Phase 1 trial of bevacizumab with concurrent chemoradiation therapy for squamous cell carcinoma of the head and neck with exploratory functional imaging of tumor hypoxia, proliferation, and perfusion.
A phase 1 trial was completed to examine the safety and feasibility of combining bevacizumab with radiation and cisplatin in patients with locoregionally advanced squamous cell carcinoma of the head and neck (HNSCC) treated with curative intent. Additionally, we assessed the capacity of bevacizumab to induce an early tumor response as measured by a series of biological imaging studies. All patients received a single induction dose of bevacizumab (15 mg/kg) delivered 3 weeks (±3 days) before the initiation of chemoradiation therapy. After the initial dose of bevacizumab, comprehensive head and neck chemoradiation therapy was delivered with curative intent to 70 Gy in 33 fractions with concurrent weekly cisplatin at 30 mg/m(2) and bevacizumab every 3 weeks (weeks 1, 4, 7) with dose escalation from 5 to 10 to 15 mg/kg. All patients underwent experimental imaging with [(18)F]fluorothymidine positron emission tomography (FLT-PET) (proliferation), [(61)Cu]Cu-diacetyl-bis(N4-methylthiosemicarbazone) PET (Cu-ATSM-PET) (hypoxia), and dynamic contrast-enhanced computed tomography (DCE-CT) (perfusion) at 3 time points: before bevacizumab monotherapy, after bevacizumab monotherapy, and during the combined therapy course. Ten patients were enrolled. All had stage IV HNSCC, all achieved a complete response to treatment, and 9 of 10 remain alive, with a mean survival time of 61.3 months. All patients experienced grade 3 toxicity, but no dose-limiting toxicities or significant bleeding episodes were observed. Significant reductions were noted in tumor proliferation (FLT-PET), tumor hypoxia (Cu-ATSM-PET), and DCE-CT contrast enhancement after bevacizumab monotherapy, with further decreases in FLT-PET and Cu-ATSM-PET during the combined therapy course. The incorporation of bevacizumab into comprehensive chemoradiation therapy regimens for patients with HNSCC appears safe and feasible. Experimental imaging demonstrates measureable changes in tumor proliferation, hypoxia, and perfusion after bevacizumab monotherapy and during chemoradiation therapy. These findings suggest opportunities to preview the clinical outcomes for individual patients and thereby design personalized therapy approaches in future trials.


Fixed Duration of Venetoclax-Rituximab in Relapsed/Refractory Chronic Lymphocytic Leukemia Eradicates Minimal Residual Disease and Prolongs Survival: Post-Treatment Follow-Up of the MURANO Phase III Study.
The MURANO study demonstrated significant progression-free survival (PFS) benefit for fixed-duration venetoclax-rituximab compared with bendamustine-rituximab in relapsed/refractory chronic lymphocytic leukemia. With all patients off treatment, we report minimal residual disease (MRD) kinetics and updated outcomes. Patients were randomly assigned to 2 years of venetoclax plus rituximab during the first six cycles, or six cycles of bendamustine-rituximab. Primary end point was PFS. Safety and peripheral blood (PB) MRD status-at cycle 4, 2 to 3 months after end of combination therapy (EOCT), and every 3 to 6 months thereafter-were secondary end points. Of 194 patients, 174 (90%) completed the venetoclax-rituximab phase and 130 (67%) completed 2 years of venetoclax. With a median follow-up of 36 months, PFS and overall survival remain superior to bendamustine-rituximab (hazard ratio, 0.16 [95% CI, 0.12 to 0.23]; and hazard ratio, 0.50 [95% CI, 0.30 to 0.85], respectively). Patients who received venetoclax-rituximab achieved a higher rate of PB undetectable MRD (uMRD; less than 10-4) at EOCT (62% v 13%) with superiority sustained through month 24 (end of therapy). Overall, uMRD status at EOCT predicted longer PFS. Among those with detectable MRD, low-level MRD (10-4 to less than 10-2) predicted improved PFS compared with high-level MRD (10-2 or greater). At a median of 9.9 months (range, 1.4 to 22.5 months) after completing fixed-duration venetoclax-rituximab, overall only 12% (16 of 130) of patients developed disease progression (11 high-level MRD, three low-level MRD). At the end of therapy, 70% and 98% of patients with uMRD remained in uMRD and without disease progression, respectively. With all patients having finished treatment, continued benefit was observed for venetoclax-rituximab compared with bendamustine-rituximab. uMRD rates were durable and predicted longer PFS, which establishes the impact of PB MRD on the benefit of fixed-duration, venetoclax-containing treatment. Low conversion to detectable MRD and sustained PFS after completion of 2 years of venetoclax-rituximab demonstrate the feasibility of this regimen.


In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice.
Mitochondria are the powerhouses of cells and produce cellular energy in the form of ATP. Mitochondrial dysfunction contributes to biological aging and a wide variety of disorders including metabolic diseases, premature aging syndromes, and neurodegenerative diseases such as Alzheimer's disease (AD) and Parkinson's disease (PD). Maintenance of mitochondrial health depends on mitochondrial biogenesis and the efficient clearance of dysfunctional mitochondria through mitophagy. Experimental methods to accurately detect autophagy/mitophagy, especially in animal models, have been challenging to develop. Recent progress towards the understanding of the molecular mechanisms of mitophagy has enabled the development of novel mitophagy detection techniques. Here, we introduce several versatile techniques to monitor mitophagy in human cells, Caenorhabditis elegans (e.g., Rosella and DCT-1/ LGG-1 strains), and mice (mt-Keima). A combination of these mitophagy detection techniques, including cross-species evaluation, will improve the accuracy of mitophagy measurements and lead to a better understanding of the role of mitophagy in health and disease.


Homo erectus calvarium from the Pleistocene of Java.
A Homo erectus calvarium [Sambungmacan 4 (Sm 4)] was recovered from Pleistocene sediments at Sambungmacan in central Java. Micro-computed tomography analysis shows a modern human-like cranial base flexion associated with a low platycephalic vault, implying that the evolution of human cranial globularity was independent of cranial base flexion. The overall morphology of Sm 4 is intermediate between that of earlier and later Javanese Homo erectus; apparent morphological specializations are more strongly expressed in the latter. This supports the hypothesis that later Pleistocene Javanese populations were substantially isolated and made minimal contributions to the ancestry of modern humans.


Acute and Long-Term Effects of Brief Sevoflurane Anesthesia During the Early Postnatal Period in Rats.
The possibility that exposure to general anesthetics during early life results in long-term impairment of neural function attracted considerable interest over the past decade. Extensive laboratory data suggest that administration of these drugs during critical stages of central nervous system development can lead to cell death, impaired neurogenesis, and synaptic growth as well as cognitive deficits. These observations are corroborated by several recent human epidemiological studies arguing that such cognitive impairment might also occur in humans. Despite the potential public health importance of this issue, several important questions remain open. Amongst them, how the duration of anesthesia exposure impact on outcome is as yet not fully elucidated. To gain insight into this question, here we focused on the short- and long-term impact of a 30-min-long exposure to clinically relevant concentrations of sevoflurane in rat pups at 2 functionally distinct stages of the brain growth spurt. We show that this treatment paradigm induced developmental stage-dependent and brain region-specific acute but not lasting changes in dendritic spine densities. Electrophysiological recordings in hippocampal brain slices from adult animals exposed to anesthesia in the early postnatal period revealed larger paired-pulse facilitation but no changes in the long-term potentiation paradigm when compared with nonanesthetized controls. 5-bromo-2-deoxyuridine pulse and pulse-chase experiments demonstrated that neither proliferation nor differentiation and survival of hippocampal progenitors were affected by sevoflurane exposure. In addition, behavioral testing of short- and long-term memory showed no differences between control and sevoflurane-exposed animals. Overall, these results suggest that brief sevoflurane exposure during critical periods of early postnatal development, although it does not seem to exert major long-term effects on brain circuitry development, can induce subtle changes in synaptic plasticity and spine density of which the physiological significance remains to be determined.


Treatment of autoimmune hemolytic anemia with Vinca-loaded platelets.
We worked with a new strategy for the treatment of autoimmune hemolytic anemia (AIHA) of the warm antibody type. Platelets were loaded with Vinca alkaloids and reacted with antiplatelet antibodies to facilitate their phagocytosis by macrophages, resulting in a measure of selective delivery of the drugs to the cells of the mononuclear phagocyte system, which destroy RBCs in AIHA. Four patients were studied. Three with AIHA refractory to splenectomy and the use of moderate doses of glucocorticoids achieved hematologic and clinical remissions. Administration of steroids was either discontinued or reduced to minimal doses after treatment. Remissions lasted three years in two patients and one year in the other. Responses were characterized by prompt prolongation of RBC survival, correction of anemia, and gradual decrease in cell-bound antibody. In one nonsplenectomized patient, the disorder did not respond. Platelet-Vinca alkaloid complex is useful in the treatment of AIHA refractory to splenectomy and use of steroids.


Different genetic characteristics of Plasmodium falciparum isolates collected during successive clinical malaria episodes in Senegalese children.
A narrow epidemiologic survey was conducted during a four-month period of intense malaria transmission in Dielmo, a holoendemic Senegalese village. Longitudinal clinical and parasitologic follow-up indicate that clinical malaria episodes always occurred after an abrupt increase in parasite densities. Polymerase chain reaction analysis of Plasmodium falciparum parasites was carried out in blood samples collected longitudinally from 10 children who had experienced several clinical episodes during this period. Our data show that the genetic diversity of the parasites circulating in this village is very large. The successive clinical episodes experienced by each child were caused by genetically distinct parasite populations that were recently inoculated and multiplied in an apparently unrestricted manner. Importantly, the genetic characteristics of the parasite populations detected during phases of asymptomatic carriage differed from those causing a clinical episode, suggesting that the various factors that control of parasite growth in these children are strain-specific.


