Measurement of renal functional response using iohexol clearance-a study of different outpatient procedures.
Glomerular filtration rate (GFR) increases after a heavy protein load; an increase termed renal functional response (RFR). Decreased RFR could be a marker of early kidney damage, but published methods are cumbersome in the outpatient setting. The present study investigates the use of iohexol clearance to measure RFR in outpatients using both one- and two-sample methods. Fourteen healthy volunteers with a mean ± SD age of 42 ± 12 years were included (six males and eight females). GFR was measured using plasma iohexol clearance with one- and two-sample methodologies. Four measurements in each individual were performed: one baseline test and three protein loading tests containing 80 g protein (commercially available protein supplementations from Myo Nutrition and Proteinfabrikken and 350 g chicken breast). RFR was calculated as percentage increase in GFR from the baseline test. Mean RFR was 11.4 ± 5.4% and 12.1 ± 6.4% using one- and two-sample methods, respectively. The three different protein loads resulted in similar mean RFR but there was considerable intra-individual variability. One- and two-sample methods for measurement of RFR showed similar results with near-identical means, but there was some intra-individual variation that was similar for different protein loads. The overall 95% limit of agreement between one- and two-sample methods for calculating RFR was -8.7 to 7.3. RFR can be investigated using plasma iohexol clearance in an outpatient setting. Protocols using commercially available protein supplementation showed a mean RFR of about 12%. One- and two-sample methods for measuring RFR yield similar results.


[Microdialysis study of tolcapone effect during blockage of neuronal dopamine reuptake caused by GBR-12909].
In vivo brain functions analysis was conducted to assess the effect of tolcapone, a novel catechol-O-methyltransferase (COMT) inhibitor on extracellular levels of dopamine (DA) and its metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the striatum of awake, freely moving eats during GBR 12909-induced blockade of DA uptake. Tolcapone administration (30 mg/kg, i.p.) failed to change the dopamine output but caused a marked and long-lasting decrease in the extracellular level of HVA and increase in that of DOPAC. In contrast, injection of the DA uptake inhibitor GBR 12909 directly into the striatum (5 microM) or i.p. (20 mg/kg) led to increase in the DA level but had no significant effect on the metabolites. Co-administration of tolcapone (30 mg/kg i.p.) and CBR 12909 (20 mg/kg) increased the DA level further, whereas the changes in HVA and DOPAC levels remained approximately the same as after injection of tolcapone alone. Behavioral observation showed GBR 12909-induced hyperactivity and stereotypy to be potentiated by tolcapone. These findings show that in the rat striatum under conditions of normal nerve activity DA uptake completely inhibits the increase in striatal DA neurotransmission induced by tolcapone (30 mg/kg i.p.) through COMT inhibition.


Relationships among wood-boring beetles, fungi, and the decomposition of forest biomass.
A prevailing paradigm in forest ecology is that wood-boring beetles facilitate wood decay and carbon cycling, but empirical tests have yielded mixed results. We experimentally determined the effects of wood borers on fungal community assembly and wood decay within pine trunks in the southeastern United States. Pine trunks were made either beetle-accessible or inaccessible. Fungal communities were compared using culturing and high-throughput amplicon sequencing (HTAS) of DNA and RNA. Prior to beetle infestation, living pines had diverse fungal endophyte communities. Endophytes were displaced by beetle-associated fungi in beetle-accessible trees, whereas some endophytes persisted as saprotrophs in beetle-excluded trees. Beetles increased fungal diversity several fold. Over forty taxa of Ascomycota were significantly associated with beetles, but beetles were not consistently associated with any known wood-decaying fungi. Instead, increasing ambrosia beetle infestations caused reduced decay, consistent with previous in vitro experiments that showed beetle-associated fungi reduce decay rates by competing with decay fungi. No effect of bark-inhabiting beetles on decay was detected. Platypodines carried significantly more fungal taxa than scolytines. Molecular results were validated by synthetic and biological mock communities and were consistent across methodologies. RNA sequencing confirmed that beetle-associated fungi were biologically active in the wood. Metabarcode sequencing of the LSU/28S marker recovered important fungal symbionts that were missed by ITS2, though community-level effects were similar between markers. In contrast to the current paradigm, our results indicate ambrosia beetles introduce diverse fungal communities that do not extensively decay wood, but instead reduce decay rates by competing with wood decay fungi.


