Limitations of DNA histogram analysis by flow cytometry as a method of predicting chemosensitivity in a rat renal cancer model.
Analysis of DNA histograms obtained from rat renal cancer cells stained with propidium iodide and submitted to flow cytometry revealed a tumor cell population with prominent 2C and 4C peaks and with usually less than 10% each of S-phase or hyper-4C cells. The presence of an increased proportion of 4C cells was found to depend on the age and size of the tumor nodule. Sampling replicate portions of the same tumor or different tumors of the same size and age frequently revealed highly variable 4C:2C ratios. Treatment of animals bearing this tumor with a single i.p. injection of cyclophosphamide (CY), under conditions known to reduce the tumor burden by 80% within 1 week, or with 5-fluorouracil (FUra), which is ineffective against this tumor, in many cases did not yield changes in DNA histograms that permitted one to distinguish the effective drug. Either no marked difference in histogram shape occurred after therapy, or FUra induced more striking differences in cell cycle position than did CY. In tumor generations with greater than 15% S-phase cells, treatment with CY resulted in multiple effects on DNA histograms. These included detecting increased numbers of moribund cells (hypo-2C), a decrease in 4C cells, and an increase in hyper-4C cells. These changes did not occur with the ineffective agent FUra. The tumors grown in vitro show no evidence of replication of 4C cells. The DNA histograms of late-log-phase cultures have a major 2C peak and a minor S plus G2 hump. Since neither the untreated tumor in vivo nor that grown in vitro has a major hyper-4C cell population, it is probable that the tumor stem cells are chiefly 2C (diploid-hyperdiploid). Treatment in vitro of late-log-phase cultures with CY or FUra produces DNA histograms which permit identification of the effective agent. After CY, a major part of the cell population was hypo-2C (moribund) cells.


Effectiveness of probiotics in irritable bowel syndrome: Updated systematic review with meta-analysis.
To investigate the efficacy of probiotics in irritable bowel syndrome (IBS) patients. PubMed, Cochrane library, Scopus, Google Scholar, and Clinicaltrial.gov databases were searched for literature published between September 2007 and December 2013. The applied Mesh terms were "probiotics," "irritable bowel syndrome," and "irritable bowel syndrome treatment." The collected data contained24 clinical trials, of which 15 were eligible for meta-analysis and nine were reviewed systematically. All studies were randomized placebo-controlled trials in patients with IBS that investigated the efficacy of probiotics in IBS improvement. The Jadad score was used to assess the methodological quality of trials. The quality scale ranges from 0 to 5 points, with a score ≤ 2 indicating a low quality report, and a score of ≥ 3 indicating a high quality report. Relative risk (RR), standardized effect size, and 95%CI were calculated using the DerSimonian-Laird method. The Cochran Q test was used to test heterogeneity with P < 0.05. Funnel plots were constructed and Egger's and Begg-Mazumdar tests were performed to assess publication bias. A total of 1793 patients were included in the meta-analysis. The RR of responders to therapies based on abdominal pain score in IBS patients for two included trials comparing probiotics to placebo was 1.96 (95%CI: 1.14-3.36; P = 0.01). RR of responders to therapies based on a global symptom score in IBS patients for two included trials comparing probiotics with placebo was 2.43 (95%CI: 1.13-5.21; P = 0.02). For adequate improvement of general symptoms in IBS patients, the RR of seven included trials (six studies) comparing probiotics with placebo was 2.14 (95%CI: 1.08-4.26; P = 0.03). Distension, bloating, and flatulence were evaluated using an IBS severity scoring system in three trials (two studies) to compare the effect of probiotic therapy in IBS patients with placebo, the standardized effect size of mean differences for probiotics therapy was -2.57 (95%CI: -13.05--7.92). Probiotics reduce pain and symptom severity scores. The results demonstrate the beneficial effects of probiotics in IBS patients in comparison with placebo.


The flux growth of single-crystalline CoTiO3 polyhedral particles and improved visible-light photocatalytic activity of heterostructured CoTiO3/g-C3N4 composites.
Highly crystalline and idiomorphic CoTiO3 single crystals with a well-defined polyhedral morphology were grown successfully for the first time by a facile flux method. Herein, the effects of the molten salt type and cobalt precursor on the phase composition, crystallization habit and morphology of the CoTiO3 products were also investigated. Importantly, using the flux-grown CoTiO3 crystal as the visible-light sensitizer due to its narrow band gap to couple with graphitic carbon nitride (g-C3N4) by a direct in situ thermal induced polycondensation route, novel CoTiO3/g-C3N4 composite photocatalysts were obtained. The as-synthesized samples were systematically characterized by XRD, EDS, SEM, TEM, SAED, HRTEM, FT-IR, XPS, DRS and PL techniques. The results revealed that CoTiO3 polyhedral crystals were closely combined with g-C3N4 nanosheets leading to the formation of a heterojunction structure at the interface between CoTiO3 and g-C3N4. Photocatalytic evaluation showed that the heterostructured CoTiO3/g-C3N4 composite exhibited much higher photocatalytic activity for the degradation of methyl orange under visible light irradiation than that of individual CoTiO3 and g-C3N4, which could be mainly ascribed to the synergistic effect between CoTiO3 and g-C3N4, including the enhanced visible-light harvesting ability and more efficient separation and longer lifetime of photogenerated charge carriers. Furthermore, the composite photocatalyst showed an excellent stability and reusability during four successive cycles. Finally, a possible mechanism responsible for the charge separation and improved photocatalytic activity was proposed.


A fully continuous system of DNA profile evidence evaluation that can utilise STR profile data produced under different conditions within a single analysis.
The introduction of probabilistic DNA interpretation systems has made it possible to evaluate many profiles that previously (under a manual interpretation system) were not. These probabilistic systems have been around for a number of years and it is becoming more common that their use within a laboratory has spanned at least one technology change. This may be a change in laboratory hardware, the DNA profiling kit used, or the manner in which the profile is generated. Up until this point, when replicates DNA profiles are generated, that span a technological change, the ability to utilise all the information in all replicates has been limited or non-existent. In this work we explain and derive the models required to evaluate (what we term) multi-kit analysis problems. We demonstrate the use of the multi-kit feature on a number of scenarios where such an analysis would be desired within a laboratory. Allowing the combination of profiling data that spans a technological change will further increase the amount of DNA profile information produced in a laboratory that can be evaluated.


Morbidity and mortality following envenomation by the common night adder (Causus rhombeatus) in three dogs.
In South Africa dogs are frequently presented to veterinarians following snakebite. The offending snakes are usually puff adders (Bitis arietans), cobras (Naja spp.) and mambas (Dendroaspis spp.). Night adder (Causus rhombeatus) bites in dogs have not yet been reported in South Africa. This article deals with three cases of dogs bitten by night adders in which extensive tissue damage was noted and one fatality occurred. Night adder bites may be indistinguishable from puff adder bites. Non-specific treatment included addressing the hypovolaemia and swelling. Specific treatment involving immunotherapy using the South African polyvalent antivenom would be ineffective as it does not contain immunoglobulins against night adder venom. Veterinarians should also include night adders as the possible cause of dogs suffering from severe and painful swellings suspected to be due to snakebites.


Collaborative Ocular Tuberculosis Study Consensus Guidelines on the Management of Tubercular Uveitis-Report 2: Guidelines for Initiating Antitubercular Therapy in Anterior Uveitis, Intermediate Uveitis, Panuveitis, and Retinal Vasculitis.
The Collaborative Ocular Tuberculosis Study (COTS), supported by the International Ocular Inflammation Society, International Uveitis Study Group, and Foster Ocular Immunological Society, set up an international, expert-led consensus project to develop evidence- and experience-based guidelines for the management of tubercular uveitis (TBU). The absence of international agreement on the use of antitubercular therapy (ATT) in patients with TBU contributes to a significant heterogeneity in the approach to the management of this condition. Consensus statements for the initiation of ATT in TBU were generated using a 2-step modified Delphi technique. In Delphi step 1, a smart web-based survey based on background evidence from published literature was prepared to collect the opinion of 81 international experts on the use of ATT in different clinical scenarios. The survey included 324 questions related to tubercular anterior uveitis (TAU), tubercular intermediate uveitis (TIU), tubercular panuveitis (TPU), and tubercular retinal vasculitis (TRV) administered by the experts, after which the COTS group met in November 2019 for a systematic and critical discussion of the statements in accordance with the second round of the modified Delphi process. Forty-four consensus statements on the initiation of ATT in TAU, TIU, TPU, and TRV were obtained, based on ocular phenotypes suggestive of TBU and corroborative evidence of tuberculosis, provided by several combinations of immunologic and radiologic test results. Experts agreed on initiating ATT in recurrent TAU, TIU, TPU, and active TRV depending on the TB endemicity. In the presence of positive results for any 1 of the immunologic tests along with radiologic features suggestive of past evidence of tuberculosis infection. In patients with a first episode of TAU, consensus to initiate ATT was reached only if both immunologic and radiologic test results were positive. The COTS consensus guidelines were generated based on the evidence from published literature, specialists' opinions, and logic construction to address the initiation of ATT in TBU. The guidelines also should inform public policy by adding specific types of TBU to the list of conditions that should be treated as tuberculosis.


Caffeine increases maximal fat oxidation during a graded exercise test: is there a diurnal variation?
There is evidence that caffeine increases the maximal fat oxidation rate (MFO) and aerobic capacity, which are known to be lower in the morning than in the afternoon. This paper examines the effect of caffeine intake on the diurnal variation of MFO during a graded exercise test in active men. Using a triple-blind, placebo-controlled, crossover experimental design, 15 active caffeine-naïve men (age: 32 ± 7 years) completed a graded exercise test four times at seven-day intervals. The subjects ingested 3 mg/kg of caffeine or a placebo at 8 am in the morning and 5 pm in the afternoon (each subject completed tests under all four conditions in a random order). A graded cycling test was performed. MFO and maximum oxygen uptake (VO2max) were measured by indirect calorimetry, and the intensity of exercise that elicited MFO (Fatmax) calculated. MFO, Fatmax and VO2max were significantly higher in the afternoon than in the morning (all P < 0.05). Compared to the placebo, caffeine increased mean MFO by 10.7% (0.28 ± 0.10 vs. 0.31 ± 0.09 g/min respectively, P < 0.001) in the morning, and by a mean 29.0% (0.31 ± 0.09 vs. 0.40 ± 0.10 g/min, P < 0.001) in the afternoon. Caffeine also increased mean Fatmax by 11.1% (36.9 ± 14.4 [placebo] vs. 41.0 ± 13.1%, P = 0.005) in the morning, and by 13.1% (42.0 ± 11.6 vs. 47.5 ± 10.8%, P = 0.008) in the afternoon. These findings confirm the previously reported diurnal variation in the whole-body fat oxidation rate during graded exercise in active caffeine-naïve men, and indicate that the acute ingestion of 3 mg/kg of caffeine increases MFO, Fatmax and VO2max independent of the time of day. NCT04320446 . Registered 25 March 2020 - Retrospectively registered.


