Deficiency in AIM2 induces inflammation and adipogenesis in white adipose tissue leading to obesity and insulin resistance.
Absent in melanoma 2 (AIM2) is a cytosolic sensor for double-stranded DNA and a tumour suppressor. Binding of double-stranded DNA to AIM2 forms the AIM2 inflammasome, leading to activation of caspase-1 and production of IL-1β and IL-18. Although inflammasome-independent effects of AIM2 have been reported, its role in energy metabolism is unknown. We aimed to evaluate the effect of AIM2 in energy metabolism and glucose homeostasis. Male and female whole body Aim2 knockout (Aim2-/-) mice were used in the current study. Body weight, food intake, body composition, energy expenditure, fasting blood glucose levels, GTT and body temperature were measured at indicated time points. RNA sequencing was carried out on gonadal white adipose tissue (gWAT) in 14-month-old female mice. mRNA and protein levels in tissues were analysed by quantitative real-time PCR and immunoblot. Immune cell infiltration in gWAT was examined by flow cytometry. Stromal vascular fractions isolated from gWAT were used to investigate adipocyte differentiation. Male and female Aim2-/- mice were obese compared with wild-type controls from 7 weeks of age until 51 weeks of age, with increased adiposity in both subcutaneous and visceral fat depots. While there were no differences in food intake, Aim2-/- mice demonstrated decreased energy expenditure and impaired brown adipose tissue function compared with wild-type controls. Fasting glucose and insulin levels were elevated, and Aim2-/- mice were glucose intolerant on intraperitoneal GTT. RNA sequencing revealed marked upregulation of the IFN-inducible gene Ifi202b, which encodes protein 202 (p202) and elevated inflammatory signalling in gWAT of Aim2-/- mice. Increased infiltration of total and Ly6Clow monocytes was noted at 8 weeks of age in gWAT, before the onset of obesity and insulin resistance. Ifi202b knockdown blocked adipogenesis in stromal vascular fractions and reduced inflammation in bone marrow-derived macrophages, demonstrating a key role of p202 in mediating the increased adipogenesis and inflammation in Aim2-/- mice. These results demonstrate a fundamental role for AIM2 in energy metabolism, inflammation and insulin resistance. Our studies establish a novel link between the innate immunity proteins, AIM2 and p202, and metabolism.


A review of the potential forensic significance of traditional herbal medicines.
Traditional herbal substances may contain highly toxic chemicals and heavy metals, in addition to naturally occurring organic toxins. These substances may cause illness, exacerbate pre-existing ill health or result in death, particularly if taken in excess or in an unusual manner (e.g., injected rather than ingested). Lack of regulation of the content and quality of herbal medicines may result in contamination and adulteration with prescription medications. As there may be no history of the specific use of these products their contribution to death may not be fully appreciated during a standard autopsy. Even when their existence is known or suspected, it may be difficult to identify these substances on standard toxicologic screening. Herbal medicines may also be responsible for a range of symptoms and signs that may confuse the clinical presentation of cases. Given these issues the role of herbal medicines in forensic practice needs to be more clearly defined as deaths may be occurring where herbal medicines have made a significant, but as-yet unrecognized, contribution.


The fate of phenothiazine-based redox shuttles in lithium-ion batteries.
The stability and reactivity of the multiple oxidation states of aromatic compounds are critical to the performance of these species as additives and electrolytes in energy-storage applications. Both for the overcharge mitigation in ion-intercalation batteries and as electroactive species in redox flow batteries, neutral, radical-cation, and radical-anion species may be present during charging and discharging processes. Despite the wide range of compounds evaluated for both applications, the progress identifying stable materials has been slow, limited perhaps by the overall lack of analysis of the failure mechanism when a material is utilized in an energy-storage device. In this study, we examined the reactivity of phenothiazine derivatives, which have found interest as redox shuttles in lithium-ion battery applications. We explored the products of the reactions of neutral compounds in battery electrolytes and the products of radical cation formation using bulk electrolysis and coin cell cycling. Following the failure of each cell, the electrolytes were characterized to identify redox shuttle decomposition products. Based on these results, a set of decomposition mechanisms is proposed and further explored using experimental and theoretical approaches. The results highlight the necessity to fully characterize and understand the chemical degradation mechanisms of the redox species in order to develop new generations of electroactive materials.


Small molecule antibody targeting HLA class I inhibits myeloma cancer stem cells by repressing pluripotency-associated transcription factors.
Cancer stem cells have been proposed to be responsible for tumorigenesis and recurrence in various neoplastic diseases, including multiple myeloma (MM). We have previously reported that MM cells specifically express HLA class I at high levels and that single-chain Fv diabody against this molecule markedly induces MM cell death. Here we investigated the effect of a new diabody (C3B3) on cancer stem cell-like side population (SP) cells. SP fraction of MM cells highly expressed ABCG2 and exhibited resistance to chemotherapeutic agents; however, C3B3 induced cytotoxicity in both SP cells and main population (MP) cells to a similar extent. Moreover, C3B3 suppressed colony formation and tumorigenesis of SP cells in vitro and in vivo. Crosslinking of HLA class I by C3B3 mediated disruption of lipid rafts and actin aggregation, which led to inhibition of gene expression of β-catenin and pluripotency-associated transcription factors such as Sox2, Oct3/4 and Nanog. Conversely, knockdown of Sox2 and Oct3/4 mRNA reduced the proportion of SP cells, suggesting that these factors are essential in maintenance of SP fraction in MM cells. Thus, our findings reveal that immunotherapeutic approach by engineered antibodies can overcome drug resistance, and provide a new basis for development of cancer stem cell-targeted therapy.


Top-down fabricated silicon nanowire sensors for real-time chemical detection.
Silicon nanowire (SiNW) sensors have been developed by using top-down fabrication that is CMOS (complementary metal-oxide-semiconductor) compatible for resistive chemical detection with fast response and high sensitivity. Top-down fabrication by electron beam lithography and reactive ion etching of a silicon on insulator (SOI) substrate enables compatibility with the CMOS fabrication process, accurate alignment with other electrical components, flexible design of the nanowire geometry and good control of the electrical characteristics. The SiNW sensors showed a large operation range for pH detection (pH = 4-10) with an average sensitivity of (Delta R/R)/pH = 2.6%/pH and a rise time of 8 s. A small pH level difference (Delta pH = 0.2) near neutral pH conditions (pH = 7) could be resolved with the SiNW sensors. The sensor response to the presence of alkali metal ions and the long term drifting effects were also investigated.


Survival of Campylobacter jejuni inoculated into ground beef.
Ground beef was inoculated with mixed cultures of Campylobacter jejuni, and the samples were subjected to various cooking and cold-storage temperatures. When samples were heated in an oven at either 190 or 218 degrees C, approximately 10(7) cells of C. jejuni per g were inactivated (less than 30 cells per g) in less than 10 min after the ground beef reached an internal temperature of 70 degrees C. When the samples were held at -15 degrees C over 14 days of storage, the numbers of C. jejuni declined by 3 log10. When inoculated samples were stored with an equal amount of Cary-Blair diluent at 4 degrees C, no changes in viability were observed over 14 days of storage. Twenty-five times as much C. jejuni was recovered from inoculated ground beef when either 10% glycerol or 10% dimethyl sulfoxide was added to an equal amount of ground beef before freezing as was recovered from peptone-diluted ground beef. Twice as much inoculated C. jejuni was recovered from ground beef plus Cary-Blair diluent as was recovered from ground beef plus peptone diluent.


Reactivity of lymphocytes from patients with syphilis towards T. pallidum antigen in the leucocyte migration and lymphocyte transformation tests.
The reactivity of lymphocytes to Treponema pallidum antigen was studied before and after treatment in nine patients with early syphilis using a leucocyte migration test and a lymphocyte transformation test. Lymphocyte reactivity was also investigated in six patients treated for syphilis within the last 4 years, and in five untreated patients with a positive result to the T. pallidum immobilization test, but negative results to other serum tests for syphilis antibodies and without any known exposure to risk of infection by syphilis. Ten seronegative patients with different dermatological disorders served as a control group. A significant increase in lymphocyte reactivity to T. pallidum antigen was recorded in both tests in vitro after treatment. There was no difference in lymphocyte reactivity to T. pallidum antigen between the other patients studied and the control group. In early syphilis the spontaneous migration was found to be inhibited before treatment. Tuberculin skin tests were also performed and found to be suppressed in patients with primary and secondary syphilis. No difference in phytohaemagglutinin response was found between any of the groups. Plasma from patients with primary and secondary syphilis was found to change the in vitro reactivity of normal lymphocytes when stimulated with different mitogens.


Synchronized quartz crystal microbalance and nanoplasmonic sensing of biomolecular recognition reactions.
We present a method providing synchronized measurements using the two techniques: quartz crystal microbalance with dissipation (QCM-D) monitoring and localized surface plasmon resonance (LSPR). This was achieved by letting a thin gold film perforated with short-ranged ordered plasmon-active nanoholes act as one of the electrodes of a QCM-D crystal. This enabled transmission-mode optical spectroscopy to be used to temporally resolve colorimetric changes of the LSPR active substrate induced upon biomolecular binding events. The LSPR response could thus be compared with simultaneously obtained changes in resonance frequency, Deltaf, and energy dissipation, DeltaD, of the QCM-D device. Since the LSPR technique is preferentially sensitive to changes within the voids of the nanoholes, while the QCM-D technique is preferentially sensitive to reactions on the planar region between the holes, a surface chemistry providing the same binding kinetics on both gold and silica was used. This was achieved by coating the substrate with poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG), which was shown to bind in the same manner on silica and gold modified with a carboxyl-terminated thiol. In this way, the combined setup provided new information about structural changes upon PLL-g-PEG adsorption. We also demonstrate subsequent binding of NeutrAvidin and an immunoreaction utilizing biotin-modified IgG. The combined information from the synchronized measurements was also used in a new way to estimate the sensing volume of the LSPR sensor.