Is it Possible to Recover Cardiac Functions After Total Knee Arthroplasty?
Patients suffering from knee osteoarthritis lead a less active life than their healthy peers. It is well known that insufficient physical activity is the most common cause of chronic diseases. However, there is not enough research to enlighten the effect of increased functional capacity on cardiac functions after Total Knee Arthroplasty (TKA). This study aimed to investigate whether the orthopedic surgeons can predict that the patients will be healthier after TKA in terms of cardiac functions or not? 109 patients who underwent TKA were prospectively followed for one year. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and short form 36 (SF-36) surveys, BMI measures, average step count per day, the six-minute walking test (6MWT), the Five-Times-Sit-to-Stand Test (FTSST) and Doppler echocardiography were performed both in the preoperative and postoperative period. After TKA, there was a substantial improvement in terms of WOMAC and SF36 survey scores. The average step count increased from 2199.6±690.8 steps/day to 4124.3±1638.8 steps/day. 6MWT and FTSST improved significantly as well. The average brisk walking time was 174.23±95.11 minutes/week. The means of early and late mitral inflow velocity ratios (E/A and Em/Am ratios) increased from 0.71±0.12 to 0.77±0.13 and from 0.66±0.13 to 0.76± 0.15 at the first year follow-up visit, respectively (p<0.001). In the first year, objective physical capacity measures increased together with the expected improvements in disease-specific and generic measures. After TKA, left ventricular diastolic functions may be considered to have recovered in the light of the healing signs via echocardiography.


Therapeutic exercise for rotator cuff tendinopathy: a systematic review of contextual factors and prescription parameters.
Exercise is widely regarded as an effective intervention for symptomatic rotator cuff tendinopathy but the prescription is diverse and the important components of such programmes are not well understood. The objective of this study was to systematically review the contextual factors and prescription parameters of published exercise programmes for rotator cuff tendinopathy, to generate recommendations based on current evidence. An electronic search of AMED, CiNAHL, CENTRAL, MEDLINE, PEDro and SPORTDiscus was undertaken from their inception to June 2014 and supplemented by hand searching. Eligible studies included randomized controlled trials evaluating the effectiveness of exercise in participants with rotator cuff tendinopathy. Included studies were appraised using the Cochrane risk of bias tool and synthesized narratively. Fourteen studies were included, and suggested that exercise programmes are widely applicable and can be successfully designed by physiotherapists with varying experience; whether the exercise is completed at home or within a clinic setting does not appear to matter and neither does pain production or pain avoidance during exercise; inclusion of some level of resistance does seem to matter although the optimal level is unclear, the optimal number of repetitions is also unclear but higher repetitions might confer superior outcomes; three sets of exercise are preferable to two or one set but the optimal frequency is unknown; most programmes should demonstrate clinically significant outcomes by 12 weeks. This systematic review has offered preliminary guidance in relation to contextual factors and prescription parameters to aid development and application of exercise programmes for rotator cuff tendinopathy.


Budding yeast Wpl1(Rad61)-Pds5 complex counteracts sister chromatid cohesion-establishing reaction.
Sister chromatid cohesion, which is mediated by the cohesin complex, is vital for faithful segregation of chromosomes in mitosis and meiosis (reviewed in). Cohesion is established during S phase, and this process requires the function of the acetyltransferase Eco1/Ctf7. The mechanism of the cohesion establishment is, however, still unclear. Here, we describe isolation and identification of genetic suppressors of budding yeast eco1-1 temperature-sensitive mutant. By using a recently described microarray-based method, we successfully mapped 11 intergenic suppressor mutations in two genes, wpl1 (also known as rad61) and pds5. Pds5 is a known accessory factor of cohesin complex, and we show that Wpl1/Rad61 protein forms a complex with Pds5 and colocalizes with cohesin on chromosomes, as its presumed human homolog Wapl. Impaired function of Wpl1-Pds5 complex makes Eco1 dispensable for cell survival. We also provide evidence that Wpl1 is required for efficient association of cohesin with G2 phase chromosomes and that Eco1 promotes dissociation of Wpl1-Pds5 from cohesin via acetylation of Smc3, a cohesin subunit. Taken together, the presented data suggest that Wpl1-Pds5 complex is inhibitory for cohesion establishment and that Eco1 establishes cohesion by hindering the function of Wpl1-Pds5 temporally in S phase.


Structure and promoter activity of an islet-specific glucose-6-phosphatase catalytic subunit-related gene.
In liver and kidney, the terminal step in the gluconeogenic pathway is catalyzed by glucose-6-phosphatase (G-6-Pase). This enzyme is actually a multicomponent system, the catalytic subunit of which was recently cloned. Numerous reports have also described the presence of G-6-Pase activity in islets, although the role of G-6-Pase in this tissue is unclear. Arden and associates have described the cloning of a novel cDNA that encodes an islet-specific G-6-Pase catalytic subunit-related protein (IGRP) (Arden SD, Zahn T, Steegers S, Webb S, Bergman B, O'Brien RM, Hutton JC: Molecular cloning of a pancreatic islet-specific glucose-6-phosphatase catalytic subunit related protein (IGRP). Diabetes 48:531-542, 1999). We screened a mouse BAC library with this cDNA to isolate the IGRP gene, which spans approximately 8 kbp of genomic DNA. The exon/intron structure of the IGRP gene has been mapped and, as with the gene encoding the liver/kidney G-6-Pase catalytic subunit, it is composed of five exons. The sizes of these exons are 254 (I), 110 (II), 112 (III), 116 (IV), and 1284 (V) bp, similar to those of the G-6-Pase catalytic subunit gene. Two interspecific backcross DNA mapping panels were used to unambiguously localize the IGRP gene (map symbol G6pc-rs) to the proximal portion of mouse chromosome 2. The IGRP gene transcription start site was mapped by primer extension analysis, and the activity of the IGRP gene promoter was analyzed in both the islet-derived HIT cell line and the liver-derived HepG2 cell line. The IGRP and G-6-Pase catalytic subunit gene promoters show a reciprocal pattern of activity, with the IGRP promoter being approximately 150-fold more active than the G-6-Pase promoter in HIT cells.


[6]-Gingerol-induced cell cycle arrest, reactive oxygen species generation, and disruption of mitochondrial membrane potential are associated with apoptosis in human gastric cancer (AGS) cells.
Ginger (Zingiber officinale Roscoe), a monocotyledonous herb, is widely used as an herbal medicine owing to the phytoconstituents it possesses. In the current study, the quantity of [6]-gingerol, the major phenolic ketone, in the fresh ginger and dried ginger rhizome was found to be 6.11 µg/mg and 0.407 µg/mg. Furthermore, [6]-gingerol was assessed for its antiapoptotic effects in human gastric adenocarcinoma (AGS) cells evidenced by acridine orange/ethidium bromide staining technique and Annexin-V assay. An increase in reactive oxygen species (ROS) generation led to a decrease in mitochondrial membrane potential (MMP) and subsequent induction of apoptosis. Results disclose that perturbations in MMP are associated with deregulation of Bax/Bcl-2 ratio at protein level, which leads to upregulation of cytochrome-c triggering the caspase cascade. These enduringly suggest that [6]-gingerol can be effectively used for targeting the mitochondrial energy metabolism to manage gastric cancer cells.


Microphthalmia resulting from MSX2-induced apoptosis in the optic vesicle.
Microphthalmia is a relatively common ocular malformation. Molecular mechanisms that lead to this dire condition are largely unknown. Msx genes have been shown to be expressed in the developing eye. In the Msx1;Msx2, double mutant mouse, eye development arrests early in embryogenesis. To investigate possible functions of Msx2 in early ocular development, we created transgenic animals that overexpress Msx2. Msx2 transgenic embryos and nontransgenic littermates were examined histopathologically. The effect of Msx2 overexpression on retinal cell proliferation was assayed by bromodeoxyuridine (BrdU) incorporation and immunohistochemical staining. Apoptosis was determined by TUNEL labeling. Expression of retina and retinal pigmented epithelium (RPE)-specific genes was investigated by performing in situ hybridization or immunohistochemical staining. Forced expression of the Msx2 gene resulted in optic nerve aplasia and microphthalmia in all transgenic animals. In developing retinas of Msx2 transgenic animals, proliferation was significantly reduced and increased numbers of retinal cells underwent apoptosis. Marker analysis showed suppression of Bmp4 and induction of Bmp7 gene expression in the optic vesicle. Ectopic concurrent expression of the RPE cell markers Cx43 and Trp-2 in the neural retinal layer suggests cell fate respecification. These results indicate that forced expression of Msx2 perturbs BMP signaling in the developing eye and is accompanied by an increase in retinal cell death and a reduction in cell proliferation. Thus, deregulated Msx2 gene expression may be a plausible genetic mechanism by which the autosomal dominant form of congenital microphthalmia may arise.


The effect of saccade metrics on the corollary discharge contribution to perceived eye location.
Corollary discharge (CD) is hypothesized to provide the movement information (direction and amplitude) required to compensate for the saccade-induced disruptions to visual input. Here, we investigated to what extent these conveyed metrics influence perceptual stability in human subjects with a target-displacement detection task. Subjects made saccades to targets located at different amplitudes (4°, 6°, or 8°) and directions (horizontal or vertical). During the saccade, the target disappeared and then reappeared at a shifted location either in the same direction or opposite to the movement vector. Subjects reported the target displacement direction, and from these reports we determined the perceptual threshold for shift detection and estimate of target location. Our results indicate that the thresholds for all amplitudes and directions generally scaled with saccade amplitude. Additionally, subjects on average produced hypometric saccades with an estimated CD gain <1. Finally, we examined the contribution of different error signals to perceptual performance, the saccade error (movement-to-movement variability in saccade amplitude) and visual error (distance between the fovea and the shifted target location). Perceptual judgment was not influenced by the fluctuations in movement amplitude, and performance was largely the same across movement directions for different magnitudes of visual error. Importantly, subjects reported the correct direction of target displacement above chance level for very small visual errors (<0.75°), even when these errors were opposite the target-shift direction. Collectively, these results suggest that the CD-based compensatory mechanisms for visual disruptions are highly accurate and comparable for saccades with different metrics.