Interviews with "vapers": implications for future research with electronic cigarettes.
Awareness and use of electronic cigarettes (e-cigs) has increased dramatically. Electronic Nicotine Delivery Devices deliver an aerosol comprised usually of water, propylene glycol and/or glycerin, nicotine, and flavorings. Scant research exists to evaluate the efficacy and safety of such devices, and only one quantitative survey of European users (N = 81) has been published. This qualitative study explores e-cig users' ("vapers") experiences. Participants attended a convention or club meeting in St. Louis, MO, and were interviewed individually or in small groups. Qualitative methods were used to analyze interview data for both deductive and emergent themes to broad research questions. Even with a relatively small sample of formal participants (N = 15), there were pervasive themes including the language and culture of vaping; social and informational support among vapers and their use of Internet resources (learning about e-cigs); the learning curve to using e-cigs and the numerous modifications ("mods") available for e-cigs and personal vaporizers; motives and perceived benefits of using e-cigs versus cigarettes including cigarette-like enjoyment, cost, restored sense of taste and smell, and improved breathing and exercise tolerance; rapidly reduced nicotine tolerance and dependence; and a strong interest in e-cig-related research and policy. The learning curve to using e-cigs has important implications for laboratory tests of these devices with novice users. Similarly, the multiple e-cig options and the use of "mods" create challenges for researchers and policy makers. Transdisciplinary research is urgently needed, and experienced "vapers" are very interested and willing research participants.


Hesperidin reduces adverse symptomatic intracerebral hemorrhage by promoting TGF-β1 for treating ischemic stroke using tissue plasminogen activator.
Treatment with recombinant tissue plasminogen activator (rt-PA) is the most effective therapeutic option against brain ischemic stroke at the present time. However, elevated incidence of symptomatic intracerebral hemorrhage (SIH) greatly hinders ideal treatment outcome of rt-PA. We sought to assess the impacts of hesperidin on SIH following rt-PA therapies. Patients with ischemic stroke were assigned into two groups in a random fashion, to receive either rt-PA + placebo (Pc) or rt-PA + hesperidin. Treatment outcome was evaluated 24 h after the initial reperfusion using the transcranial Doppler ultrasonography (TCD) and the NIH Stroke Scale (NIHSS). Further, serum concentrations of transforming growth factor (TGF)-β1, matrix metalloproteinase (MMP)-2, and MMP-9 were examined. Following the initial administration, stroke patients continued to receive either daily Pc or daily hesperidin, and the treatment outcome after 7 days was examined using the TCD, NIHSS, Glasgow Outcome Scale (GOS), and the Modified Rankin Scale (MRS). Combined treatment of rt-PA with hesperidin yielded significant improvement of outcomes, as revealed by better TCD and NIHSS scores as well as decreased SIH incidences, which could be attributable to elevation of TGF-β1 and reduction in serum levels of both MMP-2 and MMP-9 caused by hesperidin. Follow-up hesperidin treatment for 7 consecutive days also markedly enhanced the recovery of stroke patients, as indicated by TCD, MRS, GOS, and NIHSS. Findings of the present study strongly suggested potential clinical application of hesperidin supplement in rt-PA therapies to reduce SIH and thereby improve the treatment outcomes of rt-PA in patients with ischemic stroke.


Radiological and clinical predictors of scoliosis in patients with Chiari malformation type I and spinal cord syrinx from the Park-Reeves Syringomyelia Research Consortium.
Scoliosis is frequently a presenting sign of Chiari malformation type I (CM-I) with syrinx. The authors' goal was to define scoliosis in this population and describe how radiological characteristics of CM-I and syrinx relate to the presence and severity of scoliosis. A large multicenter retrospective and prospective registry of pediatric patients with CM-I (tonsils ≥ 5 mm below the foramen magnum) and syrinx (≥ 3 mm in axial width) was reviewed for clinical and radiological characteristics of CM-I, syrinx, and scoliosis (coronal curve ≥ 10°). Based on available imaging of patients with CM-I and syrinx, 260 of 825 patients (31%) had a clear diagnosis of scoliosis based on radiographs or coronal MRI. Forty-nine patients (5.9%) did not have scoliosis, and in 516 (63%) patients, a clear determination of the presence or absence of scoliosis could not be made. Comparison of patients with and those without a definite scoliosis diagnosis indicated that scoliosis was associated with wider syrinxes (8.7 vs 6.3 mm, OR 1.25, p < 0.001), longer syrinxes (10.3 vs 6.2 levels, OR 1.18, p < 0.001), syrinxes with their rostral extent located in the cervical spine (94% vs 80%, OR 3.91, p = 0.001), and holocord syrinxes (50% vs 16%, OR 5.61, p < 0.001). Multivariable regression analysis revealed syrinx length and the presence of holocord syrinx to be independent predictors of scoliosis in this patient cohort. Scoliosis was not associated with sex, age at CM-I diagnosis, tonsil position, pB-C2 distance (measured perpendicular distance from the ventral dura to a line drawn from the basion to the posterior-inferior aspect of C2), clivoaxial angle, or frontal-occipital horn ratio. Average curve magnitude was 29.9°, and 37.7% of patients had a left thoracic curve. Older age at CM-I or syrinx diagnosis (p < 0.0001) was associated with greater curve magnitude whereas there was no association between syrinx dimensions and curve magnitude. Syrinx characteristics, but not tonsil position, were related to the presence of scoliosis in patients with CM-I, and there was an independent association of syrinx length and holocord syrinx with scoliosis. Further study is needed to evaluate the nature of the relationship between syrinx and scoliosis in patients with CM-I.


Large three-dimensional poly(glycerol sebacate)-based scaffolds - a freeze-drying preparation approach.
Large three-dimensional poly(glycerol sebacate) (PGS)-based scaffolds were successfully fabricated via a freeze-drying and a subsequent curing process, in the presence of a minor structure-supporting polymer such as biocompatible poly(l-lactic acid) (PLLA). The resulting PGS/PLLA scaffolds exhibited highly interconnected open-cell structures. The presence of solid PLLA prevented the low-viscosity pre-polymer of PGS from enclosing open cells during the curing stage, thereby avoiding structural collapse of the scaffold. The ratio of PGS to PLLA affected the cell structure and mechanical properties, with a higher ratio of PGS offering a softer and more ductile scaffold in general. The PGS/PLLA scaffold containing 75 wt% PGS presented a porosity of 85%, an average pore size of 93 μm, a Young's modulus of 0.36 MPa, a tensile strength of 0.03 MPa, an elongation at yield of 13%, and good scaffold structure retention during in vitro degradation tests with weight losses of 40% and 9% respectively in 31 days with and without the presence of lipase enzyme. This novel versatile approach will allow for easy fabrication of large, optimal, interconnected porous PGS-based scaffolds for soft tissue engineering.


Minocycline-EDTA lock solution prevents catheter-related bacteremia in hemodialysis.
There is growing concern about the development of antibacterial resistance with the use of antibiotics in catheter lock solutions. The use of an antibiotic that is not usually used to treat other serious infections may be an alternative that may reduce the clinical impact should resistance develop. We conducted a randomized controlled trial to compare a solution of minocycline and EDTA with the conventional unfractionated heparin for the prevention of catheter-related bacteremia in hemodialysis patients during a period of 90 d. The study included 204 incident catheters (27.8% tunneled); 14 catheters were excluded because of early dysfunction and 3 because of protocol violations. We observed catheter-related bacteremia in 19 patients in the heparin group (4.3 per 1000 catheter-days) and in 5 patients in the minocycline-EDTA group (1.1 per 1000 catheter-days; P = 0.005). We did not detect a significant difference in the rate of catheter removal for dysfunction. Catheter-related bacteremia-free survival was significantly higher in the minocycline-EDTA group than in the heparin group (P = 0.005). In conclusion, a minocycline-EDTA catheter lock solution is effective in the prevention of catheter-related bacteremia in hemodialysis patients.


A Clinical Practice Guideline for the Management of Patients With Acute Spinal Cord Injury and Central Cord Syndrome: Recommendations on the Timing (≤24 Hours Versus >24 Hours) of Decompressive Surgery.
To develop recommendations on the timing of surgical decompression in patients with traumatic spinal cord injury (SCI) and central cord syndrome. A systematic review of the literature was conducted to address key relevant questions. A multidisciplinary guideline development group used this information, along with their clinical expertise, to develop recommendations for the timing of surgical decompression in patients with SCI and central cord syndrome. Based on GRADE, a strong recommendation is worded as "we recommend," whereas a weak recommendation is presented as "we suggest." Conclusions from the systematic review included (1) isolated studies reported statistically significant and clinically important improvements following early decompression at 6 months and following discharge from inpatient rehabilitation; (2) in one study on acute central cord syndrome without instability, a marginally significant improvement in total motor scores was reported at 6 and 12 months in patients managed with early versus late surgery; and (3) there were no significant differences in length of acute care/rehabilitation stay or in rates of complications between treatment groups. Our recommendations were: "We suggest that early surgery be considered as a treatment option in adult patients with traumatic central cord syndrome" and "We suggest that early surgery be offered as an option for adult acute SCI patients regardless of level." Quality of evidence for both recommendations was considered low. These guidelines should be implemented into clinical practice to improve outcomes in patients with acute SCI and central cord syndrome by promoting standardization of care, decreasing the heterogeneity of management strategies, and encouraging clinicians to make evidence-informed decisions.