Gastrointestinal hormones and their role in obesity.
Pandemic obesity is the most pressing health issue of this century. The most successful treatment so far is bariatric surgery, but for various reasons, surgery cannot be applied to all patients who require treatment. Gastrointestinal hormones are likely to play a key role in the success of bariatric surgery. This article examines in detail three of these gut hormones: peptide YY, oxyntomodulin and pancreatic polypeptide, and reviews how recent developments may offer new targets for therapy. Both the free fatty acid 2 and the melanocortin 4 receptors have been discovered to regulate peptide YY and glucagon-like peptide-1 secretion, and drugs targeting these may represent new antiobesity therapies. Dual agonism of glucagon-like peptide-1 and glucagon receptors, for example with oxyntomodulin, has synergistic effects in reducing appetite and increasing energy expenditure. Plasma pancreatic polypeptide concentration correlates with visceral adiposity, and may serve as a biomarker for metabolic syndrome. Gut hormones continue to show promise on an individual basis as anti-obesity treatments, but combination therapies are needed to achieve beneficial effects comparable to bariatric surgery. Innovative pathways for stimulating native gut hormone secretion may well provide an alternative method for weight loss without necessitating the administration of gut hormone analogues via injection.


Beyond hemispheric dominance: brain regions underlying the joint lateralization of language and arithmetic to the left hemisphere.
Language and arithmetic are both lateralized to the left hemisphere in the majority of right-handed adults. Yet, does this similar lateralization reflect a single overall constraint of brain organization, such an overall "dominance" of the left hemisphere for all linguistic and symbolic operations? Is it related to the lateralization of specific cerebral subregions? Or is it merely coincidental? To shed light on this issue, we performed a "colateralization analysis" over 209 healthy subjects: We investigated whether normal variations in the degree of left hemispheric asymmetry in areas involved in sentence listening and reading are mirrored in the asymmetry of areas involved in mental arithmetic. Within the language network, a region-of-interest analysis disclosed partially dissociated patterns of lateralization, inconsistent with an overall "dominance" model. Only two of these areas presented a lateralization during sentence listening and reading which correlated strongly with the lateralization of two regions active during calculation. Specifically, the profile of asymmetry in the posterior superior temporal sulcus during sentence processing covaried with the asymmetry of calculation-induced activation in the intraparietal sulcus, and a similar colateralization linked the middle frontal gyrus with the superior posterior parietal lobule. Given recent neuroimaging results suggesting a late emergence of hemispheric asymmetries for symbolic arithmetic during childhood, we speculate that these colateralizations might constitute developmental traces of how the acquisition of linguistic symbols affects the cerebral organization of the arithmetic network.


Priority setting and health policy and systems research.
Health policy and systems research (HPSR) has been identified as critical to scaling-up interventions to achieve the millennium development goals, but research priority setting exercises often do not address HPSR well. This paper aims to (i) assess current priority setting methods and the extent to which they adequately include HPSR and (ii) draw lessons regarding how HPSR priority setting can be enhanced to promote relevant HPSR, and to strengthen developing country leadership of research agendas. Priority setting processes can be distinguished by the level at which they occur, their degree of comprehensiveness in terms of the topic addressed, the balance between technical versus interpretive approaches and the stakeholders involved. When HPSR is considered through technical, disease-driven priority setting processes it is systematically under-valued. More successful approaches for considering HPSR are typically nationally-driven, interpretive and engage a range of stakeholders. There is still a need however for better defined approaches to enable research funders to determine the relative weight to assign to disease specific research versus HPSR and other forms of cross-cutting health research. While country-level research priority setting is key, there is likely to be a continued need for the identification of global research priorities for HPSR. The paper argues that such global priorities can and should be driven by country level priorities.


3-Dimensionally Printed Affordable Nose Model: A Reliable Start in Endoscopic Training for Young Neurosurgeons.
Training neurosurgical skills is one of the most important tasks of a residency program. Techniques' complexity and pathology rarity define a long learning curve for mastering different surgical skills for which simulation on anatomic samples is extremely important. For this purpose, cadaver laboratory training is the most reliable tool. However, since access to cadaveric specimens is limited, due to costs and availability, surgical skills could be developed using inanimate models. This work aimed to develop a printable 3-dimensional model of the nasal cavity and sellar floor using an open-source downloadable file, to give residents the opportunity to improve their endoscopic surgical skills in a low-risk atmosphere with little cost. The 3D model was realized taking as a sample a real-case CT scan imaging from which the sellar floor was removed. A quail egg was placed underneath the printed model covering the sellar floor opening. Under endoscopic visualization, the "sellar floor" was drilled by each participant with the goal of sparing the egg's inner membrane. Once the task was achieved, surgeons were asked to participate in a satisfaction survey. The total cost for printing was 6.31€ (6,72$). A satisfaction survey showed technical improvement (90%), increased confidence (80%), and bringing learned skills into the operating room (70%), leading to a 100% agreement in introducing this project into residency programs. Training on affordable anatomic models represents a useful tool in technical skills improvement. We believe this model could help residents bring their technical capabilities to more sophisticated levels.


Linking DNA replication to heterochromatin silencing and epigenetic inheritance.
Chromatin is organized into distinct functional domains. During mitotic cell division, both genetic information encoded in DNA sequence and epigenetic information embedded in chromatin structure must be faithfully duplicated. The inheritance of epigenetic states is critical in maintaining the genome integrity and gene expression state. In this review, we will discuss recent progress on how proteins known to be involved in DNA replication and DNA replication-coupled nucleosome assembly impact on the inheritance and maintenance of heterochromatin, a tightly compact chromatin structure that silences gene transcription. As heterochromatin is important in regulating gene expression and maintaining genome stability, understanding how heterochromatin states are inherited during S phase of the cell cycle is of fundamental importance.


Glaucoma Suspects: The Impact of Risk Factor-Driven Review Periods on Clinical Load, Diagnoses, and Healthcare Costs.
To model the healthcare impact (clinical attendance time and financial cost) and clinical outcomes (glaucoma diagnoses) of different risk factor-driven review frequencies for glaucoma suspect patients up until the point of discharge or diagnosis. Medical records of 494 glaucoma suspects were examined to extract the clinical diagnosis. Two criteria for review periods were defined, based on contrasting stringency from established clinical guidelines: American Academy of Ophthalmology (AAO), more stringent/less frequent; and the Australian National Health and Medical Research Council (NHMRC), less stringent/more frequent. We used these data to model patient outcomes and healthcare costs using a Markov model. The less stringent/more frequent criterion resulted in more high-risk glaucoma suspects requiring more frequent review compared with the more stringent/less frequent criterion. Across the 15 Markov cycles (7.5 years), the less stringent/more frequent review criterion resulted in 6.6% more diagnoses and fewer overall clinical visits (14.7%) and reduced cost per diagnosis by 12% to 32% (P < 0.0001). The number of glaucoma diagnoses made using each criterion converged at 2.5 to 3 years. The stringency of risk assessments for glaucoma suspects impacts review periods and therefore clinical load, healthcare costs, and diagnosis rates. Using current testing methods, more frequent review periods appear advantageous for diagnostic efficiency, with both lower clinic load and lower cost up until the point of discharge or glaucoma diagnosis. A less stringent criterion for assessing the risk of developing glaucoma potentially offers a more cost-effective method for reviewing glaucoma suspects, especially within the first 2.5 years.


What factors are associated with loss of alignment after open reduction and internal fixation for tibial plateau fractures? A retrospective multicenter (TRON group) study.
Tibial plateau fractures (TPFs) are one of the most challenging intra-articular fractures to treat. Along with reconstruction of the articular surfaces, appropriate alignment of the knee joints must be obtained and maintained after open reduction and internal fixation (ORIF) for TPFs because loss of alignment (LA) is associated with worse clinical outcomes. We aimed to investigate and clarify the risk factors related to LA after ORIF for TPFs. This multicenter, retrospective cohort study used data of hospitals of the Trauma Research Group (TRON group) from January 1, 2011, to December 31, 2020. Among 293 TPFs extracted from the database, we evaluated the alignment of the articular surface to the anatomical axis of the tibia in the immediate postoperative and last follow-up radiographs. We defined a change of alignment from the immediate postoperative radiograph as LA. We evaluated the risk factors of LA using univariate and multiple logistic regression analyses. LA was observed in 27 fractures (9.2%). In multiple logistic regression analyses, preoperative articular step-off and postoperative condylar widening were statistically associated with LA (OR = 1.1, 95% CI: 1.02-1.19 and P = 0.012; OR = 1.04, 95% CI: 1.00-1.08, P = 0.045, respectively). We calculated the threshold by drawing a receiver operating characteristic curve using the final regression model. The threshold of postoperative widening was 8.2 mm. We divided the 293 TPFs into two groups according to this threshold and determined differences between the two groups using Fisher's exact test. The two groups were statistically significantly different (P = 0.00502). Preoperative articular step-off and postoperative condylar widening could be associated with LA after ORIF for TPFs. We suggest that intraoperative restoration of condylar widening is important for the prevention of malalignment following ORIF for TPF.


Strong antimicrobial activity of xanthohumol and other derivatives from hops (Humulus lupulus L.) on gut anaerobic bacteria.
Anaerobic bacteria, such as Bacteroides fragilis or Clostridium perfringens, are part of indigenous human flora. However, Clostridium difficile represents also an important causative agent of nosocomial infectious antibiotic-associated diarrhoea. Treatment of C. difficile infection is problematic, making it imperative to search for new compounds with antimicrobial properties. Hops (Humulus lupulus L.) contain substances with antibacterial properties. We tested antimicrobial activity of purified hop constituents humulone, lupulone and xanthohumol against anaerobic bacteria. The antimicrobial activity was established against B. fragilis, C. perfringens and C. difficile strains according to standard testing protocols (CLSI, EUCAST), and the minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBC) were calculated. All C. difficile strains were toxigenic and clinically relevant, as they were isolated from patients with diarrhoea. Strongest antimicrobial effects were observed with xanthohumol showing MIC and MBC values of 15-107 μg/mL, which are close to those of conventional antibiotics in the strains of bacteria with increased resistance. Slightly higher MIC and MBC values were obtained with lupulone followed by higher values of humulone. Our study, thus, shows a potential of purified hop compounds, especially xanthohumol, as alternatives for treatment of infections caused by select anaerobic bacteria, namely nosocomial diarrhoea caused by resistant strains.


