Phase I evaluation of a synthetic mutant of beta-interferon.
A synthetic mutant of beta-interferon, produced by recombinant DNA technology, was prepared with serine substituted for the naturally occurring cysteine at amino acid 17. This molecule, after purification to homogeneity, was evaluated in 23 patients with cancer for tolerated doses, safety, and pharmacokinetics. Each patient was begun on twice weekly administration, one dose i.m., then an identical dose i.v. Doses, escalated weekly, were tolerated by 9 of 12 patients at 100 X 10(6) units i.m., 11 of 14 patients at 100 X 10(6) units i.v., and 8 of 10 patients receiving i.v. doses of 200 X 10(6) units. Fever (greater than or equal to 38.9 degrees C), the commonest cause for ceasing dose escalation, occurred in 11 of 13 patients who developed limiting i.v. toxicity and 6 of 11 who developed limiting i.m. toxicity. Patients who did not have progressive cancer after completion of dose escalation received five consecutive daily doses at their maximum tolerated single dose by each route, i.m. and i.v. These two 5-day treatments were given without difficulty. All patients treated with 300 X 10(6) units or less, i.m. (n = 13) or i.v. (n = 10), were able to receive five daily doses without limiting toxicity. Peak serum titers occurred immediately after i.v. administration and declined in an exponential manner thereafter. Despite absence of measurable titers in serum after i.m. injection, fever and significant (P less than 0.05) depression of WBC and platelet counts, serum calcium, and serum cholesterol occurred (prestudy to maximum tolerated dose). An immunoglobulin antibody to beta-interferon, detected by enzyme-linked immunoabsorbent assay, developed in 17 of 23 patients. Neutralizing activity (titer 10(2] was found in only 1 of 23 patients. No immune-mediated sequelae (symptomatic or renal) were identified. Further Phase I and II trials with this molecule will determine whether it will prove to have a better therapeutic index or different spectrum of therapeutic activity from alpha-interferon or gamma-interferon.


Concentration and molecular weight of sodium hyaluronate in synovial fluid from patients with rheumatoid arthritis and other arthropathies.
The molecular weight distribution of hyaluronate (HA) in synovial fluid (SF) from 10 patients with rheumatoid arthritis (RA), from six patients with other joint disorders, and from five recently deceased persons without joint affections was investigated by a gel chromatographic procedure. A new and highly specific radioassay was used for determination of the HA concentration in the effluent from the chromatographic column, and this allowed analyses on 0.5 ml or less of untreated synovial fluid. The results confirmed the findings by others that the weight-average molecular weight (Mw) of HA in SF from patients with RA (4.8 X 10(6)) was similar to that in other joint diseases (5.0 X 10(6)) and moderately but significantly (p less than 0.001) lower than that of normal SF (7.0 X 10(6)). Furthermore, the molecular weight distribution of HA in the pathological SF was generally broad and varied considerably between individuals. The HA concentration in the pathological SF varied between 0.17 and 1.32 g/l, which is in accordance with previous reports and considerably lower than that of normal SF. Neither the nature of the arthropathy and the extent of the inflammatory process nor the pharmacological treatment had a tendency to influence the HA concentration in the SF, the mean molecular weight of HA, or its molecular weight distribution. Although the concentration of HA in SF drops in joint disease, the total amount of the polysaccharide is greatly enhanced. Also the amount of high molecular weight polysaccharide (Mw greater than 6 X 10(6)) is in excess in joint disease. The pathological state is therefore characterised not by lack of high molecular weight hyaluronate but by a dilution of it.


CASA chromogenic medium for enteric Campylobacter species.
We prospectively assessed stool samples from 370 patients for Campylobacter species by comparing three selective agar media incubated at two temperatures: 42°C and 37°C. Twenty patients (5.4%) were found positive. The chromogenic medium CASA (AES Chemunex, France) proved highly efficient for C. jejuni and C. coli recovery, while lessening the workload in the lab.


Use of non-steroidal drugs for the treatment of sprains.
Further to the learning zone article, 'Care of patients with minor injuries' ( January 25 ), I would like to comment on the treatment of musculoskeletal injuries such as sprains.


Calcium-Sensing Receptor Internalization Is β-Arrestin-Dependent and Modulated by Allosteric Ligands.
G protein-coupled receptor (GPCR) internalization is crucial for the termination of GPCR activity, and in some cases is associated with G protein-independent signaling and endosomal receptor signaling. To date, internalization has been studied in great detail for class A GPCRs; whereas it is not well established to what extent the observations can be generalized to class C GPCRs, including the extracellular calcium-sensing receptor (CaSR). The CaSR is a prototypical class C GPCR that maintains stable blood calcium (Ca2+) levels by sensing minute changes in extracellular free Ca2+ It is thus necessary that the activity of the CaSR is tightly regulated, even while continuously being exposed to its endogenous agonist. Previous studies have used overexpression of intracellular proteins involved in GPCR trafficking, pathway inhibitors, and cell-surface expression or functional desensitization as indirect measures to investigate CaSR internalization. However, there is no general consensus on the processes involved, and the mechanism of CaSR internalization remains poorly understood. The current study provides new insights into the internalization mechanism of the CaSR. We have used a state-of-the-art time-resolved fluorescence resonance energy transfer-based internalization assay to directly measure CaSR internalization in real-time. We demonstrate that the CaSR displays both constitutive and concentration-dependent Ca2+-mediated internalization. For the first time, we conclusively show that CaSR internalization is sensitive to immediate positive and negative modulation by the CaSR-specific allosteric modulators N-(3-[2-chlorophenyl]propyl)-(R)-α-methyl-3-methoxybenzylamine (NPS R-568) and 2-chloro-6-[(2R)-2-hydroxy-3-[(2-methyl-1-naphthalen-2-ylpropan-2-yl)amino]propoxy]benzonitrile (NPS 2143), respectively. In addition, we provide compelling evidence that CaSR internalization is β-arrestin-dependent while interestingly being largely independent of Gq/11 and Gi/o protein signaling. SIGNIFICANCE STATEMENT: A novel highly efficient cell-based real-time internalization assay to show that calcium-sensing receptor (CaSR) internalization is β-arrestin-dependent and sensitive to modulation by allosteric ligands.


[Optimal intensity and duration of training and the duration of walks during the after-care of myocardial infarct patients at a sanatorium].
Daily exercise drills producing a heart rate increment of about 31.5% of the one recorded at first stress tolerance test and lasting for 105 min, in combination with daily 75 minutes' walks were not sufficient to increase stress tolerance. More intensive drills, producing a 50.6% heart rate increment, while lasting less than 1 hour per day, in combination with 2.5 hours' daily walks, resulted in a significant stress tolerance increase. In neither of the two groups was it associated with any significant rise of the double product, implicating extracardiac factors as the principal contributors.


Content and Feature Preferences for a Physical Activity App for Adults With Physical Disabilities: Focus Group Study.
Hundreds of thousands of mobile phone apps intended to improve health and fitness are available for download across platforms and operating systems; however, few have been designed with people with physical disabilities in mind, ignoring a large population that may benefit from an effective tool to increase physical activity. This study represents the first phase in the development process of a fitness tracking app for people with physical disabilities interested in nontraditional sport. The aim of this research was to explore user preferences for content, appearance, and operational features of a proposed physical activity app for people with physical disabilities to inform the design of a mobile phone app for increasing physical activity. Four focus groups were conducted with 15 adults with physical disabilities who currently participate in nontraditional, non-Paralympic sport. Data collected from the focus group sessions centered on content, functionality, and appearance of apps currently used by participants as well as preferences for a future app. Participants (mean age 35.7, SD 9.2 years) were mostly white (13/15, 87%), and all were currently participating in CrossFit and at least one other sport. Five main themes were identified. Themes included preferences for (1) workout-specific features that were tailored or searchable by disability, (2) user experience that was intuitive and accessible, (3) profile personalization options, (4) gamification features that allowed for competition with self and other users, and (5) social features that allowed increased interaction among users. Participants expressed a primary interest in having a fitness app that was designed for people with physical disabilities such that the features present in other fitness tracking apps were relevant to them and their community of adaptive athletes. The results showed that features related to user experience, social engagement, and gamification are considered important to people with physical disabilities. Features highlighted by participants as most desired, from a consumer perspective, were in line with research identifying attributes of quality apps that use behavior change techniques to influence positive physical activity behavior change. Such insights should inform the development of any fitness app designed to integrate users with disabilities as a primary user base.


Recruitment-to-inflation Ratio Assessed through Sequential End-expiratory Lung Volume Measurement in Acute Respiratory Distress Syndrome.
Positive end-expiratory pressure (PEEP) benefits in acute respiratory distress syndrome are driven by lung dynamic strain reduction. This depends on the variable extent of alveolar recruitment. The recruitment-to-inflation ratio estimates recruitability across a 10-cm H2O PEEP range through a simplified maneuver. Whether recruitability is uniform or not across this range is unknown. The hypotheses of this study are that the recruitment-to-inflation ratio represents an accurate estimate of PEEP-induced changes in dynamic strain, but may show nonuniform behavior across the conventionally tested PEEP range (15 to 5 cm H2O). Twenty patients with moderate-to-severe COVID-19 acute respiratory distress syndrome underwent a decremental PEEP trial (PEEP 15 to 13 to 10 to 8 to 5 cm H2O). Respiratory mechanics and end-expiratory lung volume by nitrogen dilution were measured the end of each step. Gas exchange, recruited volume, recruitment-to-inflation ratio, and changes in dynamic, static, and total strain were computed between 15 and 5 cm H2O (global recruitment-to-inflation ratio) and within narrower PEEP ranges (granular recruitment-to-inflation ratio). Between 15 and 5 cm H2O, median [interquartile range] global recruitment-to-inflation ratio was 1.27 [0.40 to 1.69] and displayed a linear correlation with PEEP-induced dynamic strain reduction (r = -0.94; P < 0.001). Intraindividual recruitment-to-inflation ratio variability within the narrower ranges was high (85% [70 to 109]). The relationship between granular recruitment-to-inflation ratio and PEEP was mathematically described by a nonlinear, quadratic equation (R2 = 0.96). Granular recruitment-to-inflation ratio across the narrower PEEP ranges itself had a linear correlation with PEEP-induced reduction in dynamic strain (r = -0.89; P < 0.001). Both global and granular recruitment-to-inflation ratio accurately estimate PEEP-induced changes in lung dynamic strain. However, the effect of 10 cm H2O of PEEP on lung strain may be nonuniform. Granular recruitment-to-inflation ratio assessment within narrower PEEP ranges guided by end-expiratory lung volume measurement may aid more precise PEEP selection, especially when the recruitment-to-inflation ratio obtained with the simplified maneuver between PEEP 15 and 5 cm H2O yields intermediate values that are difficult to interpret for a proper choice between a high and low PEEP strategy.