[Combined laser instrumentation surgery in children with anterior eye segment diseases].
A series comprising 5 combined laser instrumentation methods was developed and patented for the treatment of inborn, posttraumatic, and postuveal abnormalities of the anterior eye segments in children. Two hundred and ninety-six eyes (246 children aged 2 months to 16 years) were operated on. YAG and argon-laser phases of combined operations were made under general anesthesia in children less than 5 years of age and in older children unable to cooperate. Combined laser-instrumentation treatments for anterior eye diseases in children are safe and highly effective and may be recommended for wide use in pediatric ophthalmology as an alternative to instrumental microsurgical methods.


Spin drag of a Fermi gas in a harmonic trap.
Using a Boltzmann equation approach, we analyze how the spin drag of a trapped interacting fermionic mixture is influenced by the nonhomogeneity of the system in a classical regime where the temperature is much larger than the Fermi temperature. We show that for very elongated geometries, the spin damping rate can be related to the spin conductance of an infinitely long cylinder. We characterize analytically the spin conductance both in the hydrodynamic and collisionless limits and discuss the influence of the velocity profile. Our results are in good agreement with recent experiments and provide a quantitative benchmark for further studies of spin drag in ultracold gases.


Fecal excretion of calprotectin in colorectal cancer: relationship to tumor characteristics.
The aim of this study was to evaluate fecal calprotectin in patients treated for colorectal cancer. Furthermore, the changes in fecal calprotectin concentration from before to after surgery were investigated. In 155 patients with newly diagnosed colorectal cancer, two spot samples were taken from the same feces on two consecutive days. Three ways of evaluating calprotectin excretion were compared, (1st spot 1st stool; maximum of 1st spot 1st stool and 2nd spot 1st stool; maximum of 1st spot 1st stool and 1st spot 2nd stool) and gave similar results with median fecal calprotectin values 47 mg/l, 52 mg/l and 54 mg/l, respectively. Median calprotectin concentration did not differ significantly between different tumor stages, although the levels were slightly lower in Dukes stage A tumor than in the rest of the stages. Neither were there any differences in the concentrations related to the localization, size or the histological grading of the carcinoma. As the currently used cut-off level for fecal calprotectin is 10 mg/l, 87% of all patients had elevated fecal calprotectin. Seventy-nine percent of the patients had levels above 15 mg/l and 74% had levels above 20 mg/l (1st spot 1st stool). In patients who delivered fecal samples after the operation the calprotectin value fell significantly from a preoperative median value of 45 mg/l to 14 mg/l after the resection. The majority of patients with colorectal cancer have increased fecal concentration of calprotectin. One single fecal spot seems to be sufficient for determination of the calprotectin level. Measurement of fecal calprotectin may possibly become of value as a marker for colorectal cancer, although calprotectin, similar to fecal occult blood (FOB) tests, is a non-specific test for colorectal pathology, also being elevated in inflammatory bowel diseases. Further investigation of its specificity is therefore needed.


Endogenous CCK disrupts the MMC pattern via capsaicin-sensitive vagal afferent fibers in the rat.
A meal disrupts migrating motor complexes (MMC) in the rat intestine through stimulation of peripheral cholecystokinin (CCK)-B and central CCK-A receptors. The aim of this study was to determine pathways implicated in postprandial disruption of the MMC mediated by CCK. Sprague-Dawley rats were prepared with electrodes for electromyography in the small intestine, and ablation of vagal afferent C-fibers by capsaicin was carried out. Endogenous release of CCK was induced by oral administration of soybean trypsin inhibitor (SBTI). In control rats SBTI disrupted MMC and generated an irregular spiking activity that lasted longer than 3 h. Intravenous infusion of L-365,260 (2 x 10(-7) mol/kg) but not of L-364,718 (3 x 10(-9) mol/kg) restored the MMC pattern. In capsaicin-treated rats, SBTI did not modify fasting activity. Infusion of CCK octapeptide (CCK-8) at 3 x 10(-9) mol.kg-1.h-1 disrupted the MMC, although the response was quantitatively and qualitatively different from SBTI. The effect was reversed by intravenous infusion of L-364,718 or L-365,260 and intracerebroventricular infusion of L-364,718. In capsaicin-treated rats, the intracerebroventricular or intravenous infusion of L-364,718 inhibited CCK-8 effects. However, the intravenous infusion of L-365,260 did not reverse the MMC pattern. These results suggest that the disruption of the MMC mediated by CCK is due to stimulation of peripheral CCK-B receptors located in vagal afferent fibers. This initiates a reflex including stimulation of central CCK-A receptors. Exogenous CCK also stimulates peripheral CCK-A receptors not located in capsaicin-sensitive vagal afferent fibers.


