Lymphocyte predominant cells of nodular lymphocyte predominant Hodgkin lymphoma interact with rosetting T cells in an immunological synapse.
Nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) is a subtype of Hodgkin lymphoma with a preserved B-cell phenotype and follicular T helper (TFH ) cells rosetting around the tumor cells, the lymphocyte-predominant (LP) cells. As we recently described reactivity of the B-cell receptors of LP cells of some NLPHL cases with Moraxella spp. proteins, we hypothesized that LP cells could present peptides to rosetting T cells in a major histocompatibility complex class II (MHCII)-bound manner. Rosetting PD1+ T cells were present in the majority of NLPHL cases, both in typical (17/20) and variant patterns (16/19). In most cases, T-cell rosettes were CD69+ (typical NLPHL, 17/20; NLPHL variant, 14/19). Furthermore, both MHCII alpha and beta chains were expressed in the LP cells in 23/39 NLPHL. Proximity ligation assay and confocal laser imaging demonstrated interaction of the MHCII beta chain expressed by the LP cells and the T-cell receptor alpha chain expressed by rosetting T cells. We thus conclude that rosetting T cells in NLPHL express markers that are encountered after antigenic exposure, that MHCII is expressed by the LP cells, and that LP cells interact with rosetting T cells in an immunological synapse in a subset of cases. As they likely receive growth stimulatory signals in this way, blockade of this interaction, for example, by PD1-directed checkpoint inhibitors, could be a treatment option in a subset of cases in the future.


Altered erythrocyte C3b receptor expression, immune complexes, and complement activation in homosexual men in varying risk groups for acquired immune deficiency syndrome.
We studied levels of erythrocyte C3b receptors (E-CR1) and correlated them to the level of circulating immune complexes (CIC) and complement activation in patients with or at risk for acquired immunodeficiency syndrome (AIDS). A significant reduction was found in patients with AIDS (185 +/- 93 CR1/cell), AIDS-related complex, and generalized lymphadenopathy, whereas healthy male homosexuals or normal controls had 434 +/- 193 and 509 +/- 140 CR1/cell, respectively (P less than 0.001). Family studies indicate that this defect is acquired. Reduction in E-CR1 was associated with increased levels of CIC when assayed by binding to Raji cells, but not when tested by C1q binding. Complement activation was assessed by levels of C3bi/C3d-g in plasma, measured with a monoclonal antibody specific for a neoantigen in C3d. AIDS patients had increased C3 activation (2.68 +/- 1.67%) when compared with normal controls (0.9 +/- 0.22%) (P less than 0.01). The decreased E-CR1, the presence of CIC, and C3 activation suggest that complement activation by immune complexes may play a role in the clinical expression of the disease.


Effect of dietary supplementation with oregano essential oil on performance of broilers after experimental infection with Eimeria tenella.
A study was carried out to examine the effect of dietary supplementation of oregano essential oil on performance of broiler chickens experimentally infected with Eimeria tenella at 14 days of age. A total of 120 day-old Cobb-500 chicks separated into 4 equal groups with three replicates each, were used in this study. Two groups, one infected with 5 x 10(4) sporulated oocysts of E. tenella and the other not, were given a basal diet and served as controls. The other two groups also infected with E. tenella were administered diets supplemented with oregano essential oil at a level of 300 mg/kg, or with the anticoccidial lasalocid at 75 mg/kg. Following this infection, survival rate, bloody diarrhoea and oocysts excretion as well as lesion score were determined. Throughout the experimental period of 42 days, body weight gain and feed intake were recorded weekly, and feed conversion ratios were calculated. Two weeks after the infection with E. tenella supplementation with dietary oregano oil resulted in body weight gains and feed conversion ratios not differing from the non-infected group, but higher than those of the infected control group and lower than those of the lasalocid group. These parameters correspond with the extent of bloody diarrhoea, survival rate, lesion score and oocyst numbers and indicated that oregano essential oil exerted an anticoccidial effect against E. tenella, which was, however, lower than that exhibited by lasalocid.


Metallo-beta-lactamase-producing Pseudomonas aeruginosa in two hospitals from southern Brazil.
This study determined the prevalence of metallo-beta-lactamase (MBL)-producing Pseudomonas aeruginosa in two hospitals located in the Southern part of Brazil and compare the performance of two different phenotypic tests. Thirty-one non-repetitive Pseudomonas aeruginosa isolates from various clinical samples from patients admitted to two hospitals located in Rio Grande do Sul, Brazil (twenty-three from a hospital in Porto Alegre City and eight isolates from a hospital in Vale dos Sinos Region). All strains suggestive of possessing MBLs by phenotypic methods were included in this study. Phenotypic detection of MBLs was carried out simultaneously by using both the MBL Etest and disk approximation test using 2-mercaptopropionic acid close to a ceftazidime disk. Strains positive were further confirmed using molecular techniques for bla(VIM), bla(IMP) and bla(SPM-1). The prevalence of MBLs from samples of inpatients from the hospital located in Porto Alegre was 30.4% and that of inpatients from Vale dos Sinos hospital was only 3.1%. Only MBL type SPM-1 was detected in these samples by molecular analysis and all were detected by the Etest MBL strips. The prevalence of P. aeruginosa that produce MBLs can be markedly different in distinct geographical areas, even among different hospitals in the same area. In our study, the EDTA-based method was the only method able to detect all strains harboring the SPM-1 enzyme.


Increased stroke volume and aortic stiffness contribute to isolated systolic hypertension in young adults.
Isolated systolic hypertension is a common condition in individuals aged older than 60 years. However, isolated systolic hypertension has also been described in young individuals, although the mechanisms are poorly understood. We hypothesized that in young adults, isolated systolic hypertension and essential hypertension have different hemodynamic mechanisms and the aim of this study was to test this hypothesis in a cohort of subjects from The ENIGMA Study. Peripheral and central blood pressure, aortic pulse wave velocity, cardiac output, stroke volume, and peripheral vascular resistance were determined in 1008 subjects, aged 17 to 27 years. Compared with normotensive subjects, those with isolated systolic hypertension had significantly higher peripheral, central, and mean blood pressure, aortic pulse wave velocity, cardiac output, and stroke volume (P<0.001 for all comparisons). However, there were no differences in pulse pressure amplification, heart rate, or peripheral vascular resistance between the two groups. Compared with subjects with essential hypertension, mean pressure, heart rate, and peripheral vascular resistance were all significantly lower in isolated systolic hypertensive subjects, but pulse pressure amplification, aortic pulse wave velocity, cardiac output, and stroke volume were higher (P<0.001 for all comparisons). We have demonstrated that in young adults, isolated systolic hypertension and essential hypertension arise from different hemodynamic mechanisms. Isolated systolic hypertension appears to result from an increased stroke volume and/or aortic stiffness, whereas the major hemodynamic abnormality underlying essential hypertension is an increased peripheral vascular resistance. Long-term follow-up of these individuals is now required to determine whether they are at increased risk compared with age-matched normotensive individuals.


Rational design of Co embedded N,S-codoped carbon nanoplates as anode materials for high performance lithium-ion batteries.
The construction of specific structures and heteroatom-doped nanomaterials is essential for enhancing the electrochemical performance of lithium-ion batteries (LIBs). Herein, three structures of cobalt nanoparticle embedded N,S-codoped carbon (CNSCo-x) nanocomposites are obtained by pyrolysis of different amounts of thioacetamide (TAA) sulfuretted zeolitic imidazolate frameworks (ZIF-67) under an Ar/H2 atmosphere. With the addition of 0.5 mmol of TAA, CNSCo-0.5 with an orderly and porous nanoplate structure is obtained. The CNSCo-0.5 exhibits the best rate behavior and cycling stability with an initial discharge capacity of 1355 mA h g-1 at 200 mA g-1, and high discharge capacities of 1360.4 mA h g-1 at the 200th cycle at 200 mA g-1, and 647.9 mA h·g-1 at the 400th cycle at 1000 mA g-1. This facile synthetic strategy could be extended to other MOFs and is significant for developing new materials for LIBs.


Histone H2B monoubiquitination facilitates the rapid modulation of gene expression during Arabidopsis photomorphogenesis.
Profiling of DNA and histone modifications has recently allowed the establishment of reference epigenomes from several model organisms. This identified a major chromatin state for active genes that contains monoubiquitinated H2B (H2Bub), a mark linked to transcription elongation. However, assessment of dynamic chromatin changes during the reprogramming of gene expression in response to extrinsic or developmental signals has been more difficult. Here we used the major developmental switch that Arabidopsis thaliana plants undergo upon their initial perception of light, known as photomorphogenesis, as a paradigm to assess spatial and temporal dynamics of monoubiquitinated H2B (H2Bub) and its impact on transcriptional responses. The process involves rapid and extensive transcriptional reprogramming and represents a developmental window well suited to studying cell division-independent chromatin changes. Genome-wide H2Bub distribution was determined together with transcriptome profiles at three time points during early photomorphogenesis. This revealed de novo marking of 177 genes upon the first hour of illumination, illustrating the dynamic nature of H2Bub enrichment in a genomic context. Gene upregulation was associated with H2Bub enrichment, while H2Bub levels generally remained stable during gene downregulation. We further report that H2Bub influences the modulation of gene expression, as both gene up- and downregulation were globally weaker in hub1 mutant plants that lack H2Bub. H2Bub-dependent regulation notably impacted genes with fast and transient light induction, and several circadian clock components whose mRNA levels are tightly regulated by sharp oscillations. Based on these findings, we propose that H2B monoubiquitination is part of a transcription-coupled, chromatin-based mechanism to rapidly modulate gene expression.


Ontogeny in the visual system of Nile tilapia.
Retinal neurogenesis in fish facilitates cellular rearrangement throughout ontogeny, potentially allowing for optimization of the visual system to shifts in habitat and behaviour. To test this possibility, we studied the developmental trajectory of the photopic visual process in the Nile tilapia. We examined ontogenetic changes in lens transmission, photoreceptor sensitivity and post-receptoral sensitivity, and used these to estimate changes in cone pigment frequency and retinal circuitry. We observed an ontogenetic decrease in ultraviolet (UV) photoreceptor sensitivity, which resulted from a reduction in the SWS1 cone pigment frequency, and was associated with a reduction in lens transmission at UV wavelengths. Additionally, post-receptoral sensitivity to both UV and long wavelengths decreased with age, probably reflecting changes in photoreceptor sensitivity and retinal circuitry. This novel remodelling of retinal circuitry occurred following maturation of the visual system but prior to reaching adulthood, and thus may facilitate optimization of the visual system to the changing sensory demands. Interestingly, the changes in post-receptoral sensitivity to long wavelengths could not be predicted by the changes observed in lens transmission, cone pigment frequency or photoreceptor sensitivity. This finding emphasizes the importance of considering knowledge of visual sensitivity and retinal processing when studying visual adaptations and attempting to relate visual function to the natural environment. This study advances our understanding of ontogeny in visual systems and demonstrates that the association between different elements of the visual process can be explored effectively by examining visual function throughout ontogeny.


