered. This manuscript is the first of this series that deals with diagnosis and treatment of invasive candidiasis. Other publications in this series include: 'Recommendations for the management of candidemia in adults in Latin America', 'Recommendations for the management of candidemia in children in Latin America', and 'Recommendations for the management of candidemia in neonates in Latin America'.


Vasopressin regulates apical and basolateral Na(+)-H+ antiporters in mouse medullary thick ascending limbs.
We assessed in isolated perfused mouse medullary thick ascending limb (MTAL) segments Na(+)-H+ antiporter activity in both apical and basolateral membranes and the effects of arginine vasopressin (AVP) on the activities of these antiporters under isotonic conditions using 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein to monitor intracellular pH (pHi). When the apical Na(+)-H+ antiporter was inhibited in the absence of AVP with removal of luminal Na+ plus addition of 0.5 mM amiloride, a small but significant increase in pHi was observed after luminal NH4Cl-induced acidification of MTAL cells to pHi less than 6.7. This increase in pHi was dependent on basolateral Na+ and inhibited with 0.5 mM basolateral amiloride, consistent with the function of a basolateral Na(+)-H+ antiporter. Basolateral AVP (100 microU/ml) enhanced the rate of pHi recovery due to the basolateral Na(+)-H+ antiporter by more than twofold. In contrast, AVP decreased the apical Na(+)-H+ antiporter activity by 50%. In the absence of AVP, addition of 0.5 mM amiloride to the luminal perfusate reduced steady-state pHi by 0.40 +/- 0.07 units, whereas exposure of the basolateral membrane to the same concentration of amiloride had no effect on pHi (delta pHi = 0.01 +/- 0.01 units). AVP reduced the magnitude of cell acidification on exposure of apical membranes to amiloride (delta pHi = 0.16 +/- 0.03) but increased the pHi response to basolateral amiloride (delta pHi = 0.09 +/- 0.00). Thus Na(+)-H+ antiporters are present on both apical and basolateral membranes of the mouse MTAL in the absence of AVP. AVP stimulates the basolateral, while inhibiting the apical, Na(+)-H+ antiporter.(ABSTRACT TRUNCATED AT 250 WORDS)


Systematic review of pregnancy in women with inherited cardiomyopathies.
Pregnancy exposes women with inherited cardiomyopathies to increased risk for heart failure and arrhythmias. In this paper, we review the clinical course and management of pregnant women with the following inherited cardiomyopathies: hypertrophic cardiomyopathy, dilated cardiomyopathy, arrhythmogenic right ventricular cardiomyopathy, left ventricular non-compaction cardiomyopathy, and restrictive cardiomyopathy. We also discuss peripartum cardiomyopathy. Pregnancy is generally well tolerated in asymptomatic patients with inherited cardiomyopathies. However, worsening of the clinical condition can occur during pregnancy, despite intensive medical treatment. If prior cardiac events, poor functional class (New York Heart Association class III or IV), or advanced left ventricular systolic dysfunction are present, the risk of maternal cardiac complications during pregnancy are markedly increased. The postpartum condition is generally no worse than the antepartum condition, but no long-term follow-up studies have been reported. Preconception evaluation and counselling are important aspects of managing women with inherited cardiomyopathies. Genetic counselling and DNA testing should be offered to all women following the diagnosis of an inherited cardiomyopathy.


The clinical reasoning of an occupational therapy assistant.
This qualitative case study focused on the clinical reasoning of a certified occupational therapy assistant who had 16 years of practice experience. Observation and interview methods were used to collect data. Transcripts and field notes were coded using a priori codes of the forms of clinical reasoning of occupational therapists identified in published research. The study participant demonstrated the use of pragmatic, procedural, interactive, conditional, and narrative reasoning. We encourage further research to support these findings and to increase the understanding of the forms of clinical reasoning used by occupational therapy assistants.


Spatial regulation of an E3 ubiquitin ligase directs selective synapse elimination.
Stereotyped synaptic connectivity can arise both by precise recognition between appropriate partners during synaptogenesis and by selective synapse elimination. The molecular mechanisms that underlie selective synapse removal are largely unknown. We found that stereotyped developmental elimination of synapses in the Caenorhabditis elegans hermaphrodite-specific motor neuron (HSNL) was mediated by an E3 ubiquitin ligase, a Skp1-cullin-F-box (SCF) complex composed of SKR-1 and the F-box protein SEL-10. SYG-1, a synaptic adhesion molecule, bound to SKR-1 and inhibited assembly of the SCF complex, thereby protecting nearby synapses. Thus, subcellular regulation of ubiquitin-mediated protein degradation contributes to precise synaptic connectivity through selective synapse elimination.


Anticipating undiagnosed asthma in symptomatic adults with normal pre- and post-bronchodilator spirometry: a decision tool for bronchial challenge testing.
Some patients with asthma demonstrate normal spirometry and remain undiagnosed without further testing. To determine clinical predictors of asthma in symptomatic adults with normal spirometry, and to generate a tool to help clinicians decide who should undergo bronchial challenge testing (BCT). Using random-digit dialling and population-based case-finding, we recruited adults from the community with respiratory symptoms and no previous history of diagnosed lung disease. Participants with normal pre- and post-bronchodilator spirometry subsequently underwent BCT. Asthma was diagnosed in those with symptoms and a methacholine provocative concentration (PC20) of < 8 mg/ml. Sputum and blood eosinophils, and exhaled nitric oxide were measured. Univariate analyses identified potentially predictive variables, which were then used to construct a multivariable logistic regression model to predict asthma. Model sensitivity, specificity, and area under the receiver operating curve (AUC) were calculated. Of 132 symptomatic individuals with normal spirometry, 34 (26%) had asthma. Of those ultimately diagnosed with asthma, 33 (97%) answered 'yes' to a question asking whether they experienced cough, chest tightness or wheezing provoked by exercise or cold air. Other univariate predictors of asthma included female sex, pre-bronchodilator FEV1 percentage predicted, and percent positive change in FEV1 post bronchodilator. A multivariable model containing these predictive variables yielded an AUC of 0.82 (95% CI: 0.72-0.91), a sensitivity of 82%, and a specificity of 66%. The model was used to construct a nomogram to advise clinicians which patients should be prioritized for BCT. Four readily available patient characteristics demonstrated a high sensitivity and AUC for predicting undiagnosed asthma in symptomatic adults with normal pre- and post-bronchodilator spirometry. These characteristics can potentially help clinicians to decide which individuals with normal spirometry should be investigated with bronchial challenge testing. However, further prospective validation of our decision tool is required.