The Representation of Interaural Time Differences in High-Frequency Auditory Cortex.
Early representations of auditory features often involve neuronal populations whose tuning is substantially wider than behavioral discrimination thresholds. Although behavioral discrimination performance can be sometimes achieved by single neurons when using the appropriate part of their (wide) tuning curves, neurons that encode the resulting high-acuity representations have rarely been described. Here we demonstrate the existence of neurons with extremely narrow tuning for interaural time differences (ITDs), a major physical cue for the azimuth of sound sources. The tuning width of ITD-tuned brainstem neurons is mostly determined by the properties of their acoustic input, and may be 10-100 times wider than behavioral thresholds. In contrast, we show that tuning widths of some neurons in the primary auditory cortex in the cat high-frequency auditory cortex (measured using transposed stimulus) can be very sharp and approach behavioral thresholds. Furthermore, while best ITDs of brainstem neurons often lie outside the range of naturally encountered ITDs (the ethological range), the range of best ITDs of the narrowly tuned cortical neurons corresponds well to the ethological range. Thus, our results suggest that the auditory cortex contains a high-resolution representation of ITDs that explicitly decodes the widely tuned brainstem representations.


Undiagnosed coarctation of the aorta as a cause of aortic dissection in the young.
A 35-year-old man presented to the Emergency Department with a history of severe acute central chest pain and simultaneous bilateral paralysis the legs with double incontinence. There was no significant past medical or family history. A CT scan showed a thoracic dissection of the aorta extending from the aortic root to the aortic bifurcation and in to the common iliac arteries. The patient was consented for an axillo-femoral bypass and was taken to theatre and operated on for 7 hours. The patient unfortunately died under anaesthesia. A hospital post-mortem was requested to identify the cause of the dissection. The patient's heart was sent to a cardiac pathologist who identified an undiagnosed coarctation of the aorta. Genetic testing was negative for Marfan syndrome.As a result of the post-mortem, it was recommended that first degree relatives of the deceased undergo ultrasound examination of the cardiovascular system as appropriate to exclude coarctation of the aorta.


A longitudinal twin study on IQ, executive functioning, and attention problems during childhood and early adolescence.
Variation in human behavior may be caused by differences in genotype and by non-genetic differences ("environment") between individuals. The relative contributions of genotype (G) and environment (E) to phenotypic variation can be assessed with the classical twin design. We illustrate this approach with longitudinal data collected in 5 and 12-year-old Dutch twins. At age 5 data on cognitive abilities as assessed with a standard intelligence test (IQ), working memory, selective and sustained attention, and attention problems were collected in 237 twin pairs. Seven years later, 172 twin pairs participated again when they were 12 years old and underwent a similar protocol. Results showed that variation in all phenotypes was influenced by genetic factors. For IQ the heritability estimates increased from 30% at age 5, to 80% at age 12. For executive functioning performance genetic factors accounted for around 50% of the variance at both ages. Attention problems showed high heritabilities (above 60%) at both ages, for maternal and teacher ratings. Longitudinal analyses revealed that executive functioning during childhood was weakly correlated with IQ scores at age 12. Attention problems during childhood, as rated by the mother and the teacher were stronger predictors (r = -0.28 and -0.36, respectively). This association could be attributed to a partly overlapping set of genes influencing attention problems at age 5 and IQ at age 12. IQ performance at age 5 was the best predictor of IQ at age 12. IQ at both ages was influenced by the same genes, whose influence was amplified during development.


Data on assessment of safety and tear proteome change in response to orthokeratology lens - Insight from integrating clinical data and next generation proteomics.
Breath-O™ Correct Ortho-K lenses are newly designed ortho-K lenses which are made from a silicon and fluoride containing methacrylate compound. This compound is said to be more flexible, durable and less likely to break compared to traditional Ortho-K lenses. The special design of this Ortho-K lens can reshape the corneal profile to induce temporary myopic reduction while producing beneficial peripheral hyperopic defocus for myopia control. To evaluate the safety and ocular surface responses of overnight Ortho-K wear over 1 and 3 months using this new type of material, we evaluated the clinical parameters (corneal integrity, corneal biomechanics, corneal endothelial health, non-invasive keratographical break-up time) and profiled the change of global tear proteome on healthy young subjects using next generation proteomics (SWATH-MS). The acquired mass spectrometric data were processed and analyzed using a cloud based Oneomics™ bioinformatic platform. All raw data generated from Information-dependent acquisition (IDA) and SWATH acquisitions were accepted and published in the Peptide Atlas public repository for general release (http://www.peptideatlas.org/PASS/PASS01367).