Particulate carrier systems as adjuvants for cancer vaccines.
To overcome the immunosuppressive milieu of malignancy and lack of well-defined antigens, potent adjuvants are needed for cancer immunotherapy. Numerous small molecular immunomodulators have the potential to fulfill this role. To enhance the immune response and decrease the toxicity, particulate systems including nanoparticles and macroparticles have been increasingly proposed as carriers for cancer antigen and adjuvant delivery. These systems have the potential to co-deliver the antigens and adjuvants simultaneously in the same particle. In addition, the particles can be engineered for localized and targeted delivery, whether it be to the cellular or sub-cellular level. These properties limit systemic side effects and improve delivery efficiency, and thus enhance the vaccine's immune response. In particular, the particles can be constructed to mimic the size and surface patterns of microbes, organisms to which we have evolved a strong immune response. The release characteristics of the particles can likewise be controlled to simulate the body's response to infections. Boosting the immune response of vaccines by virtue of their intrinsic immunostimulatory properties, these particles can be dosing-sparing and have the potential to reduce production cost of vaccines. As the interest in personalized cancer vaccines increases with their encouraging outcomes in clinical trials, particulate carrier systems have the potential to play an important role in optimizing cancer vaccines.


Epigenetic regulation of glucose metabolism.
Glucose metabolism is a central process in mammalian energy homeostasis. Its deregulation is a key factor in development of metabolic disease like diabetes and cancer. In recent decades, our understanding of gene regulation at the signaling, chromatin and posttranscriptional levels has seen dramatic developments. A number of epigenetic mechanisms that do not affect the genetic code can be assessed with new technologies. However, increasing complexity becomes a major challenge for translation into clinical application. The current review provides an update of transcriptional control of glucose metabolism, focusing on epigenetic regulators, DNA-methylation, histone modifications and noncoding RNAs. Recent studies heavily support the importance of those mechanisms for future therapeutics and preventive efforts for metabolic diseases.


Clinical perspective: have the results of recent clinical trials of lipid-lowering therapies influenced the way we should practice? A Latin American perspective of current issues in clinical lipidology.
Traditional tools to evaluate cardiovascular disease risk may underestimate the risk of cardiovascular events. Although reduction of low-density lipoprotein cholesterol (LDL-C) is the mainstay of therapy to mitigate cardiovascular risk from atherosclerosis, noninvasive imaging techniques and biomarkers are now allowing us to identify subclinical atherosclerosis, or high-risk patients, and are providing clinical researchers with new target end points for randomized controlled clinical trials. Current surrogates include carotid intima-media thickness, coronary artery calcification, and high-sensitivity C-reactive protein levels. There is evidence that these biomarkers are useful in clinical practice to improve risk quantification in subjects considered at intermediate risk of coronary events according to clinical risk stratification. Some studies, but not all, have demonstrated achievement of surrogate end points with lipid-lowering therapy in addition to LDL-C reductions. A group of clinical lipidologists from Latin American countries convened to give a perspective on recent clinical trials in clinical lipidology, their designs, and data regarding currently used biomarkers. It was noted that the success of some surrogate end points as possible markers of clinical efficacy has relied heavily on patient selection and trial design. On the basis of current evidence, we believe that correcting elevated LDL-C levels should remain the primary target of therapy for patients with dyslipidemia. The group also agreed that the evidence from recent clinical trials supports the potential role of new biomarkers for the screening and identification of patients at high cardiovascular risk in the absence of overt hyperlipidemia.


An in vivo swine study for xeno-grafts of calcium sulfate-based bone grafts with human dental pulp stem cells (hDPSCs).
The purpose of this in vivo study was to evaluate the effect of human dental pulp stem cells (hDPSCs) on various resorbable calcium sulfate/calcium phosphate bone grafts in bone regeneration. Granular particles of calcium sulfate dehydrate (CSD), α-calcium sulfate hemihydrate/amorphous calcium phosphate (α-CSH/ACP), and CSD/β-tricalcium phosphates (β-TCP) were prepared for in vitro dissolution and implantation test. The chemical compositions of specimen residues after dissolution test were characterized by XRD. The ratios of new bone formation for implanted grafts/hDPSCs were evaluated using mandible bony defect model of Lanyu pig. All the graft systems exhibited a similar two-stage dissolution behavior and phase transformation of poor crystalline HAp. Eight weeks post-operation, the addition of hDPSCs to various graft systems showed statistically significant increasing in the ratio of new bone formation (p<0.05). Null hypothesis of hDPSCs showing no scaffold dependence in bone regeneration was rejected. The results suggest that the addition of hDPSCs to calcium sulfate based xenografts could enhance the bone regeneration in the bony defect.


Synovial sarcoma in the right atrium and right ventricle.
A metastatic synovial sarcoma in the right atrium and ventricle is described. A 36-year-old man was admitted to our hospital with generalized fatigue, dyspnoea, and precordial pain. Transthoracic echocardiography demonstrated a metastatic tumour in both the right atrium and right ventricle and revealed obstruction of the inflow tract of the right ventricle caused by a metastatic right atrial tumour. Thoracic computed tomography revealed a pleural-based paravertebral mass in the left intrathoracic cavity and multiple pulmonary nodules in both lungs. Cardiac surgery was performed for palliative treatment due to right cardiac failure and a risk of fatal embolization. The patient died 12 months after the cardiac surgery.


Genetic diversity and recombination in natural populations of the nematode-trapping fungus Arthrobotrys oligospora from China.
Nematophagous fungi can trap and capture nematodes and other small invertebrates. This unique ability has made them ideal organisms from which to develop biological control agents against plant- and animal-parasitic nematodes. However, effective application of biocontrol agents in the field requires a comprehensive understanding about the ecology and population genetics of the nematophagous fungi in natural environments. Here, we genotyped 228 strains of the nematode-trapping fungus Arthrobotrys oligospora using 12 single nucleotide polymorphic markers located on eight random DNA fragments. The strains were from different ecological niches and geographical regions from China. Our analyses identified that ecological niche separations contributed significantly, whereas geographic separation contributed relatively little to the overall genetic variation in our samples of A. oligospora. Interestingly, populations from stressful environments seemed to be more variable and showed more evidence for recombination than those from benign environments at the same geographic areas. We discussed the implications of our results to the conservation and biocontrol application of A. oligospora in agriculture and forestry.


Developing new SSR markers from ESTs of pea (Pisum sativum L.).
The development of expressed sequence tags (ESTs) from pea has provided a useful source for mining novel simple sequence repeat (SSR) markers. In the present research, in order to find EST-derived SSR markers, 18 552 pea ESTs from the National Center for Biotechnology Information (NCBI) database were downloaded and assembled into 10 086 unigenes. A total of 586 microsatellites in 530 unigenes were identified, indicating that merely 5.25% of sequences contained SSRs. The most abundant SSRs within pea were tri-nucleotide repeat motifs, and among all the tri-nucleotide repeats, the motif GAA was the most abundant type. In total, 49 SSRs were used for primer design. EST-SSR loci were subsequently screened on 10 widely adapted varieties in China. Of these, nine loci showed polymorphic profiles that revealed two to three alleles per locus. The polymorphism information content value ranged from 0.18 to 0.58 with an average of 0.41. Furthermore, transferable analysis revealed that some of these loci showed transferability to faba bean. Because of their polymorphism and transferability, these nine novel EST-SSRs will be valuable tools for marker-assisted breeding and comparative mapping of pea in the future.


Topological invariant for the orbital spin of the electrons in the quantum Hall regime and Hall viscosity.
Hall conductance of noninteracting fermions filling a certain number of Landau levels can be written as a topological invariant. A particular version of this invariant when expressed in terms of the single particle Green's functions directly generalizes to cases when interactions are present including those of fractional Hall states, although in those cases this invariant no longer corresponds to Hall conductance. We argue that when evaluated for both integer and fractional Hall states this invariant gives twice the total orbital spin of fermions which in turn is closely related to the Hall viscosity, a quantity characterizing the integer and fractional Hall states which recently received substantial attention in the literature.


Loading Amlodipine on Diamond Nanoparticles: A Novel Drug Delivery System.
Diamond nanoparticles (Nanodiamond) are biocompatible drug delivery platforms with outstanding surface properties. Their passage into the brain has been confirmed previously. Thus, nanodiamond could provide a drug delivery system to shuttle several drugs through the blood-brain barrier (BBB) which represents a real challenge for the effective delivery of several drugs into the brain. Amlodipine is a calcium channel blocker that cannot pass through BBB and may elicit neuroprotective effects to reverse calcium-induced excitotoxicity and mitochondrial dysfunction that underlie several neurologic disorders including Alzheimer's disease and stroke. The study aimed to investigate the loading of amlodipine on nanodiamond particles. Nanodiamond particles were oxidized in a strong oxidizing acidic mixture of sulfuric and nitric acids. Adsorption of amlodipine on nanodiamond particles was achieved in alkaline pH using various concentrations of sodium hydroxide. The loaded amlodipine was determined by high-performance liquid chromatography and confirmed by Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy. The highest percentage (41%) of loaded amlodipine onto nanodiamond particles was achieved in alkaline medium using 2 mM NaOH at a corresponding pH of 8.5. Also, characteristic FTIR bands of amlodipine and nanodiamond were shown obviously in the nanodiamond-amlodipine conjugates. Moreover, the successful loading of amlodipine on diamond nanoparticles was confirmed by transmission electron microscopy. The present study demonstrates the successful loading of amlodipine onto nanodiamond particles. These findings offer a potential for applying diamond nanoparticles as a drug delivery system to shuttle amlodipine into the brain and open the door to deliver other similar drugs into the brain.