The usefulness of immunohistochemical markers in the differential diagnosis of renal neoplasms.
Histologic subtyping of RCC has been shown to be of prognostic value; therefore, it is important to classify malignant epithelial tumors of the kidney correctly and also to differentiate them from benign ones. Overlapping morphologic features of renal tumors sometimes make histologic subtyping difficult. The accurate diagnosis and classification of RCC are based on cytoarchitectural features and require correlation with immunophenotype and cytogenetic characteristics. RCC Ma and CD10, two markers with relative renal specificity, have been used to confirm a diagnosis of suspected RCC and can facilitate the accurate diagnosis of metastatic RCC, in particular, in FNA. Although CCRCC and PRCC share most immunomarkers, CK7 and AMACR expression can be helpful in the differential diagnosis of challenging histologic variants of the two. In addition, E-cadherin aids in the distinction between types 1 and 2 PRCC. Useful markers in the differential diagnosis between ChRCC and CCRCCare CK7, RCC Ma, CD10, VIM, CD117, parvalbumin, and E-cadherin. We propose CK7/CK20/CD15 as a useful primary immunopanel to differentiate ChRCC from ONC reliably.


Quest for highly connected metal-organic framework platforms: rare-earth polynuclear clusters versatility meets net topology needs.
Gaining control over the assembly of highly porous rare-earth (RE) based metal-organic frameworks (MOFs) remains challenging. Here we report the latest discoveries on our continuous quest for highly connected nets. The topological exploration based on the noncompatibility of a 12-connected RE polynuclear carboxylate-based cluster, points of extension matching the 12 vertices of the cuboctahedron (cuo), with 3-connected organic ligands led to the discovery of two fascinating and highly connected minimal edge-transitive nets, pek and aea. The reduced symmetry of the employed triangular tricarboxylate ligand, as compared to the prototype highly symmetrical 1,3,5-benzene(tris)benzoic acid guided the concurrent occurrence of nonanuclear [RE9(μ3-OH)12(μ3-O)2(O2C-)12] and hexanuclear [RE6(OH)8(O2C-)8] carboxylate-based clusters as 12-connected and 8-connected molecular building blocks in the structure of a 3-periodic pek-MOF based on a novel (3,8,12)-c trinodal net. The use of a tricarboxylate ligand with modified angles between carboxylate moieties led to the formation of a second MOF containing solely nonanuclear clusters and exhibiting once more a novel and a highly connected (3,12,12)-c trinodal net with aea topology. Notably, it is the first time that RE-MOFs with double six-membered ring (d6R) secondary building units are isolated, representing therefore a critical step forward toward the design of novel and highly coordinated materials using the supermolecular building layer approach while considering the d6Rs as building pillars. Lastly, the potential of these new MOFs for gas separation/storage was investigated by performing gas adsorption studies of various probe gas molecules over a wide range of pressures. Noticeably, pek-MOF-1 showed excellent volumetric CO2 and CH4 uptakes at high pressures.


The chemical space project.
One of the simplest questions that can be asked about molecular diversity is how many organic molecules are possible in total? To answer this question, my research group has computationally enumerated all possible organic molecules up to a certain size to gain an unbiased insight into the entire chemical space. Our latest database, GDB-17, contains 166.4 billion molecules of up to 17 atoms of C, N, O, S, and halogens, by far the largest small molecule database reported to date. Molecules allowed by valency rules but unstable or nonsynthesizable due to strained topologies or reactive functional groups were not considered, which reduced the enumeration by at least 10 orders of magnitude and was essential to arrive at a manageable database size. Despite these restrictions, GDB-17 is highly relevant with respect to known molecules. Beyond enumeration, understanding and exploiting GDBs (generated databases) led us to develop methods for virtual screening and visualization of very large databases in the form of a "periodic system of molecules" comprising six different fingerprint spaces, with web-browsers for nearest neighbor searches, and the MQN- and SMIfp-Mapplet application for exploring color-coded principal component maps of GDB and other large databases. Proof-of-concept applications of GDB for drug discovery were realized by combining virtual screening with chemical synthesis and activity testing for neurotransmitter receptor and transporter ligands. One surprising lesson from using GDB for drug analog searches is the incredible depth of chemical space, that is, the fact that millions of very close analogs of any molecule can be readily identified by nearest-neighbor searches in the MQN-space of the various GDBs. The chemical space project has opened an unprecedented door on chemical diversity. Ongoing and yet unmet challenges concern enumerating molecules beyond 17 atoms and synthesizing GDB molecules with innovative scaffolds and pharmacophores.


Preparation and sorption properties of materials from paper sludge.
Three materials were prepared from paper sludge (PS) using different treatment processes and their sorption abilities for phosphate and methylene blue (MB) were determined. The samples were a powder sample prepared by heating PS in air (sample C), a pellet prepared by grinding, forming and heating PS in air (sample G) and a powder prepared by physical activation of PS in flowing wet nitrogen (sample A). The three samples were heated at 600-900 degrees C for 6h. On heating at 700-800 degrees C, the organic fibers, limestone (CaCO3), kaolinite (Al2Si2O5(OH)4) and talc (Mg3Si4O10(OH)2) in the original PS were converted to amorphous CaO-Al2O3-SiO2 (CAS) and talc in sample C, while CAS was formed in sample G and activated carbon, CAS and talc was formed in sample A. On heating at 900 degrees C the CAS converted to gehlenite (Ca2Al2SiO7) and anorthite (CaAl2Si2O8). The specific surface areas (SBET) of the three samples achieved maximum values of 23, 37 and 70 m2/g upon heating at 700, 600 and 600 degrees C, respectively. The SBET value of the activated sample A was distinctly lower than usually reported for activated carbon. The samples C, G and A achieved maximum phosphate sorption capacities of 2.04, 1.38 and 1.70 mmol/g, calculated from the Langmuir model, upon heating at 700, 700 and 800 degrees C, respectively. The maximum sorption capacity for phosphate in sample C is attributed to the sorption by CAS, namely, adsorption on the alumina component and precipitation as Ca-phosphates. The MB multifunctional sorption capacity of sample A was 0.11 mmol/g. The phosphate and MB sorption rates show better correlation with a pseudo-second order model than with other models.


Mixtures of octanol and an ionic liquid: Structure and transport.
Ionic liquids (ILs) with long alkyl substituents are amphiphilic, which leads to a bicontinuous liquid structure. The strongly interacting anionic and cationic head groups form a long range charge network, with the hydrocarbon tails forming a nonpolar domain. Such nonpolar domains have been shown to dissolve a variety of neutral organic solvents. In mixtures of ILs with solvents the neutral organic molecules residing in the nonpolar domains experience different environments and friction from the charged cations and anions. Thus, the neutral molecules diffuse much faster than predicted by hydrodynamic scaling using the average viscosity of the mixture. In this work, we report studies on the structure and transport properties of mixtures of 1-octanol with the IL trihexyltetradecylphosphonium bis(trifluoromethylsulfonyl)imide (P6,6,6,14 +/NTf2 -). The majority of the atom fraction in the P6,6,6,14 + cation comprises four hydrocarbon substituents. The unique amphiphilic nature of ILs with the P6,6,6,14 + cation makes 1-octanol fully miscible with the IL at ambient temperatures. X-ray scattering experiments show that the IL structure persists in the mixtures for 1-octanol mole fractions as large as xoct = 0.90. The self-diffusion coefficients of the three molecular species in the mixtures were measured by NMR experiments. The self-diffusion of the P6,6,6,14 + cation is well described by the Stokes-Einstein equation, while the diffusivity of the NTf2 - anion is slightly lower than the hydrodynamic prediction. The measured diffusivities of octanol in these mixtures are 1.3-4 times higher than the hydrodynamic predictions.