Analysis of Single-Walled Carbon Nanotubes in Estuarine Sediments by Density Gradient Ultracentrifugation Coupled to Near-Infrared Fluorescence Spectroscopy Reveals Disassociation of Residual Metal Catalyst Nanoparticles.
The continued growth of the nanotechnology industry and the incorporation of nanomaterials into consumer applications will inevitably lead to their release into environmental systems. Single-walled carbon nanotubes (SWCNTs) in particular have exhibited many attractive optical, mechanical, and electrical properties that lend themselves to new and exciting applications. Assessing their environmental impact upon release into the environment is contingent upon quantifying and characterizing SWCNTs in environmental matrixes. In this study, SWCNTs were isolated from estuarine sediments using density gradient ultracentrifugation (DGU), followed by online flow-through analysis of the density fractions via near-infrared spectroscopy. This approach yielded significant improvements in the quantitative detection limit, from 62 to 1.5 μg g-1. In addition, fractions of the density gradient were also obtained for further analysis by bulk inductively coupled plasma mass spectrometry (ICP-MS) and single-particle ICP-MS. Using fluorescent, semiconductive SWCNTs, the primary fluorescent nanotube fraction was found to be separated from the sediment matrix during DGU; however, the residual metal catalyst particles that had been assumed to be physically bound to the SWCNTs were found to form a separate band in the density gradient apart from the fluorescent SWCNTs. This result was repeated for a number of SWCNT types regardless of the metal catalyst and synthesis method, with a 0.1 g cm-3 density difference between most fractions. The apparent disconnect between the fluorescent fraction of SWCNTs and their metal-containing constituents potentially complicates CNT risk assessment as analysis techniques focusing solely on either CNT fluorescence or metal fingerprints may misrepresent exposure concentrations and their toxicological implications.


Axial rotation of the lumbar spine and the effect of flexion. An in vitro and in vivo biomechanical study.
A series of experiments were performed on eight whole, cadaveric lumbar spines and on eight male volunteers to determine whether axial rotation changed with subjects bending forward compared with being in a neutral posture and whether rotation was affected by articular tropism. Kirschner wires were inserted into the spinous processes of the eight cadaveric lumbar spines, and the axial rotation of the wires was measured while the spine was rotated in a torsion apparatus. Similarly, Steinmann pins were inserted into the spinous processes of L3, L4, and L5 of the eight volunteers, and the axial rotation of the pins was measured while the subjects rotated in a torsion apparatus. Axial rotation was found to be less when combined with forward flexion, and articular tropism did not influence the amplitude of rotation.


Pain: Is It All in the Brain or the Heart?
Scientists have reported that pain is always created by the brain. This may not be entirely true. Pain is not only a sensory experience, but also can be associated with emotional, cognitive, and social components. The heart is considered the source of emotions, desire, and wisdom. Therefore, the aim of this article was to review the available evidence about the role of the heart in pain modulation. Dr. Armour, in 1991, discovered that the heart has its "little brain" or "intrinsic cardiac nervous system." This "heart brain" is composed of approximately 40,000 neurons that are alike neurons in the brain, meaning that the heart has its own nervous system. In addition, the heart communicates with the brain in many methods: neurologically, biochemically, biophysically, and energetically. The vagus nerve, which is 80% afferent, carries information from the heart and other internal organs to the brain. Signals from the "heart brain" redirect to the medulla, hypothalamus, thalamus, and amygdala and the cerebral cortex. Thus, the heart sends more signals to the brain than vice versa. Research has demonstrated that pain perception is modulated by neural pathways and methods targeting the heart such as vagus nerve stimulation and heart-rhythm coherence feedback techniques. The heart is not just a pump. It has its neural network or "little brain." The methods targeting the heart modulate pain regions in the brain. These methods seem to modulate the key changes that occur in the brain regions and are involved in the cognitive and emotional factors of pain. Thus, the heart is probably a key moderator of pain.


Amyloid-beta dynamics are regulated by orexin and the sleep-wake cycle.
Amyloid-beta (Abeta) accumulation in the brain extracellular space is a hallmark of Alzheimer's disease. The factors regulating this process are only partly understood. Abeta aggregation is a concentration-dependent process that is likely responsive to changes in brain interstitial fluid (ISF) levels of Abeta. Using in vivo microdialysis in mice, we found that the amount of ISF Abeta correlated with wakefulness. The amount of ISF Abeta also significantly increased during acute sleep deprivation and during orexin infusion, but decreased with infusion of a dual orexin receptor antagonist. Chronic sleep restriction significantly increased, and a dual orexin receptor antagonist decreased, Abeta plaque formation in amyloid precursor protein transgenic mice. Thus, the sleep-wake cycle and orexin may play a role in the pathogenesis of Alzheimer's disease.


Outbreak of multiple clones of imipenem-resistant Acinetobacter baumannii isolates expressing OXA-58 carbapenemase in an intensive care unit.
To investigate the resistance mechanisms and the genetic relationship of imipenem-resistant Acinetobacter baumannii isolates recovered in the intensive care unit (ICU) of a tertiary care hospital. Imipenem-resistant A. baumannii clinical and environmental isolates were collected in the ICU of the Red Cross General Hospital, Athens, Greece between March and October 2002. The isolates were tested by Etest MBL, PCR, RT-PCR and sequencing for carbapenemase-encoding genes, PFGE and synergy experiments using meropenem and the efflux pump inhibitor carbonyl cyanide chlorophenylhydrazone. During the study period, 15 clinical and two environmental imipenem-resistant (MIC 8 to >128 mg/L) A. baumannii isolates were recovered. PFGE showed six different clones that included both clinical and environmental isolates. All 17 isolates were negative by Etest MBL and PCR for genes bla(IMP), bla(VIM), bla(SPM), bla(OXA-23-like) and bla(OXA-24-like). Genes bla(OXA-51-like) and bla(OXA-58-like) were amplified from 15 and 14 isolates, respectively. Sequencing of bla(OXA-51-like) amplicons identified bla(OXA-66) (nine cases) and bla(OXA-69) (six cases), whereas bla(OXA-58-like) sequences were classical bla(OXA-58). Reverse transcriptase-PCR showed that bla(OXA-51-like) genes were expressed in 12 and bla(OXA-58) in 10 isolates; in these isolates, inhibition of OXA enzymes by 200 mM of NaCl reduced carbapenem MICs by up to 4-fold. Overexpression of proton-gradient dependent efflux pumps did not contribute to carbapenem resistance in any isolate. Similarly, although AmpC expression was demonstrated in eight isolates, inhibition of AmpC with cloxacillin did not reduce the MICs of carbapenems significantly. These findings indicate wide dissemination of OXA-58 carbapenemase, which contributes, at least partially, to the imipenem resistance of unrelated A. baumannii isolates in our ICU.


Thermal boundary layer effects on the acoustical impedance of enclosures and consequences for acoustical sensing devices.
Expressions are derived for the acoustical impedance of a rectangular enclosure and of a finite annular cylindrical enclosure. The derivation is valid throughout the frequency range in which all dimensions of the enclosure are much less than the wavelength. The results are valid throughout the range from adiabatic to isothermal conditions in the enclosure. The effect is equivalent to placing an additional, frequency-dependent complex impedance in parallel with the adiabatic compliance. As the thermal boundary layer grows to fill the cavity, the reactive part of the impedance varies smoothly from the adiabatic value to the isothermal value. In some microphones, this change in cavity stiffness is sufficient to modify the sensitivity. The resistive part of the additional cavity impedance varies as the inverse square root of frequency at high frequencies where the boundary layer has not grown to fill the enclosure. The thermal modification gives rise to a thermal noise whose spectral density varies asymptotically as l/f(3/2) above the isothermal transition frequency.


Human footprints provide snapshot of last interglacial ecology in the Arabian interior.
The nature of human dispersals out of Africa has remained elusive because of the poor resolution of paleoecological data in direct association with remains of the earliest non-African people. Here, we report hominin and non-hominin mammalian tracks from an ancient lake deposit in the Arabian Peninsula, dated within the last interglacial. The findings, it is argued, likely represent the oldest securely dated evidence for Homo sapiens in Arabia. The paleoecological evidence indicates a well-watered semi-arid grassland setting during human movements into the Nefud Desert of Saudi Arabia. We conclude that visitation to the lake was transient, likely serving as a place to drink and to forage, and that late Pleistocene human and mammalian migrations and landscape use patterns in Arabia were inexorably linked.


Multimodal fluorescence microscopy of prion strain specific PrP deposits stained by thiophene-based amyloid ligands.
The disease-associated prion protein (PrP) forms aggregates which vary in structural conformation yet share an identical primary sequence. These variations in PrP conformation are believed to manifest in prion strains exhibiting distinctly different periods of disease incubation as well as regionally specific aggregate deposition within the brain. The anionic luminescent conjugated polythiophene (LCP), polythiophene acetic acid (PTAA) has previously been used to distinguish PrP deposits associated with distinct mouse adapted strains via distinct fluorescence emission profiles from the dye. Here, we employed PTAA and 3 structurally related chemically defined luminescent conjugated oligothiophenes (LCOs) to stain brain tissue sections from mice inoculated with 2 distinct prion strains. Our results showed that in addition to emission spectra, excitation, and fluorescence lifetime imaging microscopy (FLIM) can fruitfully be assessed for optical distinction of PrP deposits associated with distinct prion strains. Our findings support the theory that alterations in LCP/LCO fluorescence are due to distinct conformational restriction of the thiophene backbone upon interaction with PrP aggregates associated with distinct prion strains. We foresee that LCP and LCO staining in combination with multimodal fluorescence microscopy might aid in detecting structural differences among discrete protein aggregates and in linking protein conformational features with disease phenotypes for a variety of neurodegenerative proteinopathies.


Visualizing virus particle mobility in liquid at the nanoscale.
Currently, there remains a critical need to develop real-time imaging resources for life sciences. Here, we demonstrate the use of high resolution in situ imaging to observe biological complexes in liquid at the nanoscale. Using a model virus system, we produced the first time-resolved videos of individual biological complexes moving in solution within an electron microscope.


Pancreas storage in oxygenated perfluorodecalin does not restore post-transplant function of isolated pig islets pre-damaged by warm ischemia.
Cold storage in oxygenated perfluorodecalin (PFD) restores transplant function of ischemically damaged dog pancreata and reduces the impact of cold ischemia on recovery of isolated human islets. Whether PFD storage can improve islet isolation from pancreata exposed to significant warm ischemia (WI) is unclear yet. The present study aimed to clarify this question in adult pigs. After exsanguination, the intestine was removed immediately or left in the cavity for 30 min of WI. Resected pancreata were intraductally flushed with cold University of Wisconsin solution. Subsequently, pancreata were processed immediately by digestion-filtration (group I: 0 min WI, n=6; II: 30 min WI, n=6) or first stored for 3 h in oxygenated PFD (III: 0 min WI+PFD, n=5; IV: 30 min WI+PFD, n=6). Pancreata subjected to 30 min of WI yielded significantly less islets compared with the corresponding non-ischemic organs (I vs. II, P<0.01; III vs. IV, P<0.05). Oxygenation did not ameliorate the loss in islet yield (II vs. IV, NS). Ischemic islets were characterized by depleted ATP stores (388+/-73 (I) vs. 133+/-22 ng/1000 IEQ (II), P<0.01) and diminished insulin response to glucose calculated as stimulation index (SI; 2.47+/-0.36 (I) vs. 0.25+/-0.17 (II), P<0.05). PFD storage of ischemic organs partially restored ATP content (217+/-23 ng/1000 IEQ, II vs. IV, P<0.05) and glucose SI (1.60+/-0.09, II vs. IV, P<0.05) to a significant extent that reached the level of corresponding PFD-stored, non-ischemic pancreata (III vs. IV, NS). Sustained normoglycemia was exclusively observed in diabetic nude mice transplanted with islets isolated from non-ischemic organs. The significantly reduced graft function of ischemic islets (I vs. II, III vs. IV, P<0.001) was not increased by pancreatic oxygenation (II vs. IV, NS). The present study suggests that pancreas short-term storage in oxygenated PFD improves in vitro but not the in vivo function of ischemically damaged pig islets.