Developmental programming: maternal hypercholesterolemia and immunity influence susceptibility to atherosclerosis.
It is increasingly recognized that the in utero environment is an important determinant of adult disease, and extensive epidemiological evidence links dysmetabolic conditions during pregnancy with increased hypertension, cardiovascular disease, and diabetes later in life. The original Barker Hypothesis focused on low birth weight as the primary indicator of postnatal risk, but low birth weight may arise from other, non-metabolic conditions. This has impeded the identification of developmental programming mechanisms. More recently, the focus has shifted to the impact of specific maternal risk factors, such as obesity, metabolic syndrome, and diabetes, on cardiovascular risk in offspring. Inflammation plays a central role in these maternal conditions as well as in offspring atherogenesis, and two key factors that influence inflammation, maternal hypercholesterolemia and maternal immune mechanisms, have been shown to affect the developmental programming of atherosclerosis. Maternal hypercholesterolemia in pregnancy, even if only temporary, is associated with increased fatty streak formation in human fetal arteries and accelerated progression of atherosclerosis in normocholesterolemic children. Conversely, immunization of experimental animals with oxidized low-density lipoprotein cholesterol, an antigen prevalent in atherosclerotic lesions, inhibits the progression of atherosclerosis in the offspring of hypercholesterolemic mothers. These findings indicate it is possible, in principle, to program postnatal immune responses and to reduce atherosclerosis, and potentially other immunomodulated diseases, by targeted maternal immunomodulation.


Cytokine-induced depression: current status and novel targets for depression therapy.
Current treatments of depression include psychological, pharmacological and physical approaches. Pharmacological interventions to treat depression have previously focused on modifying dysfunctional neurotransmitter systems. Overall, these treatments have demonstrated an ability to manage major depression but otucomes continue to be poor in many patients, especially those with long term illness or with previous multiple relapses. This may be due to the fact that depression is a systemic and neuroprogressive illness involving multiple biological pathways such as immunological factors. There is substantial evidence that cytokine therapies induce depressive symptoms in clinical populations. The model of cytokine-induced depression has provided important information relative to the risk factors and biological pathways involved in the etiology of depressive symptoms and, most importantly, the identification and knowledge of these factors has allowed new treatment targets to be explored. When an exogenous cytokine such as interferon-alpha is administered, proinflammatory cytokines are activated, leading to alterations in neurotransmission and endocrine pathways and producing neurotoxicity. Several new treatments for depression acting through pathways other than amine neurotransmission have emerged in recent years. The regulation of the inflammatory response, the decrease in the activity of the hypothalamic-pituitary-adrenal axis and the prevention of neurotoxicity are potential targets for new drugs. Though these drugs are mostly at the proof-of-concept stage, some of them have already shown promising results for the treatment of depression.


Implementing a Child Mental Health Intervention in Child Welfare Services: Stakeholder Perspectives on Feasibility.
This pilot study integrated quantitative and qualitative data to examine the feasibility of implementing a modified version of a multiple family group behavioral parent training intervention (The 4Rs and 2Ss for Strengthening Families Program [4Rs and 2Ss]) in child welfare (CW) placement prevention services, from the perspectives of participating caregivers (n = 12) and CW staff (n = 12; i.e., 6 caseworkers, 4 supervisors, and 2 administrators). Quantitative surveys were administered to caregivers and CW staff followed by semi-structured interviews to examine the feasibility of implementing the modified 4Rs and 2Ss program, as well as factors impacting feasibility. Results indicated that quantitative benchmarks for high feasibility were met in all assessed areas (e.g., family recruitment, caseworker fidelity ratings, CW staff feasibility ratings) except for family attendance, which was markedly lower than desired. Factors facilitating feasibility included agency and research support, intervention ease-of-use, perceived benefits to existing CW practice, as well as logistical support (e.g., food, transportation, childcare) promoting attendance. Factors hindering feasibility included conflicts between research-based eligibility criteria and existing client population demographics, research-related processes resulting in delays, CW staff role conflicts, added workload burden, complex family issues, and power differentials inherent to CW services which complicated families' voluntary participation.


Is the whole more than the sum of its parts? Evolutionary trade-offs between burst and sustained locomotion in lacertid lizards.
Trade-offs arise when two functional traits impose conflicting demands on the same design trait. Consequently, excellence in one comes at the cost of performance in the other. One of the most widely studied performance trade-offs is the one between sprint speed and endurance. Although biochemical, physiological and (bio)mechanical correlates of either locomotor trait conflict with each other, results at the whole-organism level are mixed. Here, we test whether burst (speed, acceleration) and sustained locomotion (stamina) trade off at both the isolated muscle and whole-organism level among 17 species of lacertid lizards. In addition, we test for a mechanical link between the organismal and muscular (power output, fatigue resistance) performance traits. We find weak evidence for a trade-off between burst and sustained locomotion at the whole-organism level; however, there is a significant trade-off between muscle power output and fatigue resistance in the isolated muscle level. Variation in whole-animal sprint speed can be convincingly explained by variation in muscular power output. The variation in locomotor stamina at the whole-organism level does not relate to the variation in muscle fatigue resistance, suggesting that whole-organism stamina depends not only on muscle contractile performance but probably also on the performance of the circulatory and respiratory systems.


Dynamic epistasis under varying environmental perturbations.
Epistasis describes the phenomenon that mutations at different loci do not have independent effects with regard to certain phenotypes. Understanding the global epistatic landscape is vital for many genetic and evolutionary theories. Current knowledge for epistatic dynamics under multiple conditions is limited by the technological difficulties in experimentally screening epistatic relations among genes. We explored this issue by applying flux balance analysis to simulate epistatic landscapes under various environmental perturbations. Specifically, we looked at gene-gene epistatic interactions, where the mutations were assumed to occur in different genes. We predicted that epistasis tends to become more positive from glucose-abundant to nutrient-limiting conditions, indicating that selection might be less effective in removing deleterious mutations in the latter. We also observed a stable core of epistatic interactions in all tested conditions, as well as many epistatic interactions unique to each condition. Interestingly, genes in the stable epistatic interaction network are directly linked to most other genes whereas genes with condition-specific epistasis form a scale-free network. Furthermore, genes with stable epistasis tend to have similar evolutionary rates, whereas this co-evolving relationship does not hold for genes with condition-specific epistasis. Our findings provide a novel genome-wide picture about epistatic dynamics under environmental perturbations.


[Ultrastructural characteristics of a monolayer cell culture of the guinea pig pancreatic islet apparatus].
A-, B- and D-insulocytes were revealed when studying the monolayer cell culture of guinea pig pancreatic islands with electron microscope. With an increase in cultivation time (3 days) the cytoplasm ultrastructure in all the cell types changes, the number of secretory granules in B-cells decreases. At later periods (10 days) the signs of secretory cells degeneration and dedifferentiation are observed.


Triple Function of Amelogenin Peptide-Chitosan Hydrogel for Dentin Repair.
Biomimetic strategies like peptide-guided collagen mineralization promise to enhance the effectiveness of dentin remineralization. We recently reported that rationally designed amelogenin-derived peptides P26 and P32 promoted apatite nucleation, mineralized collagen, and showed potential in enamel regrowth and dentin remineralization. To facilitate the clinical application of amelogenin-derived peptides and to uncover their effectiveness in repairing dentin, we have now implemented a chitosan (CS) hydrogel for peptide delivery and have investigated the effects of P26-CS and P32-CS hydrogels on dentin remineralization using 2 in situ experimental models that exhibited different levels of demineralization. The efficacy of the peptide-CS hydrogels in dentin repair was evaluated by characterizing the microstructure, mineral density, mineral phase, and nanomechanical properties of the remineralized samples. The new strategy of atomic force microscopy PeakForce quantitative nanomechanical mapping was used for direct visualization and nanomechanical analysis of repaired dentin lesions across the lesion depth. Results from the 2 models indicated the potential triple functions of peptide-CS hydrogels for dentin repair: building a highly organized protective mineralized layer on dentin, occluding dentinal tubules by peptide-guided in situ mineralization, and promoting biomimetic dentinal collagen remineralization. Importantly, peptides released from the CS hydrogel could diffuse into the dentinal matrix and penetrate the dentinal tubules, leading to both surface and subsurface remineralization and tubule occlusion. Given our previous findings on peptide-CS hydrogels' potential for remineralizing enamel, we see further promise for hydrogels to treat tooth defects involving multiple hard tissues, as in the case of noncarious cervical lesions.


Molecular phylogenetics of the Macaronesian-endemic genus Bystropogon (Lamiaceae): palaeo-islands, ecological shifts and interisland colonizations.
Abstract A molecular phylogenetic study of Bystropogon L'Her. (Lamiaceae) is presented. We performed a cladistic analysis of nucleotide sequences of the internal transcribed spacers (ITS), of the nuclear ribosomal DNA, and of the trnL gene and trnL-trnF intergenic spacer of the chloroplast DNA. Bystropogon odoratissimus is the only species endemic to the Canary Islands that occurs in the three palaeo-islands of Tenerife. This species is not part of an early diverging lineage of Bystropogon and we suggest that it has a recent origin. This phylogenetic pattern is followed by most of the species endemic to the palaeo-islands of Tenerife. The two sections currently recognized in Bystropogon form two monophyletic groups. Taxa belonging to the section Bystropogon clade show interisland colonization limited to the Canary Islands with ecological shifts among three ecological zones. Taxa from the section Canariense clade show interisland colonization both within the Canary Islands and between the Canary Islands and Madeira. Speciation events within this clade are mostly limited to the laurel forest. The genus has followed a colonization route from the Canaries towards Madeira. This route has also been followed by at least five other plant genera with species endemic to Macaronesia. Major incongruences were found between the current infrasectional classification and the molecular phylogeny, because the varieties of Bystropogon origanifolius and Bystropogon canariensis do not form two monophyletic groups. The widespread B. origanifolius appears as progenitor of the other species in section Bystropogon with a more restricted distribution.


Combined in vivo molecular and anatomic imaging for detection of ovarian carcinoma-associated protease activity and integrin expression in mice.
Most patients with epithelial ovarian cancer (EOC) experience drug-resistant disease recurrence. Identification of new treatments is a high priority, and preclinical studies in mouse models of EOC may expedite this goal. We previously developed methods for magnetic resonance imaging (MRI) for tumor detection and quantification in a transgenic mouse model of EOC. The goal of this study was to determine whether three-dimensional (3D) fluorescence molecular tomography (FMT) and fluorescent molecular imaging probes could be effectively used for in vivo detection of ovarian tumors and response to therapy. Ovarian tumor-bearing TgMISIIR-TAg mice injected with fluorescent probes were subjected to MRI and FMT. Tumor-specific probe retention was identified in vivo by alignment of the 3D data sets, confirmed by ex vivo fluorescent imaging and correlated with histopathologic findings. Mice were treated with standard chemotherapy, and changes in fluorescent probe binding were detected by MRI and FMT. Ovarian tumors were detected using probes specific for cathepsin proteases, matrix metalloproteinases (MMPs), and integrin α(v)β(3). Cathepsin and integrin α(v)β(3) probe activation and retention correlated strongly with tumor volume. MMP probe activation was readily detected in tumors but correlated less strongly with tumor volume. Tumor regression associated with response to therapy was detected and quantified by serial MRI and FMT. These results demonstrate the feasibility and sensitivity of FMT for detection and quantification of tumor-associated biologic targets in ovarian tumors and support the translational utility of molecular imaging to assess functional response to therapy in mouse models of EOC.