Covariation bias in panic-prone individuals.
Covariation estimates between fear-relevant (FR; emergency situations) or fear-irrelevant (FI; mushrooms and nudes) stimuli and an aversive outcome (electrical shock) were examined in 10 high-fear (panic-prone) and 10 low-fear respondents. When the relation between slide category and outcome was random (illusory correlation), only high-fear participants markedly overestimated the contingency between FR slides and shocks. However, when there was a high contingency of shocks following FR stimuli (83%) and a low contingency of shocks following FI stimuli (17%), the group difference vanished. Reversal of contingencies back to random induced a covariation bias for FR slides in high- and low-fear respondents. Results indicate that panic-prone respondents show a covariation bias for FR stimuli and that the experience of a high contingency between FR slides and aversive outcomes may foster such a covariation bias even in low-fear respondents.


Management of erectile dysfunction and LUTS/incontinence: the two most common, long-term side effects of prostate cancer treatment.
The two major long-term concerns associated with different options for the management of prostate cancer, (including surgery, radiotherapy, brachytherapy, cryotherapy, HIFU, etc.) include difficulties with lower urinary tract symptoms (LUTS) and/or erectile dysfunction. LUTS can be in the form of stress urinary incontinence (SUI), urge urinary incontinence (UUI), frequency/urgency, and/or voiding difficulties. While surgery is mostly associated with SUI and radiation mostly results in UUI, there can be an overlap. Incontinence rates after cryotherapy and high intensity focused ultrasound (HIFU) are generally very low. Voiding difficulties can also happen after the above-mentioned options. Treatment of SUI can start with pelvic floor muscle exercises (PFME), penile clamps or urethral plugs. If these fail to provide satisfactory results the surgical options could include: urethral bulking agents, male slings, and artificial urinary sphincter (AUS). Surgical options are usually not recommended during the first 6-12 months after radical prostatectomy. Management of frequency, urgency and/or UUI can also be started with lifestyle modifications and PFME. Oral agents (anticholinergics and β3-agonists) are also considered before proceeding to third line options, such as Botox injection or sacral neuromodulation. The treatment options for ED resulting from the treatment of prostate cancer can include oral PDE5-I as the first line, local therapy as the second (such as MUSE, intracavernosal injections, and perhaps low intensity shock wave therapy) and finally surgery as the third line. Standard questionnaires and patient reported outcome measurement tools should be used for the assessment of LUTS and erectile dysfunction prior and after initiation of treatment to guide the management.


[Qualitative studies in the health field].
Qualitative investigation in the health care field is faced with the relative problems of scientific knowledge production or epistemological questions, methodological procedures in qualitative investigations, and the utility and use of qualitative findings. These questions and others are discussed, in a general manner, in this article.


The Impact of the Preceptorship Program on Self-efficacy and Learning Outcomes in Nursing Students.
This study aimed to determine the impact of the preceptorship program on self-efficacy and learning outcomes and the association between self-efficacy and learning outcomes in nursing students. This quasi-experimental study was conducted on 112 nursing students in 2012-2013. The subjects were selected through convenience sampling from the School of Nursing and Midwifery of Shiraz University of Medical Sciences, Iran. The general self-efficacy scale and the perception of overall learning outcomes scale were used to measure the students' perception of self-efficacy and achievement of learning outcomes. The data were analyzed using Pearson's correlation coefficient and linear regression analysis. The students' mean [standard deviation (SD)] scores of self-efficacy and learning outcomes were 30.69 (5.88) and 4.10 (0.82), respectively, after taking part in the preceptorship program. The results showed a significant association between the mean scores of self-efficacy and learning outcomes (r = 0.30, p = 0.002). In addition, self-efficacy was predicted by gender (β = -0.19, p = 0.030) and the components of "understand ethical aspects of patient care" (β = 0.31, p = 0.010), "work more systematically" (β = -0.44, p = 0.005), and "use of nursing research" (β = 0.34, p = 0.020). Although the nursing students who participated in the preceptorship program demonstrated high mean scores of self-efficacy and learning outcomes, the lack of control group led to the conclusion about the effectiveness of the program to be made in caution. However, this program is suggested to be retained in nursing students' curriculum.


[A novel transcutaneous vagus nerve stimulation leads to brainstem and cerebral activations measured by functional MRI].
Left cervical vagus nerve stimulation (VNS) using the implanted NeuroCybernetic Prosthesis (NCP) can reduce epileptic seizures and has recently been shown to give promising results for treating therapy-resistant depression. To address a disadvantage of this state-of-the-art VNS device, the use of an alternative transcutaneous electrical nerve stimulation technique, designed for muscular stimulation, was studied. Functional magnetic resonance imaging (MRI) has been used to test non-invasively access nerve structures associated with the vagus nerve system. The results and their impact are unsatisfying due to missing brainstem activations. These activations, however, are mandatory for reasoning, higher subcortical and cortical activations of vagus nerve structures. The objective of this study was to test a new parameter setting and a novel device for performing specific (well-controlled) transcutaneous VNS (tVNS) at the inner side of the tragus. This paper shows the feasibility of these and their potential for brainstem and cerebral activations as measured by blood oxygenation level dependent functional MRI (BOLD fMRI). In total, four healthy male adults were scanned inside a 1.5-Tesla MR scanner while undergoing tVNS at the left tragus. We ensured that our newly developed tVNS stimulator was adapted to be an MR-safe stimulation device. In the experiment, cortical and brainstem representations during tVNS were compared to a baseline. A positive BOLD response was detected during stimulation in brain areas associated with higher order relay nuclei of vagal afferent pathways, respectively the left locus coeruleus, the thalamus (left >> right), the left prefrontal cortex, the right and the left postcentral gyrus, the left posterior cingulated gyrus and the left insula. Deactivations were found in the right nucleus accumbens and the right cerebellar hemisphere. The method and device are feasible and appropriate for accessing cerebral vagus nerve structures, respectively. As functional patterns share features with fMRI BOLD, the effects previously studied with the NCP are discussed and new possibilities of tVNS are hypothesised.


Clinical evaluation of vasomotor system functionality in type 2 diabetic Patients.
Autonomic neuropathy and vital organ dysfunctions are the known complications in type 2 diabetes Mellitus (DM). Genetic endowments involving individuals make subtle differences in physiological systems, particularly at the time of sickness. Hence, the presented study was designed to evaluate the vasomotor system in healthy people and type 2 DM cases for determining any functionality differences between the mentioned groups. Sixty patients with type 2 diabetes (case group) and sixty healthy subjects (control group) matched for age and sex were enrolled in the study. Then, the performance of vasomotor system was assessed using valsalva maneuver, cold pressor, sustained hand-grip and mental arithmetic tests and the differences were determined via statistical methods. According to our findings, abnormal response to valsalva maneuver was found in the case group (P=0.028) and the same response was seen about mental arithmetic evaluations. In the case of cold pressor and sustained hand-grip tests, remarkable differences were not found in both groups. Important differences were also found among vasomotor dysfunction and the time of DM labeling. This study showed a higher incidence of vasomotor dysfunction in DM patients. However, revision in cold pressor and sustained hand-grip tests definition as well as methodology was recommended.


Translating child abuse research into action.
The Child Abuse Recognition Experience Study revealed that primary care clinicians did not always follow the legal mandate to report suspected child abuse to child protective services. National child abuse experts representing different disciplines met in a 2-day conference in January 2007 to discuss and develop new strategies that would address the barriers to reporting suspected child abuse and improve the protection of children. This article describes the rationale, structure, and goals of the conference.


In Situ Determination of the NiAs Phase of FeO at High Pressure and Temperature.
In situ synchrotron x-ray diffraction measurements of FeO at high pressures and high temperatures revealed that the high-pressure phase of FeO has the NiAs structure (B8). The lattice parameters of this NiAs phase at 96 gigapascals and 800 kelvin are a = 2.574(2) angstroms and c = 5.172(4) angstroms (the number in parentheses is the error in the last digit). Metallic behavior of the high-pressure phase is consistent with a covalently and metallically bonded NiAs structure of FeO. Transition to the NiAs structure of FeO would enhance oxygen solubility in molten iron. This transition thus provides a physiochemical basis for the incorporation of oxygen into the Earth's core.


The architecture of microtubular network and Golgi orientation in osteoclasts--major differences between avian and mammalian species.
In the present study, we analyze multinuclear osteoclasts obtained from several avian and mammalian species and describe the reorganization of their microtubular architecture and Golgi complex orientation during osteoclast differentiation and activation for bone resorption. In nonresorbing quail and chicken multinuclear osteoclasts, microtubules radiate from multiple centrosomal microtubule-organizing centers (MTOCs), whose number is equal to the number of nuclei. However, centrosomal MTOCs disappear at the time of cell activation for bone resorption and the Golgi membranes redistribute to circumscribe nuclei. In contrast to avian osteoclasts, both resorbing and nonresorbing rat, rabbit, and human osteoclasts have no or few centrosomal MTOCs. Instead, after cold-induced depolymerization, regrowing microtubules nucleate from the perinuclear area where immunofluoresce and immunoelectron scanning microscopy reveal pericentriolar matrix protein pericentrin associated with vimentin filaments. Furthermore, the circumnuclear reorganization of MTOCs and the Golgi is a result of mammalian osteoclast maturation and occur before any resorptive activity of the mononuclear osteoclasts and their fusion into multinucleated cells. Our results show that unlike previously suggested, the nuclear surfaces of mammalian osteoclasts act as the microtubule anchoring sites similarly to nuclear surfaces in multinucleated myotubes and suggest the role of perinuclear intermediate filament network in orchestrating the microtubular cytoskeleton.