Acetylsalicylic acid-dependent inhibition of collagen biosynthesis and beta1-integrin signaling in cultured fibroblasts.
Non-steroid anti-inflammatory drugs (NSAIDs) are known inhibitors of collagen biosynthesis in animal tissues and cell cultures. The mechanism of their action on collagen biosynthesis involves inactivation of prolidase, the enzyme that recovers proline from collagen degradation products for collagen re-synthesis. In fibroblasts, prolidase activity is stimulated through a signal induced by an activated b1-integrin receptor that contributes to an increase in enzyme phosphorylation. To discover whether this is an underlying mechanism of acetylsalicylic acid-dependent inhibition of collagen biosynthesis in fibroblasts was the aim of this study. Control and acetylsalicylic acid-treated confluent human skin fibroblasts were compared with respect to collagen biosynthesis ([3H]proline incorporation assay), prolidase activity (colorimetric assay) and the expressions of prolidase and some signaling proteins (Western immunoblot analysis). In cells treated with acetylsalicylic acid, a concomitant decrease in prolidase activity, prolidase phosphorylation (at the threonine residue), and collagen biosynthesis were observed. This was accompanied by a decrease in the expressions of SOS and phosphorylated MAP kinases, ERK1 and ERK2, but not FAK or Grab2. This suggests that a decrease in prolidase activity may contribute to a decrease in the biosynthesis of proline-containing proteins, such as collagen and SOS. A decrease in collagen biosynthesis decreases the interaction of the protein with the b1-integrin receptor (extracellular process), disturbing signaling (intracellular process). The inhibitory effect of acetylsalicylic acid on collagen biosynthesis in fibroblasts is coupled to the inhibition of prolidase phosphorylation (but not expression) and down-regulation of the intracellular signal transmitted by the b1-integrin receptor.


Being in-between and lost in the discharge process--an excursus of two empirical studies of older persons', their relatives', and care professionals' experience.
The discharge process (DP) is full of well-known risks, and a comprehensive and well-executed DP is especially important for older people with multiple health problems and continuing care needs, as well as for their relatives. Few studies focus on the experiences with the DP by older people in need of home care nursing and their relatives. Therefore, the aim was to deepen the understanding of the DP as a phenomenon described by older people, their relatives, and care professionals. The method is an excursus of the findings of two previously published research studies. By using the Reflective Lifeworld Research approach, the empirical findings were further interpreted with lifeworld theory. The results describe the essential meaning of the phenomenon of DP in relation to healthcare needs. The illness and the DP can be viewed as a course of action where the familiar becomes unfamiliar for older people and their relatives, entailing an insecure future existence characterized by the experience of being in-between. The DP is marked by bodily and existential needs. The older persons and their relatives are lost in the hospital context and trying to influence life and adapt to life circumstances, while being relentlessly dependent on care professionals. Care professionals work from both an organizational and a medical approach. Disharmony and disagreement seem to arise easily among the professionals regarding the planning negatively affecting the patients and their relatives. More efforts are needed in the DP to empower older people and their relatives to go on with their life at home. The caring practice needs to more clearly meet and address the individual needs of older people and their relatives and their understanding of their illness. It needs to give them lifeworld and life goals to alleviate their suffering and to help them adjust to their new situation.


Spatial patterns of male alcohol-related mortality in Belarus, Lithuania, Poland and Russia.
Eastern Europe is known to suffer from a large burden of alcohol-related mortality. However, persisting unfavourable conditions at the national level mask variation at the sub-national level. We aim to explore spatial patterns of cause-specific mortality across four post-communist countries: Belarus, Lithuania, Poland and Russia (European part). We use official mortality data routinely collected over 1179 districts and cities. The analysis refers to males aged 20-64 years and covers the period 2006-2014. Mortality variation is mainly assessed by means of the standardised mortality ratio. Getis-Ord Gi* statistic is employed to detect hot and cold spots of alcohol-related mortality. Alcohol-related mortality exhibits a gradient from very high levels in northwestern Russia to low levels in southern Poland. Spatial transitions from higher to lower mortality are not explicitly demarcated by national boundaries. Within these countries, hot spots of alcohol-related mortality dominate the territories of northwestern and western Russia, eastern and northwestern Belarus, southeastern Lithuania, and eastern and central Poland. The observed mortality gradient is likely associated with the spread of alcohol epidemics from the European part of Russia to the other countries, which appears to have started more than a century ago. Contemporary socioeconomic and demographic factors should be taken into account when developing anti-alcohol policies. The same is true for the peculiarities of culture, norms, traditions and behavioural patterns observed in specific geographical areas of the four countries. Reducing alcohol-related harm in the areas identified as hot spots should be prioritised.


The role of UDP-Glc:glycoprotein glucosyltransferase 1 in the maturation of an obligate substrate prosaposin.
An endoplasmic reticulum (ER) quality control system assists in efficient folding and disposal of misfolded proteins. N-linked glycans are critical in these events because their composition dictates interactions with molecular chaperones. UDP-glucose:glycoprotein glucosyltransferase 1 (UGT1) is a key quality control factor of the ER. It adds glucoses to N-linked glycans of nonglucosylated substrates that fail a quality control test, supporting additional rounds of chaperone binding and ER retention. How UGT1 functions in its native environment is poorly understood. The role of UGT1 in the maturation of glycoproteins at basal expression levels was analyzed. Prosaposin was identified as a prominent endogenous UGT1 substrate. A dramatic decrease in the secretion of prosaposin was observed in ugt1(-/-) cells with prosaposin localized to large juxtanuclear aggresome-like inclusions, which is indicative of its misfolding and the essential role that UGT1 plays in its proper maturation. A model is proposed that explains how UGT1 may aid in the folding of sequential domain-containing proteins such as prosaposin.


Sinhala translation of child behaviour checklist: validity and reliability.
To translate the child behaviour checklist (CBCL) into Sinhala and validate it for assessment of mental health status of children aged 5-10 years. Translation/back-translation method was used to translate the English CBCL into Sinhala. Each item in the Sinhala CBCL (CBCL-S) was rated by mental health professionals to determine semantics, content, and conceptual validity types. To ascertain criterion validity, total scores obtained for CBCL-S by administering it to parents or parent surrogates of 49 girls and 80 boys aged 5-10 years attending the specialist psychiatry clinics and 69 boys and 69 girls in the same age group from the community were compared with clinical diagnoses by a child psychiatrist. Receiver operator characteristic curves were drawn to obtain the cut-off points in CBCL-S for boys and girls separately. Semantics, content, and conceptual and criterion validity of CBCL-S were satisfactory. At the cut-off level of 39, CBCL-S had a sensitivity of 90% and a specificity of 88% for boys and a sensitivity of 89% and a specificity of 92% for girls. Internal consistency, test-retest reliability, and inter-interviewer reliability of CBCL-S were satisfactory. CBCL-S is a valid and reliable instrument to measure mental health status of Sinhalese children aged 5-10 years in Sri Lanka.


Does childhood trauma influence offspring's birth characteristics?
: A recent epigenetic hypothesis postulates that 'a sex-specific male-line transgenerational effect exists in humans', which can be triggered by childhood trauma during 'the slow growth period' just before puberty. The evidence is based on a few rather small epidemiological studies. We examine what response childhood trauma predicts, if any, in the birth size and prematurity risk of almost 800 000 offspring. Children of parity 1, 2 or 3, born 1976-2002 in Sweden, for whom we could trace both parents and all four grandparents, constituted generation 3 (G3, n = 764 569). Around 5% of their parents, G2, suffered parental (G1) death during their own childhood. The association of such trauma in G2 with G3 prematurity and birthweight was analysed, while controlling for confounders in G1 and G2. We examined whether the slow growth period was extra sensitive to parental loss. Parental (G1) death during (G2) childhood predicts premature birth and lower birthweight in the offspring generation (G3). This response is dependent on G2 gender, G2 age at exposure and G3 parity, but not G3 gender. The results are compatible with the Pembrey-Bygren hypothesis that trauma exposure during boys' slow growth period may trigger a transgenerational response; age at trauma exposure among girls seems less important, suggesting a different set of pathways for any transgenerational response. Finally, parental death during childhood was not important for the reproduction of social inequalities in birthweight and premature birth.


Prevalence of psychotic symptoms and their risk factors in urban Tanzania.
This study aimed to determine the prevalence of psychotic symptoms in urban Tanzania and their relationship with demographic, socio-economic and social factors. A random sample of 899 adults aged 15-59 was surveyed. The main outcome measure was endorsement of one or more psychotic symptoms identified by the Psychosis Screening Questionnaire. 3.9% respondents reported one or more psychotic symptoms in the preceding year. Significantly higher rates of symptoms were found in those who had recently experienced two or more stressful life events, those with CMD and people who had used cannabis in the preceding year.