[Intraocular lenses with blue light filter].
To evaluate if a SN60AT <<Natural>> IOL (Alcon) with IMPRUV filter (Alcon) offers any advantage in patients having cataract surgery, as compared with Acrysof SA60 IOL (Alcon). This was a prospective analytical study. We studied sixty-four eyes from 32 patients with cataracts. After a random assignment, phacoemulsification with the implant was performed using the Acrysof SN60AT tinted lens in one eye and Acrysof SA60 in the other. A variety of variables was subsequently assessed: contrast sensitivity with a CSV 1000E test, colour perception through a Farnsworth 25 test and subjective measures by a patient survey. Differences between the groups were assessed statistically. The contrast sensitivity results were obtained for 3, 6, 12 and 18 cycles per degree (cpd) frequencies under scotopic conditions, with better results obtained following the use of SN60AT: 0.75 (CSV 1000E test units) for 3 cpd, 0.86 for 6 cpd, 0.54 for 12 cpd and 0.35 for 18 cpd. The differences were statistically significant between the two groups for 3 cpd and 6 cpd. A total of 62.5% of patients using a tinted lens and 50% with SA60 achieved good results in the Farnsworth test; 12.5% and 21% had fair results and 25% and 28.1% respectively had poor results. We obtained an improvement in favour of the tinted lens for all frequencies, with this being statistically significant at lower frequencies (3 and 6 cpd). No significant differences in the Farnsworth test performance or in the patient survey were found. The tinting provided by the IMPRUV filter brings an increase in contrast sensitivity without affecting colour perception.


Nutritional assessment of the child with cancer.
Children with cancer are in need of nutritional support to combat the catabolic effects of malignant tumors and debilitation from surgical or other treatment and complicating infections and to maintain their immune system and other host defenses to combat cancer and infection. Also, growth must be supported in the child. Nutritional assessment should be an integral part of the care of the hospitalized and the ambulatory child. The main components of nutritional assessment are described: medical history; psychocultural history; dietary assessment; clinical examination for signs of deficiency; anthropometry; and biochemical and hematological assessment. Parameters of cell-mediated immunity, often depressed by malnutrition, are also part of the assessment. Appropriate reference data for children are included. Assessment is the first step in nutritional support.


Smooth, volume-accurate material interface reconstruction.
A new material interface reconstruction method for volume fraction data is presented. Our method is comprised of two components: first, we generate initial interface topology; then, using a combination of smoothing and volumetric forces within an active interface model, we iteratively transform the initial material interfaces into high-quality surfaces that accurately approximate the problem's volume fractions. Unlike all previous work, our new method produces material interfaces that are smooth, continuous across cell boundaries, and segment cells into regions with proper volume. These properties are critical during visualization and analysis. Generating high-quality mesh representations of material interfaces is required for accurate calculations of interface statistics, and dramatically increases the utility of material boundary visualizations.


Failure of perchlorate to inhibit Tc-99m isonitrile binding by the thyroid during myocardial perfusion studies.
The thyroid gland receives an average radiation dose of 3 rads during two Tc-99m isonitrile (MIBI) myocardial perfusion studies, if 20 mCi is administered both at rest and at peak exercise. In patients with coronary artery disease, multiple myocardial perfusion studies may be required, resulting in a high level of thyroid radiation. We attempted to reduce this radiation exposure by blocking thyroidal Tc-99m MIBI uptake with oral potassium perchlorate (KCIO4). Fourteen normal subjects received 0.6g to 0.8g KCIO4 20-25 minutes before tracer injection. Subjects who received KCIO4 at rest (n = 11) did not receive KCIO4 at their stress study, and vice versa (n = 3). Thyroid uptake values were obtained with a thyroid probe 20 minutes after injection for both rest and stress studies and were corrected for saturation effects. There was no difference between fractional thyroid uptake values with and without preceding perchlorate administration: 1.9 +/- 0.5% and 1.8 +/- 0.3% (mean +/- SD), respectively. Failure to block Tc-99m MIBI uptake after intravenous (IV) injection is probably due to high thyroidal blood flow and nonspecific tracer accumulation. The concentration of this radioisotope in adjacent muscles also contributes to the high thyroid radiation dose. In summary, administration of KCIO4 before Tc-99m MIBI studies does not reduce the thyroidal radiation dose or uptake of this tracer, suggesting that thyroidal uptake of this tracer is not mediated by the iodine trapping mechanism.


GMP-140 mediates adhesion of stimulated platelets to neutrophils.
In vivo, platelets associate with neutrophils at sites of hemorrhage or inflammation. In vitro, stimulated platelets bind to neutrophils in a Ca2(+)-dependent manner. GMP-140, an integral membrane glycoprotein found in secretory granules of platelets and endothelium, is rapidly translocated to the cell surface after cellular activation. It shares sequence similarity with two leukocyte adhesion molecules, ELAM-1 and a lymphocyte homing receptor. We have recently shown that neutrophils bind to purified GMP-140 in a Ca2(+)-dependent fashion, and that GMP-140 participates in adhesion of neutrophils to activated endothelium. In this study we demonstrate that GMP-140 also mediates adhesion of neutrophils to stimulated platelets. Fixed thrombin-activated human platelets, but not unstimulated platelets, formed rosettes around neutrophils in the presence of Ca2+. The binding of platelets to neutrophils was inhibited by a monoclonal antibody to GMP-140 and by purified GMP-140. By promoting close cell-cell contact, GMP-140 may recruit both platelets and neutrophils to sites of tissue injury as well as modulate the function of each cell type by the other.


Endothelial progenitor cells in diabetic retinopathy.
Diabetic retinopathy (DR) is a leading cause of visual impairment worldwide. Patients with DR may irreversibly lose sight as a result of the development of diabetic macular edema (DME) and/or proliferative diabetic retinopathy (PDR); retinal blood vessel dysfunction and degeneration plays an essential role in their pathogenesis. Although new treatments have been recently introduced for DME, including intravitreal vascular endothelial growth factor inhibitors (anti-VEGFs) and steroids, a high proportion of patients (~40-50%) do not respond to these therapies. Furthermore, for people with PDR, laser photocoagulation remains a mainstay therapy despite this being an inherently destructive procedure. Endothelial progenitor cells (EPCs) are a low-frequency population of circulating cells known to be recruited to sites of vessel damage and tissue ischemia where they promote vascular healing and re-perfusion. A growing body of evidence suggests that the number and function of EPCs are altered in patients with varying degrees of diabetes duration, metabolic control, and in the presence or absence of DR. Although there are no clear-cut outcomes from these clinical studies, there is mounting evidence that some EPC sub-types may be involved in the pathogenesis of DR and may also serve as biomarkers for disease progression and stratification. Moreover, some EPC sub-types have considerable potential as therapeutic modalities for DME and PDR in the context of cell therapy. This study presents basic clinical concepts of DR and combines this with a general insight on EPCs and their relation to future directions in understanding and treating this important diabetic complication.


Role of the Phagocyte in Host-Parasite Interactions XXVII. Myeloperoxidase-H(2)O(2)-Cl-Mediated Aldehyde Formation and Its Relationship to Antimicrobial Activity.
Evidence is presented which suggests that the mechanism of action of the myeloperoxidase-H(2)O(2)-Cl(-) antimicrobial system in the phagocyte is by the formation of aldehydes. Aldehyde production resulting from myeloperoxidase-mediated decarboxylation and deamination of alanine was quantitated with 20,000-g granules from guinea pig polymorphonuclear leukocytes serving as the enzyme. Equimolar quantities of acetaldehyde and CO(2) were obtained. There was an absolute requirement for both H(2)O(2) and Cl(-) for decarboxylation by the myeloperoxidase-containing granules. The myeloperoxidase-H(2)O(2)-Cl(-) system decarboxylated both d- or l-alanine equally and had a pH optimum of 5.3. Decarboxylation of l-alanine by intact guinea pig polymorphonuclear leukocytes was increased 2.5-fold by phagocytosis. Guaiacol peroxidation by the granules was inhibited 90% in the presence of Cl(-) at acid pH. Under these conditions, decarboxylation and deamination of amino acids by myeloperoxidase were significantly stimulated, resulting in aldehyde production. Taurine, a competitive inhibitor of amino acid decarboxylation, inhibited bactericidal activity of the myeloperoxidase-H(2)O(2)-Cl(-) system but had no effect on the myeloperoxidase-H(2)O(2)-I(-) bactericidal system. Since the myeloperoxidase-H(2)O(2)-I(-) system does not participate in amino acid decarboxylation, its mechanism of antimicrobial action appears to be different from that found with Cl(-).


Coupled Effects of Spreading Solvent Molecules and Electrostatic Repulsions on the Behavior of PS-b-PAA Monolayers at the Air-Water Interface.
We describe the surface behavior of PS-b-PAA monolayers at the air/water interface using N,N-dimethyformamide (DMF) as spreading solvent. At low pH, when the PAA blocks are neutral, the surface pressure versus molecular area isotherm shows a pseudoplateau associated with the presence of remaining spreading solvent molecules in the monolayer, as we described in a former study (Guennouni et al., Langmuir, 2016). We show here that the width of the plateau decreases when increasing pH up to its complete disappearance at high pH, when PAA blocks are fully charged, although two regimes of compressibilities on the isotherm still exist. A refined structural study at pH 9 combining specular neutron reflectivity (SNR), grazing-incidence small-angle X-ray scattering (GISAXS), and atomic force microscopy (AFM) in liquid measurements shows that (i) PAA blocks are stretched in solution, as expected from polyelectrolyte brushes in the osmotic regime; (ii) the system undergoes a spinodal decomposition during deposit at the air/water interface in the presence of DMF. Upon compression, the Qxy* position of the peak associated with the spinodal structure remains almost constant but its intensity evolves strongly and passes through a maximum at intermediate pressures. This reveals two operating processes in the system: strong electrostatic repulsions between chains that prevent in-plane reorganizations and force such reorganizations to occur from the surface to the volume and progressive expulsion of the DMF molecules from the monolayer. These processes have antagonist effects on the intensity of the peak: the increase of the repulsions makes it more pronounced, whereas the expulsion of solvent makes it vanish due to the loss of contrast.