Formation and cleansing performance of bicontinuous microemulsions in water/poly (oxyethylene) alkyl ether/ester-type oil systems.
Phase behaviors in water/poly(oxyethylene) dodecyl ether (C₁₂EO(n), n = 4, 6, 8)/cetyl isooctanoate (CIO) systems were studied. In the C₁₂EO₆ and C₁₂EO₈ systems, self-assembled structures with positive curvatures, such as O/W microemulsions, and micellar cubic and hexagonal phases, were observed. A wider region of a lamellar liquid-crystalline phase, which included a narrow microemulsion region joined by a miscibility gap, was observed in the C₁₂EO₄ system. The structure of the microemulsion phase in the C₁₂EO₄ system was characterized by pulsed-field-gradient NMR (PFG-NMR) and small angle X-ray scattering (SAXS) techniques. PFG-NMR measurements indicated that the structure of the microemulsion was bicontinuous; both water and oil phases were continuous within the microemulsion. Pair-distance distribution function, p (r), and structure factors obtained by Generalized Indirect Fourier Transformation (GIFT) analysis of the SAXS data showed that the microemulsion domain sizes decreased with an increase in the oil content. The structure of the bicontinuous microemulsion was consistent with the results of a detergency test, in which the microemulsion samples were applied to lipstick dirt on an artificial skin plate. Detergency was observed to be better for the microemulsion at lower oil contents because of the larger oil domain size at these low concentrations.


Clonotypic Features of Rearranged Immunoglobulin Genes Yield Personalized Biomarkers for Minimal Residual Disease Monitoring in Multiple Myeloma.
Due to improved treatment, more patients with multiple myeloma (MM) reach a state of minimal residual disease (MRD). Different strategies for MM MRD monitoring include flow cytometry, allele-specific oligonucleotide-quantitative PCR, next-generation sequencing, and mass spectrometry (MS). The last 3 methods rely on the presence and the stability of a unique immunoglobulin fingerprint derived from the clonal plasma cell population. For MS-MRD monitoring it is imperative that MS-compatible clonotypic M-protein peptides are identified. To support implementation of molecular MRD techniques, we studied the presence and stability of these clonotypic features in the CoMMpass database. An analysis pipeline based on MiXCR and HIGH-VQUEST was constructed to identify clonal molecular fingerprints and their clonotypic peptides based on transcriptomic datasets. To determine the stability of the clonal fingerprints, we compared the clonal fingerprints during disease progression for each patient. The analysis pipeline to establish the clonal fingerprint and MS-suitable clonotypic peptides was successfully validated in MM cell lines. In a cohort of 609 patients with MM, we demonstrated that the most abundant clone harbored a unique clonal molecular fingerprint and that multiple unique clonotypic peptides compatible with MS measurements could be identified for all patients. Furthermore, the clonal immunoglobulin gene fingerprints of both the light and heavy chain remained stable during MM disease progression. Our data support the use of the clonal immunoglobulin gene fingerprints in patients with MM as a suitable MRD target for MS-MRD analyses.


Antitumor activity of the dual PI3K/mTOR inhibitor gedatolisib and the involvement of ABCB1 in gedatolisib resistance in canine tumor cells.
The phosphatidylinositol 3‑kinase/mammalian target of rapamycin (PI3K/mTOR) signaling pathway is a therapeutic target for various types of human tumors, and dual PI3K/mTOR inhibitors demonstrate antitumor activities in both preclinical and clinical studies. However, resistance mechanisms limit their abilities. As the molecular mechanisms involved in the cellular resistance are not clear in any canine tumors, an understanding of resistance mechanisms would support the potential use of dual PI3K/mTOR inhibitors in canine tumors. The antitumor activity of gedatolisib on cell viability, protein phosphorylation, and cell cycle distribution was assessed using 12 canine tumor cell lines from 6 types of tumors. In addition, the molecular determinants involved in the cellular sensitivity to gedatolisib were explored by investigating the involvement of serum‑and‑glucocorticoid‑induced kinase 1 (SGK1), PIK3CA, and ATP‑binding cassette, subfamily B, member 1 (ABCB1). The results demonstrated that gedatolisib decreased cell viability in all cell lines, with IC50 values <1 µM in 10 of the 12 lines. Gedatolisib inhibited Akt and mTOR complex 1 substrate phosphorylation and induced G0/G1 cell cycle arrest. However, certain cell lines with higher IC50 values were more resistant to these effects. These cell lines exhibited higher ABCB1 activity and the ABCB1 inhibitor cyclosporin A enhanced the decrease of cell viability caused by gedatolisib. SGK1 overexpression did not confer resistance to gedatolisib. The mutations of E545K and H1047R in PIK3CA were not observed. The present results indicated that gedatolisib decreased cell viability in canine tumor cell lines and ABCB1 played an important role in gedatolisib resistance, supporting the potential use of gedatolisib for canine tumors.


Modeling back propagating action potential in weakly excitable dendrites of neocortical pyramidal cells.
Simultaneous recordings from the soma and apical dendrite of layer V neocortical pyramidal cells of young rats show that, for any location of current input, an evoked action potential (AP) always starts at the axon and then propagates actively, but decrementally, backward into the dendrites. This back-propagating AP is supported by a low density (-gNa = approximately 4 mS/cm2) of rapidly inactivating voltage-dependent Na+ channels in the soma and the apical dendrite. Investigation of detailed, biophysically constrained, models of reconstructed pyramidal cells shows the following. (i) The initiation of the AP first in the axon cannot be explained solely by morphological considerations; the axon must be more excitable than the soma and dendrites. (ii) The minimal Na+ channel density in the axon that fully accounts for the experimental results is about 20-times that of the soma. If -gNa in the axon hillock and initial segment is the same as in the soma [as recently suggested by Colbert and Johnston [Colbert, C. M. & Johnston, D. (1995) Soc. Neurosci. Abstr. 21, 684.2]], then -gNa in the more distal axonal regions is required to be about 40-times that of the soma. (iii) A backward propagating AP in weakly excitable dendrites can be modulated in a graded manner by background synaptic activity. The functional role of weakly excitable dendrites and a more excitable axon for forward synaptic integration and for backward, global, communication between the axon and the dendrites is discussed.


Phosphorus mobilization in lake sediments: Experimental evidence of strong control by iron and negligible influences of manganese redox reactions.
Iron (Fe) and manganese (Mn) reactions have been regarded as the primary factors responsible for the mobilization of phosphorus (P) in lake sediments, although their individual roles are hard to distinguish. In this study, in situ mobilization of P, Fe and Mn in sediments was assessed by high resolution spatio-temporal sampling of their labile forms using diffusive gradient in thin films (DGT) and suction device (Rhizon) techniques. It was found that the monthly concentration distributions showed greater agreement and better correlation coefficients between labile P and labile Fe, than those between labile P and labile Mn, implying that Fe plays a key role in controlling P release in sediments. Furthermore, better correlations were observed between hourly changes in concentrations of soluble reactive phosphorus (SRP) and soluble Fe(II), than those between SRP and soluble Mn. Changes were observed under simulated anaerobic incubation conditions, suggesting that P release was caused by the reductive dissolution of Fe oxides. This was supported by the lack of influences on P release from reductive dissolution of Mn oxides in the sediment-water interface and top sediment layers under the anaerobic incubations. In simulated algal bloom experiments, positive correlations and consistent changes were observed between SRP and soluble Fe(II) concentrations, but not between SRP and soluble Mn concentrations. This further demonstrated the Fe-dependent and Mn-independent release of P in sediments. Therefore, Fe redox reactions have a high impact on P mobilization in sediments, while Mn redox reactions appear to have negligible influences.


Bilateral traumatic renal artery thrombosis.
A case of bilateral traumatic renal artery thrombosis involving three renal arteries is presented. Four other cases of this entity are summarized for comparison. The recommended course of investigation is outlined, and the urgency of immediate surgical intervention is emphasized.


Three-dimensional packing with complex orbit coils for the endovascular treatment of intracranial aneurysms.
Endovascular treatment of intracranial aneurysms is too often associated with aneurysm recurrence due to coil compaction. High packing of coils prevents compaction. To increase the packing attenuation, we sought to evaluate the results of selective embolization of aneurysms with complex-shaped coils alone. Twenty consecutive patients with an intracranial aneurysm were treated by selective embolization. There were 12 women and eight men, with a mean age of 48 years. Fourteen patients presented with subarachnoid hemorrhage, whereas six were asymptomatic. Mean size of aneurysms was 5 mm (range, 3-11 mm). In all cases, we tried to pack the aneurysm with complex coils only (Orbit, Cordis, Miami Lakes, FL), delivered in a concentric fashion. Remodeling technique was used in two cases of wide-neck aneurysms. Clinical and anatomic outcome were assessed by using the modified Glasgow outcome scale and 6-month MR angiography (MRA). From two to five complex coils were delivered within the aneurysms. Occlusion by using complex coils alone was successful in 16 patients, and resulted in 14 complete occlusions and two neck remnants. In four patients, additional helical Orbit coils or Guglielmi detachable coils were required to complete aneurysm obliteration. No technical complication occurred, and clinical outcome was excellent in 14 cases and good in two. Follow-up MRAs were obtained in all patients and showed only one slight recanalization. Intracranial aneurysms may be treated by selective embolization with complex coils only. Imaging follow-up at 6 months shows a low rate of coil compaction.


Iris juvenile xanthogranuloma in an infant - spontaneous hyphema and secondary glaucoma.
Juvenile xanthogranuloma (JXG) is a benign histiocytic skin disorder mainly encountered during infancy and childhood. Although with multiple potential localizations, less than 1% of the cases exhibit ocular manifestations. Some of these might lead to serious complications, specifically, secondary glaucoma that can result in severe and blinding eye disease. The aim of the present case report was to demonstrate typical clinical features, emphasize the difficulties attributed when managing these patients and literature review. We present the case of 4 months old female baby with spontaneous hyphema and secondary unilateral glaucoma due to ocular JXG. The natural history and treatment of the condition were extremely difficult to handle due to multiple opinions in histopathology related to other severe conditions that resembled with the lesions detected in this case: myelomonocytic leukemia and Langerhans cell histiocytosis. Although a minority of patients with JXG have ocular involvement, recognition of this condition is important because a treatment delay can lead to serious complications, such as glaucoma and spontaneous hyphema, as in our case. A thorough differential diagnosis represents the key to a proper management plan in these patients, both on short and long term. "Triple disease" defined as JXG plus neurofibromatosis type 1 (NF-1) and juvenile chronic myelogenous leukemia (JCML) has been reported, but it was not confirmed in our patient.