Effectiveness of adenine arabinoside 5'-monophosphate in kidney transplant recipients with chronic active hepatitis B.
Hepatitis B virus infection is responsible for both morbidity and mortality in kidney transplant recipients. Adenine arabinoside 5'-monophosphate (ARA-AMP), a synthetic purine nucleotide with anti-viral activity, leads to a sustained interruption of HBV replication in approximately 40% of immunocompetent patients. We report the results of a pilot study using ARA-AMP to treat HBV-related chronic active hepatitis in kidney transplant recipients. Ten patients (2 females and 8 males, mean age 44 years, mean time post-transplantation 163 months) received a 28-day course of ARA-AMP intramuscularly: 5 mg/kg twice daily for the first 5 days during hospitalization and subsequently 5 mg/kg once daily at home for the remaining 23 days. Mean follow-up was 18 months, ranging from 7 to 28 months. All patients but one had biopsy-proven chronic hepatitis, including five cases of cirrhosis. All patients had been chronic HBs Ag carriers for more than 1 year and had active replication as assessed by the presence of serum HBV DNA (mean titre, 270 pg/ml, ranging from 12 to 997 pg/ml, Genostics method). HBe Ag was present in 7 of the 10 patients. Pretreatment creatinine was normal. In four of the 10 patients, HBV DNA became undetectable respectively 1, 1, 5, and 11 months after beginning ARA-AMP. In five patients, HBV DNA decreased during ARA-AMP therapy but subsequently increased although no change was noted during the follow-up period.(ABSTRACT TRUNCATED AT 250 WORDS)


Autophagy in the Vertebrate Inner Ear.
Autophagy is a conserved catabolic process that results in the lysosomal degradation of cell components. During development, autophagy is associated with tissue and organ remodeling, and under physiological conditions it is tightly regulated as it plays a housekeeping role in removing misfolded proteins and damaged organelles. The vertebrate inner ear is a complex sensory organ responsible for the perception of sound and for balance. Cell survival, death and proliferation, as well as cell fate specification and differentiation, are processes that are strictly coordinated during the development of the inner ear in order to generate the more than a dozen specialized cell types that constitute this structure. Here, we review the existing evidence that implicates autophagy in the generation of the vertebrate inner ear. At early stages of chicken otic development, inhibiting autophagy impairs neurogenesis and causes aberrant otocyst morphogenesis. Autophagy provides energy for the clearing of dying cells and it favors neuronal differentiation. Moreover, autophagy is required for proper vestibular development in the mouse inner ear. The autophagy-related genes Becn1, Atg4g, Atg5, and Atg9, are expressed in the inner ear from late developmental stages to adulthood, and Atg4b mutants show impaired vestibular behavior associated to defects in otoconial biogenesis that are also common to Atg5 mutants. Autophagic flux appears to be age-regulated, augmenting from perinatal stages to young adulthood in mice. This up-regulation is concomitant with the functional maturation of the hearing receptor. Hence, autophagy can be considered an intracellular pathway fundamental for in vertebrate inner ear development and maturation.


Evaluating the safety and efficacy of BMI-based preoperative administration of low-molecular-weight heparin in morbidly obese patients undergoing Roux-en-Y gastric bypass surgery.
One of the most concerning and potentially fatal complications of gastric bypass surgery is pulmonary embolism (PE) with published rates exceeding 1%. Although this procedure has been proven effective in reducing the morbidity and mortality of obesity and associated health care costs, it is elective and risk must be minimized. No dosing guidelines exist for pharmacologic prophylaxis in obese patients who are already at increased risk for these events. Although the current ASMBS and Chest guidelines recommend preoperative pharmacologic prophylaxis against thromboembolic events, no standard dosing protocols exist for the obese population. We propose a protocol including immediate pre followed by twice daily postoperative BMI-based dosing of low-molecular-weight heparin (LMWH), along pneumatic compression devices, and early ambulation. We retrospectively reviewed the charts of 170 patients who underwent Roux-en-Y gastric bypass surgery between March 2004 and December 2007. The incidence of deep venous thrombosis (DVT) and PE and bleeding complications associated with a BMI-based preoperative dosing protocol of LMWH was determined. All patients received LMWH preoperatively within 1 h of the incision, with doses varying between 30 and 60 mg given subcutaneously. Eleven patients received 30 mg, 145 patients received 40 mg, 9 patients received 50 mg, and 5 patients received 60 mg. None of the patients suffered from clinically significant DVT or PE during the hospital stay or in follow up (>2 years). Five patients (2.9%) were treated with discontinuation of lovenox and blood transfusion for postoperative bleeding. One of those patients returned to the operating room for exploration. We propose that immediate BMI-based preoperative dosing of LMWH along with postoperative prophylaxis is both safe and effective and should be standard for all patients undergoing Roux-en-Y gastric bypass surgery.


Relationship between head-only rotation and caloric test results in vestibular disorders.
Our purpose was to determine the possible existence of a relationship between abnormal findings from head-only rotation and those from caloric tests applied in patients with peripheral vestibular disorders. We retrospectively studied the head-only rotation and caloric test results of 75 patients experiencing dizziness and peripheral vestibular dysfunction. We assessed the relationship between abnormal test results using the Cramér coefficient. Abnormal findings in the head-only rotation test occurred in 87.5% of patients with vestibular hypoexcitability and in 94.3% of those with hyperexcitability or directional preponderance of post-caloric nystagmus. Statistical analysis of the association between the values found for the parameters of head-only rotation and caloric test results revealed a Cramér coefficient that was less than 0.70. We found no relationship between abnormal findings from head-only rotation and caloric tests applied to individuals with peripheral vestibular disorders.


Single cell profiling of circulating tumor cells: transcriptional heterogeneity and diversity from breast cancer cell lines.
To improve cancer therapy, it is critical to target metastasizing cells. Circulating tumor cells (CTCs) are rare cells found in the blood of patients with solid tumors and may play a key role in cancer dissemination. Uncovering CTC phenotypes offers a potential avenue to inform treatment. However, CTC transcriptional profiling is limited by leukocyte contamination; an approach to surmount this problem is single cell analysis. Here we demonstrate feasibility of performing high dimensional single CTC profiling, providing early insight into CTC heterogeneity and allowing comparisons to breast cancer cell lines widely used for drug discovery. We purified CTCs using the MagSweeper, an immunomagnetic enrichment device that isolates live tumor cells from unfractionated blood. CTCs that met stringent criteria for further analysis were obtained from 70% (14/20) of primary and 70% (21/30) of metastatic breast cancer patients; none were captured from patients with non-epithelial cancer (n = 20) or healthy subjects (n = 25). Microfluidic-based single cell transcriptional profiling of 87 cancer-associated and reference genes showed heterogeneity among individual CTCs, separating them into two major subgroups, based on 31 highly expressed genes. In contrast, single cells from seven breast cancer cell lines were tightly clustered together by sample ID and ER status. CTC profiles were distinct from those of cancer cell lines, questioning the suitability of such lines for drug discovery efforts for late stage cancer therapy. For the first time, we directly measured high dimensional gene expression in individual CTCs without the common practice of pooling such cells. Elevated transcript levels of genes associated with metastasis NPTN, S100A4, S100A9, and with epithelial mesenchymal transition: VIM, TGFß1, ZEB2, FOXC1, CXCR4, were striking compared to cell lines. Our findings demonstrate that profiling CTCs on a cell-by-cell basis is possible and may facilitate the application of 'liquid biopsies' to better model drug discovery.


Large-scale computational drug repositioning to find treatments for rare diseases.
Rare, or orphan, diseases are conditions afflicting a small subset of people in a population. Although these disorders collectively pose significant health care problems, drug companies require government incentives to develop drugs for rare diseases due to extremely limited individual markets. Computer-aided drug repositioning, i.e., finding new indications for existing drugs, is a cheaper and faster alternative to traditional drug discovery offering a promising venue for orphan drug research. Structure-based matching of drug-binding pockets is among the most promising computational techniques to inform drug repositioning. In order to find new targets for known drugs ultimately leading to drug repositioning, we recently developed eMatchSite, a new computer program to compare drug-binding sites. In this study, eMatchSite is combined with virtual screening to systematically explore opportunities to reposition known drugs to proteins associated with rare diseases. The effectiveness of this integrated approach is demonstrated for a kinase inhibitor, which is a confirmed candidate for repositioning to synapsin Ia. The resulting dataset comprises 31,142 putative drug-target complexes linked to 980 orphan diseases. The modeling accuracy is evaluated against the structural data recently released for tyrosine-protein kinase HCK. To illustrate how potential therapeutics for rare diseases can be identified, we discuss a possibility to repurpose a steroidal aromatase inhibitor to treat Niemann-Pick disease type C. Overall, the exhaustive exploration of the drug repositioning space exposes new opportunities to combat orphan diseases with existing drugs. DrugBank/Orphanet repositioning data are freely available to research community at https://osf.io/qdjup/.


A non-human primate model of stroke reproducing endovascular thrombectomy and allowing long-term imaging and neurological read-outs.
Stroke is a devastating disease. Endovascular mechanical thrombectomy is dramatically changing the management of acute ischemic stroke, raising new challenges regarding brain outcome and opening up new avenues for brain protection. In this context, relevant experiment models are required for testing new therapies and addressing important questions about infarct progression despite successful recanalization, reversibility of ischemic lesions, blood-brain barrier disruption and reperfusion damage. Here, we developed a minimally invasive non-human primate model of cerebral ischemia (Macaca fascicularis) based on an endovascular transient occlusion and recanalization of the middle cerebral artery (MCA). We evaluated per-occlusion and post-recanalization impairment on PET-MRI, in addition to acute and chronic neuro-functional assessment. Voxel-based analyses between per-occlusion PET-MRI and day-7 MRI showed two different patterns of lesion evolution: "symptomatic salvaged tissue" (SST) and "asymptomatic infarcted tissue" (AIT). Extended SST was present in all cases. AIT, remote from the area at risk, represented 45% of the final lesion. This model also expresses both worsening of fine motor skills and dysexecutive behavior over the chronic post-stroke period, a result in agreement with cortical-subcortical lesions. We thus fully characterized an original translational model of ischemia-reperfusion damage after stroke, with consistent ischemia time, and thrombus retrieval for effective recanalization.


Recent advances in conjugated polymers for light emitting devices.
A recent advance in the field of light emitting polymers has been the discovery of electroluminescent conjugated polymers, that is, kind of fluorescent polymers that emit light when excited by the flow of an electric current. These new generation fluorescent materials may now challenge the domination by inorganic semiconductor materials of the commercial market in light-emitting devices such as light-emitting diodes (LED) and polymer laser devices. This review provides information on unique properties of conjugated polymers and how they have been optimized to generate these properties. The review is organized in three sections focusing on the major advances in light emitting materials, recent literature survey and understanding the desirable properties as well as modern solid state lighting and displays. Recently, developed conjugated polymers are also functioning as roll-up displays for computers and mobile phones, flexible solar panels for power portable equipment as well as organic light emitting diodes in displays, in which television screens, luminous traffic, information signs, and light-emitting wallpaper in homes are also expected to broaden the use of conjugated polymers as light emitting polymers. The purpose of this review paper is to examine conjugated polymers in light emitting diodes (LEDs) in addition to organic solid state laser. Furthermore, since conjugated polymers have been approved as light-emitting organic materials similar to inorganic semiconductors, it is clear to motivate these organic light-emitting devices (OLEDs) and organic lasers for modern lighting in terms of energy saving ability. In addition, future aspects of conjugated polymers in LEDs were also highlighted in this review.