Research priorities in Maternal, Newborn, & Child Health & Nutrition for India: An Indian Council of Medical Research-INCLEN Initiative.
In India, research prioritization in Maternal, Newborn, and Child Health and Nutrition (MNCHN) themes has traditionally involved only a handful of experts mostly from major cities. The Indian Council of Medical Research (ICMR)-INCLEN collaboration undertook a nationwide exercise engaging faculty from 256 institutions to identify top research priorities in the MNCHN themes for 2016-2025. The Child Health and Nutrition Research Initiative method of priority setting was adapted. The context of the exercise was defined by a National Steering Group (NSG) and guided by four Thematic Research Subcommittees. Research ideas were pooled from 498 experts located in different parts of India, iteratively consolidated into research options, scored by 893 experts against five pre-defined criteria (answerability, relevance, equity, investment and innovation) and weighed by a larger reference group. Ranked lists of priorities were generated for each of the four themes at national and three subnational (regional) levels [Empowered Action Group & North-Eastern States, Southern and Western States, & Northern States (including West Bengal)]. Research priorities differed between regions and from overall national priorities. Delivery domain of research which included implementation research constituted about 70 per cent of the top ten research options under all four themes. The results were endorsed in the NSG meeting. There was unanimity that the research priorities should be considered by different governmental and non-governmental agencies for investment with prioritization on implementation research and issues cutting across themes.


Modulation of oxidative stress by Chlorella vulgaris in streptozotocin (STZ) induced diabetic Sprague-Dawley rats.
Chlorella vulgaris (CV), a fresh water alga has been reported to have hypoglycemic effects. However, antioxidant and anti-inflammatory effects of CV in diabetic animals have not been investigated to date. The aim of the present study was to investigate the role of CV in inflammation and oxidative damage in STZ-induced diabetic rats. Male Sprague-Dawley rats (300 - 400g) were divided into 4 groups: control, CV, STZ-induced diabetic rats, and STZ rats treated with CV (150mg/kg body wt). Blood samples were drawn from orbital sinus at 1 and 4 weeks for determination of oxidative cellular damage (DNA damage and lipid peroxidation [malondialdehyde, MDA]), inflammation (tumour necrosis factor alpha, TNF-α) and antioxidant status (catalase, CAT, and superoxide dismutase, SOD). CV did not have any effects on glucose levels in diabetic rats, over the 4 weeks of treatment. However, it reduced significantly DNA damage and blood MDA levels in STZ-induced diabetic rats compared to the control group. Plasma levels of TNF-α however did not show any significant changes in STZ-induced diabetic rats fed with CV. Antioxidant enzyme SOD showed no significant changes in all groups but CAT activity was reduced in STZ-induced diabetic rats compared to the control. CV did not have hypoglycaemic effect but it has a protective role in STZ-induced diabetic rats by reducing oxidative DNA damage, and lipid peroxidation.


CyberKnife reference dosimetry: An assessment of the impact of evolving recommendations on correction factors and measured dose.
Specialized treatment machines such as the CyberKnife, TomoTherapy, or the GammaKnife, utilize flattening filter free (FFF) photon beams and may not be able to generate a 10 cm x 10 cm reference field. A new Code of Practice has recently been published (IAEA TRS483) to give recommendations for these machines. Additionally, some standard laboratories provide measured beam quality correction factors for the user's reference chamber, which can be used instead of the published tabulated beam quality correction factors. The purpose of this study was first to assess how different recommendations, as outlined below, affect the reference dosimetry at the CyberKnife, and second, to assess the impact of using measured rather than tabulated beam quality correction factors on measured dose. Following recommendations in TRS398, three field chambers (IBA CC04, Exradin A19, and Exradin A12S) were cross-calibrated with a user's reference chamber (IBA FC65-G), which was calibrated in a Cobalt-60 (Co-60) beam by a primary standards laboratory. The chamber response was corrected for influence quantities such as temperature, pressure, ion recombination, polarity, and beam quality. Additionally, correction factors for volume averaging and differences due the FFF beam spectrum were determined for the CyberKnife beam. Three different methods were utilized - TRS398; Intermediate (i.e. TRS398 with additional published recommendations); and TRS483. The measurements were undertaken in a 10 cm × 10 cm field defined by jaws for a uniform flattened (WFF) 6 MV photon beam of a Varian TrueBeam linear accelerator (linac) with a source to detector distance (SDD) of 100 cm, and in a 60 mm diameter circular field for a 6 MV flattening filter free (FFF) Accuray CyberKnife beam with SDD of 80 cm. All measurement was performed at 10 cm deep in a full scatter phantom as defined in TRS398. Differences between the three methods in volume averaging correction factors ranged from 0.01% to 0.45% depending on the chamber assessed. As expected, an increased chamber length leads to a larger correction factor. The differences in beam spectrum correction factors range from 0.09% to 0.3%. Negligible differences in beam quality correction factors were observed; however, differences up to 1% were found between measured and tabulated values. Differences in cross-calibrated chamber calibration coefficients range from 0.05% to 0.51% depending on the chamber assessed. Differences in measured dose are up to 0.87% between Method TRS398 and Intermediate, again chamber dependent, and 0.28% between Method Intermediate and TRS483. Using chambers cross-calibrated in the linac beam can lead to differences in measured dose per Monitor Unit (MU) in the CyberKnife beam of approximately 0.5% between chambers. Using Method Intermediate vs using recommendations given in TRS483 led to a difference of 0.28% in measured dose per MU, which is due to differences in volume averaging and beam spectrum correction factors. Using TRS483 is recommended as the cross-calibration is done in the CyberKnife beam and accounts for its specific reference conditions. It will also ensure consistency between different centers. The measured beam quality correction factors agree within the uncertainties with the tabulated values.


Lyme Disease Diagnosed by Alternative Methods: A Phenotype Similar to That of Chronic Fatigue Syndrome.
A subset of patients reporting a diagnosis of Lyme disease can be described as having alternatively diagnosed chronic Lyme syndrome (ADCLS), in which diagnosis is based on laboratory results from a nonreference Lyme specialty laboratory using in-house criteria. Patients with ADCLS report symptoms similar to those reported by patients with chronic fatigue syndrome (CFS). We performed a case-control study comparing patients with ADCLS and CFS to each other and to both healthy controls and controls with systemic lupus erythematosus (SLE). Subjects completed a history, physical exam, screening laboratory tests, 7 functional scales, reference serology for Lyme disease using Centers for Disease Control and Prevention criteria, reference serology for other tick-associated pathogens, and cytokine expression studies. The study enrolled 13 patients with ADCLS (12 of whom were diagnosed by 1 alternative US laboratory), 25 patients with CFS, 25 matched healthy controls, and 11 SLE controls. Baseline clinical data and functional scales indicate significant disability among ADCLS and CFS patients and many important differences between these groups and controls, but no significant differences between each other. No ADCLS patient was confirmed as having positive Lyme serology by reference laboratory testing, and there was no difference in distribution of positive serology for other tick-transmitted pathogens or cytokine expression across the groups. In British Columbia, a setting with low Lyme disease incidence, ADCLS patients have a similar phenotype to that of CFS patients. Disagreement between alternative and reference laboratory Lyme testing results in this setting is most likely explained by false-positive results from the alternative laboratory.


Nest boxes do not cause a shift in bat community composition in an urbanised landscape.
Nest boxes are often used to provide supplementary roosts for cavity-dependent wildlife, but little is known about if they influence faunal community composition. Long-term monitoring of bat boxes in south-eastern Australia indicated that their use was dominated by one generalist species (Chalinolobus gouldii), causing concern that installing bat boxes could cause a shift toward less diverse bat communities. To test this, we conducted a large-scale before-after control-impact experiment at 18 sites, over five years. Sites were either: (1) those with existing bat boxes, (2) those where boxes were added during the study, or (3) controls without boxes. We used echolocation call data from 9035 bat detector nights to compare community composition, diversity, and species' relative activity between the sites. Chalinolobus gouldii continued to dominate the use of existing boxes, but we found little difference in community composition between sites based on the presence, absence, or addition of boxes. Our study is the first to explore the influence installing artificial hollows has on localized faunal assemblages over spatio-temporal scales relevant to management. We conclude that there is cause for optimism that bat boxes might not have perverse outcomes on local community composition in the short- to medium-term, as we had feared.


Augmentation of immune responses to HIV-1 and simian immunodeficiency virus DNA vaccines by IL-2/Ig plasmid administration in rhesus monkeys.
The potential utility of plasmid DNA as an HIV-1 vaccination modality currently is an area of active investigation. However, recent studies have raised doubts as to whether plasmid DNA alone will elicit immune responses of sufficient magnitude to protect against pathogenic AIDS virus challenges. We therefore investigated whether DNA vaccine-elicited immune responses in rhesus monkeys could be augmented by using either an IL-2/Ig fusion protein or a plasmid expressing IL-2/Ig. Sixteen monkeys, divided into four experimental groups, were immunized with (i) sham plasmid, (ii) HIV-1 Env 89.6P and simian immunodeficiency virus mac239 Gag DNA vaccines alone, (iii) these DNA vaccines and IL-2/Ig protein, or (iv) these DNA vaccines and IL-2/Ig plasmid. The administration of both IL-2/Ig protein and IL-2/Ig plasmid induced a significant and sustained in vivo activation of peripheral T cells in the vaccinated monkeys. The monkeys that received IL-2/Ig plasmid generated 30-fold higher Env-specific antibody titers and 5-fold higher Gag-specific, tetramer-positive CD8+ T cell levels than the monkeys receiving the DNA vaccines alone. IL-2/Ig protein also augmented the vaccine-elicited immune responses, but less effectively than IL-2/Ig plasmid. Augmentation of the immune responses by IL-2/Ig was evident after the primary immunization and increased with subsequent boost immunizations. These results demonstrate that the administration of IL-2/Ig plasmid can substantially augment vaccine-elicited humoral and cellular immune responses in higher primates.