A prospective investigation of injury incidence and injury risk factors among Army recruits in military police training.
United States Army military police (MP) training is a 19-week course designed to introduce new recruits to basic soldiering skills, Army values and lifestyle, and law enforcement skills and knowledge. The present investigation examined injury rates and injury risk factors in MP training. At the start of training, 1,838 male and 553 female MP recruits were administered a questionnaire containing items on date of birth, height, weight, tobacco use, prior physical activity, injury history, and menstrual history. Injuries during training were obtained from electronic medical records and the training units provided data on student graduation and attrition. Successfully graduating from the course were 94.3% of the men and 83.7% of the women. Experiencing at least one injury during training were 34.2% of the men and 66.7% of the women (risk ratio (women/men) = 1.95, 95% confidence interval = 1.79-2.13). Recruits were at higher injury risk if they reported that they were older, had smoked in the past, or had performed less frequent exercise or sports prior to MP training. Men were at higher injury risk if they reported a prior injury and women were at higher risk if they reported missing at least six menstrual cycles in the last year or had previously been pregnant. The present investigation was the first to identify injury rates and identify specific factors increasing injury risk during MP training.


Repellency of live potted plants against Anopheles gambiae from human baits in semi-field experimental huts.
The repellency of potted plants against the malaria vector Anopheles gambiae sensu stricto Giles was quantified in experimental huts under semi-field conditions inside a screen-walled greenhouse. Ocimum americanum Linnaeus (Labiatae), Lantana camara L. (Verbenaceae), and Lippia uckambensis Spreng (Verbenaceae) repelled at an average of 39.7% (95% confidence interval [CI] = 29.6-48.4%), 32.4% (95% CI = 19.7-43.1%), and 33.3% (95% CI = 21.5-43.3%) of the mosquitoes, respectively (P < 0.0001 for all treatments). This was determined by logistic regression, allowing for variations associated with different bait hosts, sampling huts, and replicate test nights. In contrast, Ocimum kilimandscharicum Guerke (Labiatae), Ocimum suave Willd. (Labiatae), Corymbia citriodora Hook (Myrtaceae), Azadirachta indica A. Juss (Meliaceae), Tagetes minuta L. (Asteraceae), and Hyptis suaveolens Poit. (Lamiaceae) did not significantly repel mosquitoes. The combination of O. americanum with either L. camara or L. uckambensis repelled 31.6% (95% CI = 19.7-41.7%) and 45.2% (95% CI = 34.7-54.0%) of the mosquitoes, respectively (P < 0.0001 for both treatments). This study is the first to show that live intact plants can reduce domestic exposure to malaria vector mosquitoes. As such, they may represent a new, sustainable and readily applicable malaria vector control tool for incorporation into integrated vector management programs.


Teaching psychomotor skills online: exploring the implications of novel coronavirus on health professions education.
The safe and effective application of psychomotor skills in the clinical environment is a central pillar of the health professions. The current global coronavirus pandemic has significantly impacted health professions education (HPE) and has been of particular consequence for routine face-to-face (F2F) skill education for health professionals and clinical students worldwide. What is being experienced on an unprecedented scale parallels a problem familiar to regional, rural and remote health professionals and students: the learners are willing, and the educational expertise exists, but the two are separated by the tyranny of distance. This article considers how the problem of physical distance might be overcome, so that quality skill education might continue. Psychomotor skills are undeniably easier to teach and learn F2F, and training schedules in tertiary, in-service and accredited professional courses reflect this. This aspect of HPE is therefore at significant risk in the context of social distancing and physical isolation. Psychomotor skills are much more complex than the physical motor outputs alone might suggest, and an F2F skill session is only one way to build the complementary aspects of new skill performance. This article argues that educators and course designers can progress with psychomotor skill education from a physical distance. Videos can be used to either passively present content to learners or actively engage them. It is the design of the educational activity, rather than the resource medium itself, that enables active engagement. Furthermore, while many training schedules have been adapted to accommodate intensive F2F skill training once it is safe to do so, distributed practice and the need for reflection during the acquisition and development of new skills may challenge the pedagogical effectiveness of this approach. Skill development can be fostered in the absence of F2F teaching, and in the absence of a shared physical space. Embracing the creative licence to do so will improve equitable access to regional, rural and remote clinicians and students well beyond the resolution of the current pandemic.


Continued analysis of optical frequency-modulated continuous-wave interference.
I continue to analyze systematically the theory of optical frequency-modulated continuous-wave (FMCW) interference. Two special cases, multiple-beam optical FMCW interference and multiple-wavelength optical FMCW interference, are discussed in detail. Multiple-beam optical FMCW interference generates a signal with multiple frequencies because of mutual interference among the waves. Multiple-wavelength optical FMCW interference produces a signal whose amplitude is modulated by a synthetic wave. The applications of both types of optical FMCW interference are also discussed.


Evaluation of systemic immunologic hyperreactivity after intradermal testing in horses with chronic laminitis.
To determine whether systemic immunologic hyperreactivity exists in horses with chronic laminitis, compared with responses for nonlaminitic horses. 7 nonlaminitic horses and 7 CL horses. In experiment 1, intradermal testing (IDT) was performed on 7 nonlaminitic and 7 CL horses to evaluate the response to a combination of 70 allergens at 15 and 30 minutes and 4 and 24 hours after injection. Three nonlaminitic and 3 CL horses used in experiment 1 were used in experiment 2 to determine whether histologic differences existed between the 2 groups. The H&E-stained tissue sections were evaluated on the basis of 3 criteria. For all analyses, 2-sample t-tests were used to determine significant differences between the groups. In experiment 1, CL horses had significantly higher total responses to IDT than nonlaminitic horses at the first 3 time periods. Also, CL horses had significantly fewer total scores of 0 than nonlaminitic horses at all time periods, except at 24 hours. In experiment 2, we did not detect significant differences between groups for any criterion. Results support the hypothesis that CL horses develop hyperreactivity to various antigenic stimuli, compared with responses for nonlaminitic horses. Therefore, the possibility that antigenic challenge may result in exacerbation of clinical signs of laminitis should be discussed with horse owners. Chronic laminitis should also be a consideration when a horse becomes lame following antigenic challenges.


Genetics of the epilepsies.
Recent molecular insights into the human idiopathic epilepsies have suggested the central role of ligand-gated and voltage-gated ion channels in their etiology. So far, genes coding for sodium and potassium channel subunits as well as a nicotinic cholinergic receptor subunit have been identified for mendelian idiopathic epilepsies. In vitro and in vivo studies of mutations demonstrate functional changes, allowing new insights into mechanisms underlying hyperexcitability. Similarly, spontaneous murine epilepsy models have been associated with calcium channel molecular defects. The major challenge before us in understanding the genetics of the epilepsies is to identify genes for common forms of epilepsy following complex inheritance. Once such genes are discovered, the gene-gene-environmental interactions producing specific epilepsy syndromes can be explored.


Molecular Properties of Astaxanthin in Water/Ethanol Solutions from Computer Simulations.
Astaxanthin (AXT) is a reference model of xanthophyll carotenoids, which is used in medicine and food industry, and has potential applications in nanotechnology. Because of its importance, there is a great interest in understanding its molecular properties and aggregation mechanism in water and mixed solvents. In this paper, we report a novel model of AXT for molecular dynamics simulation. The model is used to estimate different properties of the molecule in pure solutions and in water/ethanol mixtures. The calculated diffusion coefficients of AXT in pure water and ethanol are (3.22 ± 0.01) × 10(-6) cm(2) s(-1) and (2.7 ± 0.4) × 10(-6) cm(2) s(-1), respectively. Our simulations also show that the content of water plays a clear effect on the morphology of the AXT aggregation in water/ethanol mixture. In up to 75% (v/v) water concentration, a loosely connected network of dimers and trimers and two-dimensional array structures are observed. At higher water concentrations, AXT molecules form more compact three-dimensional structures that are preferentially solvated by the ethanol molecules. The ethanol preferential binding and the formation of a well connected hydrogen bonding network on these AXT clusters, suggest that such preferential solvation can play an important role in controlling the aggregate structure.


Colonic duplication in adults: report of two cases presenting with rectal bleeding.
Gastrointestinal duplication is an uncommon congenital abnormality in two-thirds of cases manifesting before the age of 2 years. Ileal duplication is common while colonic duplication, either cystic or tubular, is a rather unusual clinical entity that remains asymptomatic and undiagnosed in most cases. Mostly occurring in pediatric patients, colonic duplication is encountered in adults only in a few cases. This study reports two cases of colonic duplication in adults. Both cases presented with rectal bleeding on admission. The study was focused on clinical, imaging, histological, and therapeutical aspects of the presenting cases. Gastrografin enema established the diagnosis in both cases. The cystic structure and the adjacent part of the colon were excised en-block. The study implies that colonic duplication, though uncommon, should be included in the differential diagnosis of rectal bleeding.


[Manipulation in the exercise of psychiatric interviews].
The term "manipulation" is defined as "getting someone to behave otherwise than he spontaneously would". Unlike cognitive therapies, it does not involve cognitive functioning and may increase psychotherapies' efficiency. In the absence of data in the scientific literature (Medline and Web of Science), we propose a synthesis of theoretical data from social psychology with a reflection on its applications in the daily practice of psychiatry. Firstly we present auto-manipulation: the "chilling effect" is the fact that people tend to keep to a decision and to duplicate it, even if it does not work. The commitment of the patient, i.e., the degree to which he/she identifies with his/her act, will be even stronger if the patient's sense of freedom is high. The rationalization process is the ability for individuals to revisit their beliefs after being forced to issue a behavior (that he/she did not adhere to) to justify it a posteriori. In the second part, we present techniques for hetero-manipulation. Priming is to ask about a low effort to "initiate" the behavior. The lure technique is to hide convenience or invent fictitious benefits of a product, but is not ethical. The labeling technique is to assign an individual to a positive value regardless of his or her behavior, which increases the probability of emission of positive behaviors. The touch technique, whatever the cultural context, encourages a patient to have more confidence in his/her therapist and to make them friendly towards the person involved by creating a positive mood, reduces stress in patients before surgery, and improve the academic performance of students. We propose reflections on the application of these concepts in daily practice in the psychiatric interview. These techniques seem fundamental in improving therapeutic alliance and the likelihood of good compliance in our patients, and should be known to all practitioners.


Decellularized orthopaedic tissue-engineered grafts: biomaterial scaffolds synthesised by therapeutic cells.
In orthopaedic surgery, the reconstruction of musculoskeletal defects is a constant challenge. Biomaterials in tissue engineering are utilized as scaffolds which serve as templates for cell proliferation and secretion as well as guides for new tissue formation. The extracellular matrix (ECM) is a desirable biological scaffold due to its complex composition and three-dimensional ultrastructure, which drive the homeostasis and regeneration of tissues. Successfully used in a variety of regenerative medicine applications, ECM scaffolds can be achieved by decellularization of engineered tissue. In addition to using decellularized grafts directly as scaffolds, decellularized grafts can also be coated on or incorporated into synthetic biomaterials to substantially enhance their biological performance regarding integration into the surrounding tissue and bioactivity for neo-tissue generation. However, at present, the most widely adopted decellularized scaffolds are decellularized native scaffolds, which have the limitations of inherent heterogeneity, fixed shapes and insufficient sources. Decellularized tissue-engineered scaffolds are promising to avoid these restrictions and are receiving attention in the regenerative medicine field. This review describes the rationale of using decellularized tissue-engineered grafts for different regenerative purposes and details their application in the repair of orthopaedic defects.