Comparison between WHO (World Health Organization) 2010 and WHO 1999 parameters for semen analysis - interpretation of 529 consecutive samples.
To quantify how many men with normal semen according to WHO (WHO - World Health Organization) 1999 criteria, should be considered with abnormal semen according to 2010 criteria and vice versa; to study which parameter of volume, concentration, motility and morphology is the most responsible of this change. We studied, using WHO 1999 parameters, 529 consecutive semen samples from 427 men, collected in our Department from January 2008 to December 2009, then we re-evaluated those results using WHO 2010 parameters; we also studied each parameter to understand how changed the classification from normal (defined normal by all parameters) to abnormal (defined abnormal by at least one parameter) using the two WHO criteria. 3 men (0.56%) were azoospermic. Among the remaining 526 samples, 199 (37.83%) were considered normal and 246 (46.76%) abnormal both according to WHO 1999 and WHO 2010 criteria; we found that none of the samples classified normal according to the previous criteria was classified abnormal according the more recent criteria, while 82 (15.58%) evaluated as abnormal according 1999 criteria changed to normal according 2010 criteria. The concordance between 1999 and 2010 evaluation was 84.44%. In this study we noted that the changes from WHO 1999 to WHO 2010 criteria did not modify the interpretation of semen quality, because comparing the two classifications we demonstrated that there is a substantial agreement, considering the three parameters (count, motility and morphology) all together, and also considering each single parameter. Anyhow, almost 16% of the patients considered infertile according to the old criteria, should be evaluated normal by the new classification and they should not need any treatment for infertility.


Inhibition of primary and metastatic tumor growth in mice by cancer-associated galactosyltransferase acceptor.
The antitumor activity of a glycopeptide purified from human malignant effusion, termed cancer-associated galactosyltransferase acceptor (CAGA), was assessed in BALB/c mice bearing primary and metastatic tumors. Initial studies with the fast-growing KA31 and slow-growing KB521 Kirsten sarcoma-transformed mouse fibroblast cell lines confirmed their tumorigenicity and metastatic potential. Inoculation of 1 X 10(5) KA31 cells s.c. resulted in palpable tumor formation in recipient animals within 14 days and death within 42 days from primary tumor growth (mean survival, 26 days; total survival, 0%). Inoculation of the slower-growing KB521 resulted in tumor formation in 85% of recipients, and tumor-bearing animals succumbed within 56 days after primary inoculation (mean survival, 48 days; total survival, 15%). Administration of CAGA by i.p. injection as a single dose or series of five daily doses (each 50 micrograms) inhibited primary tumor growth by 35 to 68% in animals receiving KA31 cells and by 25 to 70% in animals receiving KB521 cells. CAGA increased mean survival 50% from 26 to 38 days and total survival from 0 to 27% in animals bearing KA31-derived primary tumors. In animals bearing KB521-derived tumors, CAGA increased mean survival from 48 to 90 days and total survival from 15 to 50%. Similarly, CAGA was also found to significantly inhibit formation of pulmonary metastases in animals after excision of primary tumors. CAGA administration reduced death from metastatic deposits by 55 to 66% in animals initially inoculated with the KA31 cell line and by 58 to 90% in animals initially bearing primary tumors derived from the KB521 line. There was a corresponding decrease in the number of metastatic deposits per lung after administration of CAGA. Thus, CAGA appears to have potential antitumor activity against tumors with a range of growth rates and appears to inhibit both primary and metastatic tumor growth.


Peptide inhibitor of morphine- and beta-endorphin-induced analgesia.
The synthetic beta-endorphin analogs with the omission of the NH2-terminal [Met]enkephalin segment [beta-endorphin-(6-31) and beta-endorphin-(20-31)] are shown to inhibit morphine- or beta-endorphin-induced analgesia in mice by the tail-flick test, whereas the synthetic NH2-terminal pentadecapeptide beta-endorphin-(1-15) has no inhibitory activity. This study raises the possibility that endogenous inhibiting peptides exist in the brain which play a role in the regulation of endorphin actions.


Adult Vaccination.
Adult vaccination rates are low in the United States, despite clear benefits for reducing morbidity and mortality. Vaccine science is evolving rapidly, and family physicians must maintain familiarity with the most recent guidelines. The recommended adult immunization schedule is updated annually by the Advisory Committee on Immunization Practices of the Centers for Disease Control and Prevention. All eligible patients should receive SARS-CoV-2 vaccines according to the current guidelines. Adults without contraindications should also receive an annual influenza vaccine. Hepatitis A vaccine is recommended for adults with specific risk factors. All pregnant patients, adults younger than 60 years, and those 60 years and older who have risk factors should receive a hepatitis B vaccine. A 15- or 20-valent pneumococcal conjugate vaccine is recommended for all patients who are 65 years and older. Patients who receive 15-valent pneumococcal conjugate vaccine should receive a dose of 23-valent pneumococcal polysaccharide vaccine one year later. Adults 19 to 64 years of age should receive a pneumococcal vaccination if they have medical risk factors. A single dose of measles, mumps, and rubella vaccine is recommended for adults without presumptive immunity, and additional doses are recommended for patients with HIV and postdelivery for pregnant patients who are not immune to rubella. A tetanus and diphtheria toxoids booster is recommended every 10 years. For pregnant patients and those in close contact with young infants, a tetanus toxoid, reduced diphtheria toxoid, and acellular pertussis vaccine should be administered. The human papillomavirus vaccine is recommended for all people through 26 years of age. Herpes zoster vaccine is indicated for all adults 50 years and older. .


Lycorine Suppresses Endplate-Chondrocyte Degeneration and Prevents Intervertebral Disc Degeneration by Inhibiting NF-κB Signalling Pathway.
Cartilaginous endplate (CEP) degeneration is an important cause for intervertebral disc (IVD) degeneration that leads to low-back pain. The identification of compounds that may prevent CEP degeneration is of interest for the prevention of IVD degeneration. Catabolic protease expression in the CEP of disc degeneration patients was first assessed. The toxicity, function and underlying mechanism of lycorine (LY) on CEP-derived chondrocytes degeneration were assessed in vitro by flow cytometry analysis and western blotting. The concentration and function of LY in rat-tail disc-degeneration models were also assessed by HPLC (High Performance Liquid Chromatography) quantification and histological analysis. In CEP cells, Interleukin (IL)-1β upregulated the expression of matrix metalloproteinase (MMP)-3, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4 and ADAMTS-5 that is critical for the degradation of cartilage extracellular matrix. Interestingly, LY suppressed the expression of these enzymes via the inhibition of nuclear factor-κB (NFκB) signalling and thus prevented IL-1β-induced endplate cell degeneration in vitro. More importantly, LY also reduced the expression of MMP-3, MMP-13, ADAMTS-4 and ADAMTS-5 in CEP and exerted a protective effect on both CEP and nucleus pulposus (NP) degeneration. In addition to its inhibitory effect on matrix-degrading protease expression, LY treatment also reduced positive regulators of proinflammatory cytokines, such as MIF, which can be secreted by CEP cells and subsequently target NP cells. LY could serve as a potential drug for treating IVD disease.


Preventing postpartum hemorrhage after cesarean delivery: a network meta-analysis of available pharmacologic agents.
Postpartum hemorrhage causes a quarter of global maternal deaths. The World Health Organization recommends oxytocin as the first line agent to prevent hemorrhage during cesarean delivery. However, some randomized controlled trials suggest that other uterotonics are superior. We conducted a network meta-analysis comparing the ability of pharmacologic agents to reduce blood loss and minimize the need for additional uterotonics during cesarean delivery. We searched the Cochrane Central Register of Controlled Trials, Embase, and MEDLINE databases from inception to May 2020. We included randomized controlled trials that compared oxytocin, carbetocin, misoprostol, ergometrine, carboprost, or combinations of these in the prevention of postpartum hemorrhage during cesarean delivery. We performed a systematic review followed by an NMA in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Quality of the evidence was assessed with the Confidence in Network Meta-Analysis approach and Grading of Recommendations, Assessment, Development and Evaluations tool within the summary of findings table. Our primary outcomes were the estimated blood loss and need for additional uterotonics. Secondary outcomes included nausea and postpartum hemorrhage of >1000 mL. We performed sensitivity analyses to explore the influence of surgical context and oxytocin administration strategy. A total of 46 studies with 7368 participants were included. Of those, 21 trials (6 agents and 3665 participants) formed the "estimated blood loss" network and, considering the treatment effects, certainty in the evidence, and surface under the cumulative ranking curve scores, carbetocin was assessed to probably be superior to oxytocin, but only in reducing the estimated blood loss by a clinically insignificant volume (54.83 mL; 95% confidence interval, 26.48-143.78). Misoprostol, ergometrine, and the combination of oxytocin and ergometrine were assessed to probably be inferior, whereas the combination of oxytocin and misoprostol was assessed to definitely be inferior to oxytocin. A total of 37 trials (8 agents and 6193 participants) formed the "additional uterotonic" network and, again, carbetocin was assessed to probably be superior to oxytocin, requiring additional uterotonics 185 (95% confidence interval, 130-218) fewer times per 1000 cases. Oxytocin plus misoprostol, oxytocin plus ergometrine, and misoprostol were assessed to probably be inferior, whereas carboprost, ergometrine, and the placebo were definitely inferior to oxytocin. For both primary outcomes, oxytocin administration strategies had a higher probability of being the best uterotonic, if initiated as a bolus. Carbetocin is probably the most effective agent in reducing blood loss and the need for additional uterotonics. Oxytocin appears to be more effective when initiated as a bolus.


An Organ System Approach to Explore the Antioxidative, Anti-Inflammatory, and Cytoprotective Actions of Resveratrol.
Resveratrol is a phenolic phytochemical, with a stilbene backbone, derived from edible plants such as grape and peanut. It is a bioactive molecule with physiological effects on multiple organ systems. Its effects range from the neuroprotective to the nephroprotective, including cardiovascular, neuronal, and antineoplastic responses as a part of its broad spectrum of action. In this review, we examine the effects of resveratrol on the following organ systems: the central nervous system, including neurological pathology such as Parkinson's and Alzheimer's disease; the cardiovascular system, including disorders such as atherosclerosis, ischemia-reperfusion injury, and cardiomyocyte hypertrophy; the kidneys, including primary and secondary nephropathies and nephrolithiasis; multiple forms of cancer; and metabolic syndromes including diabetes. We emphasize commonalities in extracellular matrix protein alterations and intracellular signal transduction system induction following resveratrol treatment. We summarize the known anti-inflammatory, antioxidative, and cytoprotective effects of resveratrol across disparate organ systems. Additionally, we analyze the available literature regarding the pharmacokinetics of resveratrol formulations used in these studies. Finally, we critically examine select clinical trials documenting a lack of effect following resveratrol treatment.