Nurse prescriber-patient consultations: a case study in dermatology.
This paper is a report of a study exploring the content and processes in consultations between nurse prescribers and patients with dermatological conditions. Communication skills, consultation time, information and follow-up are central to the treatment and management of patients with dermatological conditions. The contribution nurses make to the care of these patients has great potential. A multiple case study was conducted with 10 practice settings across England in which nurses prescribed medicines for patients with dermatological conditions. Data were collected between June 2006 and September 2007 using semi-structured interviews (n = 40), patient questionnaires (n = 165/200) and videotaped observations of nurse consultations (n = 40). Data analysis included thematic analysis, descriptive statistics, chi-square and non-parametric tests. Nurses believed that their holistic approach to assessment, combined with their prescribing knowledge, improved prescribing decisions. Listening and explanation of treatments were aspects of nurse communication that were rated highly by patients. Listening and dealing sensitively with emotions were also aspects of the videotaped consultations that were rated highly by assessors. Nurses were less consistent in providing information about medicines. Triangulated data from this study suggest that nurse prescribing enhances the care of patients with dermatological conditions through improved prescribing decisions. If patients are to be more involved in this decision-making, nurses must give them more information about their medicines. The benefits of prescribing were most evident in the practices of dermatology specialist nurses. Further evidence is required to identify whether prescribing by specialist nurses offers similar benefits in other therapeutic areas.


Agomelatine facilitates positive versus negative affective processing in healthy volunteer models.
Agomelatine is a new antidepressant with a novel profile of pharmacological action. The clinical efficacy of agomelatine has been established in major depression, but its actions on emotional bias are unknown. Consequently, the current experimental study assessed the effect of agomelatine on emotional processing in healthy volunteers using an Emotional Test Battery shown to be sensitive to serotonin and noradrenaline reuptake inhibitors. Volunteers were randomized to receive placebo, 25 mg or 50 mg of agomelatine over a 7-day period in a double-blind parallel groups design. Emotional processing (n = 48) was assessed on the morning of day 8 using the Emotional Test Battery which included facial expression recognition, emotional memory, attentional visual probe and emotion-potentiated startle. Mood and subjective state were monitored before and during treatment. Agomelatine (25 mg) decreased subjective ratings of sadness, reduced recognition of sad facial expressions, improved positive affective memory and reduced the emotion-potentiated startle response. The results show that agomelatine has more selective effects on the processing of social facial cues than conventional antidepressants, which could contribute to less blunting of emotional experience. The study highlights the potential value of volunteer models in drug development for screening and profiling of novel antidepressants.


Community participation in mosquito breeding site control: an interdisciplinary mixed methods study in Curaçao.
As the arboviral diseases dengue, chikungunya and Zika emerge in the Americas, so does the need for sustainable vector control policies. To successfully achieve mosquito control, joint efforts of both communities and governments are essential. This study investigates this important, but by-and-large neglected topic. In June and July 2015, a cross-sectional mixed methods study applying a survey questionnaire (response rate of 82.5%; n = 339), in-depth interviews (n = 20) and focus group discussions (n = 7; 50 participants) was performed in Curaçao. The study was designed based on an integrated theoretical framework of the Health Belief Model and the Theory of Planned Behaviour. Participants showed a good knowledge of, and a high-level performance of mosquito breeding site control (MBSC) practices. Personal protection against mosquitoes (e.g. topical repellents) was perceived as relatively less effective thus practiced to lower extent compared to MBSC practices (i.e. larval source management). A lower intention to perform MBSC was independently associated with: (i) satisfaction on governmental MBSC (P = 0.012); (ii) barriers to perform MBSC practices, i.e. 'Government doesn't control other breeding sites' (P = 0.005), 'Don't know how to control breeding sites' (P = 0.041), and 'a mosquito does not transmit dengue' (P = 0.016), (iii) attitudes towards MBSC (P = 0.001) and self-efficacy (person's perceived ability to act) to perform MBSC (P = 0.002). Mixed-methods evidence highlights three possible ways of improving community participation in MBSC. First, it highlights the need for ongoing media coverage, targeting (i) communities' perceptions on transmission routes of dengue and chikungunya, and (ii) presence of car tires in yards. Secondly, it shows that promotion of governmental activities in MBSC can enhance MBSC of communities, if people develop a sense of responsibility to perform MBSC at their own properties. Thirdly, this study describes the presence of key persons in communities, who could be engaged in mosquito control policies to improve MBSC in neighbourhoods. This study reveals gaps between policy and communities' lived realities. These gaps might be overcome with the proposed interventions, resulting in a higher performance of MBSC in the community in Curaçao. Furthermore, this study shows how interdisciplinary mixed methods research can provide important, comprehensive, and in-depth insights to inform mosquito control policies.


Legislating Patient Representation: A Comparison Between Austrian and German Regulations on Self-Help Organizations as Patient Representatives.
Governments are increasingly inviting patient organizations (POs) to participate in healthcare policymaking. By inviting POs that claim to represent patients, representation comes into being. However, little is known about the circumstances under which governments accept POs as patient representatives. Based on the analysis of relevant legislation, this article investigates the criteria that self-help organizations (SHOs), a special type of PO, must fulfil in order to be accepted as patient representatives by governments in Austria and Germany. Thereby, it aims to contribute to the discussion on the role of governments in steering SHOs. There are different degrees of regulation (very little in Austria, more in Germany). Governments in both countries not only formulate explicit criteria for SHOs with respect to patient representation but also guide SHOs representing patients through implicit criteria for associations. We discuss the findings against concepts of responsiveness, authorization, and accountability. Our findings indicate that governmental steering is not negative per se as indicated by previous research but-depending on legislative criteria-can promote transparency and democratic quality in patient representation.


Bacteremia Caused by Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae in Vientiane, Lao PDR: A 5-Year Study.
Although there has been an increasing incidence of bacteremia caused by extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae (ESBL-E) across South East Asia, there are sparse data from the Lao PDR, where laboratory capacity for antimicrobial resistance surveillance is limited. We, therefore, retrospectively reviewed bacteremia caused by ESBL-producing Escherichia coli and Klebsiella pneumoniae between 2010 and 2014 at Mahosot Hospital, Vientiane, Lao PDR. Clinical and laboratory data relating to all episodes of ESBL-E bacteremia were reviewed over the 5-year period and compared with non-ESBL-E bacteremia. Blood cultures positive for E. coli or K. pneumoniae were identified retrospectively from laboratory records. Clinical and laboratory data were extracted from research databases and case notes and analyzed using STATA. Between 2010 and 2014, we identified 360 patients with E. coli (n = 249) or K. pneumoniae (n = 111) bacteremia, representing 34.8% of all patients with clinically significant bacteremia. Seventy-two (20%) isolates produced ESBL; E. coli accounted for 15.3% (55/360) and K. pneumoniae for 4.7% (17/360), respectively. The incidence of ESBL-producing E. coli bacteremia rose during the study period. By multiple logistic analysis, reported antibiotic use in the previous week was significantly associated with ESBL positivity (P < 0.001, odds ratio 3.89). Although multiresistant, most ESBL-producing E. coli and K. pneumoniae remained susceptible to meropenem (65/65; 100%) and amikacin (64/65; 98.5%). We demonstrated an alarming increase in the incidence of ESBL-E as a cause of bacteremia in Vientiane during the study period. This has implications for empiric therapy of sepsis in Laos, and ongoing surveillance is essential.


γ oscillations in the human basal ganglia.
Interest in beta activity in the basal ganglia has mushroomed since it was first identified in the subthalamic nucleus of patients with Parkinson's disease in Jonathan Dostrovsky's landmark paper (Levy et al., 2000). Here we consider a less explored phenomenon; namely gamma frequency synchronisation of neurons in the basal ganglia. Gamma oscillations have been reported in a distributed network involving the basal ganglia, thalamus and motor cortex, and have been described in a wide range of diseases as well as during increased arousal and voluntary movement. In Parkinson's disease, gamma activity is promoted by dopaminergic therapy. These features suggest that its elevation may be involved in the production of movement and this hypothesis is supported by the correlation between the amplitude of gamma activity and limb kinematics. Here we review these data, discuss the functional anatomy of gamma activity in basal ganglia and question how closely it relates to the coding of movement parameters.


Near-infrared fluorescent probes for the detection of glutathione and their application in the fluorescence imaging of living cells and tumor-bearing mice.
Two new cyanine-based fluorescent probes 1 and 2 have been developed. Probe 1 bears two cyanine units in a single molecule, and probe 2 contains a bis(trifluoromethyl)benzenethiol moiety. Both are non-fluorescent. The addition of intracellular glutathione (GSH) significantly enhanced the NIR fluorescence of the two probes. Both probes were used to image varying amounts of GSH in living cells. In tumor bearing mice, the in vivo fluorescence intensity of both probes was higher in tumors, where GSH is overexpressed, than in normal tissues. These results suggest that these new fluorogenic probes have potential for GSH-targeting diagnostic imaging.


Cytophagic histiocytic panniculitis with haemophagocytosis in a patient with familial multiple lipomatosis and review of the literature.
We report a patient with the extremely rare familial multiple lipomatosis syndrome, who developed the uncommon autoimmune disease cytophagic histiocytic panniculitis, manifested as inflammation of preexisting lipomas. Despite his initial critical condition and unsuccessful treatment with steroids, he responded to cyclosporin and remains well 15 years after diagnosis. In contrast with most previous reports, our patient stays dependent on cyclosporin; repeated attempts of discontinuing or substituting treatment were quickly followed by relapse. Haemophagocytic panniculitis is considered as a T-cell disorder, but its exact pathophysiological mechanism has not been clarified. Differential diagnosis of cytophagic histiocytic panniculitis mainly includes malignant histiocytosis, subcutaneous panniculitis-like T-cell lymphoma (SPTCL) and lupus erythematosus panniculitis (lupus profundus). We discuss the main clinical features, diagnostic challenges and treatment issues of this usually benign, but at times life-threatening autoimmune condition.