CT of Postoperative Repair of the Ascending Aorta and Aortic Arch.
While many of the classic open surgical repairs are still used to repair the ascending aorta, management of the aortic arch has become more complex via implementation of newer open surgical and endovascular techniques. Furthermore, techniques are often combined in novel repairs or to allow extended anatomic coverage. As such, a framework that rests on understanding the expected postoperative appearance is necessary for the diagnostic radiologist to best interpret CT studies in these patients. After reviewing the imaging appearances of the common components used in proximal aortic repair, the authors present a structured approach that focuses on the key relevant questions that diagnostic radiologists should consider when interpreting CT studies in these patients. For repair of the ascending aorta, this includes determining whether the aortic valve has been repaired, whether the sinuses of Valsalva have been repaired, and how the coronary arteries were managed, when necessary. In repairs that involve the aortic arch, the relevant considerations relate to management of the arch vessels and the distal extent of the repair. In focusing on these questions, the diagnostic radiologist will be able to identify and describe the vast majority of repairs. Understanding these questions will also facilitate improved understanding of novel repairs, which often use these basic building blocks. Finally, complications-which typically involve infection, noninfectious repair breakdown, hemorrhage, problems with endografts, or disease of the remaining adjacent aorta-will be identifiable as deviations from the expected postoperative appearance. Online supplemental material is available for this article. ©RSNA, 2021.


NAADP-mediated Ca2+ signaling promotes autophagy and protects against LPS-induced liver injury.
LPS has been shown to induce hepatocyte autophagy, but little is known about how LPS is able to do this during acute toxic liver injury. Our aim was to determine the existence of any selective Ca2+ signaling coupling to hepatocyte autophagy in response to LPS. LPS increased the autophagic process in hepatocytes, and CD38 knockdown prevented this response. Ned19, a specific inhibitor for nicotinic acid adenine dinucleotide phosphate (NAADP), prevented LPS-mediated Ca2+ signaling and autophagosome formation in hepatocytes. CD38 overexpression protected the liver from LPS/d-galactosamine (GalN)-induced injury, and NAADP administration promoted autophagosome formation and protected hepatocytes from injury induced by LPS/GalN. Autophagy was promoted by the up-regulation of autophagy-related gene expression via NAADP-mediated Ca2+ signaling in response to LPS. However, CD38-knockout mice displayed down-regulation in hepatocyte gene expression. Ned19 also inhibited the NAADP-stimulated induction of gene expression by inhibiting the LPS-induced nuclear translocation of transcription factor EB (TFEB). Hepatocyte autophagy protects against LPS-induced liver injury via the CD38/NAADP/Ca2+/TFEB pathway. The role of NAADP-mediated Ca2+ signaling in the autophagic process will help elucidate the complexities of autophagy regulation, which is essential toward the discovery of new therapeutic tools against acute liver injury.-Rah, S.-Y., Lee, Y.-H., Kim, U.-H. NAADP-mediated Ca2+ signaling promotes autophagy and protects against LPS-induced liver injury.


Anopheles gambiae genome reannotation through synthesis of ab initio and comparative gene prediction algorithms.
Complete genome annotation is a necessary tool as Anopheles gambiae researchers probe the biology of this potent malaria vector. We reannotate the A. gambiae genome by synthesizing comparative and ab initio sets of predicted coding sequences (CDSs) into a single set using an exon-gene-union algorithm followed by an open-reading-frame-selection algorithm. The reannotation predicts 20,970 CDSs supported by at least two lines of evidence, and it lowers the proportion of CDSs lacking start and/or stop codons to only approximately 4%. The reannotated CDS set includes a set of 4,681 novel CDSs not represented in the Ensembl annotation but with EST support, and another set of 4,031 Ensembl-supported genes that undergo major structural and, therefore, probably functional changes in the reannotated set. The quality and accuracy of the reannotation was assessed by comparison with end sequences from 20,249 full-length cDNA clones, and evaluation of mass spectrometry peptide hit rates from an A. gambiae shotgun proteomic dataset confirms that the reannotated CDSs offer a high quality protein database for proteomics. We provide a functional proteomics annotation, ReAnoXcel, obtained by analysis of the new CDSs through the AnoXcel pipeline, which allows functional comparisons of the CDS sets within the same bioinformatic platform. CDS data are available for download. Comprehensive A. gambiae genome reannotation is achieved through a combination of comparative and ab initio gene prediction algorithms.


Glucagon-like peptide 2: a new treatment for chemotherapy-induced enteritis.
Glucagon-like peptide 2 (GLP-2) is a recently identified intestinal epithelium-specific growth factor that has been shown to reduce the severity of inflammatory disorders of the intestine in rodent models. We hypothesized that GLP-2 administration would be beneficial in chemotherapy-induced enteritis either by preventing injury or by promoting recovery. Rats received no drug (control), chemotherapy alone [5-fluorouracil (5-FU), 190 mg/kg, ip] (Chemo), 5-FU followed by 3 days of GLP-2 analog (ALX-0600, 0.1 microg, sc twice daily) (CH-G), or GLP-2 analog for 6 days prior to 5-FU and for 3 days afterward (G-CH-G). Animals were pair fed. Rats received 5-bromo-2-deoxyuridine (Br-dU, 50 mg/kg, 2.5 h prior to sacrifice on Day 3 postchemotherapy) for immunohistochemical assessment of cellular proliferation. Chemotherapy induced significant reductions in body weight, villus height, and crypt depth compared with controls. Intestinal wet weight, villus height, and crypt depth were significantly higher for the CH-G group compared with the Chemo group. The CH-G group also showed a significant improvement in villus height compared with the G-CH-G group. Crypt depth, but not jejunal wet weight or villus height, was significantly improved in the G-CH-G group compared with the Chemo group. The percentage of Br-dU-labeled cells in the intestinal crypts did not differ among the groups. These results suggest, for the first time, that GLP-2 treatment initiated after chemotherapy administration enhances intestinal recovery. In contrast, GLP-2 treatment initiated prior to chemotherapy administration to prevent injury has less beneficial effect. GLP-2 administration may be beneficial to patients suffering from chemotherapy-induced enteritis.


[The innervation of anterior rudimentary limbs in snake embryos].
In comparison to the innervation of the well developed anterior limb-bud of Lacerta viridis or Emys or bicularis we have observed in embryos of Reptiles with rudimentary limb-buds (Anguis fragilis, Scelotes inornatus, Scelotes brevipes), a reduction in the number of spinal nerves entering the anterior limb-bud; this reduction is parallel to that of the number of spinal nerves entering the anterior limb-bud; this reduction is parallel to that of the number of somites involved in the limb-bud.


Engineering of an E. coli outer membrane protein FhuA with increased channel diameter.
Channel proteins like FhuA can be an alternative to artificial chemically synthesized nanopores. To reach such goals, channel proteins must be flexible enough to be modified in their geometry, i.e. length and diameter. As continuation of a previous study in which we addressed the lengthening of the channel, here we report the increasing of the channel diameter by genetic engineering. The FhuA Δ1-159 diameter increase has been obtained by doubling the amino acid sequence of the first two N-terminal β-strands, resulting in variant FhuA Δ1-159 Exp. The total number of β-strands increased from 22 to 24 and the channel surface area is expected to increase by ~16%. The secondary structure analysis by circular dichroism (CD) spectroscopy shows a high β-sheet content, suggesting the correct folding of FhuA Δ1-159 Exp. To further prove the FhuA Δ1-159 Exp channel functionality, kinetic measurement using the HRP-TMB assay (HRP = Horse Radish Peroxidase, TMB = 3,3',5,5'-tetramethylbenzidine) were conducted. The results indicated a 17% faster diffusion kinetic for FhuA Δ1-159 Exp as compared to FhuA Δ1-159, well correlated to the expected channel surface area increase of ~16%. In this study using a simple "semi rational" approach the FhuA Δ1-159 diameter was enlarged. By combining the actual results with the previous ones on the FhuA Δ1-159 lengthening a new set of synthetic nanochannels with desired lengths and diameters can be produced, broadening the FhuA Δ1-159 applications. As large scale protein production is possible our approach can give a contribution to nanochannel industrial applications.


Domain specific self-efficacy mediates the impact of pain catastrophizing on pain and disability in overweight and obese osteoarthritis patients.
This study examined whether self-efficacy mediated the relationship between pain catastrophizing and pain and disability. Participants were 192 individuals diagnosed with osteoarthritis (OA) of the knees who were overweight or obese. Multiple mediator analyses were conducted to simultaneously test self-efficacy for pain control, physical function, and emotional symptoms as mediators while controlling for demographic and medical status variables. Higher pain catastrophizing was associated with lower self-efficacy in all 3 domains (Ps < .05). Self-efficacy for pain control fully mediated the relationship between pain catastrophizing and pain (beta = .08, Sobel test Z = 1.97, P < .05). The relationship between pain catastrophizing and physical disability was fully mediated by self-efficacy for physical function (beta = .06, Sobel test Z = 1.95, P = .05). Self-efficacy for emotional symptoms partially mediated the relationship between pain catastrophizing and psychological disability (beta = .12, Sobel test Z = 2.92, P < .05). These results indicate that higher pain catastrophizing contributed to greater pain and disability via lower domain-specific self-efficacy. Efforts to reduce pain and improve functioning in OA patients should consider addressing pain catastrophizing and domain specific self-efficacy. Pain catastrophizing may be addressed through cognitive therapy techniques and self-efficacy may be enhanced through practice of relevant skills and personal accomplishments. This study found that higher pain catastrophizing contributed to greater pain and disability via domain specific self-efficacy. These results suggest that treatment efforts to reduce pain and improve functioning in OA patients who are overweight or obese should consider addressing both pain catastrophizing and self-efficacy.