Cell fusion-induced activation of interferon-stimulated genes is not required for restriction of a herpes simplex virus VP16/ICP0 mutant in heterokarya formed between permissive and restrictive cells.
Herpes simplex virus VP16 and ICP0 mutants replicate efficiently in U2OS osteosarcoma cells but are restricted in other cell types. We previously showed that the restrictive phenotype is dominant in a transient cell fusion assay, suggesting that U2OS cells lack an antiviral mechanism present in other cells. Recent data indicate that unscheduled membrane fusion events can activate the expression of interferon-stimulated genes (ISGs) in fibroblasts, raising the possibility that our earlier results were due to a fusion-induced antiviral state. However, we show here that the permissive phenotype is also extinguished following fusion with Vero cells in the absence of ISG induction.


Tissue-Resident Memory T Cells in Skin Diseases: A Systematic Review.
In health, the non-recirculating nature and long-term persistence of tissue-resident memory T cells (TRMs) in tissues protects against invading pathogens. In disease, pathogenic TRMs contribute to the recurring traits of many skin diseases. We aimed to conduct a systematic literature review on the current understanding of the role of TRMs in skin diseases and identify gaps as well as future research paths. EMBASE, PubMed, SCOPUS, Web of Science, Clinicaltrials.gov and WHO Trials Registry were searched systematically for relevant studies from their inception to October 2020. Included studies were reviewed independently by two authors. This study was conducted in accordance with the PRISMA-S guidelines. This protocol was registered with the PROSPERO database (ref: CRD42020206416). We identified 96 studies meeting the inclusion criteria. TRMs have mostly been investigated in murine skin and in relation to infectious skin diseases. Pathogenic TRMs have been characterized in various skin diseases including psoriasis, vitiligo and cutaneous T-cell lymphoma. Studies are needed to discover biomarkers that may delineate TRMs poised for pathogenic activity in skin diseases and establish to which extent TRMs are contingent on the local skin microenvironment. Additionally, future studies may investigate the effects of current treatments on the persistence of pathogenic TRMs in human skin.


Active ankylosing spondylitis increases blood loss during total hip arthroplasty for a stiff hip joint.
Total hip arthroplasty (THA) has been highlighted as the best treatment option for ankylosing spondylitis (AS) patients with advanced hip involvement. The huge blood loss associated with THA is a common concern of postoperative complications. Disease activity is a specific reflection of systematic inflammation of AS. The purpose of this study was to determine the effect of disease activity on blood loss during THA in patients with AS. Forty-nine patients with AS who underwent unilateral THAs were retrospectively studied. Ankylosing Spondylitis Disease Activity Score (ASDAS) was employed to evaluate the disease activity. Orthopedic Surgery Transfusion Hemoglobin European Overview (OSTHEO) formula was used to assess the surgical blood loss. The patients were divided into active group (ASDAS≥1.3; n = 32) and stable groups (ASDAS< 1.3; n = 17) based on the ASDAS. Peri-operative laboratory values, plain radiographs, intra-operative data, transfusion volume, and use of hemostatic agents were recorded and statistically analyzed. The ASDAS, pre-operative C-reactive protein level, erythrocyte sedimentation rate, and fibrinogen concentration in the active group were higher than the stable group (all P < 0.05); however, the pre-operative hemoglobin concentration and albumin level were higher in the stable group (both P < 0.05). The total blood loss during THA in stable patients was 1415.31 mL and 2035.04 mL in active patients (P = 0.006). The difference between the two groups was shown to be consistent after excluding the gender difference (P = 0.030). A high transfusion rate existed in both groups (stable group, 76.47% with an average of 1.53 units; active group, 84.37% with an average of 2.31 units), but there was no significant difference between the two groups (both P > 0.05). Compensated blood loss, corresponding to transfusion, was noted significantly more in the active group compared to the stable group (P = 0.027). There was no significant difference with regard to functional recovery (P > 0.05). Active AS patients are at high risk for increased blood loss during THA compared to stable patients. The underlying mechanism includes disorders of the coagulation and fibrinolytic systems, poor nutrition status, osteoporosis, imbalance of oxidative-antioxidative status and local inflammatory reaction. It is strongly recommended to perform THA in AS patients with stable disease.


[Early childhood prevention: implications for applied research and quality management].
The importance of prevention has received increasing recognition the world over. In Germany, laws are being enacted and initiatives are taken to make prevention a firmly anchored and integral part of public health care. Preventive measures in early childhood encompass a great breadth and variety of resources. However, any specific preventive measure can only be justified to the extent that it has been proven to be efficient and effective. In Germany small collection of studies have shown the efficacy of psychosocial prevention and early intervention programs. Quality standards can only be determined when empirical evidence has demonstrated the prerequisites for ensuring successful treatment. Criteria for success must include life quality or academic success (school achievement). The stability of the family must also be assessed both before and after the intervention.


[Impaired cognitive functioning in low-grade glioma patients: relationship to tumor localisation, radiotherapy and the use of anticonvulsants].
To determine the effects of radiotherapy and other medical interventions on cognitive functioning in patients with a low-grade glioma (LGG). Cross-sectional study. A total of 195 LGG patients, of whom 104 had received radiotherapy 1-22 years previously, were compared to 100 patients with a low-grade haematological malignancy and 195 healthy controls. The analysis was aimed at differentiating between the effects of the tumour (disease duration, lateralisation) and treatment effects (neurosurgery, radiotherapy, use of anticonvulsants) on cognitive function and the relative risk of cognitive disability. LGG patients had lower performance levels in all cognitive domains than haematological patients and performed even worse when they were compared to healthy controls. Radiotherapy was associated with poorer cognitive functioning; however, cognitive disability was found only in patients receiving fractional doses exceeding 2 Gy. The use of anticonvulsants was strongly associated with disorders in the area of attention and planning functions. In this study, the tumour itself was the most damaging factor with respect to cognitive function and radiotherapy was associated with cognitive disability only if elevated fractional doses were used. Epilepsy or the use of anticonvulsants was also associated with diminished cognitive functioning.