The composition of cartilage proteoglycans. An investigation using high- and low-inonic-strength extraction procedures.
1. A proteoglycan fraction (the proteoglycan subunit fraction) was prepared from extracts, with 0.15m-KCl (low-ionic-strength) and 0.5m-LaCl(3), 2.0m-CaCl(2) and 4.0m-guanidinium chloride (high-ionic-strength), of bovine nasal cartilage by equilibrium-density-gradient centrifugation, essentially as described by Hascall & Sajdera (1969). 2. The use of different centrifugation times showed that near-equilibrium conditions were reached by 48h for the fractions prepared from the high-ionic-strength extracts. The fraction isolated from the low-ionic-strength extract required a longer centrifugation time to reach equilibrium conditions. 3. The composition of the proteoglycan fractions from the various extracts was compared by analyses of their carbohydrate and amino acid contents. Difference indices were calculated from the amino acid analysis to compare the degree of compositional relationship between the protein components of the proteoglycans. 4. Small compositional differences were found between the proteoglycans isolated from the various high-ionic-strength extracts. The protein content of the fractions from the CaCl(2) extract and the guanidinium chloride extract showed the greatest difference in this respect, although their amino acid analysis was similar. 5. The proteoglycan fraction isolated from the low-ionic-strength extract shows marked differences in composition from the fractions isolated from the high-ionic-strength extracts. Its protein and glucosamine contents were lower whereas its hexuronic acid and galactosamine contents were higher than those of the latter. It also exhibits major differences in its amino acid composition. The glucosamine:galactosamine ratio of the fraction from the low-ionic-strength extract indicates that it may be an almost exclusively chondroitin sulphate-proteoglycan. Its analysis correlates closely with that of a low-molecular-weight proteoglycan isolated from pig laryngeal cartilage by Tsiganos & Muir (1969). 6. The proteoglycan fractions from both the low- and high-ionic-strength extracts migrate as a single band in zone electrophoresis carried out in a sucrose-density gradient at both pH3.0 and pH7.0, although each showed evidence of band widening during the electrophoresis. All the proteoglycan fractions migrated with the same electrophoretic mobility at pH3.0, irrespective of the differences in composition between them. 7. The differences between the proteoglycans from the low- and high-ionic-strength extracts are discussed and the view is advanced that they may be due to association between predominantly chondroitin sulphate-proteoglycans and a keratan sulphate-enriched proteoglycan species.


Visual working memories are abstractions of percepts.
Pioneering studies demonstrating that the contents of visual working memory (WM) can be decoded from the patterns of multivoxel activity in early visual cortex transformed not only how we study WM, but theories of how memories are stored. For instance, the ability to decode the orientation of memorized gratings is hypothesized to depend on the recruitment of the same neural encoding machinery used for perceiving orientations. However, decoding evidence cannot be used to test the so-called sensory recruitment hypothesis without understanding the underlying nature of what is being decoded. Although unknown during WM, during perception decoding the orientation of gratings does not simply depend on activities of orientation tuned neurons. Rather, it depends on complex interactions between the orientation of the grating, the aperture edges, and the topographic structure of the visual map. Here, our goals are to 1) test how these aperture biases described during perception may affect WM decoding, and 2) leverage carefully manipulated visual stimulus properties of gratings to test how sensory-like are WM codes. For memoranda, we used gratings multiplied by radial and angular modulators to generate orthogonal aperture biases despite having identical orientations. Therefore, if WM representations are simply maintained sensory representations, they would have similar aperture biases. If they are abstractions of sensory features, they would be unbiased and the modulator would have no effect on orientation decoding. Results indicated that fMRI patterns of delay period activity while maintaining the orientation of a grating with one modulator (eg, radial) were interchangeable with patterns while maintaining a grating with the other modulator (eg, angular). We found significant cross-classification in visual and parietal cortex, suggesting that WM representations are insensitive to aperture biases during perception. Then, we visualized memory abstractions of stimuli using a population receptive field model of the visual field maps. Regardless of aperture biases, WM representations of both modulated gratings were recoded into a single oriented line. These results provide strong evidence that visual WM representations are abstractions of percepts, immune to perceptual aperture biases, and compel revisions of WM theory.


Demand-side challenges to increase sales of new maize hybrids in Kenya.
In Eastern and Southern Africa, as the public sector has retreated from maize seed production and the private sector has emerged to fill the resulting void, a key issue for researchers, governments and private donors has been the capacity of privately owned, typically small scale, seed businesses to effectively produce and distribute hybrid seed. Roughly two decades on, research and development programming continues to focus on supply side issues in supporting the maize seed industry to bring new varieties to farmers. Motivated by thinking on agri-food value chains, this article explores the potential for achieving a stronger demand orientation in programming to support the maize seed industry, a requirement for varietal turnover. In 2019 data were collected in Kenya from i) 80 agro-dealers on their relations with seed businesses and their marketing of maize seed, ii) 466 farmers on their seed choice and engagement with agro-dealers and iii) 8 seed companies on their distribution and sales strategies. Results confirmed the overarching supply-push orientation of the industry, characterized by limited innovation and risk taking, weak collaboration between actors, low margins for retailers, and limited investments in seed marketing. Farmers showed weak appetite for acquiring new seed products, preferring instead to purchase seeds that they knew from experience. Better strategies for building seed value chains will require deeper insights on stakeholders needs and strategies, to include the capacity of seed businesses and retailers to innovate in business management and marketing.


Antibiotic Use and Fatal Outcomes among Critically Ill Patients with COVID-19 in Tacna, Peru.
Overuse of antibiotics during the Coronavirus Disease 2019 (COVID-19) pandemic could increase the selection of extensively resistant bacteria (XDR). However, it is unknown what impact they could have on the evolution of patients, particularly critically ill patients. This study aimed to evaluate the characteristics and impact of ICU-acquired infections in patients with COVID-19. A retrospective cohort study was conducted, evaluating all patients with critical COVID-19 admitted to the intensive care unit (ICU) of a hospital in Southern Peru from 28 March 2020 to 1 March 2021. Of the 124 patients evaluated, 50 (40.32%) developed a healthcare-associated infection (HAI), which occurred at a median of 8 days (IQR 6-17) after ICU admission. The proportion of patients with HAI that required ceftriaxone was significantly higher; the same was true for the use of dexamethasone. Forty bacteria isolations (80%) were classified as XDR to antibiotics, with the most common organisms being Acinetobacter baumannii (54%) and Pseudomonas aeruginosa (22%); 33% (41/124) died at the ICU during the follow-up. In the adjusted analysis, healthcare-associated infection was associated with an increased risk of mortality (aHR= 2.7; 95% CI: 1.33-5.60) and of developing acute renal failure (aRR = 3.1; 95% CI: 1.42-6.72). The incidence of healthcare infection mainly by XDR pathogens is high in critically ill patients with COVID-19 and is associated with an increased risk of complications or death.


Muscle subcellular localization and recruitment by insulin of glucose transporters and Na+-K+-ATPase subunits in transgenic mice overexpressing the GLUT4 glucose transporter.
Insulin-stimulated glucose uptake in skeletal muscle is mediated through the GLUT4 glucose transporter. Transgenic (TG) mice overexpressing human GLUT4 in skeletal muscle show an increased ability to handle a glucose load. Here, the participation of the overexpressed GLUT4 in the response to insulin was examined. In TG mouse muscle, the GLUT4 protein content was 10-fold higher in crude membrane (CM), sevenfold higher in internal membrane (IM), and 15-fold higher in a plasma membrane (PM)-rich fraction, relative to non-TG littermates. This suggested partial saturation of the normal sorting mechanisms. The distribution and abundance of the GLUT1 glucose transporter was not affected. Insulin injection (4.3 U/kg body wt) increased GLUT4 in the PM-rich fraction; the increase was threefold higher in TG than in non-TG mice. Insulin decreased the GLUT4 content of the IM in both animal groups and of a second, heavier intracellular membrane fraction only in TG mice. The net content of Na+-K+-pump subunits was 40-65% lower in CM from TG compared with non-TG littermates. In spite of this, insulin caused a three- to sixfold higher translocation of the alpha2 and beta1 subunits of the Na+-K+-pump in TG compared with non-TG animals. The results suggest that overexpression of GLUT4 confers to the muscle increased ability to translocate subunits of the Na+-K+-pump either as a direct consequence of the recruitment of glucose transporters or as an adaptation to the more demanding metabolic state.


[The diseases of circulatory system in employees of railway transport].
The article presents the epidemiologic and medical social aspects of diseases of circulatory system in employees of railway transport in 2000-2010 exemplified by Privolzhskiy railroad. The established tendencies in prevalence of pathology of cardio-vascular system in railroad workers makes the issues of practical implementation of priority of prevention in the organization of medical care to this group of patients to come to foreground. The main directions for complex prevention of diseases of circulatory system in employees of railway transport are presented.


Current understanding of multi-species biofilms.
Direct observation of a wide range of natural microorganisms has revealed the fact that the majority of microbes persist as surface-attached communities surrounded by matrix materials, called biofilms. Biofilms can be formed by a single bacterial strain. However, most natural biofilms are actually formed by multiple bacterial species. Conventional methods for bacterial cleaning, such as applications of antibiotics and/or disinfectants are often ineffective for biofilm populations due to their special physiology and physical matrix barrier. It has been estimated that billions of dollars are spent every year worldwide to deal with damage to equipment, contaminations of products, energy losses, and infections in human beings resulted from microbial biofilms. Microorganisms compete, cooperate, and communicate with each other in multi-species biofilms. Understanding the mechanisms of multi-species biofilm formation will facilitate the development of methods for combating bacterial biofilms in clinical, environmental, industrial, and agricultural areas. The most recent advances in the understanding of multi-species biofilms are summarized and discussed in the review.


The Hog1 MAP kinase controls respiratory metabolism in the fungal pathogen Candida albicans.
Signal transduction pathways mediated by mitogen-activated protein kinases (MAPKs) play crucial roles in eukaryotic cells. In the pathogenic fungus Candida albicans the HOG MAPK pathway regulates the response to external stresses (osmotic and oxidative among others) and is involved in morphogenesis and virulence. We show here that the lack of the Hog1 MAPK increases the sensitivity of this fungus to inhibitors of the respiratory chain. hog1 mutants also show an enhanced basal respiratory rate compared to parental strains, and higher levels of intracellular reactive oxygen species despite an increased expression of detoxifying enzymes. We also demonstrate that although oxidative phosphorylation is essentially unaffected, hog1 mutants have an altered mitochondrial membrane potential. Data indicate that hog1-defective mutants are more dependent on mitochondrial ATP synthesis, probably due to an increased cellular ATP demand. Our results therefore link a MAPK pathway with respiratory metabolism in pathogenic fungi.