Cummings Memorial Lecture - 1975. The market basket: food for thought.
The world food crisis is as critical today as when it was debated at the 1974 World Food Conference in Rome. Since the United States and Canada-and to a lesser extent, Australia and New Zealand-lead in the production of corn, wheat and soybeans, the North American "bread basket" has become the "market basket" of the world. For welfare, economic, and political reasons, our energies, resources, and deliberations must be expanded toward optimum production of wholesome food products. I do not recommend that we permit food additives in "questionably" safe or excessive concentrations in our agricultural products. I do recommend, however, that tolerance limits for food additives be established based on a comprehensive review of all contributing factors-the world food crisis and the rational interpretation of both positive and negative animal data as they relate to man. As Dr. Herbert Stokinger put it so aptly: "Avoid the establishment of unnecessarily severe standards." 2. Funds for research and teaching of food and nutrition should be greatly increased, so that all who can read and write may be made aware of the daily dietary requirements for the maintenance of good health. 3. Unsubstantiated scare tactics in publications of the scientific and lay press can only lead to well-intended but often emotionally-inspired restrictions, ordinances, and laws. Such decisions are likely to either under- or over-define the requirements and standards for food additives and other chemicals which are important to the well-being of the populace.


Measles virus infection of epithelial cells in the macaque upper respiratory tract is mediated by subepithelial immune cells.
Measles virus (MV), one of the most contagious viruses infecting humans, causes a systemic infection leading to fever, immune suppression, and a characteristic maculopapular rash. However, the specific mechanism or mechanisms responsible for the spread of MV into the respiratory epithelium in the late stages of the disease are unknown. Here we show the crucial role of PVRL4 in mediating the spread of MV from immune to epithelial cells by generating a PVRL4 "blind" recombinant wild-type MV and developing a novel in vitro coculture model of B cells with primary differentiated normal human bronchial epithelial cells. We utilized the macaque model of measles to analyze virus distribution in the respiratory tract prior to and at the peak of MV replication. Expression of PVRL4 was widespread in both the lower and upper respiratory tract (URT) of macaques, indicating MV transmission can be facilitated by more than only epithelial cells of the trachea. Analysis of tissues collected at early time points after experimental MV infection demonstrated the presence of MV-infected lymphoid and myeloid cells contacting respiratory tract epithelium in the absence of infected epithelial cells, suggesting that these immune cells seed the infection in vivo. Thereafter, lateral cell-to-cell spread of MV led to the formation of large foci of infected cells in the trachea and high levels of MV infection in the URT, particularly in the nasal cavity. These novel findings have important implications for our understanding of the high transmissibility of measles.


Human Immunodeficiency Virus type 1 reverse transcriptase.
The Human Immunodeficiency Virus type 1 (HIV-1) is a retrovirus and a causative agent of the Acquired Immuno Deficiency Syndrome (AIDS). Retroviruses are distinct from other viruses in their ability to encode an enzyme called reverse transcriptase (RT). The RT is the enzyme mainly involved in replication. It performs RNA- as well as DNA-dependent DNA synthesis in order to convert the single-stranded viral RNA genome into double-stranded DNA. The double-stranded DNA is stably integrated into the host cell genome and is used as a template for the production of a new viral generation. The HIV-1 RT is partially encoded by the POL open reading frame of the HIV-1 genome and consists of two subunits of 66 kDa (p66) and 51 kDa (p51). The p66 polypeptide encodes the reverse transcriptase and the RNase H domain. Half of the p66 molecules are further processed to generate the p51 protein with an identical N-terminus, but lacking the C-terminus which encodes the RNase H domain. In vivo both polypeptides are found in equimolar amounts thus forming a heterodimer. This dimerization is critical for the enzymatic activity. In this review we summarize (i) the replication cycle of HIV-1, (ii) the enzymatic properties of HIV-1 RT and (iii) the structure-function relationship of the HIV-1 RT in view of the known three dimensional structure.


Synthesis and receptor binding activity of elephant beta-endorphin, a beta-endorphin homolog with highly potent analgesic activity.
Elephant beta-endorphin and its analog, elephant beta-endorphin(6-31) were synthesized by standard solid phase method. Receptor binding activity showed that elephant beta-endorphin was five to six times more potent than human beta-endorphin in its ability to bind to opiate receptors on rat brain membrane. In a previous study (Wong, C.-L., Wai, M.-K., Cheng, H.-C., Chung, D. & Yamashiro, D (1990) Clinical and Experimental Pharmacology and Physiology 16, 33-37), tail flick test for intracerebroventricularly administered beta-endorphin showed that the antinociceptive potency of elephant beta-endorphin was seven to eight times higher than that of human beta-endorphin in mice. Results from both studies suggest that elephant beta-endorphin was a much more potent antinociceptive agent than human beta-endorphin in tail flick test and its higher analgesic activity might be due to its higher affinity for opiate receptors in the brain.


Optical tuning of dielectric properties of La0.7Sr0.3MnO3/SrTiO3 superlattices in the terahertz range.
Two (La0.7Sr0.3MnO3)n/(SrTiO3)m superlattices with different superlattice period but the same total thickness were deposited on LaAlO3 substrates by pulsed laser deposition. Dielectric properties of these samples were investigated by means of terahertz time-domain spectroscopy (THz-TDS) under external continuous wave green laser excitation and optical-pump terahertz-probe spectroscopy (OPTP) at room temperature. Experimental results show that the real part of the permittivity for both superlattices increases significantly with increasing green laser pump power, which indicates the decrease of the plasma frequency, along with the increase of the electron scattering rate, soft mode eigenfrequency and oscillator strength in the Drude-Lorentz model. Furthermore, it's observed that the insulating superlattice exhibits a more significant dielectric tunability than the metallic superlattice. Besides, the carrier lifetime of superlattices is much shorter than the La0.7Sr0.3MnO3 thin film in the OPTP measurements, indicating that the electrons excited in the La0.7Sr0.3MnO3 layers may be trapped by the defects located in the interfaces of La0.7Sr0.3MnO3 and SrTiO3 or the SrTiO3 layers. With the optical field-induced tunability of dielectric properties, (La0.7Sr0.3MnO3)n/(SrTiO3)m superlattices show great potential in the actively tunable devices in the THz range.


Purification and properties of arginase of rat kidney.
l-Arginase from rat kidney was partially purified and some properties were compared with those of l-arginase of rat liver. The kidney enzyme was firmly bound to the mitochondrial fraction and after solubilization required arginine or an unknown factor in tissue extracts for stabilization after dialysis. The two enzymes differed also in stability with respect to acetone treatment, heating or freezing. In further contrast with liver arginase, arginase from kidney was not adsorbed to CM-cellulose at pH7.5 and its activity was not increased by incubation with Mn(2+). Other differences were seen in relative specificities for substrates, ratio of hydrolysis rates with high and low concentrations of arginine and effects of certain inhibitors. Antisera prepared to pure liver arginase did not cross-react with partially purified kidney arginase.


Community-based SARS-CoV-2 testing in low-income neighbourhoods in Rotterdam: Results from a pilot study.
High incidence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and low testing uptake were reported in low-income neighbourhoods in Rotterdam. We aimed to improve willingness and access to testing by introducing community-based test facilities, and to evaluate the effectiveness of a rapid antigen detection test (RDT). Two to eleven test facilities operated consecutively in three low-income neighbourhoods in Rotterdam, offering the options of walk-in or appointments. Background characteristics were collected at intake and one nasopharyngeal swab was taken and processed using both RDT and reverse transcription polymerase chain reaction (RT-PCR). Visitors were asked to join a survey for evaluation purposes. In total, 19 773 visitors were tested - 9662 (48.9%) without an appointment. Walk-in visitors were older, lived more often in the proximity of the test facilities, and reported coronavirus disease (COVID-19)-related symptoms less often than by-appointment visitors. For 67.7% of the visitors, this was the first time they got tested. A total of 1211 (6.1%) tested SARS-CoV-2-positive with RT-PCR, of whom 309 (25.5%) were asymptomatic. Test uptake increased among residents of the pilot neighbourhoods, especially in the older age groups, compared to people living in comparable neighbourhoods without community-based testing facilities. RDT detected asymptomatic individuals with 71.8% sensitivity, which was acceptable in this high prevalence setting. Visitors reported positive attitudes towards the test facilities and welcomed the easy access. Offering community-based SARS-CoV-2 testing seems a promising approach for increasing testing uptake among specific populations in low-income neighbourhoods.


Islet cell and extrapancreatic expression of the LIM domain homeobox gene isl-1.
The phenotypically distinct cell types of the islets of Langerhans express a number of different hormone genes in a regulated and cell-specific manner. Analysis of the molecular factors important for the control of insulin gene transcription has led to the identification of a number of insulin gene-binding proteins. The cDNAs for several of these proteins have been isolated, including isl-1, a LIM domain homeobox protein reported to be preferentially expressed in islet cells. We now report that isl-1 mRNA transcripts are present not only in islet cells, but also in pheochromocytoma (PC-12) cell lines. Isl-1 mRNA transcripts were also detected in several different regions of rat brain as well as in rat kidney. Analysis of the patterns of gene expression in rat islet cell lines and human pancreatic endocrine tumors demonstrated a variable relationship between the hormonal phenotype of the tumor and expression of the isl-1 and insulin genes. Immunocytochemical studies demonstrated both cytoplasmic and nuclear staining for isl-1 in human pancreatic endocrine tumors, in normal pancreatic A, B, D, and PP cells, and in distal tubular cells of the kidney. Isl-1 immunoreactivity was also widely distributed in neurons in both the central and peripheral nervous system, pituitary, and several human pituitary tumors and pheochromocytomas. The results of these studies implicate a potential role for isl-1 outside the endocrine pancreas in the nervous system and kidney.


Indican excretion in the elderly.
It has been suggested that up to a half of elderly hospitalized patients excrete an abnormally high concentration of indican compared with young individuals. Thus, the commonly used colorimetric qualitative test for urinary indican excess may be invalid in the aged. However, of 21 young control subjects, 18 healthy elderly subjects, and 23 disabled, elderly, long-stay patients, no individual had levels of indican detectable by the standard colorimetric qualitative test. The use of a quantitative assay demonstrated that urinary indican concentration is similar in young and elderly individuals. A diurnal variation in urinary indican concentration was discovered which may affect the interpretation of the qualitative test.