Repression of both isoforms of disproportionating enzyme leads to higher malto-oligosaccharide content and reduced growth in potato.
Two glucanotransferases, disproportionating enzyme 1 (StDPE1) and disproportionating enzyme 2 (StDPE2), were repressed using RNA interference technology in potato, leading to plants repressed in either isoform individually, or both simultaneously. This is the first detailed report of their combined repression. Plants lacking StDPE1 accumulated slightly more starch in their leaves than control plants and high levels of maltotriose, while those lacking StDPE2 contained maltose and large amounts of starch. Plants repressed in both isoforms accumulated similar amounts of starch to those lacking StDPE2. In addition, they contained a range of malto-oligosaccharides from maltose to maltoheptaose. Plants repressed in both isoforms had chlorotic leaves and did not grow as well as either the controls or lines where only one of the isoforms was repressed. Examination of photosynthetic parameters suggested that this was most likely due to a decrease in carbon assimilation. The subcellular localisation of StDPE2 was re-addressed in parallel with DPE2 from Arabidopsis thaliana by transient expression of yellow fluorescent protein fusions in tobacco. No translocation to the chloroplasts was observed for any of the fusion proteins, supporting a cytosolic role of the StDPE2 enzyme in leaf starch metabolism, as has been observed for Arabidopsis DPE2. It is concluded that StDPE1 and StDPE2 have individual essential roles in starch metabolism in potato and consequently repression of these disables regulation of leaf malto-oligosaccharides, starch content and photosynthetic activity and thereby plant growth possibly by a negative feedback mechanism.


The Legacy of Recurrent Introgression during the Radiation of Hares.
Hybridization may often be an important source of adaptive variation, but the extent and long-term impacts of introgression have seldom been evaluated in the phylogenetic context of a radiation. Hares (Lepus) represent a widespread mammalian radiation of 32 extant species characterized by striking ecological adaptations and recurrent admixture. To understand the relevance of introgressive hybridization during the diversification of Lepus, we analyzed whole exome sequences (61.7 Mb) from 15 species of hares (1-4 individuals per species), spanning the global distribution of the genus, and two outgroups. We used a coalescent framework to infer species relationships and divergence times, despite extensive genealogical discordance. We found high levels of allele sharing among species and show that this reflects extensive incomplete lineage sorting and temporally layered hybridization. Our results revealed recurrent introgression at all stages along the Lepus radiation, including recent gene flow between extant species since the last glacial maximum but also pervasive ancient introgression occurring since near the origin of the hare lineages. We show that ancient hybridization between northern hemisphere species has resulted in shared variation of potential adaptive relevance to highly seasonal environments, including genes involved in circadian rhythm regulation, pigmentation, and thermoregulation. Our results illustrate how the genetic legacy of ancestral hybridization may persist across a radiation, leaving a long-lasting signature of shared genetic variation that may contribute to adaptation. [Adaptation; ancient introgression; hybridization; Lepus; phylogenomics.].


Role of NeuroD1 on the negative regulation of Pomc expression by glucocorticoid.
The mechanism of the negative regulation of proopiomelanocortin gene (Pomc) by glucocorticoids (Gcs) is still unclear in many points. Here, we demonstrated the involvement of neurogenic differentiation factor 1 (NeuroD1) in the Gc-mediated negative regulation of Pomc. Murine pituitary adrenocorticotropic hormone (ACTH) producing corticotroph tumor-derived AtT20 cells were treated with dexamethasone (DEX) (1-100 nM) and cultured for 24 hrs. Thereafter, Pomc mRNA expression was studied by quantitative real-time PCR and rat Pomc promoter (-703/+58) activity was examined by luciferase assay. Both Pomc mRNA expression and Pomc promoter activity were inhibited by DEX in a dose-dependent manner. Deletion and point mutant analyses of Pomc promoter suggested that the DEX-mediated transcriptional repression was mediated via E-box that exists at -376/-371 in the promoter. Since NeuroD1 is known to bind to and activate E-box of the Pomc promoter, we next examined the effect of DEX on NeuroD1 expression. Interestingly, DEX dose-dependently inhibited NeuroD1 mRNA expression, mouse NeuroD1 promoter (-2.2-kb) activity, and NeuroD1 protein expression in AtT20 cells. In addition, we confirmed the inhibitory effect of DEX on the interaction of NeuroD1 and E-box on Pomc promoter by chromatin immunoprecipitation (ChIP) assay. Finally, overexpression of mouse NeuroD1 could rescue the DEX-mediated inhibition of Pomc mRNA expression and Pomc promoter activity. Taken together, it is suggested that the suppression of NeuroD1 expression and the inhibition of NeuroD1/E-box interaction may play an important role in the Gc-mediated negative regulation of Pomc.


Examination of key intermediates in the catalytic cycle of aspartate-beta-semialdehyde dehydrogenase from a gram-positive infectious bacteria.
Aspartate-beta-semialdehyde dehydrogenase (ASADH) catalyzes a critical branch point transformation in amino acid bio-synthesis. The products of the aspartate pathway are essential in microorganisms, and this entire pathway is absent in mammals, making this enzyme an attractive target for antibiotic development. The first structure of an ASADH from a Gram-positive bacterium, Streptococcus pneumoniae, has now been determined. The overall structure of the apoenzyme has a similar fold to those of the Gram-negative and archaeal ASADHs but contains some interesting structural variations that can be exploited for inhibitor design. Binding of the coenzyme NADP, as well as a truncated nucleotide analogue, into an alternative conformation from that observed in Gram-negative ASADHs causes an enzyme domain closure that precedes catalysis. The covalent acyl-enzyme intermediate was trapped by soaking the substrate into crystals of the coenzyme complex, and the structure of this elusive intermediate provides detailed insights into the catalytic mechanism.


How first-time mothers perceive and deal with teething symptoms: a randomized controlled trial.
Teething, especially in their first child, continues to be a daunting problem for parents. The objective of this paper was to assess the effects of providing first-time mothers with information about symptoms commonly associated with teething and ways to manage these. In a randomized controlled trial to decrease the incidence of early childhood caries, we included information on teething as another issue in a child's oral health. Mothers in the intervention group received three rounds of printed information: at enrolment during pregnancy and when the child was 6 and 12 months old. Information on teething arrived when a child reached 6 months of age. Outcome assessment was at 20 ± 2.5 months of age. Data were complemented with a systematic search for evidence on teething symptoms and how to alleviate them in other populations. Of 649 expectant mothers enrolled in the study, 441 completed the 'Child's oral health' questionnaire. There were no significant differences in teething symptoms reported by mothers in the intervention (n = 232) and control (n = 209) groups. However, mothers in the intervention group were less likely to use topical and oral medications to manage teething problems (P < 0.03) and relied more on rubbing the gums to ease discomfort (P < 0.005) than mothers in the control group. Providing mothers with information on how to address teething symptoms markedly reduced the use of medications for symptom relief. There is still need for better evidence, first, on what symptoms can or cannot be attributed to teething and, second, on what is effective in alleviating them.