Theories of endometrial carcinogenesis: a multidisciplinary approach.
Historical observations have suggested that endometrial carcinomas vary in histopathologic appearance and clinical features. More recent, systematic studies have provided epidemiologic, clinicopathologic, and molecular support for these observations. Specifically, studies suggest that the most common type of endometrial carcinoma, endometrioid adenocarcinoma, develops from endometrial hyperplasia in the setting of excess estrogen exposure and usually pursues an indolent clinical course. In contrast, a minority of endometrial carcinomas, best represented by serous carcinoma, do not seem to be related to estrogenic risk factors or elevated serum hormone levels, and these tumors seem to develop from atrophic rather than hyperplastic epithelium. We have proposed that serous carcinomas develop from "endometrial intraepithelial carcinoma," a lesion representing malignant transformation of the endometrial surface epithelium. Whereas endometrioid carcinoma and endometrial hyperplasia are associated with microsatellite instability and ras and PTEN mutations, serous carcinoma and endometrial intraepithelial carcinoma are associated with p53 mutations and abnormal accumulation of p53 protein. Based on these data regarding the pathogenesis of endometrioid and serous carcinoma, we have proposed a dualistic model of endometrial carcinogenesis incorporating a "classic" estrogen-driven pathway and an "alternative" pathway seemingly unrelated to hormones. It is hoped that further studies may permit the extension and modification of this model and that these advances will lead to improved diagnosis, management, and prevention.


Structural and mechanistic aspects of evolution of beta-lactamases and penicillin-binding proteins.
Penicillin-binding proteins (PBPs) and b-lactamases are related enzymes, the former are the targets for b-lactam antibiotics and the latter are resistance enzyme to these antibiotics. The two families of enzymes share structural topologies and certain mechanistic features. However, these classes of enzymes have diversified substantially and have broadened the reaction repertoire for their catalytic properties. This report addresses the issues of the evolution of function of these proteins.


Pathogenesis of necrotising enteritis with special reference to intestinal hypersensitivity reactions.
The aetiological aspects of 83 sporadic cases of necrotising enteritis (NE) have been studied. Of 56 cases in which histology of the intestine was possible, eight showed appearances (oedema and local eosinophilia) suggestive of a type I hypersensitivity reaction, while in 37 the appearances were suggestive of a type III reaction. We suggest that these reactions, which were more common in children and young adults, were initiating factors in the intestinal necrosis. The type III reactions (submucosal arteritis, fibrinoid necrosis of arteriolar walls, intramural and perivascular infiltration with polymorphonuclear, mononuclear, and eosinophil cells, and submucous oedema) were in seven cases accompanied by extraintestinal lesions (hypercellularity of glomeruli, amorphous material in the Bowman's capsular space, tubular casts, mononuclear cell infiltration into the hepatic portal tracts, congestion and oedema of the lung) which were compatible with systemic immune complex disease. The mesenteric lymph nodes in 12 out of 15 cases with intestinal arteritis showed appearances indicative of a humoral immune response. We suggest that NE is a two-stage process. In stage 1, a necrotic focus is established in the intestinal mucosa-submucosa by 'initiating' factors of vascular (functional or organic) or microbial (exotoxic, endotoxic, or Shwartzman) origin. Functional circulatory insufficiency in the intestine is of particular relevance to necrotising enteritis in neonates and in adults with traumatic shock or cardiac insufficiency. The jejunal and--to a lesser extent--the ileal microcirculation appear to be particularly vulnerable to microcirculatory insufficiency. Ninety-seven per cent of our cases were of NE of the small intestine of which 76% involved the jejunum alone or as a part of a jejunoileitis. These 'initiating' factors act either singly or synergistically with 'promoting' factors (changes in the volume, composition, or pH of the diet, intestinal stasis, or bacterial factors) in the establishment of necrotic foci in the intestine. Stage 2 results from the colonisation of the necrotic foci by intestinal clostridia, the toxigenic capacity of which will determine the progress of the intestinal lesion. Clinically established NE is essentially gas gangrene of the intestinal wall. Our bacteriological findings (microscopic, cultural, and serological) support a pathogenetic role of Cl. welchii in the established stages of necrotising enteritis. Strains of Cl. welchii from NE cases had significantly higher histidine decarboxylase activity than strains from control sources; it is possible that the resultant histamine production could act as a promoting factor in stage 1. Neutralising antibody against the Wanowrie virus, an Asian arbovirus which produces haemorrhagic enteritis in mice, was absent in the paired sera from 10 cases examined.


Association between diabetes and increased prevalence of paranasal sinus disease: a cross-sectional study in Japanese adults.
The association between diabetes and paranasal sinus disease has not been thoroughly investigated. We cross-sectionally investigated the association between diabetes and the presence of paranasal sinus disease, which was confirmed by a head MRI scan in 1350 adults who underwent a health screening program focusing on brain diseases and metabolic syndrome. Logistic regression, which was adjusted for age, sex, body mass index, waist-to-hip ratio, hypertension, smoking status, alcohol intake, and white blood cell count, was performed to calculate the odds ratio (OR) of having paranasal sinus disease among adults with diabetes in relation to those without. The dose-response relationship between hemoglobin A1c (HbA1c) levels and the presence of paranasal sinus disease was also investigated. Of the 1350 participants (mean age, 61.6 ± 10.0 years; 71.6% men), 220 diabetes cases were identified. Paranasal sinus disease was diagnosed in 151 adults. The adjusted OR of having paranasal sinus disease was 1.74 (95% confidence interval [CI], 1.12-2.71) in those with diabetes. The odds of having paranasal sinus disease increased with HbA1c levels. Compared to those with HbA1c of ≤5.4%, those with HbA1c of 5.5%-6.4%, 6.5%-7.9%, and ≥8.0% were more likely to have paranasal sinus disease, with adjusted ORs of 1.32 (95% CI, 0.88-1.98), 1.63 (95% CI, 0.86-3.09) and 2.71 (95% CI, 1.12-6.61), respectively (P for trend = 0.019). Diabetes may be significantly associated with higher prevalence of paranasal sinus disease in Japanese adults. We should keep this increased risk in mind when a diabetic patient is suspected of having paranasal sinus disease.


CD4+25+ regulatory T cells limit Th1-autoimmunity by inducing IL-10 producing T cells following human lung transplantation.
Chronic human lung allograft rejection is manifested by bronchiolitis obliterans syndrome (BOS). BOS has a multifactorial etiology. Previous studies have indicated that both cellular and humoral alloimmunity play a significant role in the pathogenesis of BOS. Recently, autoimmunity has also been demonstrated to contribute to lung allograft rejection in animal models. However, the significance of autoimmunity in BOS remains unknown. In this report, we investigated the role of naturally occurring CD4(+)CD25(+) regulatory T cells (T-regs) in modulating cellular autoimmunity to collagen type V (col-V), a 'sequestered' yet immunogenic self-protein present in the lung tissue, following lung transplantation (LT). We demonstrated that col-V reactive CD4(+) T cells could be detected in the peripheral blood of lung transplant recipients. There was a predominance of IL-10 producing T cells (T(IL-10)) reactive to col-V with significantly lower levels of IFN-gamma and IL-2 producing T cells (Th1 cells). The col-V specific T(IL-10) cells suppressed the proliferation and expansion of col-V specific Th1 cells by IL-10-dependent and contact-independent pathways. The T(IL-10) cells were distinct but their development was dependent on the presence of T-regs. Furthermore, during chronic lung allograft rejection there was a significant decline of T(IL-10) cells with concomitant expansion of col-V-specific IFN-gammaproducing Th1 cells.


Non-invasive stress evaluation in domestic horses (Equus caballus): impact of housing conditions on sensory laterality and immunoglobulin A.
The study aimed to evaluate sensory laterality and concentration of faecal immunoglobulin A (IgA) as non-invasive measures of stress in horses by comparing them with the already established measures of motor laterality and faecal glucocorticoid metabolites (FGMs). Eleven three-year-old horses were exposed to known stressful situations (change of housing, initial training) to assess the two new parameters. Sensory laterality initially shifted significantly to the left and faecal FGMs were significantly increased on the change from group to individual housing and remained high through initial training. Motor laterality shifted significantly to the left after one week of individual stabling. Faecal IgA remained unchanged throughout the experiment. We therefore suggest that sensory laterality may be helpful in assessing acute stress in horses, especially on an individual level, as it proved to be an objective behavioural parameter that is easy to observe. Comparably, motor laterality may be helpful in assessing long-lasting stress. The results indicate that stress changes sensory laterality in horses, but further research is needed on a larger sample to evaluate elevated chronic stress, as it was not clear whether the horses of the present study experienced compromised welfare, which it has been proposed may affect faecal IgA.


The inclusion of women, girls and gender equality in National Strategic Plans for HIV and AIDS in southern and eastern Africa.
The global HIV and AIDS epidemics disproportionately affect women, particularly young women in southern and eastern Africa. UNAIDS, amongst other actors, has singled out National Strategic Plans for HIV and AIDS (NSPs) as a critical platform for ensuring that women and girls are meaningfully included in national HIV and AIDS responses. Despite this, there is little evidence as to how or whether NSPs integrate responses to women and girls. Using a collaboratively developed framework, we assessed how 20 countries in southern and eastern Africa integrated women and girls in their NSPs. We identified that in general there is poor inclusion, apart from access to post-exposure prophylaxis in the case of sexual violence and access to vertical transmission services. Drawing on Moser's distinction between women's practical and strategic interests, we suggest that overall women and girls are poorly included in NSPs, and where there are policies and programmes, there is an overwhelming focus on women's practical interests, without any consideration of women's strategic interests. We argue that this limits the potential of NSPs to be platforms for national responses that meaningfully seek to transform gender relations.