Pharmacophore-Based Virtual Screening, Quantum Mechanics Calculations, and Molecular Dynamics Simulation Approaches Identified Potential Natural Antiviral Drug Candidates against MERS-CoV S1-NTD.
Middle East respiratory syndrome coronavirus (MERS-CoV) is a highly infectious zoonotic virus first reported into the human population in September 2012 on the Arabian Peninsula. The virus causes severe and often lethal respiratory illness in humans with an unusually high fatality rate. The N-terminal domain (NTD) of receptor-binding S1 subunit of coronavirus spike (S) proteins can recognize a variety of host protein and mediates entry into human host cells. Blocking the entry by targeting the S1-NTD of the virus can facilitate the development of effective antiviral drug candidates against the pathogen. Therefore, the study has been designed to identify effective antiviral drug candidates against the MERS-CoV by targeting S1-NTD. Initially, a structure-based pharmacophore model (SBPM) to the active site (AS) cavity of the S1-NTD has been generated, followed by pharmacophore-based virtual screening of 11,295 natural compounds. Hits generated through the pharmacophore-based virtual screening have re-ranked by molecular docking and further evaluated through the ADMET properties. The compounds with the best ADME and toxicity properties have been retrieved, and a quantum mechanical (QM) based density-functional theory (DFT) has been performed to optimize the geometry of the selected compounds. Three optimized natural compounds, namely Taiwanhomoflavone B (Amb23604132), 2,3-Dihydrohinokiflavone (Amb23604659), and Sophoricoside (Amb1153724), have exhibited substantial docking energy >-9.00 kcal/mol, where analysis of frontier molecular orbital (FMO) theory found the low chemical reactivity correspondence to the bioactivity of the compounds. Molecular dynamics (MD) simulation confirmed the stability of the selected natural compound to the binding site of the protein. Additionally, molecular mechanics generalized born surface area (MM/GBSA) predicted the good value of binding free energies (ΔG bind) of the compounds to the desired protein. Convincingly, all the results support the potentiality of the selected compounds as natural antiviral candidates against the MERS-CoV S1-NTD.


Development and characterization of murine models of medulloblastoma extraneural growth in bone.
Medulloblastoma is a malignant pediatric brain neoplasm with an unusual predilection for metastasis to the skeleton. The objective of this study was to generate and characterize murine models of medulloblastoma extraneural growth in bone as 'discovery tools' for the identification of unrecognized signal transduction pathways and factors driving metastatic bone disease. To this end, the human Daoy and D283 medulloblastoma cell lines were inoculated into the intratibial medullary space of athymic nude mice. Daoy injected mice developed a primarily osteolytic radiographic and histological phenotype. In contrast, both areas of osteolytic and osteosclerotic activity were evident in D283 inoculated bones. D283 and Daoy cell conditioned media increased in vitro osteoblast differentiation and is consistent with the enhanced bone turnover characteristic of bone metastases. Daoy cells also significantly increased bone marrow osteoclast formation, consistent with the robust in vivo osteolytic phenotype. A survey of secreted factors implicated in bone metastasis and expressed by D283 and Daoy was performed. High expression of the bone-homing factor, CXCR4, was observed in both Daoy and D283 tissues. Consistent with the skeletal phenotypes, Daoy cells, while secreting the osteoblastic factor ET-1, abundantly produced the osteolytic factors RANKL, PTHrP and TNFα. D283 cells produced high levels of both RANKL and ET-1. These newly described animal models of medulloblastoma bone metastasis are expected to serve as platforms to aid in the elucidation of novel bone metastasis signaling cascades and to test therapeutics that target both medulloblastoma metastasis and the primary tumor.


Severe imported falciparum malaria among adults requiring intensive care: a retrospective study at the hospital for tropical diseases, London.
Malaria is the commonest imported infection in the UK. Malaria requiring ICU admission has a reported mortality of up to 25%. The relationship between ethnicity, immunity, and risk of malaria is complex. The Malaria Score for Adults (MSA) and Coma Acidosis Malaria (CAM) score have recently been proposed to risk stratify patients with malaria. Retrospective study of patients with WHO severe falciparum malaria admitted to ICU at the Hospital for Tropical Diseases, London, UK. The relationship between clinical variables and risk of death or a prolonged ICU stay were examined with logistic regression. The predictive value of the MSA and CAM score were calculated. 124 patients were included. Cerebral malaria and acute kidney injury occurred earlier (median day 1) than acute respiratory distress syndrome (median day 3). Six patients had community acquired bacterial co-infection. Eight patients were co-infected with HIV, five of whom were newly diagnosed. The positive predictive value of a CAM score ≥2 or an MSA ≥5 for death were 12% and 22% respectively. Five patients died. No variable was significantly associated with risk of death. There were no significant differences between individuals raised in endemic countries compared to non-endemic countries. Mortality in patients managed in a specialist centre was low. Patients who died succumbed to complications associated with a prolonged stay on ICU rather than malaria per se. The clinical usefulness of the MSA and CAM score was limited. Co-infection with HIV was relatively common but compared to studies in children, bacteraemia was uncommon. The relationship between ethnicity and immunity to severe disease is complex.


Insomnia and cow's milk allergy in infants.
A group of eight infants (six boys and two girls, 7 to 46 weeks of age) is reported, in whom a causal relationship between cow's milk allergy and chronic sleeplessness was suspected. They were referred because of waking and crying episodes that had occurred since the early days of life during sleep hours. During an average night, they slept about 4.5 hours and woke their parents about five times. They cried a lot during the day and were described as fussy. Two infants had been treated with phenothiazine without improvement. No cause for chronic insomnia was found during a standard medical and psychologic workup. An all-night polygraphic recording confirmed the disrupted sleep pattern of these infants, as compared with that of normal infants, and excluded further causes of arousals. Due to a clinical suspicion of atopy, the infants were further subjected to a series of allergy tests. IgE levels were shown to be elevated in each child, and radioallergosorbent tests were positive for cow's milk protein. The infants were than fed exclusively with a hydrolyzed milk protein mixture for 4 weeks. Sleep normalized within 2 weeks in every infant: night sleep increased to a median of 10 hours, and the awakenings only occurred occasionally. In four infants less than 6 months of age, cow's milk was reintroduced in the diet, and within 1 week all four became severely sleepless. Cow's milk was again excluded from the diet and the babies' sleep behaviors were again normalized. It is concluded that, when no evident cause for sleeplessness can be found in an infant, the possibility of milk allergy should be given serious consideration.


Cellular ADP-ribosyltransferase with the same mechanism of action as diphtheria toxin and Pseudomonas toxin A.
An ADP-ribosyltransferase was found in elongation factor 2 (EF-2) preparations from polyoma virus-transformed baby hamster kidney (pyBHK) cells. Like fragment A of diphtheria toxin and Pseudomonas toxin A, this eukaryotic cellular enzyme transfers [14C]adenosine from NAD+ to EF-2. However, the cellular transferase is immunologically distinct from fragment A. The transferase also can be distinguished from fragment A and Pseudomonas toxin A by the inhibition of the activity of the former by cytoplasmic extracts and by histamine. Snake venom phosphodiesterase digestion of the [14C]adenosine-labeled EF-2 product of the cellular transferase reaction yielded [14C]AMP, indicating that the cellular enzyme is a mono(ADP-ribosyl)transferase. The forward ADP-ribosylation reaction catalyzed by the cellular enzyme is reversed by fragment A, yielding [14C]NAD+. The results strongly suggest that the cellular transferase is a mono(ADP-ribosyl)transferase, which ADP-ribosylates the same diphthamide residue of EF-2 as does fragment A and Pseudomonas toxin A.


Redox Active Colloids as Discrete Energy Storage Carriers.
Versatile and readily available battery materials compatible with a range of electrode configurations and cell designs are desirable for renewable energy storage. Here we report a promising class of materials based on redox active colloids (RACs) that are inherently modular in their design and overcome challenges faced by small-molecule organic materials for battery applications, such as crossover and chemical/morphological stability. RACs are cross-linked polymer spheres, synthesized with uniform diameters between 80 and 800 nm, and exhibit reversible redox activity as single particles, as monolayer films, and in the form of flowable dispersions. Viologen-based RACs display reversible cycling, accessing up to 99% of their capacity and 99 ± 1% Coulombic efficiency over 50 cycles by bulk electrolysis owing to efficient, long-distance intraparticle charge transfer. Ferrocene-based RACs paired with viologen-based RACs cycled efficiently in a nonaqueous redox flow battery employing a simple size-selective separator, thus demonstrating a possible application that benefits from their colloidal dimensions. The unprecedented versatility in RAC synthetic and electrochemical design opens new avenues for energy storage.


Hormonal stimulation of avian embryonic cartilage growth in vitro: histologic and ultrastructural features.
We studied the histologic and ultrastructural features of embryonic chick cartilage after the cartilage had been incubated in serum-free medium that contained hormones and growth factors known to stimulate in vitro cartilage growth. Pelvic cartilages from 9 d chick embryos were incubated in BGJb ( Fitton -Jackson modification) medium alone (control) or medium containing one of the following: N6 monobutyryl cyclic AMP 0.5 mM, forskolin 100 microM, triiodothyronine (T3) 10 nM, insulin 45 nM, or somatomedin C 0.67 nM. At the end of 3 d of incubation the cartilages were fixed in buffered formalin. Significant growth (increases in size, wet and dry weight) was seen with each treatment group. N6-Monobutyryl cAMP treated cartilage had an increased number of flattened immature chondrocytes with large nuclei and prominent nucleoli. The histologic and ultrastructural features of forskolin treated cartilage were indistinguishable from N6-monobutyryl cAMP treatment. The T3 treated cartilage contained large hypertrophic chondrocytes with prominent lacunar typical of mature cartilage. T3 treated cartilage had considerable vacuole formation and dilated endoplasmic reticulum. Insulin and somatomedin treated cartilage had histologic appearance similar to control cartilage. Thus, the effects of various hormones on embryonic cartilage growth in vitro can be separated as to whether growth is the result of chondrocytic hyperplasia (cyclic AMP mediated), chondrocytic hypertrophy with maturation (T3), or a combination of both hyperplasia and hypertrophy (insulin and somatomedin-C).