In vitro Assessment of Pathogen Effector Binding to Host Proteins by Surface Plasmon Resonance.
The mechanisms of virulence and immunity are often governed by molecular interactions between pathogens and host proteins. The study of these interactions has major implications on understanding virulence activities, and how the host immune system recognizes the presence of pathogens to initiate an immune response. Frequently, the association between pathogen molecules and host proteins are assessed using qualitative techniques. As small differences in binding affinity can have a major biological effect, in vitro techniques that can quantitatively compare the binding between different proteins are required. However, these techniques can be manually intensive and often require large amounts of purified proteins. Here we present a simplified Surface Plasmon Resonance (SPR) protocol that allows a reproducible side-by-side quantitative comparison of the binding between different proteins, even in cases where the binding affinity cannot be confidently calculated. We used this method to assess the binding of virulence proteins (termed effectors) from the blast fungus Magnaporthe oryzae, to a domain of a host immune receptor. This approach represents a rapid and quantitative way to study how pathogen molecules bind to host proteins, requires only limited quantities of proteins, and is highly reproducible. Although this method requires the use of an SPR instrument, these can often be accessed through shared scientific services at many institutions. Thus, this technique can be implemented in any study that aims to understand host-pathogen interactions, irrespective of the expertise of the investigator.


Regulation of renal CYP4A expression and 20-HETE synthesis by nitric oxide in pregnant rats.
20-Hydroxyeicosatetraenoic acid (20-HETE), which promotes renal vasoconstriction, is formed in the rat kidney primarily by cytochrome P-450 (CYP) 4A isoforms (4A1, 4A2, 4A3, 4A8). Nitric oxide (NO) has been shown to bind to the heme moiety of the CYP4A2 protein and to inhibit 20-HETE synthesis in renal arterioles of male rats. However, it is not known whether NO interacts with and affects the activity of CYP4A1 and CYP4A3, the major renal CYP4A isoforms in female rats. Incubation of recombinant CYP4A1 and 4A3 proteins with sodium nitroprusside (SNP) shifted the absorbance at 440 nm, indicating the formation of a ferric-nitrosyl-CYP4A complex. The absorbance for CYP4A3 was about twofold higher than that of CYP4A1. Incubation of SNP or peroxynitrite (PN; 0.01-1 mM) with CYP4A recombinant membranes caused a concentration-dependent inhibition of 20-HETE synthesis, with both chemicals having a greater inhibitory effect on CYP4A3-catalyzed activity. Moreover, incubation of CYP4A1 and 4A3 proteins with PN (1 mM) resulted in nitration of tyrosine residues in both proteins. In addition, PN and SNP inhibited 20-HETE synthesis in renal microvessels from female rats by 65 and 59%, respectively. We previously showed that microvessel CYP4A1/CYP4A3 expression and 20-HETE synthesis are decreased in late pregnancy. Therefore, we investigated whether such a decrease is dependent on NO, the synthesis of which has been shown to increase in late pregnancy. Administration of NG-nitro-l-arginine methyl ester (l-NAME) to pregnant rats for 6 days (days 15-20 of pregnancy) caused a significant increase in systolic blood pressure, which was prevented by concurrent treatment with the CYP4A inhibitor 1-aminobenzotriazole (ABT). Urinary NO2/NO3 excretion decreased by 40 and 52% in l-NAME- and l-NAME + ABT-treated groups, respectively. Interestingly, renal microvessel 20-HETE synthesis showed a marked increase following l-NAME treatment, and this increase was diminished with coadministration of ABT. These results demonstrate that NO interacts with CYP4A proteins in a distinct manner and it interferes with renal microvessel 20-HETE synthesis, which may play an important role in the regulation of blood pressure and renal function during pregnancy.


Surface aligned magnetic moments and hysteresis of an endohedral single-molecule magnet on a metal.
The interaction between the endohedral unit in the single-molecule magnet Dy_{2}ScN@C_{80} and a rhodium (111) substrate leads to alignment of the Dy 4f orbitals. The resulting orientation of the Dy_{2}ScN plane parallel to the surface is inferred from comparison of the angular anisotropy of x-ray absorption spectra and multiplet calculations in the corresponding ligand field. The x-ray magnetic circular dichroism is also angle dependent and signals strong magnetocrystalline anisotropy. This directly relates geometric and magnetic structure. Element specific magnetization curves from different coverages exhibit hysteresis at a sample temperature of ∼4 K. From the measured hysteresis curves, we estimate the zero field remanence lifetime during x-ray exposure of a submonolayer to be about 30 seconds.


Diet and obstructive lung diseases.
The results presented in this review suggest that the impact of nutrition on obstructive lung disease is most evident for antioxidant vitamins, particularly vitamin C and, to a lesser extent, vitamin E. By decreasing oxidant insults to the lung, antioxidants could modulate the development of chronic lung diseases and lung function decrement. Antioxidant vitamins could also play an important role in gene-environment interactions in complex lung diseases such as childhood asthma. Data also suggest that omega-3 fatty acids may have a potentially protective effect against airway hyperreactivity and lung function decrements; however, relevant data are still sparse. Although epidemiologic data suggest that consumption of fresh fruit may reduce risk of noncarcinogenic airway limitation, there are no clear data on which nutrients might be most relevant. While some studies evaluate daily intake of vitamin C, other studies use fruit consumption as a surrogate for antioxidant intake. Given the dietary intercorrelations among antioxidant vitamins, particularly vitamin C, beta-carotene, and flavonoids, as well as other micronutrients, it may be difficult to isolate a specific effect. Some population subgroups with higher levels of oxidative stress, such as cigarette smokers, may be more likely to benefit from dietary supplementation, since some studies have suggested that antioxidant intake may have a greater impact in this group. Studies of lung function decrement and COPD in adults suggest that daily intake of vitamin C at levels slightly exceeding the current Recommended Dietary Allowance (60 mg/day among nonsmokers and 100 mg/day among smokers) may have a protective effect (20). In the Schwartz and Weiss (85) and Britton et al. (87) studies, an increase of 40 mg/day in vitamin C intake led to an approximate 20-ml increase in FEV1. Daily mean vitamin C intakes in these studies were 66 mg and 99.2 mg, respectively, and the highest intake level (178 mg/day) was approximately three times the Recommended Dietary Allowance. Although the amplitude of the effect was modest, if these effects accumulate over 20-30 years, they could have a meaningful impact on the rate at which pulmonary function declines, particularly in symptomatic subjects (85). Longitudinal data support the hypothesis that fresh fruit consumption has a beneficial impact on the lung (95). Among children, consumption of fresh fruit, particularly fruit high in vitamin C, has been related to a lower prevalence of asthma symptoms and higher lung function (64). This effect was observed event at low levels of fruit consumption (one or two servings per week vs. less than one serving per week), which suggests that a small increase in dietary intake could have a beneficial effect. Consumption of fish has also been related to lower airway hyperreactivity among children (75) and higher lung function in adults (100); however, longitudinal data do not provide evidence that increased omega-3 fatty acid intake protects against lung disease (101). Experimental studies of persons with asthma suggest that magnesium infusion may have a place in the acute treatment of asthma, but it does not seem to have long-term benefits. The studies of sodium, selenium, and fish oils do not show convincing evidence of clinical benefits. Studies of vitamin C supplementation suggest a short-term protective effect on airway responsiveness and pulmonary function. It remains to be proven whether consistent use of vitamin C would have a protective effect on the evolution of chronic asthma. Results from supplementation studies conducted among subjects exposed to high levels of oxidants (57-60) suggest that daily intake of antioxidant vitamins exceeding the Recommended Dietary Allowance may have a beneficial effect on lung airways and that intake higher than the Recommended Dietary Allowance should be recommended for populations chronically exposed to photooxidant air pollutants (such as ozone), cigarette smoking, or vigorous exercise. It is difficult to determine the amounts of antioxidant vitamins that people should consume. In particular, although vitamin C was shown to have maximum bioavailability when given in a single dose of 200 mg (102), experiments on which this finding was based were conducted under normal conditions. Guidelines from the US National Cancer Institute (103) recommend consumption of five servings of fruit and vegetables daily, corresponding to a vitamin C intake exceeding 200 mg. Dietary surveys carried out in the US population indicate that less than 12 percent of US children and adults meet this recommended level of intake (104). Diet appears to be an important cofactor in the development of obstructive lung disease, although data are still sparse. There is a need for further research in experimental and epidemiologic settings to better understand the physiologic effects of antioxidant vitamins, omega-3 fatty acids, and other nutrients on lung tissues. The impact of diet on the incidence and evolution of asthma and COPD should be investigated using a cohort design that accounts for known risk factors. This will allow researchers to evaluate the exposure-disease relation over an adequate time frame and obtain insight into the causality of the relation. Some of these studies should enroll infants and young children to determine the impact of early diet on respiratory health. Research should also focus on the equally challenging policy issues--namely, finding effective methods of convincing people to increase their daily consumption of fresh fruits and vegetables, to stop smoking cigarettes, and to minimize their environmental and occupational exposure to pollutants and other agents that cause respiratory disease.


Epidemiology of Viral Hepatitis and Liver Diseases in Pakistan.
Pakistan is among ten most populous countries in world. All sorts of hepatitis are highly prevalent in Pakistan. Due to lack of proper mechanisms to handle sewerage, disinfection and lack of clean water supply across the country, hepatitis A and E are endemic in Pakistan. Moreover, Pakistan ranked in intermediate prevalence zone for hepatitis B and C. Also, prevailing socioeconomic conditions, political statuses and unstable healthcare delivery system are not suitable for tackling further progression of hepatitis viruses and its complications. The notable factors related to epidemiology of different hepatitis viruses have been described in this communication. Butt AS. Epidemiology of Viral Hepatitis and Liver Diseases in Pakistan. Euroasian J Hepato-Gastroenterol 2015;5(1):43-48.