Water is a preservative of microbes.
Water is the cellular milieu, drives all biochemistry within Earth's biosphere and facilitates microbe-mediated decay processes. Instead of reviewing these topics, the current article focuses on the activities of water as a preservative-its capacity to maintain the long-term integrity and viability of microbial cells-and identifies the mechanisms by which this occurs. Water provides for, and maintains, cellular structures; buffers against thermodynamic extremes, at various scales; can mitigate events that are traumatic to the cell membrane, such as desiccation-rehydration, freeze-thawing and thermal shock; prevents microbial dehydration that can otherwise exacerbate oxidative damage; mitigates against biocidal factors (in some circumstances reducing ultraviolet radiation and diluting solute stressors or toxic substances); and is effective at electrostatic screening so prevents damage to the cell by the intense electrostatic fields of some ions. In addition, the water retained in desiccated cells (historically referred to as 'bound' water) plays key roles in biomacromolecular structures and their interactions even for fully hydrated cells. Assuming that the components of the cell membrane are chemically stable or at least repairable, and the environment is fairly constant, water molecules can apparently maintain membrane geometries over very long periods provided these configurations represent thermodynamically stable states. The spores and vegetative cells of many microbes survive longer in the presence of vapour-phase water (at moderate-to-high relative humidities) than under more-arid conditions. There are several mechanisms by which large bodies of water, when cooled during subzero weather conditions remain in a liquid state thus preventing potentially dangerous (freeze-thaw) transitions for their microbiome. Microbial life can be preserved in pure water, freshwater systems, seawater, brines, ice/permafrost, sugar-rich aqueous milieux and vapour-phase water according to laboratory-based studies carried out over periods of years to decades and some natural environments that have yielded cells that are apparently thousands, or even (for hypersaline fluid inclusions of mineralized NaCl) hundreds of millions, of years old. The term preservative has often been restricted to those substances used to extend the shelf life of foods (e.g. sodium benzoate, nitrites and sulphites) or those used to conserve dead organisms, such as ethanol or formaldehyde. For living microorganisms however, the ultimate preservative may actually be water. Implications of this role are discussed with reference to the ecology of halophiles, human pathogens and other microbes; food science; biotechnology; biosignatures for life and other aspects of astrobiology; and the large-scale release/reactivation of preserved microbes caused by global climate change.


Small deletion at the 7q21.2 locus in a CCM family detected by real-time quantitative PCR.
Cerebral cavernous malformations (CCMs) represent a common autosomal dominant disorder that predisposes patients to haemorrhagic strokes and focal neurological signs. About 56% of the hereditary forms of CCMs have been so far associated with mutations in the KRIT1 (Krev Interaction Trapped 1) gene, located at 7q21.2 (CCM1 locus). We described the complete loss of 7q21.2 locus encompassing the KRIT1 gene and 4 flanking genes in a CCM family by using a dense set of 12 microsatellite markers. The complete loss of the maternal copy of KRIT1 gene region was confirmed by Real-Time Quantitative Polymerase Chain Reaction (RT-QPCR) and the same approach was used for expression analysis. Additional RT-QPCR analysis showed the extension of the deletion, for a total of 700 kb, to the adjacent downstream and upstream-located genes, MTERF, AKAP9, CYP51A1, as well as a partial loss of the ANKIB1 gene. Here we report the molecular characterization of an interstitial small genomic deletion of the 7q21.2 region in a CCMs affected family, encompassing the KRIT1 gene. Our findings confirm the loss of function mechanism for the already known CCM1 locus, without any evident involvement of the other deleted genes. Moreover, our investigations highlight the usefulness of the RT-QPCR to the molecular characterization of the breakpoints genomic deletions and to the identification of internal deleted genes involved in the human genetic diseases.


[Gouty arthritis in a total knee prosthesis].
This article describes a 69-year-old man with a bilateral total knee prosthesis in whom both knees were swollen and painful. Fever was present and the levels of inflammation markers were markedly elevated. Gout was diagnosed in the right knee and haemarthros during calcium carbasalate use in the left knee. Gout in a knee with a total knee prosthesis is rare and can mimic septic arthritis. Early diagnosis is of particular importance as the therapy of the two conditions differs considerably. Gout is treated with medication while surgical treatment is necessary in a case of septic arthritis, which may even involve removal of the prosthesis.


Influence of obesity and malnutrition on the metabolism of some cancer-related hormones.
It has been suggested that the well-documented relationship of dietary composition to the incidence of human breast cancer is mediated by the effects of dietary constituents on hormone levels. There is fairly good evidence for diet-hormone relationships in animals, but the evidence in humans is unconvincing. In this paper, we describe three of our findings relating nutrition to hormone levels: (a) that obesity causes retention of a tracer of estradiol in women but not in men, a finding we attribute to the presence of specific estrogen receptor in the adipose tissue of women but not men; (b) that obese men have elevated plasma estrone and estradiol levels but obese women do not, a finding we attribute to greater androstenedione-to-estrone conversion in the adipose tissue of men than in that of women; and (c) that cachectic girls with anorexia nervosa fail to have the normal nocturnal surge of prolactin secretion, a finding that we attribute to deficiency of tryptophan, which is an adequate stimulus for prolactin secretion. These findings give support to the concept that dietary factors affect hormone secretion and/or metabolism in humans.


Osmium Complex-Chromophore Conjugates with Both Singlet-to-Triplet Absorption and Long Triplet Lifetime through Tuning of the Heavy-Atom Effect.
Os(II) complexes showing singlet-to-triplet absorption are of growing interest as a new class of triplet sensitizers that circumvent energy loss during intersystem crossing, and they enable effective utilization of input photon energy in various applications, such as photoredox catalysis, photodynamic therapy, and photon upconversion. However, triplet excited-state lifetimes of Os(II) complexes are often too short (τ < 1 μs) to transfer their energy to neighboring molecules. While the covalent conjugation of chromophores has been known to extend the net excited-state lifetimes through an intramolecular triplet energy transfer (IMET), heavy-atom effects of the central metals on the attached chromophore units have rarely been discussed. Here, we investigate the relationship between the spin-density contribution of the heavy metals and the net triplet excited-state lifetimes for a series of Os(II) and Ru(II) bis(terpyridine) complexes modified with perylene units. Phosphorescence lifetimes of these compounds strongly depend on the lifetimes of the perylenyl group-localized excited states that are shortened by the heavy-atom effect. The degree of heavy-atom effect can be largely circumvented by introducing meta-phenylene bridges, where the perylene unit retains its intrinsic long excited-state lifetime. The thermal activation to the short-lived excited states is suppressed, thanks to sufficient but still small energy losses during the IMET process. Involvement of the metal center was also confirmed by the prolonged lifetime by replacing Os(II) with Ru(II) that possesses a smaller spin-orbit coupling constant. These results indicate the importance of ligand structures that give a minimum heavy-atom effect as well as the sufficient energy gap among the excited states and fast IMET for elongating the triplet excited-state lifetime without sacrificing the excitation energy.


Antimicrobial triterpenoid glycosides from Cephalaria scoparia.
Five new oleanane-type triterpenoid glycosides (1-5), named scoposides A-E, along with one known bisdesmosidic triterpene glycoside, were isolated from the aerial parts of Cephalaria scoparia. Three prosapogenins (2a-4a) were also obtained after alkaline hydrolysis of the bisdesmosidic compounds 2-4. The structures of compounds 1-5 were determined by spectroscopic (1D and 2D NMR, HRESIMS) and chemical methods. The antimicrobial effects of compounds 1-5 were evaluated against a small panel of Gram-positive and Gram-negative bacteria.


Chemogenomic profiling of endogenous PARK2 expression using a genome-edited coincidence reporter.
Parkin, an E3 ubiquitin ligase, is a central mediator of mitochondrial quality control and is linked to familial forms of Parkinson's disease (PD). Removal of dysfunctional mitochondria from the cell by Parkin is thought to be neuroprotective, and pharmacologically increasing Parkin levels may be a novel therapeutic approach. We used genome-editing to integrate a coincidence reporter into the PARK2 gene locus of a neuroblastoma-derived cell line and developed a quantitative high-throughput screening (qHTS) assay capable of accurately detecting subtle compound-mediated increases in endogenous PARK2 expression. Interrogation of a chemogenomic library revealed diverse chemical classes that up-regulate the PARK2 transcript, including epigenetic agents, drugs controlling cholesterol biosynthesis, and JNK inhibitors. Use of the coincidence reporter eliminated wasted time pursuing reporter-biased false positives accounting for ∼2/3 of the actives and, coupled with titration-based screening, greatly improves the efficiency of compound selection. This approach represents a strategy to revitalize reporter-gene assays for drug discovery.


A positive feedback loop between alpha1-adrenoceptors and inflammatory cytokines in keratinocytes.
A positive feedback loop between inflammatory cytokines and alpha1-adrenoceptors (α1-AR) (a target of the sympathetic nervous system neurotransmitter norepinephrine) influences inflammatory responses in immune cells. This cross-talk between the sympathetic nervous system and immune system may play a role in promoting chronic inflammation. Emerging evidence shows that α1-AR interact with inflammatory cytokines in keratinocytes, and this epidermal adrenergic signalling may contribute to skin inflammatory responses following injury, disease or stress. In this study, utilizing an in vitro approach, we hypothesized that α1-AR interact in a positive feedback loop with inflammatory mediators in keratinocytes. The pro-inflammatory cytokine tumor necrosis factor α (TNFα) was used to induce an inflammatory state in cultured keratinocytes. TNFα increased interleukin (IL)-1β, IL-6, IL-8 and nerve growth factor (NGF) production and gene expression levels of α1-AR subtype B (α1B-AR). Additional stimulation of α1-AR further increased IL-6 levels, while maintaining high levels of IL-8 and decreasing levels of IL-1β and NGF. Our results suggest that reciprocal influences between α1-ARs and inflammatory cytokines may play a role in normal inflammatory responses. However, if unchecked, this cycle could contribute to pathology (e.g. chronic inflammatory diseases, chronic pain conditions, and stress-induced cancer progression).


ADH genotype does not modify the effects of alcohol on high-density lipoprotein.
Alcohol consumption has beneficial effects on mortality which are mainly due to reduction in cardiovascular disease. These are believed to be due, at least in part, to the increase in plasma high-density lipoprotein (HDL) which is associated with alcohol consumption. It has been proposed that ADH3 genotype modifies the relationships between alcohol intake and cardiovascular disease by altering the HDL response to alcohol. The aim of this paper was to test for effects of ADH2 and ADH3 genotypes on the response of HDL components to habitual alcohol consumption. Adult male and female subjects were genotyped for ADH2 and ADH3; and plasma HDL cholesterol, apolipoprotein A-I, and apolipoprotein A-II were measured. Nine hundred one subjects had both ADH2 and ADH3 genotypes and HDL cholesterol results, while 753 had both genotypes and all three lipid results. The effect of alcohol intake on the three measured HDL components, and a factor score derived from them, was estimated for each of the ADH2 and ADH3 genotype groups. All the measured components of HDL increased with increasing alcohol consumption over the range of intakes studied, 0-4 drinks per day. There were no significant interactions between alcohol consumption and ADH2 or ADH3 genotypes. The concept that alcohol dehydrogenase genotype and alcohol metabolic rate modify the effects of alcohol on plasma HDL concentration is not supported by our results.