Electromechanical failure of MoS2 nanosheets.
Molybdenum disulfide (MoS2) has attracted particular attention as a promising electronics and optoelectronics material due to its significant physical properties. In this research, the electromechanical properties of MoS2 nanosheets are systematically investigated with the conductive AFM nanoindentation method. The suspended MoS2 nanosheets with the thickness of tens of nanometers can sustain external applied load (∼4.3 μN) until the bias increases to a critical value (+4 V). Small external load (∼400 nN) may also lead to a failure when the bias (+7 V) overcomes the contact Schottky barrier and generates high electric current. The extent of destruction of MoS2 nanosheets is related to the external applied load and the bias. Besides, the MoS2 nanosheets suspended on the holes are more likely to be damaged than those supported on the substrate under the same conditions. The volcanic volume expansion profiles of the damaged area after the electromechanical failure are caused by electric current-induced local heating anodic oxidation and buckling-induced structural instability. The emergence of gas bubbles in the damaged area proves the strong oxidation process. These experiments have proved that the electric current can promote the mechanical failure of MoS2 nanosheets. The findings can also provide beneficial guidance for the electromechanical applications of MoS2 nano-devices.


Osteomyelitis Infection of Mycobacterium marinum: A Case Report and Literature Review.
Mycobacterium marinum (M. marinum) is a ubiquitous waterborne organism that grows optimally at temperatures around 30°C. It is a nontuberculous Mycobacterium found in nonchlorinated water with worldwide prevalence. It is the most common atypical Mycobacterium that causes opportunistic infection in humans. M. marinum can cause superficial infections and localized invasive infections in humans, with the hands being the sites most frequently affected. It can cause skin lesions, which are either single, papulonodular lesions, confined to an extremity, or may resemble cutaneous sporotrichosis. This infection can also cause deeper infections including tenosynovitis, bursitis, arthritis, and osteomyelitis. Disseminated infections and visceral involvements have been reported in immunocompromised patients. We here report a case of severe deep soft tissue infection with necrotizing fasciitis and osteomyelitis of the left upper extremity (LUE) caused by M. marinum in an immunocompromised patient.


Alternative splicing of 3-hydroxy-3-methylglutaryl coenzyme A reductase is associated with plasma low-density lipoprotein cholesterol response to simvastatin.
HMGCR(3-Hydroxy-3-methylglutaryl coenzyme A reductase), the direct target of statin inhibition, undergoes alternative splicing of exon 13, which encodes part of the statin-binding domain of the enzyme. We hypothesized that HMGCR alternative splicing might be related to the interindividual variation in plasma low-density lipoprotein cholesterol response to statin treatment. We measured mRNA expression of both the full-length and the alternatively spliced HMGCR transcript lacking exon 13 (HMGCRv_1) in 170 simvastatin-incubated immortalized lymphocyte cell lines derived from participants in the Cholesterol and Pharmacogenetics (CAP) study who were treated with simvastatin 40 mg/d for 6 weeks. Greater upregulation of HMGCRv_1 in vitro was significantly correlated (P<or=0.0001) with smaller in vivo reductions of plasma total cholesterol, low-density lipoprotein cholesterol, apoprotein B, and triglycerides and explained 6% to 15% of the variation in their response to treatment. In contrast, no significant relationship was found between expression of the full-length HMGCR transcript and in vivo response. By siRNA knockdown of the full-length transcript, we found that HMGCR enzyme activity measured in cells enriched in HMGCRv_1 was relatively resistant to statin inhibition, consistent with the association of increased alternative splicing with reduced statin response in the CAP study. In addition, we found that a common HMGCR single-nucleotide polymorphism (rs3846662) located within intron 13 was associated with variation in the proportion of HMGCR mRNA that is alternatively spliced. Variation in the production of an HMGCR isoform with reduced statin sensitivity is a determinant of interindividual differences in low-density lipoprotein cholesterol, apolipoprotein B, and triglyceride response to statin treatment.