Circadian rhythms in mitochondrial respiration.
Many physiological processes are regulated with a 24-h periodicity to anticipate the environmental changes of daytime to nighttime and vice versa. These 24-h regulations, commonly termed circadian rhythms, among others control the sleep-wake cycle, locomotor activity and preparation for food availability during the active phase (daytime for humans and nighttime for nocturnal animals). Disturbing circadian rhythms at the organ or whole-body level by social jetlag or shift work, increases the risk to develop chronic metabolic diseases such as type 2 diabetes mellitus. The molecular basis of this risk is a topic of increasing interest. Mitochondria are essential organelles that produce the majority of energy in eukaryotes by converting lipids and carbohydrates into ATP through oxidative phosphorylation. To adapt to the ever-changing environment, mitochondria are highly dynamic in form and function and a loss of this flexibility is linked to metabolic diseases. Interestingly, recent studies have indicated that changes in mitochondrial morphology (i.e., fusion and fission) as well as generation of new mitochondria are dependent on a viable circadian clock. In addition, fission and fusion processes display diurnal changes that are aligned to the light/darkness cycle. Besides morphological changes, mitochondrial respiration also displays diurnal changes. Disturbing the molecular clock in animal models leads to abrogated mitochondrial rhythmicity and altered respiration. Moreover, mitochondrial-dependent production of reactive oxygen species, which plays a role in cellular signaling, has also been linked to the circadian clock. In this review, we will summarize recent advances in the study of circadian rhythms of mitochondria and how this is linked to the molecular circadian clock.


Trends in procedures for infertility and caesarean sections: was NICE disinvestment guidance implemented? NICE recommendation reminders.
National Institute for Health and Clinical Excellence (NICE) clinical guidelines and subsequent NICE issued 'recommendation reminders' advocate discontinuing two fertility procedures and caesarean sections in women with hepatitis. We assess whether NICE guidance in 2004 and recommendation reminders were associated with a change in the rate of clinical procedures performed. Routine inpatient Hospital Episode Statistics (HES) data were extracted from the HES database for 1st April 1998 to 31st March 2010 using OPCS procedure codes for varicocele operations in infertile men, endometrial biopsies in infertile women and caesarean sections in women with hepatitis B or C. We used Joinpoint regression to identify points in time when the trend in procedure rates changed markedly, to identify any influence of the release of NICE guidance. Between 1998-2010, planned caesarean sections in women with and without hepatitis B or C increased yearly (annual percentage change (APC) 4.9%, 95% CI 2.1% to 7.7%) in women with hepatitis, compared with women without (APC 4.0% [95% CI 2.7% to 5.3%] up to 2001, APC -0.6% [95% CI -2.8% to 1.8%] up to 2004 and 1.3% [95% CI 0.8% to 1.8%] up to 2010). In infertile women under 40 years of age, endometrial biopsies for investigation of infertility increased, APC 6.0% (95% CI 3.6% to 8.4%) up to 2003, APC 1.5% (95% CI -4.3% to 7.7%) to 2007 followed by APC 12.8% (95% CI 1.0% to 26.0%) to 2010. Varicocele procedures remained relatively static between 1998 and 2010 (APC -0.5%, 95% CI -2.3% to 1.3%). There was no decline in use of the three studied procedures, contrary to NICE guidance, and no change in uptake associated with the timing of NICE guidance or recommendation reminders. 'Do not do' recommendation reminders may be ineffective at improving clinical practice or achieving disinvestment.


Phenothiazine-linked nucleosides and nucleotides for redox labelling of DNA.
Nucleosides and 2'-deoxyribonucleoside triphosphates (dNTPs) bearing phenothiazine (PT) attached to a nucleobase (cytosine or 7-deazaadenine) either directly or through an acetylene linker were prepared through Suzuki or Sonogashira cross-coupling and triphosphorylation, and were studied as building blocks for polymerase construction of modified DNA. The directly PT-substituted dNTPs were better substrates for polymerases than the alkyne-linked dNTPs but all of them were used in enzymatic synthesis of DNA using primer extension, nicking enzyme amplification, PCR or 3'-tail labelling by terminal deoxynucleotidyl transferase. The phenothiazine served as an oxidizable redox label (giving two analytically useful signals of oxidation on electrode) for nucleosides and DNA and was also used in orthogonal combination with previously developed benzofurazane or nitrophenyl labels for redox coding of DNA bases. Therefore, the title PT-linked dNTPs are useful additions to the portfolio of nucleotides for enzymatic synthesis of redox-labelled DNA for electrochemical analysis.


Evaluation of Bacillus sphaericus bioinsecticide produced with white soybean meal as culture medium for the control of Culex (Culex) quinquefasciatus.
Bioinsecticides are shown to be useful in control programs to prevent several diseases, based on their specificity and efficiency against insect vectors. In the current study a bioinsecticide based on Bacillus sphaericus was produced using a white soybean culture medium and applied to larvae of Culex quinquefasciatus, the susceptible species, and Aedes aegypti, the refractory species used as the negative control. Efficacy was compared with that of the product fermented with the Luria Bertani (LB) reference medium. The experiments showed that C. quinquefasciatus was highly susceptible to the product prepared with white soybean meal, reaching 100% larval mortality even at 10mg/L, while A. aegypti failed to reach 70% mortality at a concentration of 1g/L. By comparison with the reference medium, the proposed culture medium showed high larvicidal power, reaching a LD90 of 2.26 mg/L, while 4.37 mg/L was needed for the LB medium to achieve the same mortality rate. Cost comparison between the formulations favored the use of the bioinsecticide produced with white soybean meal. After factoring in the LD90 value, the cost ratio favored the new raw material by nearly 1:220.


Daphnia magna negatively affected by chronic exposure to purified Cry-toxins.
Cry-toxin genes originating from Bacillus thuringiensis are inserted into genetically modified (GM) plants, often called Bt-plants, to provide insect resistance to pests. Significant amounts of Bt-plant residues, and thus Cry-toxins, will be shed to soil and aquatic environments. We exposed Daphnia magna to purified Cry1Ab and Cry2Aa toxins for the full life-span of the animals. We used single toxins in different doses and combinations of toxins and Roundup(®), another potential stressor on the rise in agricultural ecosystems. Animals exposed to 4.5 mg/L (ppm) of Cry1Ab, Cry2Aa and the combination of both showed markedly higher mortality, smaller body size and very low juvenile production compared to controls. Animals exposed to 0.75 mg/L also showed a tendency towards increased mortality but with increased early fecundity compared to the controls. Roundup(®) stimulated animals to strong early reproductive output at the cost of later rapid mortality. We conclude that i) purified Cry-toxins in high concentrations are toxic to D. magna, indicating alternative modes-of-action for these Cry-toxins; ii) Cry-toxins act in combination, indicating that 'stacked events' may have stronger effects on non-target organisms; iii) further studies need to be done on combinatorial effects of multiple Cry-toxins and herbicides that co-occur in the environment.


Results of surgical repair of abductor avulsion after primary total hip arthroplasty.
Abductor avulsion after primary total hip arthroplasty (THA) is an uncommon event and is associated with the lateral approach. Results of surgical repair have only been reported in one previous study. In a retrospective review of 2657 primary THAs, we identified 19 (0.7%) patients operated upon for repair of abductor avulsion on average 19 months after primary THA. The study's aim was to assess improvement in pain and limp, patient satisfaction, and functional outcome (Harris Hip Score) after surgical repair. Eighteen patients were evaluated at a mean follow-up of 38 months. Overall, only half of the patients had substantial improvement of both limp and pain. Limping and functional outcome markedly improved with early repair (<15 months), whereas obesity was associated with worse functional results.


Flow cytometry, a new approach to detect anti-live trypomastigote antibodies and monitor the efficacy of specific treatment in human Chagas' disease.
Sera from patients chronically infected with Trypanosoma cruzi display antibodies that bind to epitopes of living trypomastigotes, known as lytic antibodies (LA), and are detected by a complement-mediated lysis test. Conventional serology antibodies (CSA) are also present in sera from patients with chronic infections but, in contrast to LA, are unable to recognize viable trypomastigotes. The presence of LA has been used as an important element in the criterion of cure in human Chagas' disease. Using flow cytometry technology, we introduced a new and sensitive immunomethod for the detection of anti-live trypomastigote membrane-bound antibodies. On the basis of serological tests (LA and CSA detection) and parasitological assays such as hemoculture (HE), patients were classified into the following groups: chronically infected untreated patients (NT) and treated not-cured patients (TNC), with positive HE and both LA and CSA in their sera; "dissociated" HE-negative patients (DIS), in whom LA was not detected whereas CSA were present; a group of cured HE-negative patients (CUR), who were both LA and CSA negative; and, as control, a group of non-chagasic individuals (NC). Sera from these patients were assayed by incubation with live bloodstream trypomastigotes, which were subsequently exposed to fluorescein isothiocyanate-conjugated anti-human immunoglobulin G. The parasites were then fixed, run in the cytometer, and identified on basis of their size and granularity gain adjustments. On the basis of experience with the complement-mediated lysis test, a level of 20% of parasites being fluorescein isothiocyanate fluorescence positive was used as a cutoff between effective and ineffective treatments. With this criterion, our results indicated that sera from NT and TNC patients were antibody positive whereas all sera from DIS, CUR, and NC patients did not contain membrane-bound antibodies. This new approach is a tool to easily identify anti-live T. cruzi membrane-bound antibodies that can be used to monitor the efficacy of Chagas' disease treatment.


Cannabis treatment outcomes among legally coerced and non-coerced adults.
Treatment seeking for cannabis dependence in general, and particularly the number of criminal justice referrals to cannabis treatment, has increased over the past decade. This study aims to compare the characteristics, psychosocial functioning and treatment outcome of those legally coerced into cannabis treatment compared to those entering treatment without legal coercion. This study is a retrospective audit of the administrative clinical records of 27,198 adults presenting to public Texas treatment programs with cannabis as their primary drug problem between 2000 and 2005. Of the 69% legally coerced into treatment, there was less psychological distress and greater likelihood of having completed treatment compared with non-coerced clients. Participants who were legally coerced into treatment were also more likely to have received less intensive forms of treatment and to have not used cannabis in the month prior to 90-day post-treatment follow-up. More public health information is needed on cannabis dependence and increased availability of subsidised early and brief interventions in a variety of primary health care settings would reduce the late presentations of the more severely impaired voluntary clients. The limitations of this dataset are discussed.