[A Japanese version of the FLANDERS handedness questionnaire].
Quantitative assessment of handedness is required in various clinical and research settings in psychology, neuroscience, and medicine. In the present study we tested the reliability and validity of a Japanese version of the FLANDERS handedness questionnaire, which was a new measure of skilled hand preference originally reported by Nicholls, Thomas, Loetscher, and Grimshaw (2013). Participants (N=431) completed three types of handedness questionnaires: the FLANDERS handedness questionnaire, Edinburgh Handedness Inventory, and H · N handedness test. Factor analysis revealed that the Japanese version of FLANDERS handedness questionnaire had a single-factor structure and high internal consistency. This questionnaire also posssed high test-retest reliability and criterion-referenced validity. These results indicate that the Japanese version of the FLANDERS handedness questionnaire is a valid and useful measure of skilled hand preference for Japanese participants.


IDDM in BB rats. Enhanced MHC class I heavy-chain gene expression in pancreatic islets.
Modulation in major histocompatibility complex (MHC) gene expression correlates with the inflammatory reactions that occur during graft rejection and autoimmune disease. We analyzed the expression of class I and II MHC genes in the pancreatic islets of prediabetic and newly diabetic BB rats by immunohistochemistry of tissue sections and Northern blotting of RNA extracted from isolated islets. We show that enhanced levels of MHC class I heavy-chain RNA are present in pancreatic islets before overt inflammation and the onset of insulin-dependent diabetes mellitus (IDDM) in the spontaneously diabetic BB rat. Immunohistochemical analysis revealed enhanced class I antigen expression throughout the pancreatic islets of newly diabetic animals but no induction of class II antigen on endocrine cells within the islet. Varying degrees of inflammatory infiltrate were observed in the sections exhibiting enhanced class I antigen expression or in nearby serial sections. Southern blot analysis revealed no restriction-fragment-length polymorphism or amplification of the endogenous class I heavy-chain genes compared with those of seroidentical disease-resistant Wistar-Furth rats. I-A alpha and I-E alpha hybridizing RNA appeared de novo before overt diabetes, although concomitantly with T-lymphocyte-receptor beta-chain and interferon-gamma gene hybridizing RNA and after MHC class I heavy-chain RNA enhancement was observed. These data indicate the possibility that enhanced class I heavy-chain gene expression plays a role in the progression of IDDM.


The Driver Behaviour Questionnaire in Arab Gulf countries: Qatar and United Arab Emirates.
Manchester Driver Behaviour Questionnaire (DBQ) is one of the most widely used instruments for measuring self-reported driving style and investigating the relationship between driving behaviour and accident involvement. In spite of the fact that Arab Gulf countries have a higher road accident fatality rate compared to European countries and USA, the DBQ has not been used in Arab countries so far. The aim of the present study was to investigate the factor structure of the DBQ, then to examine the relationships between the factors of the DBQ and accident involvement, and finally to compare DBQ scores between the two gulf countries: Qatar and United Arab Emirates (UAE). In this study, 1110 Qatari (263 females and 847 males) and 1286 UAE drivers (294 females and 992 males) filled a survey questionnaire including the DBQ and background information. The results showed that UAE drivers scored higher on almost all DBQ items than Qatari drivers. Surprisingly, only very small differences between men and women on the DBQ item scores were found in UAE. Factor analysis resulted in four factors, which were named as errors, pushing-speeding violations, lapses, and aggression-speeding violations. However, there were a number of differences in the factor structure of the DBQ in UAE and Qatar when compared to the theoretical four-factor structure of the DBQ. Reliabilities of some subscales were also questionably lower than in the original British data. Logistic regression analyses showed that errors, lapses, and aggression-speeding violations predicted accident involvement in Qatar but not in UAE after controlling the effect of the demographic variables (age, sex, and annual mileage).


The septum as origin of a lordosis-inhibiting influence in female rats: effect of neural transection.
The septum as the origin of a lordosis-inhibiting influence in female rats has been studied. Prior to testing, brain surgery was performed. Some rats were given a ventromedial cut (VMC) just above the anterior commissure, other rats, a posterior transverse cut (PC) or a dorsal horizontal cut (DC) just below the corpus callosum, and still other rats, a bilateral ventrolateral cut (VLC) in the ventrolateral septum and the bed nucleus of the stria terminalis (BST). Additionally, castrated females and those with a VLC sham operation served as castrated controls and sham-operated controls, respectively. All rats were given a daily dose of 0.2 micrograms of estradiol benzoate (EB) for three days and 0.5 mg of progesterone just prior to testing. The castrated controls evidenced no lordotic response. Lordotic activities also were very low in the surgically operated groups, with the exception of the VLC rats who demonstrated significantly high lordotic scores following the administration of 0.2 micrograms of EB. Two further testings, with the EB increased to 0.5 micrograms, were conducted, the interval between these tests being 2 weeks, and the VLC rats again demonstrated significantly higher lordotic activity than did the other rats. These results suggest that the septum exerts a lordosis-inhibiting influence, and that the ventrolateral outputs play a critical role in sending this inhibitory signal.


Putative mechanisms of cough after treatment with angiotensin converting enzyme inhibitors.
A dry cough is the most common adverse effect of angiotensin converting enzyme (ACE) inhibitors, even if it is not particularly serious. Controlled studies have suggested that the incidence may eventually be as high as 20% and the problem seems to occur much more often in women. The mechanism of the cough associated with ACE inhibitor treatment is unrelated to inhibition of the renin-angiotensin system because treatment with angiotensin receptor blockers and renin inhibitors has not caused similar problems. Other substrates of ACE have been implicated, and the side effect has been linked in particular to an accumulation of bradykinin or tachykinins in the airways with consequent stimulation of vagal afferents that subserve the cough reflex, particularly the non-myelinated or C fibres. Some of the effects of bradykinin in particular may be due to secondary activation of prostaglandins (PG), especially PGE2 and PGI2. Anecdotal evidence that cyclooxygenase inhibitors prevent the cough has not been sustained in well controlled clinical trials, but recent evidence suggests that inhaled cromoglycate may have a significant inhibitory effect. The mechanism of action of cromoglycate is not well understood but evidence of inhibition of local neural reflexes has been inferred, mostly from animal studies. The observation seems unlikely to have much practical benefit for it is difficult to envisage routine use of an inhaled agent to prevent a drug side effect. The question of whether ACE inhibitors are safe in patients with asthma is still open, and most rechallenge studies have shown little effect on lung function. Data from one large-scale surveillance study suggest that a few individuals may experience dyspnoea and wheezing but no causal relationship has been established. Delineation of the mechanism of the ACE inhibitor cough may lead to a better understanding of the mechanism of cough in inflammatory airways disease.


Chronic tricuspid regurgitation and bifascicular block due to blunt chest trauma.
Closed-chest trauma in a young man was followed by rupture of a right ventricular papillary muscle and bifascicular block. This produced signs and symptoms of tricuspid regurgitation and recurrent syncope. Treatment by valve replacement and pacemaker implantation was successful. Review of 30 cases of traumatic tricuspid regurgitation reveals that this patient had characteristic findings: adult onset of isolated tricuspid regurgitation, a history of trauma, right bundle branch block, and cardiomegaly without signs of left ventricular failure. In addition, right atrial hypertension of longstanding may produce cyanosis because of right-left shunting through a patent foramen ovale.


Beyond dopamine: glutamate as a target for future antipsychotics.
The dopamine hypothesis of schizophrenia remains the primary theoretical framework for the pharmacological treatment of the disorder. Despite various lines of evidence of dopaminergic abnormalities and reasonable efficacy of current antipsychotic medication, a significant proportion of patients show suboptimal treatment responses, poor tolerability, and a subsequent lack of treatment concordance. In recent decades, intriguing evidence for the critical involvement of other neurotransmitter systems in the pathophysiology of schizophrenia has emerged, most notably of dysfunctions within the glutamate pathways. Consequently, the glutamate synapse has arisen as a promising target for urgently needed novel antipsychotic compounds-particularly in regards to debilitating negative and cognitive symptoms poorly controlled by currently available drugs. In this paper, recent findings integrating glutamatergic and dopaminergic abnormalities in schizophrenia and their implications for novel pharmacological targets are discussed. An overview of compounds in various stages of development is given: drugs enhancing NMDA receptor function as well as metabotropic glutamate receptor (mGluR) agonist and positive allosteric modulators (PAMs) are emphasised. Together with other agents more indirectly affecting glutamatergic neurotransmission, their potential future role in the pharmacotherapy of schizophrenia is critically evaluated.


NMDA receptors in GABAergic synapses during postnatal development.
GABA (gamma-aminobutyric-acid), the main inhibitory neurotransmitter in the adult brain, exerts depolarizing (excitatory) actions during development and this GABAergic depolarization cooperates with NMDARs (N-methyl-D-aspartate receptors) to drive spontaneous synchronous activity (SSA) that is fundamentally important for developing neuronal networks. Although GABAergic depolarization is known to assist in the activation of NMDARs during development, the subcellular localization of NMDARs relative to GABAergic synapses is still unknown. Here, we investigated the subcellular distribution of NMDARs in association with GABAergic synapses at the developmental stage when SSA is most prominent in mice. Using multiple immunofluorescent labeling and confocal laser-scanning microscopy in the developing mouse hippocampus, we found that NMDARs were associated with both glutamatergic and GABAergic synapses at postnatal day 6-7 and we observed a direct colocalization of GABA(A)- and NMDA-receptor labeling in GABAergic synapses. Electron microscopy of pre-embedding immunogold-immunoperoxidase reactions confirmed that GluN1, GluN2A and GluN2B NMDAR subunits were all expressed in glutamatergic and GABAergic synapses postsynaptically. Finally, quantitative post-embedding immunogold labeling revealed that the density of NMDARs was 3 times higher in glutamatergic than in GABAergic synapses. Since GABAergic synapses were larger, there was little difference in the total number of NMDA receptors in the two types of synapses. In addition, receptor density in synapses was substantially higher than extrasynaptically. These data can provide the neuroanatomical basis of a new interpretation of previous physiological data regarding the GABA(A)R-NMDAR cooperation during early development. We suggest that during SSA, synaptic GABA(A)R-mediated depolarization assists NMDAR activation right inside GABAergic synapses and this effective spatial cooperation of receptors and local change of membrane potential will reach developing glutamatergic synapses with a higher probability and efficiency even further away on the dendrites. This additional level of cooperation that operates within the depolarizing GABAergic synapse, may also allow its own modification triggered by Ca(2+)-influx through the NMDA receptors.