PRKCZ methylation is associated with sunlight exposure in a North American but not a Mediterranean population.
Sunlight exposure has been shown to alter DNA methylation patterns across several human cell-types, including T-lymphocytes. Since epigenetic changes establish gene expression profiles, changes in DNA methylation induced by sunlight exposure warrant investigation. The purpose of this study was to assess the effects of sunlight exposure on CD4+ T-cell methylation patterns on an epigenome-wide scale in a North American population of European origin (n=991). In addition, we investigated the genetic contribution to epigenetic variation (methylQTL). We used linear regression to test the associations between methylation scores at 461,281 cytosine-phosphate-guanine (CpG) sites and sunlight exposure, followed by a genome-wide association analysis (methylQTL) to test for associations between methylation at the top CpG locus and common genetic variants, assuming an additive genetic model. We observed an epigenome-wide significant association between sunlight exposure and methylation status at cg26930596 (p=9.2×10(-8)), a CpG site located in protein kinase C zeta (PRKCZ), a gene previously shown to be entrained by light. MethylQTL analysis resulted in significant associations between cg26930596 and two intergenic single nucleotide polymorphisms on chromosome 3, rs4574216 (p=1.5×10(-10)) and rs4405858 (p=1.9×10(-9)). These common genetic variants reside downstream of WWTR1, a transcriptional co-activator of PRKCZ. Associations observed in the North American population, however, did not replicate in an independent Mediterranean cohort. Our preliminary results support the role of sunlight exposure in epigenetic processes, and lay the groundwork for future studies of the molecular link between sunlight and physiologic processes such as tumorigenesis and metabolism.


Scale-dependent bi-trophic interactions in a semi-arid savanna: how herbivores eliminate benefits of nutrient patchiness to plants.
The scale of resource heterogeneity may influence how resources are locally partitioned between co-existing large and small organisms such as trees and grasses in savannas. Scale-related plant responses may, in turn, influence herbivore use of the vegetation. To examine these scale-dependent bi-trophic interactions, we varied fertilizer [(nitrogen (N)/phosphorus (P)/potassium (K)] applications to patches to create different scales of nutrient patchiness (patch size 2 × 2 m, 10 × 10 m, or whole-plot 50 × 50 m) in a large field experiment in intact African savanna. Within-patch fertilizer concentration and the total fertilizer load per plot were independently varied. We found that fertilization increased the leaf N and P concentrations of trees and grasses, resulting in elevated utilization by browsers and grazers. Herbivory off-take was particularly considerable at higher nutrient concentrations. Scale-dependent effects were weak. The net effect of fertilization and herbivory was that plants in fertilized areas tended to grow less and develop smaller rather than larger standing biomass compared to plants growing in areas that remained unfertilized. When all of these effects were considered together at the community (plot) level, herbivory completely eliminated the positive effects of fertilization on the plant community. While this was true for all scales of fertilization, grasses tended to profit more from coarse-grained fertilization and trees from fine-grained fertilization. We conclude that in herbivore-dominated communities, such as the African savanna, nutrient patchiness results in the herbivore community profiting rather more than the plant community, irrespective of the scale of patchiness. At the community level, the allometric scaling theory's prediction of plant-and probably also animal-production does not hold or may even be reversed as a result of complex bi-trophic interactions.


Alpha-1 Antitrypsin and Lung Cell Apoptosis.
Discovery of alpha-1 antitrypsin (A1AT) as the principal circulating inhibitor of neutrophil elastase was critical to the appreciation of protease/antiprotease imbalance involvement in the pathogenesis of emphysema. Additional targets of A1AT have been uncovered, along with their contribution to alveolar wall destruction induced by cigarette smoke exposure. We highlight in this report mechanisms of A1AT antiapoptotic effects on structural lung endothelial cells. This function was largely dependent on uptake of the protein from the circulation via clathrin- and, in part, caveolae-mediated endocytosis and on specific interactions with cysteine proteases such as capsase-3, -6, and -7. Exposures to cigarette smoke diminished A1AT intracellular uptake and its anticaspase action, suggesting that even in A1AT-suficient individuals, cigarette smoke may weaken the serpin's endothelial prosurvival effect. In addition, cigarette smoke exposure or genetic mutations known to induce posttranslational modifications such as oxidation or polymerization may alter A1AT bidirectional intracellular traffic in endothelial cells and thus determine its functional bioavailability in certain lung compartments. Uncovering and harnessing the A1AT canonical and noncanonical mechanisms will advance our understanding of the pathogenesis of emphysema and may provide means to improve the effectiveness of therapies in both A1AT-sufficient and A1AT-deficient individuals.


tRNA insulator function: insight into inheritance of transcription states?
DNA in eukaryotes is invariably present as a complex with histone and non-histone proteins called chromatin. These proteins play an important role in the proper regulation of genes during development and differentiation. Transcription factors and the covalent modifications of DNA, histone and non-histone proteins establish an epigenetic state that is heritable and which does not involve a change in genotype. The heritability of transcription states through cell division brings up specific questions: How are epigenetic marks established and re-established in the daughter cells following DNA replication and mitosis? In this article we explore what is known of the cell cycle dependence of epigenetic inheritance with particular emphasis on yeast loci and discuss the role of specific proteins responsible for the establishment and maintenance of these states.


[Bone and skin vascularization of the lower 4th of the leg].
The authors have studied the vascular network of bone and skin in the lower third of the leg by injection of methyl methacrylate in 31 cadavers. The vascular supply of the bone is given by metaphyseal and epiphyseal arteries branching from the main trunks. Different types of vascular network are described. The vascular supply of the skin is given by a subaponeurotic plexus with branches crossing in areas which can be considered as radiating vascular hili. These hili should be preserved during surgical operations. Conclusions are given concerning the security of the surgical approaches at the lower third of the leg.


Fatty acids and hypolipidemic drugs regulate peroxisome proliferator-activated receptors alpha - and gamma-mediated gene expression via liver fatty acid binding protein: a signaling path to the nucleus.
Peroxisome proliferator-activated receptor alpha (PPARalpha) is a key regulator of lipid homeostasis in hepatocytes and target for fatty acids and hypolipidemic drugs. How these signaling molecules reach the nuclear receptor is not known; however, similarities in ligand specificity suggest the liver fatty acid binding protein (L-FABP) as a possible candidate. In localization studies using laser-scanning microscopy, we show that L-FABP and PPARalpha colocalize in the nucleus of mouse primary hepatocytes. Furthermore, we demonstrate by pull-down assay and immunocoprecipitation that L-FABP interacts directly with PPARalpha. In a cell biological approach with the aid of a mammalian two-hybrid system, we provide evidence that L-FABP interacts with PPARalpha and PPARgamma but not with PPARbeta and retinoid X receptor-alpha by protein-protein contacts. In addition, we demonstrate that the observed interaction of both proteins is independent of ligand binding. Final and quantitative proof for L-FABP mediation was obtained in transactivation assays upon incubation of transiently and stably transfected HepG2 cells with saturated, monounsaturated, and polyunsaturated fatty acids as well as with hypolipidemic drugs. With all ligands applied, we observed strict correlation of PPARalpha and PPARgamma transactivation with intracellular concentrations of L-FABP. This correlation constitutes a nucleus-directed signaling by fatty acids and hypolipidemic drugs where L-FABP acts as a cytosolic gateway for these PPARalpha and PPARgamma agonists. Thus, L-FABP and the respective PPARs could serve as targets for nutrients and drugs to affect expression of PPAR-sensitive genes.


Information and knowledge in biology: time for reappraisal.
The second law of thermodynamics accounts for irreversibility of processes in the universe. As a statement about increasing disorder, it also plays a central role in creating order. Structuring is a way of how to increase the rate of dissipation of matter and energy. This is the reason why chemical reactions on Earth have produced a profusion of structures. Chemical structures with particularly high stability, maintained by continual dissipation, are designated, somewhat arbitrarily, as living systems. To preserve stability, organisms are unceasingly performing ontic work, assisted by epistemic work. Biological evolution is a progressing process of knowledge acquisition (cognition) and, correspondingly, of growth of complexity. The acquired knowledge represents epistemic complexity. Biological species are the main "bookkeepers" of acquired knowledge, with individual members of the species functioning as "explorers" of novelty. Science, a human species-specific mode of acquiring knowledge, abounds in metaphors no less than art. In the postgenomic era, the metaphor of information, along with the related metaphor of selfish genes, may need reconsideration and/or complementation. The world of great complexity, which is becoming the focus of studies of contemporary biology, may require-similarly as is the case of quantum physics-descriptions based on the principle of complementarity. Embodied knowledge, molecular engine, ontic and epistemic work, and triggering may become parts of a new conceptual armory.


Developing EDI messages and supporting data models: a "bottom up" approach based on "single business purpose" messages meets user needs.
Electronic Data Interchange (EDI) is increasingly being used for the exchange of patient, administrative, and clinical information. There is work in Europe and the United States directed at describing a preferred approach to the task of defining and modeling the information content of standard EDI messages. The literature is currently silent about the criteria for selecting the scope of an individual message. This paper argues the case for developing a larger number of discrete, simpler single purpose messages, rather than "superset" framework messages. The paper also questions the validity of the currently prevailing view that EDI messages should draw their initial content and business purpose from an already predefined model purporting to be "the model of the healthcare delivery system." The paper also examines the ongoing work on defining EDI scenarios of the Open EDI Group [1], which is developing an Open EDI Reference Model Standard on experience gained in using the approach to message development advocated in CR 1350:1993 [1] and the early experience in gaining user acceptance of the first messages developed using this approach.


Manganese-enhanced magnetic resonance imaging for early detection and characterization of breast cancers.
Very early cancer detection is the key to improving cure. Our objective was to investigate manganese (Mn)-enhanced magnetic resonance imaging (MRI) for very early detection and characterization of breast cancers. Eighteen NOD scid gamma mice were inoculated with MCF7, MDA, and LM2 breast cancer cells and imaged periodically on a 3 T scanner beginning on day 6. T1-weighted imaging and T1 measurements were performed before and 24 hours after administering MnCl2. At the last imaging session, Gd-DTPA was administered and tumors were excised for histology (hematoxylin-eosin and CD34 staining). All mice, except for two inoculated with MCF7 cells, developed tumors. Tumors enhanced uniformly on Mn and showed clear borders. Early small tumors (≤ 5 mm3) demonstrated the greatest enhancement with a relative R1 (1/T1) change of 1.57 ± 0.13. R1 increases correlated with tumor size (r = -.34, p = .04). Differences in R1 increases among the three tumor subtypes were most evident in early tumors. Histology confirmed uniform cancer cell distribution within tumor masses and vasculature in the periphery, which was consistent with rim-like enhancement on Gd-DTPA. Mn-enhanced MRI is a promising approach for detecting very small breast cancers in vivo and may be valuable for very early cancer detection.