Safety of the application of nanosilver and nanogold in topical cosmetic preparations.
The safety of the use of cosmetic preparations with silver or gold nanoparticles was assessed. This study describes the methodology and results of research involving the generation of suspensions of silver and gold nanoparticles and creams with added silver or gold at concentrations of 20, 65, 110, 155, and 200 mg/kg. The silver nanoparticles ranged from 8 to 140 nm, and the gold nanoparticles, measured 13-99 nm. The sizes were determined using dynamic light scattering. The presence of metallic nanoparticles in the obtained oil-in-water emulsions was confirmed with UV-vis spectroscopy and transmission electron microscopy with an X-ray scattering spectrometer (TEM-EDX). Additionally, based on the TEM-EDX results, it was possible to analyse the distributions of the silver nanoparticles in the tested cosmetic emulsions. Microbiological tests showed that both the silver and gold nanoparticle emulsion possessed satisfactory fungicidal properties. Based on viscosity curves, the lowest estimated yield limits were achieved by the reference cream and the creams with the gold and silver nanoparticles at concentrations of 20 and 65 mg/kg, respectively, which improved their consistencies and distributions on the skin. The best appraisals from the respondents in terms of consistency, absorption, oiling, colour, and smell were received for the emulsion containing 200 mg/kg gold nanoparticles. The worst assessment in terms of uniformity, colour, and smell were obtained for the emulsion with 200 mg/kg silver nanoparticles. However, the most important aspect of this study was the assessment of the permeabilities of the metallic nanoparticles through imitation skin in the form of dermal membranes. The highest permeabilities were confirmed for the creams with metallic nanoparticles present at 110--200 mg/kg. This permeability is an issue of concern given the toxic properties of metallic nanoparticles for living organisms.


Bacopa monniera ameliorates amnesic effects of diazepam qualifying behavioral-molecular partitioning.
Benzodiazepines are known to produce amnesia by involvement of GABAergic system and by interference of long term potentiation (LTP). In this study, we examined effect of Bacopa monniera on downstream molecules of LTP after diazepam-induced amnesia in mice. We used a Morris water maze scale for evaluating the effect of Bacopa monniera after screening for muscle coordination by rota rod. The index of acquisition and retrieval was recorded as escape latency time (ELT). Behavioral results showed that Bacopa monniera (120 mg kg(-1) oral) significantly reversed diazepam- (1.75 mg kg(-1) i.p.) induced amnesia in Morris water maze task. The molecular studies revealed that diazepam upregulated mitogen activated protein kinase (MAP kinase), phosphorylated CREB (pCREB) and inducible nitric oxide synthase (iNOS), while it downregulated nitrite, nitrate, total nitrite, cAMP response element binding protein (CREB) expression, phosphodiesterase, cyclic adenosine monophosphate (cAMP) without affecting calmodulin levels. Bacopa monniera suppressed the diazepam induced upregulation of MAP kinase, pCREB and iNOS and attenuated the downregulation of nitrite. It did not affect the cAMP, PDE, nitrate, total nitrite, total CREB level. These behavioral findings displayed the reversal of diazepam-induced amnesia by Bacopa monniera without qualifying the molecular details although some downstream molecules of LTP may be involved.


[Automation of the technological medical process in ambulatory polyclinic and hospital establishments].
Now, the creation of automated instruments, apparatus and systems providing complex instrumentation of medical technological processes in ambulant-polyclinical and hospital centers became the main line of the medical technique's development. Primary problems and objects of automation are determined from the proposed models of managing the treatment and diagnostic processes in these establishments.


Variation in resource limitation of plant reproduction influences natural selection on floral traits of Asclepias syriaca.
The availability of both pollen and resources can influence natural selection on floral traits, but their relative importance in shaping floral evolution is unclear. We experimentally manipulated pollinator and resource (fertilizer and water) availability in the perennial wildflower Asclepias syriaca L. Nine floral traits, one male fitness component (number of pollinia removed), and two female fitness components (number of pollinia inserted and number of fruits initiated) were measured for plants in each of three treatments (unmanipulated control, decreased pollinator access, and resource supplementation). Although decreasing pollinators' access to flowers did result in fewer pollinia inserted and removed, fruit set and phenotypic selection on floral traits via female and male fitness did not differ from the control. In contrast, resource supplementation increased fruit set, and phenotypic selection on seven out of nine floral traits was stronger via female than male fitness, consistent with the prediction that selection via female fitness would be greater when reproduction was less resource-limited. Our results support the hypothesis that abiotic resource availability can influence floral evolution by altering gender-specific selection.


[In-vitro and in-vivo activities of moxifloxacin and garenoxacin against Mycobacterium leprae].
Moxifloxacin(MFLX) and garenoxacin(GRNX), new synthetic antibacterial agents, were assessed for in vitro anti-M. leprae activities. The anti-bacterial activities of these two drugs were compared to those of sparfloxacin (SPFX), gatifloxacin(GFLX), levofloxacin(LVFX) and rifampicin (RFP). The anti-M leprae activity obtained by Buddemeyer system was stronger in order of RFP, MFLX, SPFX, GFLX and GRNX and LVFX. The anti-M. leprae activity of MFLX or GRNX was also examined by the nude mouse footpad method. MFLX completely inhibited the growth of M. leprae inoculated into nude mouse footpads, when given orally at a daily dose of 10 mg/kg , while GRNX completely inhibited at a daily dose of 60 mg/kg. Both in vitro and in vivo tests indicated that MFLX was equal or superior to SPFX, but GRNX was equipotent to LVFX, in terms of anti-M. leprae activities.


Effect of nitrate on sulfur transformations in sulfidogenic sludge of a marine aquaculture biofilter.
The effect of NO(3)(-) addition on dissimilatory SO(4)(2-) reduction and sulfide conversion in organic-rich sludge from the digestion basin of a recirculating marine aquaculture system was studied. SO(4)(2-) reduction could only explain a minor fraction (up to 4-9%) of the observed total sulfide production (up to 35 mmol L(-1) day(-1)), indicating that the main source of sulfide in the sludge was not SO(4)(2-) reduction, but desulfuration during the decomposition of organic matter. Although NO(3)(-) inhibited SO(4)(2-) reduction, but not desulfuration, the primary NO(3)(-) mitigation effect was the onset of NO(3)(-)-mediated sulfide oxidation (up to 75 mmol L(-1) day(-1)), partially to elemental sulfur (S(0)). Above NO(3)(-) concentrations of 0.6 mM in the bulk water, the net sulfide production and oxidation zones were moved deeper into flocs and sludge cores, which effectively prevented sulfide from entering the water column. However, the sulfide efflux from the sludge instantly recovered after NO(3)(-) depletion. Thus, the NO(3)(-) level in the water column controls the zonation and magnitude of sulfur transformations in the sludge. The effect of NO(3)(-) relies therefore on its sustained presence in the water column, which in turn depends on a well-functioning nitrification in the mariculture system.