Object-finding skill created by repeated reward experience.
For most animals, survival depends on rapid detection of rewarding objects, but search for an object surrounded by many others is known to be difficult and time consuming. However, there is neuronal evidence for robust and rapid differentiation of objects based on their reward history in primates (Hikosaka, Kim, Yasuda, & Yamamoto, 2014). We hypothesized that such robust coding should support efficient search for high-value objects, similar to a pop-out mechanism. To test this hypothesis, we let subjects (n = 4, macaque monkeys) view a large number of complex objects with consistently biased rewards with variable training durations (1, 5, or 30 + days). Following training, subjects searched for a high-value object (Good) among a variable number of low-value objects (Bad). Consistent with our hypothesis, we found that Good objects were accurately and quickly targeted, often by a single and direct saccade with a very short latency (<200 ms). The dependence of search times on display size reduced significantly with longer reward training, giving rise to a more efficient search (40 ms/item to 16 ms/item). This object-finding skill showed a large capacity for value-biased objects and was maintained in the long-term memory with no interference from reward learning with other objects. Such object-finding skill, and in particular its large capacity and long term retention, would be crucial for maximizing rewards and biological fitness throughout life where many objects are experienced continuously and/or intermittently.


[Prostatic specific antigen (PS), pro-inflammatory cytokines, and prostatic pathology (benign prostatic hyperplasia and cancer). Relationship with malignancy].
Cancer is a complex process in which cytokines play an important role. Cytokines are low-molecular weight soluble proteins involved in cellular transmission signals and several disorders. Pro-inflammatory cytokines (IL-1, TNF-alpha and IL-6) are involved in prostate cancer development. The aim of this study was to relate the expression (analyzed by Western blot and immuno-histochemistry) of several pro-inflammatory cytokines (IL-1, TNF-alpha and IL-6) with serum levels of prostate-specific antigen (PSA) in normal (no pathological samples) as pathological samples (hyperplasia and cancer), in order to elucidate their possible role in tumor progression. We are also discussing the possible use of these cytokines as a potential therapeutic target. This study was carried out in 5 normal, 25 benign prostatic hyperplastic (BPH) and 17 prostate cancer (PC) human prostates. Immunohistochemical and Western blot analysis were performed. Serum levels of PSA were assayed by a PSA DPC immulite assays (Diagnostics Products Corporation, Los Angeles, CA). In BPH, IL-1alpha, IL-6 and TNF-alpha were only expressed in patients with PSA serum levels of 0-4 ng/ml or 4-20 ng/ml, but not in the group >20 ng/ml. In PC these cytokines were only expressed in patients with PSA serum levels >4 ng/ml, although the expression of these cytokines was elevated when PSA levels were >20 ng/ml. In PC there might be an association between high expression of pro-inflammatory cytokines (IL-1, TNFalpha and IL-6), elevated serum levels of PSA and cancer progression. A better understanding of the biologic mechanism of this association may improve the finding of new targets for therapy in these patients.


Use of high-throughput screening results to prioritize chemicals for potential adverse biological effects within a West Virginia watershed.
Organic chemicals from industrial, agricultural, and residential activities can enter surface waters through regulated and unregulated discharges, combined sewer overflows, stormwater runoff, accidental spills, and leaking septic-conveyance systems on a daily basis. The impact of point and nonpoint contaminant sources can result in adverse biological effects for organisms living in or near surface waters. Assessing the adverse or toxic effects that may result when exposure occurs is complicated by the fact that many commonly used chemicals lack toxicity information or water quality standards. To address these challenges, an exposure-activity ratio (EAR) screening approach was used to prioritize environmental chemistry data in a West Virginia watershed (Wolf Creek). Wolf Creek is a drinking water source and recreation resource with documented water quality impacts from point and nonpoint sources. The EAR screening approach uses high-throughput screening (HTS) data from ToxCast as a method of integrating environmental chemical occurrence and biological effects data. Using water quality schedule 4433, which targets 69 organic waste compounds typically found in domestic and industrial wastewater, chemicals were screened for potential adverse biological affects at multiple sites in the Wolf Creek watershed. Cumulative EAR mixture values were greatest at Sites 2 and 3, where bisphenol A (BPA) and pentachlorophenol exhibited maximum EAR values of 0.05 and 0.002, respectively. Site 2 is downstream of an unconventional oil and gas (UOG) wastewater disposal facility with documented water quality impacts. Low-level organic contaminants were found at all sample sites in Wolf Creek, except Site 10, where Wolf Creek enters the New River. The application of an EAR screening approach allowed our study to extend beyond traditional environmental monitoring methods to identify multiple sites and chemicals that warrant further investigation.


Rates and determinants of virologic and immunological response to HAART resumption after treatment interruption in HIV-1 clinical practice.
To describe CD4 and HIV RNA changes during treatment resumption (TR) after treatment interruption (TI) compared with response to first highly active antiretroviral therapy (HAART) and to investigate predictors. Using Concerted Action on SeroConversion to AIDS and Death in Europe (CASCADE) data, we identified subjects who interrupted first HAART, not initiated during primary infection. We estimated rate of CD4 change during TR and time from TR to HIV RNA<500 copies per milliliter and subsequent rebound and factors associated with these outcomes. Of 281 persons treated for median 18.4 months before interrupting, 259 resumed HAART. CD4 increases in the first 3 months on HAART were similar pre-TI and post-TI but after 3 months were significantly higher during pre-TI HAART, with median +106 and +172 cells per microliter at 3 and 18 months, respectively, during initial HAART compared with +99 and +142 cells per microliter during post-TI HAART, respectively. Subjects with lower CD4 counts at TI, aged older than 40 years, and those resuming the same HAART as their pre-TI regimen had lower CD4 increases during the first 3 months of TR. The majority (86%) of individuals reinitiating therapy achieved HIV RNA<500 copies per milliliter. Immune reconstitution after TI is generally poorer than after first HAART, particularly for patients aged older than 40 years at TI and those with poorer immunological responses to pre-TI HAART. Reinitiation of the same HAART regimen as pre-TI also seems to have unfavorable outcomes.


Evolution of sex-biased maternal effects in birds: II. Contrasting sex-specific oocyte clustering in native and recently established populations.
In species that produce broods of multiple offspring, parents need to partition resources among simultaneously growing neonates that often differ in growth requirements. In birds, multiple ovarian follicles develop inside the female at the same time, resulting in a trade-off of resources among them and potentially limiting maternal ability for sex-specific allocation. We compared resource acquisition among oocytes in relation to their future sex and ovulation order in two populations of house finches with contrasting sex-biased maternal strategies. In a native Arizona population, where mothers do not bias offspring sex in relation to ovulation order, the male and female oocytes did not show sex-specific trade-offs of resources during growth and there was no evidence for spatial or temporal segregation of male and female oocytes in the ovary. In contrast, in a recently established Montana population where mothers strongly bias offspring sex in relation to ovulation order, we found evidence for both intra-sexual trade-offs among male and female oocytes and sex-specific clustering of oocytes in the ovary. We discuss the importance of sex-specific resource competition among offspring for the evolution of sex-ratio adjustment and sex-specific maternal resource allocation.


An epidemiological study of sheep pox infection in Karnataka State, India.
Analysis of retrospective quantitative sheep pox epidemiological data from the Government Animal Husbandry Department, Karnataka, India, covering 24 years revealed significant information on sheep pox. The state has a dense sheep population including some valuable breeds. Data revealed the endemicity of the disease: there were a considerable number of outbreaks and attacks, high mortality and case fatality rates and low immunisation coverage. None of the years studied were free from infection. Temporally, the disease was most prevalent between November and May. Spatially, the disease was recorded in 19 out of 27 districts; in some of these districts sheep pox was highly endemic, in some it was endemic at low levels and in the remaining districts outbreaks occurred sporadically. Environmental factors influenced disease occurrence. Vaccine production met only one tenth of the requirement, and its peak utilisation was in the dry season.


Estrogen receptor-α directly regulates the hypoxia-inducible factor 1 pathway associated with antiestrogen response in breast cancer.
A majority of breast cancers are driven by estrogen via estrogen receptor-α (ERα). Our previous studies indicate that hypoxia-inducible factor 1α (HIF-1α) cooperates with ERα in breast cancer cells. However, whether ERα is implicated in the direct regulation of HIF-1α and the role of HIF-1α in endocrine therapy response are unknown. In this study we found that a subpopulation of HIF-1α targets, many of them bearing both hypoxia response elements and estrogen response elements, are regulated by ERα in normoxia and hypoxia. Interestingly, the HIF-1α gene itself also bears an estrogen response element, and its expression is directly regulated by ERα. Clinical data revealed that expression of the HIF-1α gene or a hypoxia metagene signature is associated with a poor outcome to endocrine treatment in ERα(+) breast cancer. HIF-1α was able to confer endocrine therapy resistance to ERα(+) breast cancer cells. Our findings define, for the first time to our knowledge, a direct regulatory pathway between ERα and HIF-1α, which might modulate hormone response in treatment.


Diffractive imaging for periodic samples: retrieving one-dimensional concentration profiles across microfluidic channels.
A technique has been developed that allows determination of the concentration profiles of colloidal solutions or any kind of fluid under confinement. Currently, submicrometre-wide channels are sampled with a resolution in the 10 nm range. The method comprises regular arrays of microfluidic channels and one-dimensional X-ray phase-retrieval techniques for the analysis of small-angle X-ray diffraction from the array structures. Recording the X-ray diffraction data requires a low dose on each individual channel since the sum of the signals from all channels is detected. The determined concentration profiles represent the ensemble average rather than individual entities and are obtained in a model-independent way. As an example, amplitude and phase of the exit field and concentration profiles for a colloidal fluid within confining channels of different widths are shown.


Molecular characterization of PAHs based on land use analysis and multivariate source apportionment in multiple phases of the Yangtze estuary, China.
Spatial-temporal distributions, source identification, and possible effects of land use patterns on PAHs were studied in overlying water and surface sediments in Yangtze estuarine and nearby coastal areas. The concentrations of PAHs ranged from 172.6 ± 9.3 to 5603.7 ± 548.7 ng L-1, 841.0 ± 36.6 to 31 190.5 ± 3711.5 ng g-1 and 312.3 ± 7.6 to 9081.1 ± 862.3 ng g-1, with mean contents of 1042.9 ng L-1, 8922.9 ng g-1 and 2400.2 ng g-1 in water, suspended particulate matter (SPM) and sediments. PAH concentrations in three phases were higher in dry seasons (January and April) than in rainy seasons (July and October), and showed a distribution pattern as freshwater area > transition area > seawater area. Water and urban (especially for industrial) land uses contributed significantly to the PAH content in water and sediments. Urban and agricultural land uses were identified as important factors for regulating PAH concentrations in the SPM. TOC and BC were crucial controlling factors for the sorption and distribution of PAHs. The stable carbon isotope signatures of TOC and BC indicated the terrestrial origin of PAHs. Multivariate statistical and toxicity assessments revealed that LHK and SDK were contamination hotspots. Traffic emissions contributed 63% of PAH loadings in the three phases, yet biomass/coal combustion contributed over 85% of the toxicity caused by PAHs. Potential ecological and health risks of PAHs were low. We thus conclude that PAH pollution in the study area was primarily of terrestrial origin. BC and TOC were important predictors for PAH fate in the estuary. Urban and agricultural land use and river systems along the estuary were the major input pathways for the PAH loadings.