Local and regional control of calcium dynamics in the pancreatic islet.
Ca2+ is the key intracellular regulator of insulin secretion, acting in the β-cell as the ultimate trigger for exocytosis. In response to high glucose, ATP-sensitive K+ channel closure and plasma membrane depolarization engage a sophisticated machinery to drive pulsatile cytosolic Ca2+ changes. Voltage-gated Ca2+ channels, Ca2+ -activated K+ channels and Na+ /Ca2+ exchange all play important roles. The use of targeted Ca2+ probes has revealed that during each cytosolic Ca2+ pulse, uptake of Ca2+ by mitochondria, endoplasmic reticulum (ER), secretory granules and lysosomes fine-tune cytosolic Ca2+ dynamics and control organellar function. For example, changes in the expression of the Ca2+ -binding protein Sorcin appear to provide a link between ER Ca2+ levels and ER stress, affecting β-cell function and survival. Across the islet, intercellular communication between highly interconnected "hubs," which act as pacemaker β-cells, and subservient "followers," ensures efficient insulin secretion. Loss of connectivity is seen after the deletion of genes associated with type 2 diabetes (T2D) and follows metabolic and inflammatory insults that characterize this disease. Hubs, which typically comprise ~1%-10% of total β-cells, are repurposed for their specialized role by expression of high glucokinase (Gck) but lower Pdx1 and Nkx6.1 levels. Single cell-omics are poised to provide a deeper understanding of the nature of these cells and of the networks through which they communicate. New insights into the control of both the intra- and intercellular Ca2+ dynamics may thus shed light on T2D pathology and provide novel opportunities for therapy.


Cigarette smoking and serum bilirubin subtypes in healthy Korean men: the Korea Medical Institute study.
Cigarette smoking is a modifiable risk factor for cardiovascular disease. Bilirubin is a potent antioxidant and its concentration decreases in smokers. However, studies about the association between cigarette smoking and bilirubin are scarce and most are limited to total bilirubin. Additionally, bilirubin is highly related to hemoglobin. Therefore, this study evaluates the association between bilirubin subtypes and cigarette smoking in healthy Korean men independently of hemoglobin. This study included 48 040 Korean men aged 30 to 87 years who visited the Korea Medical Institute for routine health examinations from January to December, 2007. The association of smoking with total, direct, and indirect bilirubin was assessed by logistic regression analysis taking into consideration differences in subjects and smoking characteristics. Current smokers had lower bilirubin concentrations than never-smokers and ex-smokers. Smoking amount and duration were inversely significantly associated with total, direct, and indirect bilirubin. In a multivariable adjusted model, compared to never-smokers, the odds ratios (ORs) and 95% confidence intervals (CIs) of current smokers with the highest number of pack-years were 1.7 (1.6 to 1.9) for total, 1.5 (1.4 to 1.6) for direct, and 1.7 (1.6 to 1.9) for indirect bilirubin. After further adjustment for hemoglobin, this association became stronger (OR [95% CI], 2.1 [1.9 to 2.2] for total; 1.9 [1.8 to 2.0] for direct; 2.0 [1.9 to 2.2] for indirect bilirubin). In this study, bilirubin subtypes are inversely associated with smoking status, smoking amount, and smoking duration in healthy Korean men independently of hemoglobin. Further studies are needed to investigate this association in healthy Korean women.


Television viewing and its association with overweight in Colombian children: results from the 2005 National Nutrition Survey: a cross sectional study.
There has been an ongoing discussion about the relationship between time spent watching television and childhood obesity. This debate has special relevance in the Latin American region were the globalization process has increased the availability of screen-based entertainment at home. The aim of this study is to examine the association between television viewing and weight status in Colombian children. This cross sectional investigation included children aged 5 to12 yrs from the National Nutrition Survey in Colombia (ENSIN 2005). Weight and height were measured in 11,137 children in order to calculate body mass index. Overweight was defined by international standards. Time spent viewing television was determined for these children through parental reports. Multiple logistic regression analyses were conducted for different subgroups and adjusted for potential confounders in order to study the association between television viewing and weight status in this population. Among the surveyed children, 41.5% viewed television less than two hours/day; 36.8% between two and 3.9 hours/day and 21.7% four or more hours/day. The prevalence of overweight (obesity inclusive) in this population was 11.1%. Children who were classified as excessive television viewers (between two and 3.9 hours/day or 4 or more hours/day) were more likely to be overweight (OR: 1.44 95% CI: 1.41-1.47 and OR: 1.32 95% CI: 1.30-1.34, respectively) than children who reported to watch television less than 2 hours/day. Stratified analyses by age, gender and urbanization levels showed similar results. Television viewing was positively associated with the presence of overweight in Colombian children. A positive association between urbanization level and television viewing was detected. Considering that the majority of Colombian children lives in densely populated cities and appear to engage in excessive television viewing these findings are of public health relevance for the prevention of childhood obesity.


Pediatric asthma care in the emergency department: measuring the quality of history-taking and discharge planning.
The National Asthma Education and Prevention Program NAEPP Guidelines include recommendations for history-taking and discharge planning during an asthma visit, but there are no tools to measure performance. The objectives of this study were to define and operationalize key elements of history-taking and discharge planning, to develop a tool for measuring these elements, and to evaluate the quality of history-taking and discharge planning in the emergency department (ED) during visits for asthma using the new tool. Expert opinion and extensive literature review were used to develop a 13-item checklist containing items that should be documented during history-taking and provided during discharge planning for an ED visit for an acute asthma exacerbation by children. A convenience sample of 90 pediatric emergency medicine physicians and allergists rated each item in the checklist. The checklist was used to score audiotapes of asthma visits in the ED. Subjects were 154 parents of asthmatic children aged 4-9 years seeking care in nine inner-city EDs affiliated with asthma centers participating in the National Cooperative Inner-City Asthma Study and the physician/providers who delivered care. Seven of the 13 items on the checklist were rated as required to be performed by more than 90% of the allergist/pediatric emergency medicine physicians. Only 10% of the 154 visits included all seven of the highly rated items, whereas 19% of the visits included three or fewer. Only 7 of the 13 items (54%) were performed in more than 50% of the visits, and 4 items were performed in fewer than 25% of visits. Based on expert ratings, the checklist for measuring elements of history-taking and discharge planning during asthma visits appears to have considerable face validity. In the visits studied, the overall performance of these elements was low. Interventions to improve performance on the checklist might lead to improved care for children with asthma who frequent the ED.


Ensuring Universal Design of ICT: Triggering the Triggers!
Some ICT projects manage to create award winning, inclusive solutions, while others fail. Previous research has gathered data from 34 informants across 23 ICT-projects that have achieved universal design (UD). Their reasons for success are complex, but 15 Critical Success Criteria (CSC) can be identified. This article asks: How can we utilize these insights to promote UD efforts in the ICT-industry? The article proposes a way to model the empirical data for societal utilization; supporting future efforts to promote UD. First, we analyze the relationships between personal, processual, organizational and societal factors, and how the different critical success criteria work together to positively influence the projects in our sample. Next, we apply Hertzberg and Fogg's theories to classify the CSC as triggers, facilitators, motivators or hygiene factors. Based on this deeper understanding, we model the data and propose 3 trigger factors for UD of ICT. Using our model, we propose the following three strategies, which have a high effect potential for "triggering the triggers": 1) Legal interventions, 2) Awareness interventions, and 3) Training interventions. The contribution of the article is theoretical: a) providing richer insights into empirical data, by modeling their relationships, and b) predicting the impact of future interventions on the ICT-industry based on our modeled findings.


Association between dental caries and adverse pregnancy outcomes.
Poor oral health is not only associated with diabetes and cardiovascular disease but adverse pregnancy outcomes. However the influence of dental caries on pregnancy is unknown. The aim of this study was to evaluate the association between dental caries and adverse pregnancy outcomes and the effect of treatment for dental caries on adverse pregnancy outcomes. Primiparas who delivered a singleton between January 1, 2010 and December 31, 2014 and underwent both general health examination and oral health examination during a National Korea Health Screening Examination within 1 year of pregnancy were eligible. The data of the women who met the inclusion criteria were linked to the data of their offspring contained within the National Korea Health Screening Program for Infants and Children database. Among 120,622 women who delivered during the study period, 28,623 (23.7%) women had dental caries. Among them, 4,741 (16.6%) women were treated for dental caries after diagnosis. In a multivariable analysis, women with dental caries had an increased risk of delivering large-for-gestational-age infants (odds ratio, 1.15; 95% confidence interval, 1.07, 1.23) compared to those without dental caries. When women with dental caries were divided on the basis of the treatment of dental caries, women with dental caries but no treatment had an increased risk of delivering large-for-gestational-age infants (odds ratio, 1.15; 95% confidence interval, 1.06, 1.24); conversely, there was no increased risk in women with dental caries and treatment compared with those without. Dental caries and its treatment were not associated with preterm birth and preeclampsia. Untreated dental caries was not associated with preterm birth or preeclampsia but with the risk of delivering large-for-gestational-age infants. These whole observation may be attributed to the various characteristics of mothers who develop dental caries are not treated.


Hyperimmune anti-HBs plasma as alternative to commercial immunoglobulins for prevention of HBV recurrence after liver transplantation.
Hepatitis B immune globulins (HBIG) in combination with nucleos(t)ide analogues (NA) are effectively used for the prevention of hepatitis B virus (HBV) recurrence after liver transplantation (LT). However, associated treatment costs for HBIG are exceedingly high. Fresh frozen plasma obtained from blood donors with high anti-HBs levels (hyperimmune plasma, HIP) containing at least 4,500 IU anti-HBs was used as alternative treatment for HBV recurrence prophylaxis post-LT. Twenty-one HBV-related LT recipients received HIP starting at transplantation, followed by long-term combination treatment with NA. Mean follow-up time was 4.5 years (range 0.5-12.6) and each patient received on average 8.2 HIP per year (range 5.8-11.4). Anti-HBs terminal elimination kinetic after HIP administration was 20.6 days (range 13.8-30.9), which is comparable to values reported for commercial HBIG products. All 21 patients remained free of HBV recurrence during follow-up and no transfusion-transmitted infection or other serious complication was observed. Seven patients developed reversible mild transfusion reactions. The cost for one HIP unit was US$140; average yearly HBIG treatment cost was US$1,148 per patient, as compared to US$25,000-100,000 for treatment with commercial HBIG. The results of this study suggest that the use of HIP may be a useful and economical approach for the prevention of HBV recurrence post-LT if used in combination with NA. Additional prospective controlled studies in larger populations are needed to confirm these results.