Mechanisms behind intrauterine device-induced luteal persistence in mares.
Intrauterine glass balls are used to prevent oestrous signs in sports mares, but the mechanism of action is unknown. It has been suggested that the glass ball can mimic an embryo or act via an induced chronic uterine inflammation and absent or continuous low-grade prostaglandin (PG) release. The purpose of this study was to induce prolonged luteal function in mares using a small intrauterine device (IUD) and to study the mechanisms behind prolonged IUD-induced luteal function. A uterine swab and a biopsy specimen were obtained in early oestrus. A water-filled plastic ball, diameter 20mm and weight 3.6g, was inserted into the uterus 2-4 days after ovulation; the control mares underwent similar cervical manipulation without ball insertion. The mares were examined three times per week until day 23 and twice weekly thereafter until they returned to oestrus (transrectal palpation, ultrasonography and progesterone determination). The location of the IUD was recorded and ultrasound scans were video-recorded to assess the frequency of uterine contractions. When the mare returned to oestrus, a uterine swab and biopsy specimen were obtained and the bacteriological, cytological and histological (inflammation and glandular dilation) results compared with the samples obtained before the IUD insertion. The PG F(2alpha) metabolite levels were measured in the plasma of four control mares and eight IUD mares on days 11-16. The IUD induced a prolonged luteal phase in 75% of the mares (9/12; IUD-P); the mean dioestrous length was 57.0 days. The three mares that did not respond to the IUD (IUD-N) showed a mean dioestrous length of 15.7 days and the 12 control mares 16.1 days. The inflammation and glandular dilation scores were not significantly different in pre- and post-manipulation biopsy specimens. Although locational changes of the IUD were observed, they occurred over very small distances and were mostly limited within the body-bifurcation area. The IUD-N and control mares showed increased uterine contractility 11-16 days post-ovulation, whereas the IUD-P mares did not. The control mares (n=4) and IUD-N mares (n=2) showed increased PG levels from day 14 post-ovulation, while the IUD-P mares (n=6) showed basal levels only. We concluded that the IUD did not cause continuous PG release and suggest that close contact of the IUD with the endometrium may prevent the endometrial cells from releasing PGF(2alpha).


Identifying monitoring information needs that support the management of fish in large rivers.
Management actions intended to benefit fish in large rivers can directly or indirectly affect multiple ecosystem components. Without consideration of the effects of management on non-target ecosystem components, unintended consequences may limit management efficacy. Monitoring can help clarify the effects of management actions, including on non-target ecosystem components, but only if data are collected to characterize key ecosystem processes that could affect the outcome. Scientists from across the U.S. convened to develop a conceptual model that would help identify monitoring information needed to better understand how natural and anthropogenic factors affect large river fishes. We applied the conceptual model to case studies in four large U.S. rivers. The application of the conceptual model indicates the model is flexible and relevant to large rivers in different geographic settings and with different management challenges. By visualizing how natural and anthropogenic drivers directly or indirectly affect cascading ecosystem tiers, our model identified critical information gaps and uncertainties that, if resolved, could inform how to best meet management objectives. Despite large differences in the physical and ecological contexts of the river systems, the case studies also demonstrated substantial commonalities in the data needed to better understand how stressors affect fish in these systems. For example, in most systems information on river discharge and water temperature were needed and available. Conversely, information regarding trophic relationships and the habitat requirements of larval fishes were generally lacking. This result suggests that there is a need to better understand a set of common factors across large-river systems. We provide a stepwise procedure to facilitate the application of our conceptual model to other river systems and management goals.


Examining the Nocebo Effect in Trials of Neuromodulators for Use in Disorders of Gut-Brain Interaction.
Nocebo effects are believed to influence the rate of reported adverse events (AE) and subject withdrawal in both the treatment and placebo groups of randomized clinical trials (RCT). Neuromodulators are commonly prescribed to treat disorders of gut-brain interaction (DGBI), but adherence to these medications is often limited by side effects such as headache, dry mouth, fatigue, and altered bowel habits. We performed a systematic review and meta-analysis to assess the proportion and risk difference of patients who experienced side effects leading to withdrawal in the placebo arm vs the treatment arm of RCT of neuromodulators for DGBI. We also sought to estimate the risk of developing any AE in the placebo arm of these studies and the rate of specific individual AEs. We searched MEDLINE, Embase, Web of Science Core Collection, and the Cochrane Central Register of Controlled Trials Searches to identify RCT that included terms for DGBI and for commonly prescribed neuromodulators. We calculated pooled proportions of patients experiencing an AE leading to withdrawal in the active treatment group vs the placebo group with 95% confidence intervals (CI), the pooled proportions of patients experiencing any AE, the pooled proportions of patients experiencing specific AE such as dizziness and headache, the pooled proportions of patients experiencing severe AE, and corresponding pooled risk differences with 95% CI. There were 30 RCT included representing 2,284 patients with DGBI. Twenty-seven RCT reported data on AE leading to withdrawal. The pooled proportion of total patients with AE leading to withdrawal in the placebo group was 4% (95% CI 0.02-0.04). The pooled proportion of patients with AE leading to withdrawal who received neuromodulators was 9% (95% CI 0.06-0.13). In the 12 studies reporting data on patients experiencing at least 1 AE, the pooled proportion of patients experiencing any AE in the placebo group was 18% (95% CI 0.08-0.30), compared with 43% (95% CI 0.24-0.63) in the neuromodulator group. Thus, approximately 44% of the rate of withdrawal (0.04/0.09) and 42% of the rate reporting any side effects (0.18/0.43) in the neuromodulator group may be attributed to nocebo effects in the right context. Subgroup analysis by sex, medication class, risk of bias, and specific DGBI revealed differing withdrawal rates. There was no statistically significant difference in patients experiencing individual AE of dizziness, headache, or diarrhea. Rates of dry mouth, fatigue, and constipation were higher in treatment groups compared with those in placebo groups. Patients with DGBI in RCT randomized to placebo groups frequently experience AE and AE that lead to withdrawal consistent with a strong nocebo effect. Nonspecific AE such as dizziness, headaches, and diarrhea occurred similarly in patients receiving placebo compared with those receiving neuromodulators.


Structural basis for the helical filament formation of Escherichia coli glutamine synthetase.
Escherichia coli glutamine synthetase (EcGS) spontaneously forms a dodecamer that catalytically converts glutamate to glutamine. EcGS stacks with other dodecamers to create a filament-like polymer visible under transmission electron microscopy. Filamentous EcGS is induced by environmental metal ions. We used cryo-electron microscopy (cryo-EM) to decipher the structure of metal ion (nickel)-induced EcGS helical filament at a sub-3Å resolution. EcGS filament formation involves stacking of native dodecamers by chelating nickel ions to residues His5 and His13 in the first N-terminal helix (H1). His5 and His13 from paired parallel H1 helices provide salt bridges and hydrogen bonds to tightly stack two dodecamers. One subunit of the EcGS filament hosts two nickel ions, whereas the dodecameric interface and the ATP/Mg-binding site both host a nickel ion each. We reveal that upon adding glutamate or ATP for catalytic reactions, nickel-induced EcGS filament reverts to individual dodecamers. Such tunable filament formation is often associated with stress responses. Our results provide detailed structural information on the mechanism underlying reversible and tunable EcGS filament formation.