7-Methylguanine specific tRNA-methyltransferase from Escherichia coli.
A 7-methylguanine (m7G) specific tRNA methyltransferase from E. coli MRE 600 was purified about 1000 fold by affinity chromatography on Sepharose bound with normal E. coli tRNA. The purified enzyme catalyzes exclusively the formation of m7G in submethylated bulk tRNA of E. coli K12 met- rel-. The purified enzyme transfers the methyl group from S-adenosyl-methionine to initiator tRNA of B. subtilis and 0.8 moles m7G residues are formed per mole tRNA. It is suggested that the enzyme specifically recognizes the extra arm unpaired guanylate residue.


Functional domains of the penicillinase repressor of Bacillus licheniformis.
The penicillinase repressor (PENI) negatively regulates expression of the penicillinase gene (penP) in Bacillus licheniformis by binding to its operators located within the promoter region of penP.penI codes for a protein with 128 amino acids. Filter-binding analyses suggest that the active form of the repressor is a dimer. Genetic analyses of PENI derivatives showed that the repressor carrying either a 6-amino-acid deletion near the N terminus or a 14-amino-acid deletion at the C terminus was functionally inactive in vivo. A repressor derivative carrying a 6-amino-acid deletion within its N-terminal region was extensively purified and used in DNA footprinting and subunit cross-linking analyses. The results of these studies showed that the repressor derivative had lost its ability to bind operator specifically even though it could dimerize effectively. In similar studies, we demonstrated that an N-terminal portion of PENI with a molecular mass of 10 kDa derived by digestion with papain was able to bind operator specifically but with reduced affinity and had completely lost its ability to dimerize. These data suggest that the repressor has two functional and separable domains. The amino-terminal domain of the repressor is responsible for operator recognition, and the carboxyl-terminal domain is involved in subunit dimerization.


Comparison of laser versus sclerotherapy in the treatment of lower extremity telangiectases: a prospective study.
Lower extremity telangiectasia affects approximately 40% of women. The demand for aesthetic treatment of these veins continues to grow. Few studies have compared laser and sclerotherapy to treat leg telangiectasias. To compare the efficacy of conventional sclerotherapy and neodymium-doped yttrium aluminum garnet (Nd:YAG) laser in the treatment of leg telangiectasias. Thirty women were enrolled in the study. One leg was randomly assigned laser treatment and the other sclerotherapy with 75% glucose solution. All patients were photographed before and after treatment. The applying physician and two independent observers rated photographic improvement of the treated areas. Complications and adverse effects were noted during follow-up. Patients answered a questionnaire that addressed pain, clearing of the vessels, and satisfaction with the results. There was a significant difference between the modes of treatment regarding pain. Twelve patients using laser and 16 using sclerotherapy considered the clearing of the vessels to be good to excellent after three sessions of both laser and scleratherapy. Mean scores after photographic assessment were 7.9 for laser and 7.0 for sclerotherapy. Lower extremity telangiectases may be treated equally well using Nd:YAG 1064-nm laser or conventional sclerotherapy.


Three-arm, placebo-controlled, randomized clinical trial evaluating the metabolic effect of a combined nutraceutical containing a bergamot standardized flavonoid extract in dyslipidemic overweight subjects.
Our double-blind, placebo-controlled, parallel-group, dose-escalation, clinical trial aimed to test the effect of a combined nutraceutical containing bergamot extract (120-mg flavonoids), phytosterols, vitamin C, and chlorogenic acid from dry artichoke extract on 90 overweight dyslipidemic subjects. Participants were randomly allocated to treatment with two pills of either active treatment or placebo, or a combination of both (a pill per treatment). After 8 weeks, all active-treated groups experienced a significant improvement in triglycerides (TG) versus placebo and in low-density lipoprotein cholesterol (LDL-C) versus baseline and placebo treatments. In the high-dose-treated group, also total cholesterol (TC), nonhigh-density lipoprotein cholesterol (non-HDL-C), γ-glutamil transpeptidasi, high-sensitivity C-reactive protein (hs-CRP), and tumor necrosis factor-α (TNF-α) significantly decreased. At 24-week follow-up, TG levels maintained lower than baseline in all groups. All patients allocated to either low-dose or high-dose active treatment experienced a significant decrease in TG, LDL-C, and homeostatin model assessment of insulin resistance. In subjects taking high-dose active treatment, adiponectin significantly increased, whereas TC, non-HDL-C, insulin (fasting plasma insulin), leptin, leptin/adiponectin ratio, hs-CRP, and TNF-α were significantly reduced. The tested nutraceutical showed to improve lipid and glucose metabolism, adipokines pattern, and systemic inflammation in dyslipidemic overweight subjects.


Microbiological effects in patients with leg ulcers and diabetic foot treated with Lucilia sericata larvae.
Lucilia sericata bottle fly worms can be used to heal infected, chronic, or necrotic wounds, including those associated with ulceration and diabetic foot. The study aimed to evaluate changes in the microflora in patients treated with L sericata larvae due to leg ulcers and diabetic foot. One hundred twenty-nine patients diagnosed with lower limb ulceration and diabetic foot were enrolled in the study, of which 80 of them met the eligibility criteria for maggot debridement therapy (MDT). On the contrary, 49 unqualified patients were offered ozone therapy (22 with leg ulcers; 27 with diabetic foot). In each of these patients, a microbiological swab was performed before and after the start of therapy. The group of 80 patients was further divided into four equal groups in terms of the treated area (lower leg vs foot) and the number of larvae/cm2 (5 vs 10). Twenty-three particular species of bacteria in the infected wound were studied microbiologically in terms of presence/absence within the wound environment before and after treatment of patients with diabetic foot and lower limb ulceration. It was noted that there was a more intensive bacterial accumulation in the feet of patients compared to legs; furthermore, this applies to almost all analysed species. Diabetes status is also a clinical factor that generates a lower chance of bacterial appearance in the wound environment. Densification of MDT larvae per wound area unit also reduced the chance of the presence of Corynebacterium species, Enterobacteriaceae, Pseudomonas aeruginosa, Staphylococcus aureus MSSA, and Streptococcus coagulase negativa; however, it increased the likelihood of occurrence for Proteus mirabilis and the Proteus species. A microbiological analysis in this non-reference study shows the efficacy of larval therapy for leg and foot ulcers. Rearrangement of the microflora within the wound has been reported as a result of the therapy.


Is there convergence in the molecular pathways underlying the repeated evolution of sociality in African cichlids?
Despite wide variation in the complexity of social interactions across taxa, the basic behavioral components of sociality appear to be modulated by conserved hormone pathways. Specifically, the nonapeptide hormones oxytocin and vasopressin and their receptors have been implicated in regulating diverse social behaviors across vertebrates. Here, we took advantage of the repeated evolution of cooperative breeding in African cichlids to investigate whether there are consistent brain gene expression patterns of isotocin and arginine vasotocin (teleost homologues of oxytocin and vasopressin), as well as their receptors, between four closely related pairs of social (cooperative) and non-social (non-cooperative) species. We first found that the coding sequences for the five genes studied were highly conserved across the eight species. This is the first study to examine the expression of both isotocin receptors, and so we performed a phylogenetic analysis that suggests that these two isotocin receptors are paralogues that arose during the teleost genome duplication. When we then examined brain gene expression patterns relative to social system, we found that there were whole-brain gene expression differences between the social and non-social species in many of the species pairs. However, these relationships varied in both the direction and magnitude among the four species pairs. In conclusion, our results suggest high sequence conservation and species-specific gene expression patterns relative to social behavior for these candidate hormone pathways in the cichlid fishes.


An online survey of informal caregivers' unmet needs and associated factors.
The purpose of this study was to assess the frequency of unmet needs of carers among a convenience sample of carers, and the participant factors associated with unmet needs, to inform the development of interventions that will support a range of caregivers. The aims of this study were to: (1) assess the most frequently reported moderate-high unmet needs of caregivers; and (2) examine the age, gender, condition of the care recipient, and country variables associated with types of unmet needs reported by informal caregivers. An online cross-sectional survey among informal caregivers in English-speaking countries was conducted. Self-reported unmet needs were assessed using an unmet needs measure with the following five unmet needs domains: (1) Health information and support for care recipient; (2) Health service management; (3) Communication and relationship; (4) Self-care; and (5) Support services accessibility. Informal caregivers were asked "In the last month, what was your level of need for help with…", and the ten highest ranked moderate-high unmet needs presented as ranked proportions. Logistic regression modelling examined the factors associated with types of unmet needs. Overall, 457 caregivers were included in the final analysis. Seven of the ten highest ranked unmet needs experienced by caregivers in the last month were in the Self-care domain, including "Reducing stress in your life" (74.1%). Significant associations were found between younger caregiver age (18-45 years) and reporting moderate-high unmet needs in Health Information and support for care recipient, Health service management, and Support services accessibility (all p's = <0.05). Caregivers are not experiencing significant differences in unmet needs between countries and caree/care recipient conditions, suggesting that general interventions could be developed to support a range of caregivers across countries. Increased awareness of informal caregivers' unmet needs, particularly for younger caregivers, among health care providers may improve support provision to caregivers.


Pulmonary veno-occlusive disease: antemortem diagnosis from roentgenographic and hemodynamic findings.
A 17-year-old boy with pulmonary veno-occlusive disease underwent Swan--Ganz catheterization. A normal pulmonary capillary wedge pressure was recorded in the presence of severe pulmonary arterial hypertension and roentgenographic evidence of pulmonary venous congestion. This triad of findings permitted an unequivocal diagnosis of pulmonary veno-occlusive disease, which was later confirmed at autopsy. The hemodynamics of this condition and of others included in the differential diagnosis are presented schematically.