Effects of removal of the adventitia of the descending aorta and structural alterations in the tunica media in pigs.
To investigate the effects of removal of the adventitia on the tunica media in a pig model. The experiment was performed in eight pigs. The adventitia of the descending aorta was removed. Following euthanasia, at two, four, six and eight weeks, the aortic segment was removed. Next, slices of the aorta were stained with hematoxylin and eosin (HE) and Weigert-Van Gieson. After two weeks there was a slight cellular breakdown in the outer third of the media. After four weeks structural breakdown of elastic fibers was observed in the outer two thirds of the same layer. In six weeks, several areas of necrosis and almost complete disruption of elastic fibers were identified. Finally, after eight weeks, there was fibrosis of the entire wall with disruption of the internal elastic lamina. The removal of the adventitia leads to degeneration of the media, determining loss of the normal structure of the aortic wall that is variable in its location, intensity and shape, depending on the length and duration of the ischemic insult.


Exosome miR-335 as a novel therapeutic strategy in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is a common and deadly cancer. Most cases of HCC arise in a cirrhotic/fibrotic liver, indicating that environment may play a paramount role in cancer genesis. Previous studies from our group and others have shown that, in desmoplastic cancers, there is a rich intercellular communication between activated, cancer-associated fibroblasts and cancer cells. Moreover, extracellular vesicles (EVs), or exosomes, have been identified as an important arm of this intercellular communication platform. Finally, these studies have shown that EVs can carry microRNA (miR) species in vivo and deliver them to desmoplastic cancers. The precise role played by activated liver fibroblasts/stellate cells in HCC development is insufficiently known. Based on previous studies, it appears plausible that activated fibroblasts produce signals carried by EVs that promote HCC genesis. In the current study, we first hypothesized and then demonstrated that stellate cell-derived EVs 1) can be loaded with an miR species of choice (miR-335-5p); 2) are taken up by HCC cells in vitro and more importantly in vivo; 3) can supply the miR-335-5p cargo to recipient HCC cells in vitro as well as in vivo; and 4) inhibit HCC cell proliferation and invasion in vitro as well as induce HCC tumor shrinkage in vivo. Finally, we identified messenger RNA targets for miR-335 that are down-regulated after treatment with EV-miR-335-5p. This study informs potential therapeutic strategies in HCC, whereby stellate cell-derived EVs are loaded with therapeutic nucleic acids and delivered in vivo. (Hepatology 2018;67:940-954).


Suitability of poroelastic and viscoelastic mechanical models for high and low frequency MR elastography.
Descriptions of the structure of brain tissue as a porous cellular matrix support application of a poroelastic (PE) mechanical model which includes both solid and fluid phases. However, the majority of brain magnetic resonance elastography (MRE) studies use a single phase viscoelastic (VE) model to describe brain tissue behavior, in part due to availability of relatively simple direct inversion strategies for mechanical property estimation. A notable exception is low frequency intrinsic actuation MRE, where PE mechanical properties are imaged with a nonlinear inversion algorithm. This paper investigates the effect of model choice at each end of the spectrum of in vivo human brain actuation frequencies. Repeat MRE examinations of the brains of healthy volunteers were used to compare image quality and repeatability for each inversion model for both 50 Hz externally produced motion and ≈1 Hz intrinsic motions. Additionally, realistic simulated MRE data were generated with both VE and PE finite element solvers to investigate the effect of inappropriate model choice for ideal VE and PE materials. In vivo, MRE data revealed that VE inversions appear more representative of anatomical structure and quantitatively repeatable for 50 Hz induced motions, whereas PE inversion produces better results at 1 Hz. Reasonable VE approximations of PE materials can be derived by equating the equivalent wave velocities for the two models, provided that the timescale of fluid equilibration is not similar to the period of actuation. An approximation of the equilibration time for human brain reveals that this condition is violated at 1 Hz but not at 50 Hz. Additionally, simulation experiments when using the "wrong" model for the inversion demonstrated reasonable shear modulus reconstructions at 50 Hz, whereas cross-model inversions at 1 Hz were poor quality. Attenuation parameters were sensitive to changes in the forward model at both frequencies, however, no spatial information was recovered because the mechanisms of VE and PE attenuation are different. VE inversions are simpler with fewer unknown properties and may be sufficient to capture the mechanical behavior of PE brain tissue at higher actuation frequencies. However, accurate modeling of the fluid phase is required to produce useful mechanical property images at the lower frequencies of intrinsic brain motions.


Localization of a yeast early Golgi mannosyltransferase, Och1p, involves retrograde transport.
To analyze the mechanism of integral membrane protein localization in the early Golgi apparatus of Saccharomyces cerevisiae, we have used Och1p, a cis-Golgi mannosyltransferase. A series of influenza virus hemagglutinin (HA) epitope-tagged fusion proteins was constructed in which invertase is appended to the Golgi-luminal carboxy terminus of full-length Och1p. Several constructs included a Kex2p cleavage site between the Och1p and invertase moieties to monitor transit to the Kex2p-containing TGN. Cells expressing an Och1p-invertase fusion do not secrete invertase, but those expressing an Och1p-Kex2p site-invertase fusion protein secrete high levels of invertase in a Kex2p-dependent manner. The Och1p-Kex2p site-invertase fusion protein is cleaved with a half-time of 5 min, and the process proceeds to completion. Before cleavage the protein receives glycosyl modifications indicative of passage through the medial- and trans-Golgi, therefore cleavage occurs after ordered anterograde transport through the Golgi to the TGN. Transit to distal compartments is not induced by the invertase moiety, since noninvertase fusion constructs encounter the same glycosyltransferases and Kex2p as well. The Och1p-HA moiety, irrespective of whether it is generated by cleavage of the fusion protein in the TGN or synthesized de novo, is degraded with a half-time of about 60 min. Thus, the half-time of degradation is 12-fold longer than the time required to reach the TGN. At steady state, de novo-synthesized and TGN-generated HA epitope-tagged Och1p reside in a compartment with a buoyant density identical to that of wild-type Och1p and distinct from that of the vacuole or the TGN. Finally, och1 null cells that express an Ochlp fusion construct known to rapidly encounter the TGN glycosylate invertase to the same extent as wild-type cells, indicating that they have phenotypically wild-type Och1p activity. These results lead us to propose a model for Och1p-HA localization that involves movement to distal compartments, at least as far as the TGN, followed by retrieval to the cis compartment, presumably by vesicular transport.


IgA-mediated phagocytosis by mouse alveolar macrophages.
Ingestion of antibody-opsonized sheep red blood cells by murine alveolar and peritoneal exudate macrophages was studied. Alveolar macrophages or peritoneal macrophages were isolated by lavage and incubated with TNP-SRBC, which had been preincubated with anti-DNP IgG, IgA, and IgE. Significant enhancement of phagocytosis by alveolar macrophages of TNP-SRBC was mediated by all classes of antibody examined, most markedly with IgG and less so with IgA and IgE. Enhancement of phagocytosis by peritoneal macrophages was mediated by IgG and IgE, but not by IgA. These results suggest that the interaction of IgA and Fc receptors for IgA on alveolar macrophages may increase the level of this cell's function in the mammalian lung to clear pathogens and immune complexes from the alveolar spaces.


The contribution of anulus fibers to torque resistance.
Anulus fibers of the intervertebral disc oriented in one direction were dissected, and oppositely directed fibers were left intact as a result of a newly developed dissection method. Motion segments were dissected by this way, and motion segments after bilateral facetectomy were loaded in torsion and compared with each other. To assess the contribution of facets and anulus fibers to torque resistance. Mathematical models predict that torsional stress is transmitted only to those collagene fibers of the anulus that are angled in the direction of the applied torque. Torsion and forward bending or torsion and compression are likely to cause anulus damage. No experimental study that we are aware of has confirmed that anulus fibers are the main structure to resist against torque. Pure axial rotation moments were applied on 12 lumbar motion segments. The six components of motion were recorded. Six motion segments were investigated intact after dissection of anulus fibers directed in one direction and after additional bilateral facetectomy. In six motion segments, bilateral facetectomy was performed before anulus dissection. With the application of an axial rotation moment of 8.5 Nm to the left, axial rotation increased 2 degrees after dissection of disc fibers in one direction, and 1.2 degrees after bilateral facetectomy (P = 0.002). In the opposite direction, there were no differences. After both injuries, axial rotation was 7.6 degrees to the left (direction of fiber dissection) and -3.3 degrees to the right (P = 0.0005). In lumbar motion segments without degeneration, anulus fibers restrict axial rotation more than the facets.


Use of Amberlite Macroporous Resins To Reduce Bitterness in Whole Olives for Improved Processing Sustainability.
Olives are inedible because of high levels of bitter phenolics (e.g., oleuropein) which are removed during commercial olive processing. Current commercial processing methods are highly water-intensive, produce toxic wastewater, and are environmentally unsustainable. To address this, macroreticular polymeric resins were used to assist debittering and decrease water use. Amberlite resins XAD4, XAD16N, XAD7HP, and FPX66 were evaluated for the ability to adsorb bitter and/or high-value phenolic compounds (i.e., oleuropein, ligstroside, oleuropein aglycone, ligstroside aglycone, oleocanthal, oleacein, and hydroxytyrosol) from whole olives during typical brine storage. All resins effectively adsorbed oleuropein and ligstroside. FPX66 reduced oleuropein in whole olives suspended in a 1.0% acetic acid brine to 0.635 mg/kg wet weight in 2.5 months with no further processing. This concentration is below levels measured in commercial California-style black ripe olives (0.975 mg/kg wet weight). Resins in storage brines effectively decrease levels of bitter phenolic compounds without additional lye processing. Excellent recoveries of high-value phenolic compounds are obtained from resins (e.g., 80.2 ± 3.3% to 89.4 ± 8.9% hydroxytyrosol).