Novel Dissolution Method for Oral Film Preparations with Modified Release Properties.
Oromucosal film preparations have gained popularity in pharmaceutical research and development. Therefore, oral films have been integrated into the monograph "oromucosal preparations" of the European Pharmacopeia in 2012. Regulatory authorities explicitly demand dissolution studies for films, but neither refer to suitable methods nor established specifications. Test methods described in the literature are often limited to immediate release formulations or not applicable to investigate the drug release of films with prolonged release profiles considering the different stages of gastrointestinal transit. The aims of this study were to develop a dissolution test method, which is suitable to investigate the drug release of film preparations with immediate as well as modified release profiles and to explore the potential of the test setup considering some physiological characteristics. Therefore, a conventional flow-through cell was equipped with in-house built sample holders. Three-dimensional printing technology was used for prototyping one of the sample holders. Four different types of films were investigated, such as ODFs with immediate (ODFIR) and prolonged release (ODFPR) characteristics as well as a double-layer film (ODFDL), produced with a water-insoluble shielding layer. Anhydrous theophylline was used as a model drug for all film types. Introducing special fixtures for oral films to a conventional flow-through cell enables successful determination of the drug release behavior of oral film preparations with immediate as well as modified release properties. Investigating ODFDL, the application of film sample holders with backing plates such as film sample holder with backing plate (FHB) and 3D printed film sample holder (FH3D) showed prolonged release profiles with 14.6 ± 1.30% theophylline dissolved within 2 h for FHB compared to 92.9 ± 3.33% for the film sample holder without backing plate (FH). This indicates their suitability to examine the integrity of the shielding layer. The application of the backing plate further decreased the drug release of ODFPR < 315 to 61.0 ± 1.69% dissolved theophylline within 2 h using FHB compared to 82.3 ± 0.74% using FH, due to a reduced ODF surface exposed to the dissolution medium. The potential of the dissolution test setup to consider physiological conditions of the human gastrointestinal transit was investigated by applying different flow rates and media compositions to simulate conditions within the oral cavity, stomach, and intestine. For the application of a low flow rate of 1 ml/min, comparable to the salivary flow within the oral cavity, decreased theophylline release was observed, while similar release profiles were obtained for flow rates between 2 and 8 ml/min. Substantial impact on the theophylline release was exerted by varying the composition of the dissolution medium. Since the drug release from ODFPR is controlled by diffusion through a water-insoluble matrix, ion species and concentration strongly affect the release behavior. In the future, IVIVC studies have to be performed to explore, whether obtained data can be used to predict drug release behavior of ODFs during the human gastrointestinal transit.


Role of vesicles during adenovirus 2 internalization into HeLa cells.
In this investigation, the early period of adenovirus type 2 (Ad2)-HeLa cell interaction was analyzed by electron microscopy and biochemical techniques. Events observed in this period ranged from the disappearance of virions from the cell surface to their subsequent association with the cell nucleus. Destabilization of the virions attached to the intact cell was necessary for virions to escape from intracellular vesicles. Strong temperature dependence and rapid escape from a vesicular compartment were shown in temporal kinetic experiments. These vesicles appeared to be acidic, since lysosomotropic agents partly inhibited the release of virions from vesicles. Studies of Ad2 binding to cells in buffers of different pH values suggested that adenovirus binds to cells by two different mechanisms. At low pH the binding was most probably mediated by the penton base and at neutral pH by the fiber protein. The number of receptor sites per cell was 25,000 and 6,000 at low and neutral pH, respectively. This study suggests that the low-pH affinity between the penton base and a vesicular membrane is important inside acid vesicles when Ad2 quickly enters the cytoplasm. However, a significant fraction of the virions was possibly internalized by a pathway not requiring a passage through such vesicles.


Endothelin Receptors Expressed by Immune Cells Are Involved in Modulation of Inflammation and in Fibrosis: Relevance to the Pathogenesis of Systemic Sclerosis.
Endothelin-1 (ET-1) plays a pivotal role in vasoconstriction, fibrosis, and inflammation, the key features of systemic sclerosis (SSc). ET-1 receptors (ETA and ET(B)) are expressed on endothelial cells, smooth muscle cells, and fibroblasts, but their presence on immune cells has not been deeply investigated so far. Endothelin receptors antagonists such as bosentan have beneficial effects on vasoconstriction and fibrosis, but less is known about their potential anti-inflammatory effects. We studied the expression of ET-1 receptors on immune cells (T and B lymphocytes, monocytes, and neutrophils) and the link between ET-1 and inflammation in patients with SSc. We show here that ET-1 exerts a proinflammatory effect in CD4+ T cells, since it induces an increased IFN-γ production; preincubation with antagonists of both receptors reduces IFN-γ production. Moreover, following ET-1 stimulation, neutrophils produce proinflammatory mediators, thus amplifying the effects of activated CD4+ T cells. Our data indicate that ET-1 system is involved in the pathogenesis of inflammation and fibrosis typical of SSc, through the activation of T lymphocytes and neutrophils and the consequent release of proinflammatory and profibrotic cytokines. These findings suggest that dual ET-1 receptors antagonist therapy, besides its effect on vasculopathy, has a profound impact on the immune system favouring antiinflammatory and antifibrogenic effects.


Kawase Approach for Dolichoectactic Basilar Artery Macrovascular Decompression in a Patient With Trigeminal Neuralgia: Case Report.
Trigeminal neuralgia (TN) secondary to a dolichoectatic basilar artery (DBA) is an extremely rare phenomenon. The Kawase approach for macrovascular decompression of this rare pathology been used rarely. This report describes macrovascular decompression and basilar artery transposition in a 69-yr-old male presenting with progressively worsening left-sided typical TN secondary to a DBA compression. The DBA was successfully decompressed off of the trigeminal nerve via a pterional craniotomy and anterior petrosectomy. The patient had immediate improvement in TN symptoms postoperatively. The patient remained symptom free with nonbothersome facial numbness in the V3 segment at 8-mo postoperative follow-up in clinic. The patient suffered a sixth nerve palsy following surgery, which was later corrected by strabismus surgery. The natural history and epidemiology of TN, results of macrovascular decompression secondary to DBA compression via a traditional suboccipital retrosigmoid approach, and potential advantages of the Kawase approach are also discussed. The macrovascular decompression strategy succeeded because the compressive force was applied by the DBA to the nerve in a superolateral direction, and the decompressive sling pulled the DBA away from the nerve in an inferomedial direction. The working space and access to the clival dura through the Kawase approach allowed proper corrective pull with a sling.


Comparison of apomorphine and levodopa infusions in four patients with Parkinson's disease with symptom fluctuations.
Motor fluctuations in patients with advanced Parkinson's disease may be successfully treated with subcutaneous apomorphine infusion or intraduodenal levodopa/carbidopa infusion. No comparative trials of these two alternatives were performed. We present a subanalysis from a randomized crossover clinical trial where levodopa infusion as monotherapy was compared with any other combination of pharmacotherapy in fluctuating patients. Four patients used apomorphine infusion and oral levodopa in the comparator arm. The results of these four patients are presented in detail. The duration of the trial was 3 + 3 weeks. Patients were video-recorded half-hourly on two non-consecutive days of both treatment arms. Blinded video ratings were used. Patient self-assessments of motor function and quality-of-life (QoL) parameters were captured using an electronic diary. Ratings in moderate to severe "off" state ranged 0-44% on apomorphine infusion and 0-6% on levodopa infusion. Moderate to severe dyskinesias were not recorded in any of the treatments. QoL was reported to be improved in all patients on duodenal levodopa infusion. Monotherapy with duodenal infusion of levodopa was more efficacious and brought greater QoL than combination therapy with apomorphine infusion in these fluctuating patients.


[Gel-producing properties of exopolysaccharide acrylamide in composition with agar-agar].
A possibility to use a new polymer--exopolysaccharide acrylamide (EPAA)--for partial substitution of agar-agar in solid nutrient media in microbiology has been studied. It has been established than the EPAA in concentration 6.0-12.0 g/l with agar in concentration 10.0 g/l produce gel which is so strong as agar in concentration 15.0-20.0 g/l. Quality of gels produced depends an the method of EPAA mixing with agar. It has been shown that usage of EPAA in nutrient media does not affect growth, colony and cell morphology and biochemical properties of microorganisms. The use of EPAA to prepare nutrient media permits us to reduce expenditures for agar.


Drotrecogin alfa Eli Lilly.
Drotrecogin alfa (Xigris, recombinant activated protein C) is an anticoagulant developed and launched by Eli Lilly & Co for the treatment of sepsis [333781], [339372], [430133], [436271]. The FDA and the EMEA accepted the brand name Xigris for drotrecogin alfa in June 2001. This trade name had been proposed by Lilly in place of the previous brand name, Zovant, which was deemed unacceptable by the EMEA due to concerns that the name could be confused with hospital-based drugs [412512]. Filings for sepsis were made in the US, EU and Australia in February 2001 [398514], [447870] and in March 2001, the US FDA assigned drotrecogin alfa Priority Review status [403435]. The FDA extended the action date from July 27 to October 27, 2001 for completion of its review of the biologics license application (BLA) for drotrecogin alfa to assess further supplementary data submitted by Lilly [412512]. At the October 16, 2001 meeting (postponed from September 12), the FDA Advisory Committee on Anti-Infective Drugs split 10 to 10 over whether to recommend approval [425873], [425940]. In late October 2001, Lilly received an approvable letter from the FDA for the treatment of severe sepsis. Approval was contingent upon successful negotiation of labeling, agreement on post-approval clinical trials, and successful completion of manufacturing inspections [427301]. In November 2001, the FDA approved drotrecogin alfa for the reduction of mortality in adult patients with severe sepsis who have a high risk of death [430133]; the product was launched onto the US market days later [436271]. Following the FDA committee's split decision in October 2001, Credit Suisse First Boston, which expected mid-2002 approval but with restrictive labeling, revised its predictions from $1.265 billion in 2004 sharply downwards to $543 million [425929].


The effects of low-fat diets rich in arachidonic acid on the composition of plasma fatty acids and bleeding time in Australian aborigines.
In the present study we measured the bleeding times in fourteen Aborigines (10 diabetic, 4 non-diabetic) before and after 2 weeks on a diet of tropical seafood (rich in both arachidonic acid and the omega 3 PUFA), followed by 3 weeks on a diet in which kangaroo and freshwater fish (linoleic and arachidonic acid-rich) were the major fat sources. Both diets were very low in fat. Bleeding times increased in all subjects after the 2 weeks of tropical seafood and continued to rise on the mixed diet. The increase over 5 weeks from 4.1 +/- 0.4 to 5.9 +/- 0.4 min was highly significant (p less than 0.01). Due to the extreme isolation of the study location it was only possible to measure the plasma fatty acid composition at the beginning and end of the study. The concentration of arachidonic acid in the plasma lipids doubled whereas that of linoleic acid was almost halved, despite the fact that the diet in the second part of the study contained considerably more linoleic than arachidonic acid. That there appeared to be preferential incorporation of arachidonic acid into the plasma lipids is further supported by the observation that the rise in arachidonic acid in the cholesterol ester and phospholipid fractions was almost exactly counter-balanced by the fall in linoleic acid. In conclusion, the present study demonstrated a rise in bleeding time associated with an increased concentration of arachidonic acid and decreased concentration of linoleic acid in plasma lipids, and suggests that the mechanism by which diet modulates haemostatic function may be more complex than currently assumed.