Substrate specificity of the cdk-activating kinase (CAK) is altered upon association with TFIIH.
The transcription/DNA repair factor TFIIH consists of nine subunits, several exhibiting known functions: helicase/ATPase, kinase activity and DNA binding. Three subunits of TFIIH, cdk7, cyclin H and MAT1, form a ternary complex, cdk-activating kinase (CAK), found either on its own or as part of TFIIH. In the present work, we demonstrate that purified human CAK complex (free CAK) and recombinant CAK (rCAK) produced in insect cells exhibit a strong preference for the cyclin-dependent kinase 2 (cdk2) over a ctd oligopeptide substrate (which mimics the carboxy-terminal domain of the RNA polymerase II). In contrast, TFIIH preferentially phosphorylates the ctd as well as TFIIE alpha, but not cdk2. TFIIH was resolved into four subcomplexes: the kinase complex composed of cdk7, cyclin H and MAT1; the core TFIIH which contains XPB, p62, p52, p44 and p34; and two other subcomplexes in which XPD is found associated with either the kinase complex or with the core TFIIH. Using these fractions, we demonstrate that TFIIH lacking the CAK subcomplex completely recovers its transcriptional activity in the presence of free CAK. Furthermore, studies examining the interactions between TFIIH subunits provide evidence that CAK is integrated within TFIIH via XPB and XPD.


Fractional photothermolysis for photoaging of hands.
Laser treatment for photoaging of the hands should ideally address pigmentary alteration as well as associated skin roughness and wrinkling. Fractional resurfacing has been previously shown to effectively treat facial rhytids and dyschromia. We examined the effect of fractional resurfacing for photoaging of the hands. Ten patients (skin phototypes II to IV) with hand photodamage were randomized to receive five treatments with a 1,550-nm diode-pumped erbium fiber laser (Fraxel SR, Reliant Technologies) laser on either the right or left hand. Treatments were performed at settings of 8 to 9 mJ/microscopic treatment zone and density of 2,500 microscopic treatment zones/cm2. Subjective assessments by the patients and investigator were performed for skin roughness, wrinkling, and pigmentation using a 5-point scale. Skin biopsies were taken at baseline and at 1 and 3 months. Patient subjective assessment and physician clinical assessment at 1 and 3 months revealed a mean 51% to 75% improvement in skin pigmentation and 25% to 50% improvement in skin roughness and wrinkling. Biopsies of the skin showed increased density of dermal collagen. Patients experienced transient erythema and edema and none had scarring or other adverse effects. This was a small study. Fractional resurfacing appears to be an effective and safe treatment modality for correcting both the pigmentary and the textural aspects of photoaging of the hand.


Mitotic spindle morphogenesis: Ran on the microtubule cytoskeleton and beyond.
Assembly and disassembly of the mitotic spindle are essential for both chromosome segregation and cell division. The small G-protein Ran has emerged as an important regulator of spindle assembly. In this review, we look at the role of Ran in different aspects of spindle assembly, including its effects on microtubule assembly dynamics and microtubule organization. In addition, we examine the possibility of a spindle matrix and the role Ran might play in such a structure.


Role of the Srs2-Rad51 Interaction Domain in Crossover Control in Saccharomyces cerevisiae.
Saccharomyces cerevisiae Srs2, in addition to its well-documented antirecombination activity, has been proposed to play a role in promoting synthesis-dependent strand annealing (SDSA). Here we report the identification and characterization of an SRS2 mutant with a single amino acid substitution (srs2-F891A) that specifically affects the Srs2 pro-SDSA function. This residue is located within the Srs2-Rad51 interaction domain and embedded within a protein sequence resembling a BRC repeat motif. The srs2-F891A mutation leads to a complete loss of interaction with Rad51 as measured through yeast two-hybrid analysis and a partial loss of interaction as determined through protein pull-down assays with purified Srs2, Srs2-F891A, and Rad51 proteins. Even though previous work has shown that internal deletions of the Srs2-Rad51 interaction domain block Srs2 antirecombination activity in vitro, the Srs2-F891A mutant protein, despite its weakened interaction with Rad51, exhibits no measurable defect in antirecombination activity in vitro or in vivo Surprisingly, srs2-F891A shows a robust shift from noncrossover to crossover repair products in a plasmid-based gap repair assay, but not in an ectopic physical recombination assay. Our findings suggest that the Srs2 C-terminal Rad51 interaction domain is more complex than previously thought, containing multiple interaction sites with unique effects on Srs2 activity.


An ethnographic study of main events during hospitalisation: perceptions of nurses and patients.
To explore nurses' and patients' perceptions of care during main events of hospitalisation. Main events during hospitalisation such as admission, transfer to the operation room and preparation for discharge have been identified as times when there is significant patient-caregiver interaction. It is during these interactions that there is an opportunity for the nurse to have a positive influence on the patient's satisfaction with care. The patient's perception of care has been studied but not qualitatively. Perceptions of care from the nurses' perspectives have not been well studied. This study explored the patient's perception of care as well as the nurse's perception of the care he provided. This study used an ethnographic methodology that included participant observation and unstructured interviews. The research was conducted in two phases. First, participant observation was chosen to observe and understand nurse-patient and nurse-family behaviour. The second phase was an unstructured interview to elicit both the patient's and the nurse's views about the care experience. Two major findings were the patient and the nurse had different perceptions of the care experience and the presence of family or a support person influenced the patient's perception of care. The use of ethnography proved to be a valuable methodology for studying the interactions of patients, families and nurses. Qualitative methods such as ethnography can yield significant findings on perceptions that are unable to be gleaned by traditional quantitative methods but can serve to provide hypotheses for further study. Relevance to clinical practice. This study suggests that to maintain quality and patient satisfaction scores, hospitals will need to focus on the difference between the perceived care given and the perceived care received particularly during main events. The role of families and visitors supports positive perceptions of care.


Chondroitin sulfate proteoglycans in the developing cerebral cortex: the distribution of neurocan distinguishes forming afferent and efferent axonal pathways.
The first thalamocortical axons to arrive in the developing cerebral cortex traverse a pathway that is separate from the adjacent intracortical pathway for early efferents, suggesting that different molecular signals guide their growth. We previously demonstrated that the intracortical pathway for thalamic axons is centered on the subplate (Bicknese et al. [1994] J. Neurosci. 14:3500-3510), which is rich in chondroitin sulfate proteoglycans (CSPGs; Sheppard et al. [1991] J. Neurosci. 11:3928-3942), whereas efferent axons cross the subplate to exit in a zone containing much less CSPG. To define the molecular composition of the subplate further, we used antibodies against CSPG core proteins and chondroitin sulfate disaccharides in an immunohistochemical analysis of their distribution in the developing neocortex of the rat. Immunolabeling for neurocan, a central nervous system-specific CSPG (Rauch et al. [1992] J. Biol. Chem. 267:19537-19547), and for chondroitin 6-sulfate and unsulfated chondroitin becomes prominent in the subplate before the arrival of thalamic afferents. Immunolabeling is initially sparse in the cortical plate but appears later in maturing cortical layers. A postnatal decline in immunolabeling occurs uniformly for most proteoglycans, but, in the somatosensory cortex, labeling for neurocan, phosphacan, and chondroitin 4- and 6-sulfate declines in the centers of the whisker barrels before the walls. In contrast to neurocan, immunolabeling for other proteoglycans is either uniformly distributed (syndecan-1, N-syndecan, 5F3, phosphacan, chondroitin 4-sulfate), restricted to axons (PGM1), distributed exclusively on nonneuronal elements (2D6, NG2, and CD44), or undetectable (9.2.27, aggrecan, decorin). Thus, neurocan is a candidate molecule for delineating the intracortical pathway of thalamocortical axons and distinguishing it from that of cortical efferents.


Catch and Patch: A Pipette-Based Approach for Automating Patch Clamp That Enables Cell Selection and Fast Compound Application.
Manual patch clamp, the gold standard of electrophysiology, represents a powerful and versatile toolbox to stimulate, modulate, and record ion channel activity from membrane fragments and whole cells. The electrophysiological readout can be combined with fluorescent or optogenetic methods and allows for ultrafast solution exchanges using specialized microfluidic tools. A hallmark of manual patch clamp is the intentional selection of individual cells for recording, often an essential prerequisite to generate meaningful data. So far, available automation solutions rely on random cell usage in the closed environment of a chip and thus sacrifice much of this versatility by design. To parallelize and automate the traditional patch clamp technique while perpetuating the full versatility of the method, we developed an approach to automation, which is based on active cell handling and targeted electrode placement rather than on random processes. This is achieved through an automated pipette positioning system, which guides the tips of recording pipettes with micrometer precision to a microfluidic cell handling device. Using a patch pipette array mounted on a conventional micromanipulator, our automated patch clamp process mimics the original manual patch clamp as closely as possible, yet achieving a configuration where recordings are obtained from many patch electrodes in parallel. In addition, our implementation is extensible by design to allow the easy integration of specialized equipment such as ultrafast compound application tools. The resulting system offers fully automated patch clamp on purposely selected cells and combines high-quality gigaseal recordings with solution switching in the millisecond timescale.


Managing side effects of the novel taxane cabazitaxel in castrate-resistant prostate cancer.
Cabazitaxel, a novel taxane, was approved in June 2010 by the U.S. Food and Drug Administration for treatment of metastatic castrate-resistant prostate cancer (mCRPC) in men previously treated with docetaxel. In TROPIC (N = 755), an open-label, randomized, phase III trial, cabazitaxel (plus prednisone) was associated with improvement in median overall survival compared with mitoxantrone plus prednisone (15.1 versus 12.7 months, p < 0.0001) in patients with mCRPC who had progressed following docetaxel-based regimens. That corresponds to a 30% relative reduction in risk of death compared with the mitoxantrone regimen. In addition, significant benefit existed in median progression-free survival with cabazitaxel versus the mitoxantrone regimen (2.8 versus 1.4 months, p < 0.0001). Most common adverse events (AEs) associated with cabazitaxel were hematologic; the rates (all grade) of neutropenia, leukopenia, and anemia were greater than 90%. Diarrhea, fatigue, asthenia, and back pain were the most common grade 3 or higher nonhematologic AEs. Because expected AEs from cabazitaxel therapy can delay or even interrupt treatment, oncology nurses need to be aware of those risks and their management. This article reviews the vital role of nurses in identifying patients at high risk for AEs associated with cabazitaxel therapy and reviews strategies for prevention and management of symptoms.