Characterization of periodontitis in people with type 1 diabetes of 50 years or longer duration.
Periodontitis is more common and severe in people with diabetes than the general population. We have reported in the Joslin Medalist Study that people with type 1 diabetes of ≥50 years (Medalists) may have endogenous protective factors against diabetic nephropathy and retinopathy. In this cross-sectional study, the prevalence of periodontitis according to the Centers for Disease Control/American Academy of Periodontology classification in a subset (n = 170, mean age = 64.6 ± 6.9 years) of the Medalist cohort, and its associations to various criteria of periodontitis and diabetic complications were assessed. The prevalence of severe periodontitis in Medalists was only 13.5% which was lower than reported levels in diabetic patients of similar ages. Periodontal parameters, including bleeding on probing, plaque index, gingival index, and demographic traits, including male sex, chronological age, and age at diagnosis were significantly associated with severity of periodontitis, which did not associate with diabetes duration, hemoglobin A1c (HbA1c), body mass index, and lipid profiles. Random serum C-peptide levels inversely associated with severity of periodontitis (P = 0.03), lower probing depth (P = 0.0002), and clinical attachment loss (P = 0.03). Prevalence of cardiovascular diseases (CVD) and systemic inflammatory markers, plasma interleukin-6 (IL-6), and serum immunoglobulin G titer against Porphyromonas gingivalis positively associated with severity of periodontitis (P = 0.002 and 0.02, respectively). Antibody titer to P. gingivalis correlated positively and significantly with CVD, serum IL-6, and high-sensitivity C-reactive protein. Some Medalists could be protected from severe periodontitis even with hyperglycemia. Endogenous protective factors for periodontitis could possibly be related to residual insulin production and lower levels of chronic inflammation.


[Priapism. Our experience].
We report our experience in the diagnosis, medical management and surgical treatment of priapism. The clinical records of 67 patients that had been treated for priapism at our hospital from 1978-1994 were analyzed for age, race, occupation, marital status, symptoms, treatment, complications and time to remission of symptoms following treatment. Sixty-four patients were treated surgically and 3 patients were successfully treated medically. We performed 52 cavernospongiosal shunts, 14 sapheno-cavernous shunts and in 10 patients corpora cavernosa puncture was performed. The complications were sepsis, thrombosis, hematoma, phlebitis and urethral fistula. The complication rate was low for the cavernospongiosal shunt and corpora cavernosa puncture with phenylephrine. The highest incidence was observed in patients aged 15-40 years, with a prevalence in the black race. The principal presenting feature was a painful erection without sexual contact. Idiopathic priapism was the most prevalent, although the sickle cell anemia was an important etiological factor. The surgical technique most commonly used was the cavernospongiosal shunt. Intracavernosal injection of phenylephrine was the treatment of choice for pharmacological priapism. Seven patients had erectile dysfunction.


[Folacin determination in the blood by a fluorimetric method].
To determine the blood folacin a fluorometric method is proposed. This includes the following stage-wise operations: liberation of combined forms of folacin under the effect of enzymes contained in the blood investigated; quantification of tyrosine to make a correction for changed fluorescence intensity of this amino acid during oxilation of the solution; measurement of the fluorescence of the substances present in the solution before and after their oxidation with potassium permanganate and calculation of the folacin content. The fluorometric method enables it to greatly speed up the assay by comparison with the microbiological method of determination with the Lactobac. casei as a test-organism, the assay time being cut down from 4-5 days to 4-5 hours without prejudicing the accuracy of the determination.


Abundance of Diaprepes abbreviatus (L.) (Coleoptera: Curculionidae) neonates falling to the soil under tree canopies in Florida citrus groves.
The purpose of these experiments was to estimate the number and distribution of Diaprepes abbreviatus (L.) neonate larvae dropping from the canopy of infested citrus trees. The number of neonates was monitored in the field using passive funnel traps in two simultaneous experiments and a separate experiment for an additional year. In one experiment, traps were placed from trunk to dripline in the cardinal directions under each of five trees (132 traps total). In a second experiment, eight traps were placed under each tree in the cardinal directions, one trap 30 cm from the trunk and one trap 30 cm from the dripline/direction for 25 trees (200 traps total). Larvae were collected weekly for 50 wk in conical tubes containing ethylene glycol as a preservative. Traps closer to the tree trunk captured more larvae than traps nearer the dripline. The area under the tree canopy was positively correlated with the total estimated number of larvae captured per tree. The estimated number of total larvae/tree over the course of our experiments ranged from 955 to 7,290. The highest number of neonate larvae observed in 1 wk was 67 +/- 6/m2. There was an inverse relationship between the number of traps beneath a tree and the number of trees that needed to be sampled to estimate mean population density with a given precision. However, there was a direct relationship between number of traps/tree and the total number of traps needed for a given precision. This passive technique could be used to quantify the destructive larval stage and to assess D. abbreviatus management strategies.


Novel vaccine antigen combinations elicit protective immune responses against Escherichia coli sepsis.
Systemic infections caused by extraintestinal pathogenic Escherichia coli (ExPEC) have emerged as the most common community-onset bacterial infections and are major causes of nosocomial infections worldwide. The management of ExPEC infections has been complicated by the heterogeneity of ExPEC strains and the emergence of antibiotic resistance, thus their prevention through vaccination would be beneficial. The protective efficacy of four common ExPEC antigen candidates composed of common pilus antigens EcpA and EcpD and iron uptake proteins IutA and IroN, were tested by both active and passive immunization in lethal and non-lethal murine models of sepsis. Additionally, antibody raised to a synthetic form of a conserved surface polysaccharide, β-(1-6)-linked poly-N-acetylglucosamine (dPNAG) containing 9 monomers of (non-acetylated) glucosamine (9GlcNH2) conjugated to tetanus toxoid TT (9GlcNH2-TT) was tested in passive immunization protocols. Active immunization of mice with recombinant antigens EcpA, EcpD, IutA, or IroN elicited high levels of total IgG antibody of IgG1/IgG2a isotypes, and were determined to be highly protective against E. coli infection in lethal and non-lethal sepsis challenges. Moreover, passive immunization against these four antigens resulted in significant reductions of bacteria in internal organs and blood of the mice, especially when the challenge strain was grown in iron-restricted media. Inclusion of antibodies to PNAG increased the efficacy of the passive immunization under conditions where the challenge bacteria were grown in LB medium but not in iron-restricted media. The information and data presented are the first step toward the development of a broadly protective vaccine against sepsis-causing E. coli strains.


Structural basis of biological nitrile reduction.
The enzyme QueF catalyzes the reduction of the nitrile group of 7-cyano-7-deazaguanine (preQ(0)) to 7-aminomethyl-7-deazaguanine (preQ(1)), the only nitrile reduction reaction known in biology. We describe here two crystal structures of Bacillus subtilis QueF, one of the wild-type enzyme in complex with the substrate preQ(0), trapped as a covalent thioimide, a putative intermediate in the reaction, and the second of the C55A mutant in complex with the substrate preQ(0) bound noncovalently. The QueF enzyme forms an asymmetric tunnel-fold homodecamer of two head-to-head facing pentameric subunits, harboring 10 active sites at the intersubunit interfaces. In both structures, a preQ(0) molecule is bound at eight sites, and in the wild-type enzyme, it forms a thioimide covalent linkage to the catalytic residue Cys-55. Both structural and transient kinetic data show that preQ(0) binding, not thioimide formation, induces a large conformational change in and closure of the active site. Based on these data, we propose a mechanism for the activation of the Cys-55 nucleophile and subsequent hydride transfer.


Sodium-, chloride-, and mibefradil-sensitive calcium channels in intestinal pacing in wild-type and W/WV mice.
Pacing of intestinal smooth muscle is driven by a network of cells found in the myenteric plexus called the interstitial cells of Cajal (ICC-MP), which produce a rhythmic pacemaker current. Using intact segments of circular (CM) and longitudinal (LM) muscle from wild-type and W/WV mice, we found that sodium-, chloride-, and mibefradil-sensitive ion channel currents are required for normal pacing to occur. Application of 30 micromol/L and 300 micromol/L lidocaine, 1 mmol/L 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), 50 nmol/L and 500 nmol/L mibefradil, or low sodium Krebs significantly reduced pacing frequency in LM and CM. However, simultaneously applying DIDS and lidocaine or low sodium Krebs solution did not completely block pacing nor did it have an additive effect. Lidocaine and low sodium Krebs solution also abolished the gradient of pacing frequencies (higher proximally) found throughout the intestine, resulting in a uniform contraction frequency of 30-40/min. In W/WV mice, which lack ICC-MP, application of DIDS and lidocaine had no effect on the robust pacing in LM segments. In conclusion we found that sodium-, chloride-, and mibefradil-sensitive channel activities were required for normal pacing and to maintain the pacing gradient found throughout the intestines in wild-type but not W/WV mice.


Acetal Formation of Flavoring Agents with Propylene Glycol in E-Cigarettes: Impacts on Indoor Partitioning and Thirdhand Exposure.
The widespread use of flavored e-cigarettes has led to a significant rise in teenage nicotine use. In e-liquids, the flavor carbonyls can form acetals with unknown chemical and toxicological properties. These acetals can cause adverse health effects on both smokers and nonsmokers through thirdhand exposure. This study aims to explore the impacts of these acetals formed in e-cigarettes on indoor partitioning and thirdhand exposure. Specifically, the acetalization reactions of commonly used flavor carbonyls in laboratory-made e-liquids were monitored using proton nuclear magnetic resonance (1H NMR) spectroscopy. EAS-E Suite and polyparameter linear free energy relationships (PP-LFERs) were employed to estimate the partitioning coefficients for species. Further, a chemical two-dimensional partitioning model was applied to visualize the indoor equilibrium partitioning and estimate the distribution of flavor carbonyls and their acetals in the gas phase, aerosol phase, and surface reservoirs. Our results demonstrate that a substantial fraction of carbonyls were converted into acetals in e-liquids and their chemical partitioning was significantly influenced. This study shows that acetalization is a determinant factor in the exposure and toxicology of harmful carbonyl flavorings, with its impact extending to both direct exposure to smokers and involuntary exposure to nonsmokers.