Prodrugs of quercetin and resveratrol: a strategy under development.
The biochemical activities of plant flavonoids and stilbenoids point to many health-related applications, hampered however by a low bioavailability associated with rapid metabolic modification. A possible approach to overcome this obstacle is the development of prodrugs. In this review we provide some background information and summarize the efforts made so far to obtain suitable precursors of the two best known model polyphenols belonging to the classes just mentioned, quercetin and resveratrol. Prodrug design needs to take into account two key aspects: the nature of the chemical bond linking the core molecule to the protecting substituent, and the substituent itself, which can impart desirable physico-chemical properties. Only recently a systematic study of the several possible combinations has begun. Most bond systems tested so far appear to be either too stable or too unstable under physiological conditions. A range of substituent moieties is available, allowing the modulation of properties such as water solubility and the ability to permeate biomembranes. Work so far has been largely performed in vitro, and more in vivo experiments are definitely needed for a reliable assessment of the potentialities of the classes of prodrugs produced so far and of those still awaiting creation.


Detecting patchy nanoparticle assembly at the single-particle level.
Patchy colloidal particles offer a unique platform to explore and investigate spatially inhomogeneous colloidal interactions. In the present work we provide direct, in situ experimental observation at the single particle level on the colloidal interaction driven formation of a heterodimer, consisting of a patchy nanorod and a sphere. The gold nanorods employed during the experiments feature positively charged tip regions, while the side of the rods is covered by surface grafted PEG. The driving force of the assembly is the electric double layer interaction between the nanorod and the nanosphere possessing opposite surface charge. The great advantage of using gold heterodimers is that their optical scattering spectrum characteristics are known to be very sensitive to the actual structure of the assembly due to plasmon hybridization. This was exploited to determine the structure of the heterodimer in situ upon formation in aqueous medium. The results indicate, that despite the attractive patches being located at the nanorod tips, for the given particle pairs the spheres readily bind to the side region of the nanorods in the aqueous medium during the assembly. Finite element simulations of the electric double layer interaction reveal that this is the energetically favorable configuration for the given heterodimer. This work demonstrates the potential of in situ experiments performed at single particle level for the characterization of self-assembling structures, which can contribute to the development of rationally designed nanoscale building blocks.


Photorespiratory metabolism and nodule function: behavior of Lotus japonicus mutants deficient in plastid glutamine synthetase.
Two photorespiratory mutants of Lotus japonicus deficient in plastid glutamine synthetase (GS(2)) were examined for their capacity to establish symbiotic association with Mesorhizobium loti bacteria. Biosynthetic glutamine synthetase (GS) activity was reduced by around 40% in crude nodule extracts from mutant plants as compared with the wild type (WT). Western blot analysis further confirmed the lack of GS(2) polypeptide in mutant nodules. The decrease in GS activity affected the nodular carbon metabolism under high CO(2) (suppressed photorespiration) conditions, although mutant plants were able to form nodules and fix atmospheric nitrogen. However, when WT and mutant plants were transferred to an ordinary air atmosphere (photorespiratory active conditions) the nodulation process and nitrogen fixation were substantially affected, particularly in mutant plants. The number and fresh weight of mutant nodules as well as acetylene reduction activity showed a strong inhibition compared with WT plants. Optical microscopy studies from mutant plant nodules revealed the anticipated senescence phenotype linked to an important reduction in starch and sucrose levels. These results show that, in Lotus japonicus, photorespiration and, particularly, GS(2) deficiency result in profound limitations in carbon metabolism that affect the nodulation process and nitrogen fixation.


Membrane fluidity is related to the extent of glycation of proteins, but not to alterations in the cholesterol to phospholipid molar ratio in isolated platelet membranes from diabetic and control subjects.
Platelets from diabetic subjects are hypersensitive to aggregating agents in vitro. Membrane fluidity modulates cell function and we previously reported reduced membrane fluidity associated with hypersensitivity to thrombin in intact platelets from diabetic subjects. Reduced membrane fluidity and hypersensitivity to agonists has also been reported in platelets from non-diabetic subjects whose platelets have an increased cholesterol/phospholipid molar ratio. Glycation of platelet membrane proteins is enhanced in diabetic subjects, and could contribute to the decreased membrane fluidity in these platelets. We examined the relation among fluidity, cholesterol/phospholipid molar ratio, and glycation of proteins in isolated platelet membranes from diabetic and control subjects. Seven poorly controlled diabetic subjects were compared with 7 age- and sex-matched control subjects. The mean steady-state fluorescence polarization value in 1,6-diphenyl-1,3,5-hexatriene-labeled isolated platelet membranes from diabetic subjects (0.184 +/- 0.004) was significantly greater than from control subjects (0.171 +/- 0.004, p less than 0.01); thus, fluidity in platelet membranes from diabetic subjects is decreased. Reduced fluidity in platelet membranes from diabetic subjects could not be attributed to changes in the cholesterol/phospholipid molar ratio. Total or very low density (VLDL), low density (LDL), or high density (HDL3) lipoprotein cholesterol concentration in plasma was not significantly different between groups, but the ratio of VLDL+LDL to HDL2 + HDL3 cholesterol was significantly greater in diabetic subjects (4.79 +/- 0.73) than in control subjects (2.54 +/- 0.30, p less than 0.02). Proteins were glycated significantly more extensively in platelet membranes from diabetic subjects (25.5 +/- 0.9 nmol glucose/mg protein) than those from control subjects (21.0 +/- 0.6 nmol glucose/mg protein, p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


Perspective: detecting adaptive molecular polymorphism: lessons from the MHC.
In the 1960s, when population geneticists first began to collect data on the amount of genetic variation in natural populations, balancing selection was invoked as a possible explanation for how such high levels of molecular variation are maintained. However, the predictions of the neutral theory of molecular evolution have since become the standard by which cases of balancing selection may be inferred. Here we review the evidence for balancing selection acting on the major histocompatibility complex (MHC) of vertebrates, a genetic system that defies many of the predictions of neutrality. We apply many widely used tests of neutrality to MHC data as a benchmark for assessing the power of these tests. These tests can be categorized as detecting selection in the current generation, over the history of populations, or over the histories of species. We find that selection is not detectable in MHC datasets in every generation, population, or every evolutionary lineage. This suggests either that selection on the MHC is heterogeneous or that many of the current neutrality tests lack sufficient power to detect the selection consistently. Additionally, we identify a potential inference problem associated with several tests of neutrality. We demonstrate that the signals of selection may be generated in a relatively short period of microevolutionary time, yet these signals may take exceptionally long periods of time to be erased in the absence of selection. This is especially true for the neutrality test based on the ratio of nonsynonymous to synonymous substitutions. Inference of the nature of the selection events that create such signals should be approached with caution. However, a combination of tests on different time scales may overcome such problems.


Enhanced sensitivity to hydrogen peroxide-induced apoptosis in Evi1 transformed Rat1 fibroblasts due to repression of carbonic anhydrase III.
EVI1 is a nuclear zinc finger protein essential to normal development, which participates in acute myeloid leukaemia progression and transforms Rat1 fibroblasts. In this study we show that enforced expression of Evi1 in Rat1 fibroblasts protects from paclitaxel-induced apoptosis, consistent with previously published studies. Surprisingly, however, these cells show increased sensitivity to hydrogen peroxide (H(2)O(2))-induced apoptosis, demonstrated by elevated caspase 3 catalytic activity. This effect is caused by a reduction in carbonic anhydrase III (caIII) production. caIII transcripts are repressed by 92-97% by Evi1 expression, accompanied by a similar reduction in caIII protein. Reporter assays with the rat caIII gene promoter show repressed activity, demonstrating that Evi1 either directly or indirectly modulates transcription of this gene in Rat1 cells. Targeted knockdown of caIII alone, with Dicer-substrate short inhibitory RNAs, also increases the sensitivity of Rat1 fibroblasts to H(2)O(2), which occurs in the absence of any other changes mediated by Evi1 expression. Enforced expression of caIII in Evi1-expressing Rat1 cells reverts the phenotype, restoring H(2)O(2) resistance. Together these data show that Evi1 represses transcription of caIII gene expression, leading to increased sensitivity to H(2)O(2)-induced apoptosis in Rat1 cells and might suggest the basis for the development of a novel therapeutic strategy for the treatment of leukaemias and solid tumours where EVI1 is overexpressed.


Survival and Time to Initiation of Adjuvant Chemotherapy Among Breast Cancer Patients in Uruguay.
Increases in disease-free survival and overall survival (OS) with the use of adjuvant chemotherapy in early breast cancer (BC) are widely known; however, the optimal time to initiate treatment with adjuvant chemotherapy remains controversial. To evaluate the time elapsed between surgery and the initiation of adjuvant chemotherapy and its possible impact on OS in patients diagnosed with BC stages I-III. This retrospective study included 112 patients diagnosed with BC stages I-III who received adjuvant chemotherapy at the Mastology Unit of the Hospital de Clínicas in Uruguay from 2009 to 2019. OS was estimated using the Kaplan-Meier method, and a Cox proportional hazards model was used to estimate hazard ratios (HRs) and 95% confidence intervals. No statistically significant association was found between the time from surgery to the initiation of chemotherapy and the described variables. OS was worse for patients initiating chemotherapy more than 90 days after breast surgery (n = 19) (HR 7.63; p = 0.004) and between 61 and 90 days after surgery (n = 46) (HR 4.58; p = 0.040) compared to those who started before 30 days (n = 23). Controlling by type of surgery and stage, the prognosis of patients who started chemotherapy between 61 and 90 days after surgery was similar to that of patients who underwent chemotherapy within the first 30 days, controlling for surgery (HR 4.10; p = 0.056) and controlling for stage (HR 3.76; p = 0.075). Prognosis was worse for patients with stage III disease (p = 0.022) who underwent a mastectomy and/or axillary lymph node dissection (p = 0.025). Patients who started chemotherapy more than 90 days following surgery and those with stage III disease or underwent mastectomy and/or axillary lymph node dissection who initiated it between 61 and 90 days had a worse OS. Multiple factors are involved in the time between surgery and the initiation of chemotherapy, and further studies are needed to evaluate which of these factors influence the delay of chemotherapy in order to design strategies to avoid such delays and their negative impact on survival.