Heinz body anaemia in lambs with deficiencies of copper or selenium.
1. The progression of Heinz body anaemia was studied in groups of lambs of low- and high-copper status, produced through breeding or Cu supplementation, when they were transferred from improved pasture to rape (Brassica napus L.) in autumn. Some lambs had previously received selenium by injection. The Cu and Se supplements markedly increased superoxide dismutase (EC 184.108.40.206; SOD) and glutathione peroxidase (EC 126.96.36.199; GSHPx) activities respectively in the erythrocytes, and both supplements had elicited growth responses at pasture. 2. At the time of transfer to rape, lambs not treated with Cu had lower whole-blood haemoglobin (Hb) concentrations and a higher percentage of erythrocytes containing Heinz bodies (6.6 v. 3.7%, P less than 0.01) than Cu-treated lambs: the corresponding effects of Se treatment were similar in direction but lower in magnitude (P less than 0.05). 3. After grazing rape for 2 weeks the mean Hb concentration had fallen by 30 g/l while Heinz body count had increased from 5 to 25%. However, counts were negatively correlated with the initial values and were unaffected by the Cu and Se treatments which maintained high plasma Cu concentrations and SOD and GSHPx activities. 4. The results provide the first evidence that Cu deficiency can induce Heinz body formation and the anaemia in grazing Cu-deficient lambs may be partly haemolytic in origin. The concomitant Se deficiency added marginally to the problem but neither the separate nor combined deficiencies increased the susceptibility of lambs to brassica anaemia.


Dry eye disease is associated with deterioration of mental health in male Japanese university staff.
Dry eye disease (DED) is defined as a multifactorial disease of the tears and ocular surface that results in symptoms of discomfort, visual disturbance, and tear-film instability, with potential damage to the ocular surface. It is thought to be associated with reduced quality of life (QOL). The aim of the present study was to investigate the effects of DED on health-related QOL in Japanese university sedentary office workers who are daily users of visual display terminal. In this study, 163 university staff (99 male and 64 female), aged 23-69 years, served as study subjects. Subjects were asked to answer the following three questions. (1) How often do your eyes feel dry? (2) How often do your eyes feel irritated? (3) Have you ever been diagnosed by a clinician as having dry eye syndrome? Sixty-eight subjects who answered "constantly," "often", or "sometimes" to both questions 1 and 2 were classified as the DED Group, and the remaining 95 were defined as the Non-DED Group. QOL was assessed by the SF-36 questionnaire, which consisted of 36 items to produce three summary scores, namely, mental, physical, and role/social component summary scores. For males, the DED Group had significantly lower scores than the Non-DED Group for mental component summary (MCS) (P = 0.005). In multiple regression analysis, MCS scores were adversely related to DED in males (P = 0.015). DED was associated with worsened QOL. DED should be regarded as a factor that can lead to deterioration of mental health.


[The work of I.I. Schmalhousen on comparative morphology of vertebrates: Significance for paleontological reconstructions (data on the seismo-sensory system of agnathans)].
The morphology and function of the seismo-sensory system of agnathans (Agnatha: Heterostraci) are analyzed. This system has been reconstructed in Cyathaspidiformes and Amphiaspidiformes. The seismo-sensory system of agnathans was compared with the lateral line system of fishes: the difference between these systems in the density of seismo-sensory lines and the contribution level of these systems to orientation in the water was detected. The possibility of reconstruction of the length and direction of heterostracans cranial nerves has been shown using the position ofseismo-sensory lines and departments of the brain.


Umbilical hernia repair increases the rate of functional gastrointestinal disorders in children.
To hypothesize that hernia repair would not change the incidence of functional gastrointestinal disorders (FGIDs) due to the benign and limited nature of the procedure. This cohort study assessed a randomized selection of children aged 4-18 years who underwent hernia repair more than 4 years prior at Ann and Robert H. Lurie Children's Hospital of Chicago. Controls were siblings who had not undergone surgery previously. Parents completed the Questionnaire on Pediatric Gastrointestinal Symptoms-Rome III Version by telephone for subjects and controls. The primary outcome was the presence of FGIDs. Fifty children with hernia repair and 43 sibling controls were identified. At the time of survey, subjects with hernia repair were average age 12.9 years (range 5-18 years, 60% male) and controls were average age 12.2 years (range 4-18 years, 49% male). Average age at surgical repair was 5.2 years (median 5.2 years, range 0.2-10.4 years) and average time since surgical repair was 7.8 years (range 4.8-13.7 years). FGIDs were diagnosed in 10/50 (20%) cases of hernia repair and 2/43 (5%) controls (P = .033, Fisher 2-tailed test). Umbilical hernia repair increases the likelihood of FGIDs in childhood. Additional studies are needed to identify aspects of surgery that may be associated with development of FGIDs.