Identification and Massively Parallel Characterization of Regulatory Elements Driving Neural Induction.
Epigenomic regulation and lineage-specific gene expression act in concert to drive cellular differentiation, but the temporal interplay between these processes is largely unknown. Using neural induction from human pluripotent stem cells (hPSCs) as a paradigm, we interrogated these dynamics by performing RNA sequencing (RNA-seq), chromatin immunoprecipitation sequencing (ChIP-seq), and assay for transposase accessible chromatin using sequencing (ATAC-seq) at seven time points during early neural differentiation. We found that changes in DNA accessibility precede H3K27ac, which is followed by gene expression changes. Using massively parallel reporter assays (MPRAs) to test the activity of 2,464 candidate regulatory sequences at all seven time points, we show that many of these sequences have temporal activity patterns that correlate with their respective cell-endogenous gene expression and chromatin changes. A prioritization method incorporating all genomic and MPRA data further identified key transcription factors involved in driving neural fate. These results provide a comprehensive resource of genes and regulatory elements that orchestrate neural induction and illuminate temporal frameworks during differentiation.


Operative repair of symptomatic spondylolysis following a positive response to diagnostic pars injection.
From 1989 to 1996, 275 patients (245 male and 20 female) with back pain symptoms and spondylolysis of the lumbar spine were reviewed. All patients were evaluated by a protocol that included nonoperative treatment, bone scan, and pars injection. Only those whose symptoms failed nonoperative measures, showing negative bone scan and positive pars injection, were regarded as candidates for surgical management. Pars injection with Marcaine was done, and there were 93 cases with a positive response of reproducing symptoms and symptom relief. Patients then received autogenous bone grafting and internal fixation of the pars interarticularis defect. The internal fixation devices used included translaminar screws (AO 3.5 cancellous screw) for the most frequent level of L5, hook screws for levels above L4, and augmentation with wire for cases with concomitant spinal bifida occulta. The average age of the surgical group (85 male and 8 female) was 23 years (range: 19-35 years). After a follow-up averaging 30.4 months (range: 24-48 months), fusion results were 87%. Clinical results of 85 cases (91.3%) were excellent to good; 8 cases were fair; there were no poor cases. Direct repair of the pars defect by internal fixation and bone grafting was done to preserve involved motion segment and to prevent abnormal stresses at adjacent levels. These procedures seemed to be clinically effective.


Effect of referral strategies on access to cardiac rehabilitation among women.
Despite its proven benefits and need, women's access to cardiac rehabilitation (CR) is suboptimal. Referral strategies, such as systematic referral, have been advocated to improve access to CR. This study examined sex differences in CR referral and enrollment by referral strategies; and the impact of referral strategies for referral and enrollment concordance among women. Prospective cohort study. This prospective study included 2635 coronary artery disease inpatients from 11 Ontario hospitals that utilized one of four referral strategies. Participants completed a sociodemographic survey, and clinical data were extracted from charts. One year later, 1809 participants (452 (25%) women) completed a mailed survey that assessed CR utilization. Referral strategies were compared among women using generalized estimating equations to control for the effect of hospital. Overall, significantly more men than women were referred (67.2% and 57.8% respectively, p < 0.001), and enrolled in CR (58.6% and 49.3% respectively, p = 0.001). Of the retained women, combined systematic and liaison-facilitated referral resulted in significantly greater CR referral (OR 10.3, 95% CI 4.11-25.58) and enrollment (OR 6.6, 95% CI 4.34-9.92) among women when compared with usual referral. Conversely, concordance between referral and enrollment was greatest following usual referral (K = 0.85), and decreased with referral intensity. While a lower proportion of referred patients enroll, systematic and liaison-facilitated inpatient referral strategies result in the greatest CR enrollment rates among women. Such strategies have the potential to improve access among women, and reduce 'cherry picking' of patients for referral.


Perspective on CETSA Literature: Toward More Quantitative Data Interpretation.
The cellular thermal shift assay (CETSA) was introduced in 2013 to investigate drug-target engagement inside live cells and tissues. As with all thermal shift assays, the response measured by CETSA is not simply governed by ligand affinity to the investigated target protein, but the thermodynamics and kinetics of ligand binding and protein unfolding also contribute to the observed protein stabilization. This limitation is commonly neglected in current applications of the method to validate the target of small-molecule probes. Instead, there is an eagerness to make direct comparisons of CETSA measurements with functional and phenotypic readouts from cells at 37 °C. Here, we present a perspective of the early CETSA literature and put the accumulated data into a quantitative context. The analysis includes annotation of ~270 peer-reviewed papers, the majority of which do not consider the underlying biophysical basis of CETSA. We also detail what future technology developments are needed to enable CETSA-based optimization of structure-activity relationships and more appropriate comparisons of these data with functional or phenotypic responses. Finally, we describe ongoing developments in assay formats that allow for CETSA measurements at single-cell resolution, with the aspiration to allow differentiation in cellular target engagement between cells in co-cultures and more complex models, such as organoids and potentially even tissue.


A locally initiated and executed measles outbreak response immunization campaign in the nylon health district, Douala Cameroon 2011.
The Cameroon health system is divided into central, intermediate and peripheral levels. Of the 43 health districts with a measles outbreak in Cameroon in 2011, only the Nylon Health District organized a documented outbreak response immunization. We present the methods and results of the response campaign solely shouldered by the district and intermediate level. The risk group, targets and neighborhoods to be vaccinated were identified after a detailed analysis of initial cases. The intermediate level defined strategies, provided logistics, capacity building and 41% of the operational budget while 59% was completed by the peripheral level. Microsoft Office Excel 2007 was used to estimate coverage rates and to draw an epidemic curve. The response immunization campaign was organized on the 14th epidemiological week, 10 weeks after the onset of the outbreak which ended 11 weeks thereafter. A total of 15867(108.5%) children aged 9-59 months were vaccinated in five health areas at a direct cost (vaccines excluded) of 71.34FCFA ($0.143) per vaccinated child. An additional 824 children aged 9-59 months were vaccinated around the residence of notified cases in neighborhoods which were not involved in the response campaign. The number of cases after the response campaign was lower than before. Once vaccines are available, prompt outbreak response campaigns can be organized at operational level to obtain commendable results instead of depending solely on international organizations or central levels. Decision makers at the intermediate and operational levels should redeploy available funds during emergencies to prevent the development of extreme public health conditions.