The Identity Threat of Weight Stigma in Adolescents.
Obesity remains a serious public health issue in adolescents, who may be subjected to weight stigma leading to increased stress and poor health outcomes. Stigma can be detrimental to adolescents during self-identity formation. The purpose of this study was to examine weight stigma in adolescents in light of the Identity Threat Model of Stigma. A cross-sectional correlational design was used to examine the relationships among the variables of weight stigma, psychosocial stress, coping styles, disordered eating, and physical inactivity. Regression modeling and path analysis were used to analyze the data. Over 90% of the sample had scores indicating weight stigma or antifat bias. Avoidant coping style and psychosocial stress predicted disordered eating. The strongest path in the model was from avoidant coping to disordered eating. The Identity Threat Model of Stigma partially explained adolescents' weight stigma. Nursing practice implications are discussed.


GRR1 of Saccharomyces cerevisiae is required for glucose repression and encodes a protein with leucine-rich repeats.
Growth of the yeast Saccharomyces cerevisiae on glucose leads to repression of transcription of many genes required for alternative carbohydrate metabolism. The GRR1 gene appears to be of central importance to the glucose repression mechanism, because mutations in GRR1 result in a pleiotropic loss of glucose repression (R. Bailey and A. Woodword, Mol. Gen. Genet. 193:507-512, 1984). We have isolated the GRR1 gene and determined that null mutants are viable and display a number of growth defects in addition to the loss of glucose repression. Surprisingly, grr1 mutations convert SUC2, normally a glucose-repressed gene, into a glucose-induced gene. GRR1 encodes a protein of 1,151 amino acids that is expressed constitutively at low levels in yeast cells. GRR1 protein contains 12 tandem repeats of a sequence similar to leucine-rich motifs found in other proteins that may mediate protein-protein interactions. Indeed, cell fractionation studies are consistent with this view, suggesting that GRR1 protein is tightly associated with a particulate protein fraction in yeast extracts. The combined genetic and molecular data are consistent with the idea that GRR1 protein is a primary response element in the glucose repression pathway and is required for the generation or interpretation of the signal that induces glucose repression.


DIFFEOMORPHIC POINT SET REGISTRATION USING NON-STATIONARY MIXTURE MODELS.
This paper investigates a diffeomorphic point-set registration based on non-stationary mixture models. The goal is to improve the non-linear registration of anatomical structures by representing each point as a general non-stationary kernel that provides information about the shape of that point. Our framework generalizes work done by others that use stationary models. We achieve this by integrating the shape at each point when calculating the point-set similarity and transforming it according to the calculated deformation. We also restrict the non-rigid transform to the space of symmetric diffeomorphisms. Our algorithm is validated in synthetic and human datasets in two different applications: fiber bundle and lung airways registration. Our results shows that non-stationary mixture models are superior to Gaussian mixture models and methods that do not take into account the shape of each point.


Phylogenetic relationships and character evolution in Heuchera (Saxifragaceae) on the basis of multiple nuclear loci.
• The use of multiple genetic regions from the nuclear genome, including low-copy markers, has long been recognized as essential to robust phylogenetic construction, addressing gene tree incongruence, and allowing increased resolution to test current taxonomy and resolve basic hypotheses about character evolution, biogeography, and other organismal traits of interest to biologists. Heuchera, the largest genus of Saxifragaceae endemic to North America, has presented an unusually difficult case for systematists with limited sampling in previous molecular studies. We used morphological and multilocus molecular phylogenetic data to test the monophyly of Heuchera, better resolve hypotheses of relationships, and test hypotheses of character evolution, biogeography, and diversification rates.• Phylogenetic relationships were inferred using sequences from six nuclear loci and 39 morphological characters using concatenation and coalescent analysis. Ancestral state reconstruction, diversification, and correlated evolution of morphological traits were performed using parsimony, BiSSE, and Pagel's method, respectively.• The concatenation and species tree analyses gave confident support to the monophyly of Heuchera and suggested several well-supported subclades. The addition of morphological data significantly improved support numbers. Ancestral character reconstruction suggested frequent homoplasy and reversal of floral characters and a complex biogeographical history.• Heuchera is a natural genus; however, the current subgeneric classification is artificial and requires revision. Biogeographic reconstructions suggest a Pacific Northwest origin for the genus; morphological shifts in stamen exsertion and hypanthium length are hypothesized to have driven diversification in Heuchera in concert with possible pollinator shifts.


Effect of aminophylline on transpulmonary passage of venous air emboli in pigs.
Aminophylline has been shown to dramatically reduce the filtering capacity of the lung in dogs during venous air embolism. Similarities have been pointed out between the cardiovascular and respiratory systems of the pig and of humans. We therefore wanted to find out whether aminophylline also modifies the transpulmonary spillover of microbubbles to the arterial circulation of the pig. Twenty-eight pigs were anesthetized with pentobarbital sodium and mechanically ventilated. Aminophylline was injected intravenously into 10 of the pigs before the introduction of air bubbles into the right ventricle, while the other 18 pigs served as controls. A transesophageal echocardiographic probe was used to detect eventual air bubbles in the left atrium or in the aorta. Pigs received either air infusion, at rates varying from 0.05 to 0.20 ml.kg-1.min-1, or calibrated microbubbles, 5-300 microns diam. We found that aminophylline-treated pigs did not show any change in spillover incidence compared with controls. Furthermore, in both groups the spillover during continuous air infusion seemed to be a preterminal event, because the pigs had very low arterial pressure when arterial bubbles were observed. Finally, there was an increase in mean pulmonary arterial pressure from 18 +/- 3.4 to 26 +/- 2.2 (SD) mmHg (n = 4, P less than 0.01) in aminophylline-treated pigs after a bolus injection of microbubbles (less than or equal to 50 microns, total volume less than 0.5 ml). Our results suggest that aminophylline does not modify the transpulmonary passage of microbubbles in this porcine model. In addition, it would seem that the pulmonary circulation of the pig is sensitive to very small volumes of air, when injected as microbubbles.


Magnetic resonance imaging of great vessel, myocardial, and pericardial disease.
The outstanding ability of magnetic resonance imaging to provide anatomic information over a large field of view in any plane in patients of all sizes and body habitus has made it the current modality of choice for evaluating the thoracic aorta and for differentiating constrictive pericarditis from restrictive cardiomyopathy. The lack of radiation exposure makes it particularly advantageous for patients requiring serial studies. It serves as an important clinical adjunct to ultrasound in evaluating diseases of the abdominal aorta, other great vessels, the myocardium, and the pericardium. Myocardial function, thickness, and mass can be assessed without geometric assumptions. Myocardial tagging offers a unique way to study cardiac biomechanics. The combination of anatomic and quantitative functional information becoming available in clinically reasonable acquisition times makes magnetic resonance imaging an increasingly attractive noninvasive tool. The contribution of tissue characterization has been disappointing, but may improve with utilization of contrast agents; that of in vivo spectroscopy remains investigational.


Malaria and the nervous system.
The neurological complications of malaria--an almost universal problem although it occurs mainly in the tropics--constitute the commonest cause of death and morbidity. These complications usually occur in P. falciparum infections and in those who are non-immune, such as children, pregnant women and visitors to areas endemic for malaria. They are rare in children suffering from protein-calorie malnutrition. Febrile convulsions, the commonest childhood emergency in the African and cerebral malaria are the commonest neurological complications. Spinal cord disorders and polyneuropathy have been described but are rare. It has been suggested that various forms of mental illness and psychoses may be related to malarial infections, but much research is needed to establish cause and effect relationship. The neuropathology and pathogenesis, clinical presentations, treatment and mortality of the neurological complications are reviewed.


Turbinaria ornata (Phaeophyceae) varies size and strength to maintain environmental safety factor across flow regimes.
Water motion in coastal areas can produce hydrodynamic forces that damage or dislodge benthic macroalgae if the tissues of macroalgae are not sufficiently strong. Some macroalgae vary their morphology and strength in response to ambient water motion, but little is known of how morphology and strength of macroalgae change relative to one another across flow regimes. Here, we use Turbinaria ornata, an ecologically important macroalga, to study how both the morphology and strength of macroalgae vary with ambient water motion. Typically, T. ornata exhibits weakening of its stipe when sexually mature, leading to breakage from the substratum and dispersal, which is beneficial for reproduction. Across three flow regimes, adult T. ornata increased its size but decreased its strength as water motion decreased. However, the strength of T. ornata relative to the maximum hydrodynamic forces it is expected to encounter (the environmental safety factor) did not differ between flow regimes. Our results showed that T. ornata can conform to its local flow habitat by varying both size and strength, similar to other macroalgae. Varying multiple traits between flow regimes suggested that T. ornata is capable of surviving a wide range of flow conditions, which may permit more control over the timing of its weakening, breakage from the substratum, and dispersal, even with future increases in flow velocities (e.g., large waves from storms) that are expected to occur frequently with climate change.


Cytochrome P450 pharmacogenetics in drug development: in vitro studies and clinical consequences.
Members of the human cytochrome P450 (CYP) superfamily play a role in the metabolism of many drugs and several of them, CYP2D6, CYP2C9 and CYP2C19, have been shown to be polymorphic as a result of single nucleotide polymorphisms (SNPs), gene deletions, and gene duplications. These polymorphisms can impact the pharmacokinetics (PK), metabolism, safety and efficacy of drugs, and because of the availability of automation, genotyped human tissue, recombinant CYP preparations (rCYPs) and reagents, most pharmaceutical companies have increasingly screened out compounds that are metabolized solely by polymorphic CYPs. In the absence of suitable animal models, it has been widely accepted that such in vitro data are useful because one can obtain information prior to dosing in man and select the most appropriate clinical studies with prospectively genotyped and phenotyped subjects. Overall, current trends in the industry have been fueled by increased managed healthcare, the desire to minimize the need for therapeutic drug monitoring and CYP genotyping in medical practice, and a very competitive market place. In the past, such paradigms have not been as influential and there are numerous examples of marketed drugs that are metabolized by polymorphic CYPs.