Upregulation and nuclear translocation of testicular ghrelin protects differentiating spermatogonia from ionizing radiation injury.
Proper control of apoptotic signaling is important for maintenance of testicular homeostasis after ionizing radiation (IR). Herein, we challenged the hypothesis that ghrelin, a pleiotropic modulator, is potentially involved in IR-induced germ cell injury. Lower body exposure to 2 Gy of IR induced a notable increase of ghrelin expression in the nuclear of differentiating spermatogonia at defined stages, with an impairment in the Leydig cells (LCs)-expressing ghrelin. Unexpectedly, inhibition of the ghrelin pathway by intraperitoneal injection of a specific GHS-R1α antagonist enhanced spermatogonia elimination by apoptosis during the early recovery following IR, and thereafter resulted in impaired male fertility, suggesting that the anti-apoptotic effects of evoked ghrelin, although transient along testicular IR injury, have a profound influence on the post-injury recovery. In addition, inhibition of ghrelin signaling resulted in a significant increase in the intratesticular testosterone (T) level at the end of 21 days after IR, which should stimulate the spermatogenic recovery from surviving spermatogonia to a certain extent during the late stage. We further demonstrated that the upregulation and nuclear trafficking of ghrelin, elaborately regulated by IR-elicited antioxidant system in spermatogonia, may act through a p53-dependent mechanism. The elicitation of ghrelin expression by IR stress, the regulation of ghrelin expression by IR-induced oxidative stress and the interaction between p53 and ghrelin signaling during IR injury were confirmed in cultured spermatogonia. Hence, our results represent the first evidence in support of a radioprotective role of ghrelin in the differentiating spermatogonia. The acutely, delicate regulation of local-produced ghrelin appears to be a fine-tune mechanism modulating the balance between testicular homeostasis and early IR injury.


Antitumour and immunological activity of a beta 1----3/1----6 glucan from Glomerella cingulata.
The in vivo antitumour activity of a beta 1----3/1----6 glucan from the fungus Glomerella cingulata was investigated in vivo. The glucan exhibited a strong inhibition of tumour growth of the allogeneic Sarcoma-180 as well as the syngeneic DBA/2-MC.SC-1 fibrosarcoma with inhibition ratios up to 100%. Against the hormone sensitive Noble-Nb-R prostate carcinoma the glucan alone showed a moderate antitumour effect, whereas in combination with diethylstilbestrol an almost complete regression of the tumour could be achieved. It could be demonstrated that a highly ordered structure of the glucan is not essential for the antitumour activity. Since the glucan expressed no direct cytotoxic effects, the immunomodulating activity was investigated in vitro in order to get an indication for a possible mode of action. In the lymphocyte transformation assay the glucan at a dose of 100 micrograms/ml caused a fourfold increase in the proliferation of murine spleen lymphocytes. Moreover, the glucan stimulated the phagocytosis of zymosan by bone marrow macrophages up to 100%. However, the glucan was not able to render macrophages cytotoxic against P-815 mastocytoma cells.


Prevalence of diabetes treatment effect modifiers: the external validity of trials to older adults.
Potential treatment effect modifiers (TEMs) are specific diseases or conditions with a well-described mechanism for treatment effect modification. The prevalence of TEMs in older adults with type 2 diabetes mellitus (DM) is unknown. Objectives were to 1) determine the prevalence of pre-specified potential TEMs; 2) demonstrate the potential impact of TEMs in the older adult population using a simulated trial; 3) identify TEM combinations associated with number of hospitalizations to test construct validity. Data are from the nationally-representative United States National Health and Examination Survey, 1999-2004: 8646 Civilian, non-institutionalized adults aged 45-64 or 65+ years, including 1443 with DM. TEMs were anemia, congestive heart failure, liver inflammation, polypharmacy, renal insufficiency, cognitive impairment, dizziness, frequent mental distress, mobility difficulty, and visual impairment. A trial was simulated to examine prevalence of potential TEM impact. The cross-sectional association between TEM patterns and number of hospitalizations was estimated to assess construct validity. The prevalence of TEMs was substantial such that 19.0% (95% CI 14.8-23.2) of middle-aged adults and 38.0% (95% CI 33.4-42.5) of older adults had any two. A simulated trial with modest levels of interaction suggested the prevalence of TEMs could nullify treatment benefit in 3.9-27.2% of older adults with DM. Compared to having DM alone, hospitalization rate was increased by several combinations of TEMs with substantial prevalence. We provide national benchmarks that can be used to evaluate TEM prevalence reported by clinical trials of DM, and correspondingly their external validity to older adults.