Generation of infectious feline immunodeficiency virus (FIV) encoding FIV/human immunodeficiency virus chimeric protease.
Feline immunodeficiency virus (FIV) and human immunodeficiency virus type 1 (HIV-1) proteases (PRs) share only 23% amino acid identity and exhibit distinct specificities yet have very similar 3-dimensional structures. Chimeric PRs in which HIV residues were substituted in structurally equivalent positions in FIV PR were prepared in order to study the molecular basis of PR specificity. Previous in vitro analyses showed that such substitutions dramatically altered the inhibitor specificity of mutant PRs but changed the rate and specificity of Gag cleavage so that chimeric FIVs were not infectious. Chimeric PRs encoding combinations of the I37V, N55M, M56I, V59I, L97T, I98P, Q99V, and P100N mutations were cloned into FIV Gag-Pol, and those constructs that best approximated the temporal cleavage pattern generated by wild-type FIV PR, while maintaining HIV-like inhibitor specificity, were selected. Two mutations, M56I and L97T, were intolerant to change and caused inefficient cleavage at NC-p2. However, a mutant PR with six substitutions (I37V, N55M, V59I, I98P, Q99V, and P100N) was selected and placed in the context of full-length FIV-34TF10. This virus, termed YCL6, had low-level infectivity ex vivo, and after passage, progeny that exhibited a higher growth rate emerged. The residue at the position of one of the six mutations, I98P, further mutated on passage to either P98H or P98S. Both PRs were sensitive to the HIV-1 PR inhibitors lopinavir (LPV) and darunavir (DRV), as well as to the broad-based inhibitor TL-3, with 50% inhibitory concentrations (IC(50)) of 30 to 40 nM, consistent with ex vivo results obtained using mutant FIVs. The chimeras offer an infectivity system with which to screen compounds for potential as broad-based PR inhibitors, define structural parameters that dictate specificity, and investigate pathways for drug resistance development.


Culture nutrition and physiology impact the inhibitor tolerance of the yeast Pichia stipitis NRRL Y-7124.
Pichia stipitis NRRL Y-7124 is one of the natural yeasts best able to utilize biomass because it is able to ferment hexoses and the pentose, xylose, to economically recoverable concentrations of ethanol. To test the impact of culture conditions on inhibitor tolerance, inhibitors were spiked to growing or stationary-phase P. stipitis supplied either glucose or xylose and varying nitrogen and mineral compositions; then the ensuing specific death rate response was measured. Resistance of glucose- or xylose-grown cells to inhibitors was generally greater in stationary-phase cells than log-phase cells, despite a greater exposure of stationary cells to ethanol. Consistent with this, the specific productivity of detoxification products, furan methanol or furan-2,5-dimethanol, from respective spikes of furfural or HMF increased as cultures progressed into stationary phase. However, when xylose was the substrate, ethanol resistance behaved uniquely and was greater for log- than stationary-phase cells. Amino acid enrichment of the growth medium significantly enhanced ethanol tolerance if xylose was the carbon source, but had no impact if glucose supplied carbon. Regardless of the carbon source, amino acid enrichment of the culture medium enhanced the ability of cells to resist furfural and HMF exposure. Mineral compositions tested had little impact on inhibitor resistance except stationary-phase xylose-grown cells were more susceptible to inhibitor exposure when magnesium sulfate was excessive. Observed tolerance optimization based on specific death rate as a function of culture physiological state, carbon source, nitrogen source and mineral composition provides new knowledge supporting process designs to convert biomass to ethanol using P. stipitis.


Hyaluronan concentration and molecular mass in psoriatic arthritis: biomarkers of disease severity, resistance to treatment, and outcome.
Objective: Low molecular mass hyaluronan causes inflammatory processes and can act as a pro-inflammatory cytokine in skin and other sites of activity in psoriatic arthritis (PsA). This study investigated whether the molecular mass distribution of hyaluronan (HA) in skin and the quantity of circulating HA are related to the clinical inflammatory picture in PsA with active disease and to the effect of treatment with anti-tumour necrosis factor-α (anti-TNF-α) adalimumab. Methods: Twenty patients with TNF-α-naïve active polyarticular PsA were included in this prospective clinical trial of treatment with 40 mg s.c. adalimumab according to standard procedure. Clinical activity, patients' assessments, and skin biopsies were captured at inclusion and at the 12 week follow-up. Ten healthy individuals were recruited for comparison of HA analyses. Histochemistry of skin inflammation, serum HA, and molecular mass of HA were determined. Results: Overall improvements in clinical parameters were observed. Eight of 18 patients reached minimum disease activity after 12 weeks and disease activity was significantly reduced (p < 0.0001). Patients with elevated serum HA values were significantly older, had later onset of arthritis and more deformed joints, still had swollen joints after treatment, and had more circulating inflammatory biomarkers. More severe disease pathology showed a wide spectrum of high-molecular-mass HA accompanied by low mass HA. The treatment appears partly to normalize the HA mass distribution. Conclusion: HA concentration and mass seem to be two possible factors in the inflammatory pathology of PsA acting as biomarkers for disease severity, resistance to treatment, and worse outcome.


The role of hybrid procedures in the management of peripheral vascular disease.
This paper provides a summary of up-to-date information and experience with the combined treatment of patients suffering from peripheral arterial disease (PAD) with endovascular and open surgery, performed simultaneously and in a single operating room. Hybrid intervention is reported to have good results in well-indicated groups of patients with acute and chronic limb ischemia, even with older, high-risk patients. The indications for the use of this technique remain unclear with inconsistent opinions among vascular surgeons. The indications for treatment were divided into three main groups: 1) Patients with chronic limb ischemia, 2) acute limb ischemia, and 3) occlusion of a previous vascular reconstruction. The operating techniques for the most commonly used combinations are described. In conclusion, hybrid operating techniques are often useful when treating complex problems and multilevel disease in patients with chronic or acute lower limb ischemia. Modern vascular surgeons need to master both open and endovascular techniques, and to combine them in a creative fashion to the benefit of our patients.


Erectile dysfunction precedes and is associated with severity of coronary artery disease among Asian Indians.
Erectile dysfunction (ED) and coronary artery disease (CAD) often share common risk factors, and there is growing evidence that ED might serve as a clinical marker for cardiovascular disease. Despite rising trends of CAD in Asian Indians, limited data are available on the prevalence of ED and its correlation with CAD severity in such patients. To study the prevalence of ED in Asian Indian patients undergoing coronary angiography and to assess if the severity of ED correlates with angiographic severity of CAD. In all patients undergoing coronary angiography, ED was assessed using the International Index of Erectile Function-5 questionnaire. Among 175 male patients, ED was present in 70%; patients with ED had a higher incidence of multivessel CAD (80% vs. 36%, P 0.001), diffuse CAD (81% vs. 34%, P 0.001), and higher number of mean coronary vessels involved compared with those without ED. Those with severe ED had higher prevalence of multivessel CAD and higher number of mean coronary vessels involved compared with those with milder grades of ED. Onset of symptoms of ED preceded symptoms of CAD by a mean of 24.6 months in 84% of patients. The presence of severe ED was associated with a 21-fold higher risk of having triple-vessel disease (odds ratio [OR] 21.94, 95% confidence interval [CI] 3.41-141.09, P = 0.001) and an 18-fold higher risk of having diffuse angiographic CAD (OR 17.91, 95% CI 3.11-111.09, P = 0.001). Asian Indians with angiographic CAD frequently have ED; symptoms of ED precede that of CAD in most patients. Incidence of multivessel and diffuse CAD is significantly more common in patients with ED. It is important for physicians to be aware of the close relationship between the two conditions so that patients with ED can have optimal risk stratification for concomitant CAD whenever required.