Derivation of a clinical prediction model for the emergency department diagnosis of ectopic pregnancy.
To derive a clinical prediction model for estimating the pretest probability of ectopic pregnancy in ED patients with first-trimester abdominal pain or vaginal bleeding. All hemodynamically stable first-trimester patients presenting to the ED of a tertiary care military teaching hospital over a 14-month period with a chief complaint of abdominal pain and/or vaginal bleeding had clinical data coded prior to determining outcome. They were then followed longitudinally until a criterion standard pregnancy outcome was established. Of the 486 patients enrolled, 280 (58%) had viable intrauterine pregnancies, 167 (34%) had nonviable intrauterine pregnancies, and 39 (8%) had ectopic pregnancies. Using a recursive partitioning model, a high-risk group was derived (that was separated from intermediate and low-risk groups), consisting of patients with abdominal peritoneal signs or definite cervical motion tenderness, with a sensitivity of 31% (95% CI: 17-48%), a specificity of 93% (95% CI: 90-95%), a positive likelihood ratio of 4.3, and a negative likelihood ratio of 0.74. A low-risk group, consisting of patients with either fetal heart tones or tissue at the cervical os, or the absence of pain other than midline menstrual-like cramping and lacking any pelvic tenderness, was differentiated from an intermediate-risk group, with a sensitivity of 96% (95% CI: 81-100%), a specificity of 22% (95% CI: 18-26%), a positive likelihood ratio of 1.2, and a negative likelihood ratio of 0.17. A clinical prediction model for estimating the probability of ectopic pregnancy in ED patients has been derived. It may prove to have practical clinical application for estimating pretest probability of ectopic pregnancy as well as assisting in medical decision making when laboratory and ultrasonographic findings are nondiagnostic. Clinical application should await prospective validation in an independent sample.


Multicentre, randomised, double blind, placebo controlled, phase III study of weekly, low dose, subcutaneous interferon beta-1a in secondary progressive multiple sclerosis.
Interferon (IFN) beta has repeatedly shown benefit in multiple sclerosis (MS) in reducing the rate of relapse, the disease activity as shown with magnetic resonance imaging and, to some degree, the progression of disability; however, it is unknown how much the therapeutic response depends on the dose, the subgroup involved, and the disease stage. This multicentre, double blind, placebo controlled study explored the dose-response curve by examining the clinical benefit of low dose IFN beta-1a (Rebif), 22 micro g subcutaneously once weekly, in patients with secondary progressive MS. A total of 371 patients with clinically definite SPMS were randomised to receive either placebo or subcutaneous IFN beta-1a, 22 micro g once weekly, for 3 years. Clinical assessments were performed every 6 months. The primary outcome was time to sustained disability, as defined by time to first confirmed 1.0 point increase on the Expanded Disability Status Scale (EDSS). Secondary outcomes included a sensitive disability measure and relapse rate. Treatment had no beneficial effect on time to confirmed progression on either the EDSS (hazard ratio (HR) = 1.13; 95% confidence interval (CI) 0.82 to 1.57; p = 0.45 for 22 micro g v placebo) or the Regional Functional Status Scale (HR = 0.93; 95% CI 0.68 to 1.28; p = 0.67). Other disability measures were also not significantly affected by treatment. Annual relapse rate was 0.27 with placebo and 0.25 with IFN (rate ratio = 0.90; 95% CI 0.64 to 1.27; p = 0.55). The drug was well tolerated with no new safety concerns identified. No significant gender differences were noted. This patient population was less clinically active than SPMS populations studied in other trials. Treatment with low dose, IFN beta-1a (Rebif) once weekly did not show any benefit in this study for either disability or relapse outcomes, including a subgroup with preceding relapses. These results add a point at one extreme of the dose-response spectrum of IFN beta therapy in MS, indicating that relapses in this phase may need treatment with higher doses than in the initial phases.


Flagellar motors of marine bacteria Halomonas are driven by both protons and sodium ions.
Bacterial cells in aquatic environments are able to reach or stay near nutrient patches by using motility. Motility is usually attained by rotating flagellar motors that are energized by electrochemical potential of H+ or Na+. In this paper, the ion specificity for flagellar rotation of two marine isolates Halomonas spp. strains US172 and US201 was investigated. Both isolates require sodium for growth and possess a respiratory-driven primary sodium pump. They are motile because of lateral flagella regardless of the presence of sodium ions. Their swimming speed under various concentrations of sodium ions with and without carbonylcyanide m-chlorophenylhydrazone, a proton conductor, and with and without phenamil, a specific inhibitor for the sodium-driven flagellar motors, was examined. The effect of carbonylcyanide m-chlorophenylhydrazone on the transmembrane proton gradient was also determined. Our results showed that the flagellar motors of the Halomonas strains were energized by both H+ and Na+ in one cell. The bimodal nature of Halomonas spp. motility with respect to the driving energy source may reflect ecophysiological versatility to adapt to a wide range of salt conditions of the marine environment.


Performance of a diffusional clearance model for beta-lactam antimicrobial agents as influenced by extravascular protein binding and interstitial fluid kinetics.
A physiological model based on diffusional clearance (CLD) of drug between plasma and interstitial fluid (IF) was used to describe the disposition of beta-lactam antimicrobial agents. The CLD represents the movement of drug in and out of physiological spaces and is dependent only on the transfer properties of the drug. Estimates of CLD obtained by fitting model equations to plasma concentration-time data for 11 cephalosporin studies in human subjects fell in a fairly narrow range, with a mean value of 1,604 ml/min. Estimates of the CLD between plasma and blister fluid for three of the cephalosporins were five orders of magnitude smaller than the CLD. These observations are explained in terms of diffusion principles. Computer simulations with this model were used to assess the effect of changes in IF protein binding on antimicrobial distribution. Increases in the bound fraction of drug in IF enhanced the penetration of total (bound and unbound) drug into IF, but had no effect on the amount of unbound, active antimicrobial agent reaching the IF. The time course of unbound drug in IF was altered, however, by changes in IF protein binding. This model may also be used to predict changes in the IF distribution of beta-lactam antimicrobial agents in disease states, particularly those in which the relative distribution of albumin between plasma and IF has been altered.


Subsurface landfill leachate contamination affects microbial metabolic potential and gene expression in the Banisveld aquifer.
Microbial communities in groundwater ecosystems can develop the capacity to degrade complex mixtures of chemicals resulting from pollution by landfill leachate. Monitoring this natural attenuation requires insight into the metabolic potential and activity of microbial communities. We contrasted the metagenomes and metatranscriptomes from a leachate-polluted aquifer downstream of the Banisveld (the Netherlands) landfill with uncontaminated groundwater, which revealed changes in microbial genomic content and activity. Banisveld landfill leachate contains mono-aromatic hydrocarbons and the assessment of natural attenuation of these compounds in the aquifer had been a focal point of research. In the contaminated groundwater, active microbial functions were the ones involved in degradation of complex carbon substrates and organic pollutants. We found that benzylsuccinate synthase genes-involved in the catabolism of toluene-were highly expressed close to the source of contamination, confirming the ongoing natural attenuation of organic mono-aromatic hydrocarbon pollution in this aquifer. Additionally, metatranscriptomes were indicative of phosphorus limitation that can constrain total microbial activity and agree with the low phosphate concentrations (<0.4 μmol/L) in this aquifer. Through the application of metagenomics and metatranscriptomics, we were able to determine functional potential and expression patterns to assess the natural attenuation processes and constraints on microbial communities.


Lateral tarsoconjunctival onlay flap lower eyelid suspension in facial nerve paresis.
To evaluate the efficacy of a lateral tarsoconjunctival flap suspension procedure to improve paralytic eyelid malposition. Retrospective chart review (with photograph and videographic data) of consecutive patients between 2008 and 2013 with permanent unilateral paralytic eyelid malposition treated with a far lateral tarsoconjunctival flap lower eyelid suspension alone or in conjunction with lateral canthoplasty. Upper and lower eyelid position, lagophthalmos, ocular surface disease, patient satisfaction, and cosmesis were recorded before and after intervention. Postoperative complications and subsequent management were also recorded. A total of 110 patients were identified. Lagophthalmos, ocular surface exposure, exposure keratopathy, and eyelid retraction (both upper and lower) improved in all cases. All patients described an improvement in eye irritation, epiphora, and cosmesis. Patients with a good Bell's reflex exhibited some dynamic function to the lower eyelid on videography. Three (2.7%) eyelids had flap dehiscence that was successfully treated with repeat suturing. Seven (6.4%) patients had symptomatic temporal peripheral vision requiring partial flap takedown. Five (4.5%) patients had pyogenic granulomas excised. Seven (6.4%) patients later had a supplemental medial tarsorrhaphy for medial lagophthalmos. The lateral tarsoconjunctival flap suspension was highly effective in this series of patients with paralytic eyelid malposition. Improved eyelid position and cosmesis were universal although some patients benefitted from an additional medial tarsorrhaphy. Return of natural upward lower eyelid movement upon eyelid closure was a further benefit in some patients. Temporal vision obstruction was an infrequent side effect and flap dehiscence and granulomas were rare complications.


Antifungal Activity of a Novel Triazole, Efinaconazole and Nine Comparators against 354 Molecularly Identified Aspergillus Isolates.
Management of superficial aspergillosis is a major challenge owing to the frequent relapses and treatment failure, which may pose a potential risk, thereby gradually developing resistant species. Therefore, necessitating the development of new antifungals with higher potency should be considered as alternative strategies for efficient management of infections. We aimed to investigate the susceptibility of Aspergillus isolates toward a novel triazole, efinaconazole, in comparison with various classes of antifungal drugs. Antifungal susceptibility testing was performed according to the Clinical and Laboratory Standards Institute M38-A2 guidelines. Efinaconazole exhibited poor activity against mutant A. fumigatus strains, A. niger sensu stricto, and A. tubingensis with GM MIC values of 3.62, 1.62, and 2 μg/ml, respectively; however, surprisingly, it efficiently inhibited the growth of A. terreus sensu stricto, followed by wild-type A. fumigatus and A. flavus with GM MIC values of 0.29, 0.42, and 0.52 μg/ml, respectively. Presumably, efinaconazole is inefficient in aspergillosis treatment due to the low susceptibility of A. niger sensu stricto, A. tubingensis, and mutant A. fumigatus; however, it may be effective in treating superficial aspergillosis caused by wild-type A. fumigatus, A. terreus sensu stricto, and A. flavus. Further studies are needed to determine how these findings may translate into in vivo efficacy.