Uptake of tetracycline by membrane preparations from Escherichia coli.
1. A membrane fraction from Escherichia coli has been prepared essentially free from ribosomes by treatment of the membranes with Triton X-100 at 0 degrees C followed by differential centrifugation. 2. The ribosome-free membrane vesicles absorbed tetracycline by a reversible temperature-dependent process with an apparent K(m) of 0.029mm at pH7.5 and 37 degrees C. 3. The absorption process was negligible below 25 degrees C and had an optimum at 40 degrees C; a pH optimum at 7.5 was observed. 4. The absorption of tetracycline was strongly inhibited by EDTA and ATP; ADP inhibited less strongly and AMP had no effect. 5. There was no significant difference in the rates or extent of uptake of tetracycline by membranes prepared from tetracycline-sensitive and tetracycline-resistant, R-factor-bearing E. coli.


Myocardial injury and related mortality in hospitalized patients with COVID-19 during the Omicron pandemic: new perspectives and insights.
Myocardial injury is one of the most common comorbidity in SARS-CoV-2 infected patients, and has poor prognosis. However, the incidence of myocardial injury in patients with SARS-CoV-2 infection has not been sufficiently investigated during the Omicron wave. We conducted a retrospective study of 2690 patients with confirmed SARS-CoV-2 Omicron infection from Tongji Hospital. The results indicated that the myocardial injury accounted for 30.8% of the total patients with SARS-CoV-2 infection and was associated with higher in-hospital mortality than those without injury before and after propensity score matching (PSM) [adjusted hazard ratio (HR), 10.61; 95% confidence interval (CI), 7.76-14.51; P < 0.001; adjusted HR, 2.70; 95% CI, 1.86-3.93; P < 0.001; respectively]. Further, the levels of cytokines (IL-1β, IL-6, IL-10, and TNF-α) in patients with myocardial injury were higher than those without injury, and the higher levels of cytokines in the myocardial injury group were associated with increased mortality. Administration of angiotensin-converting enzyme inhibitors or angiotensin receptor blockers (ACEI/ARB) could significantly reduce the mortality in patients with myocardial injury (adjusted HR, 0.52; 95% CI, 0.38-0.71; P < 0.001). Additionally, the level of angiotensin II increased in patients with SARS-CoV-2 infection was even higher in myocardial injury group compared to those without injury. Collectively, the study summarized the clinical characteristic and outcome of SARS-CoV-2 infected patients with myocardial injury during the Omicron wave in China, and validated the protective role of ACEI/ARB in improving the survival of those with myocardial injury.


Endometriosis in rhesus monkeys (Macaca mulatta) following chronic exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin.
The incidence of the reproductive disease endometriosis was determined in a colony of rhesus monkeys chronically exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD or dioxin) for a period of 4 years. Ten years after termination of dioxin treatment, the presence of endometriosis was documented by surgical laparoscopy and the severity of disease was assessed. The incidence of endometriosis was directly correlated with dioxin exposure and the severity of disease was dependent upon the dose administered (p < 0.001). Three of 7 animals exposed to 5 ppt dioxin (43%) and 5 of 7 animals exposed to 25 ppt dioxin (71%) had moderate to severe endometriosis. In contrast, the frequency of disease in the control group was 33%, similar to an overall prevalence of 30% in 304 rhesus monkeys housed at The Harlow Primate Center with no dioxin exposure. This 15-year study indicates that latent female reproductive abnormalities may be associated with dioxin exposure in the rhesus. Therefore, the effects of this toxin may be more diverse than previously recognized.


Nanoparticle-nanoparticle vs. nanoparticle-substrate hot spot contributions to the SERS signal: studying Raman labelled monomers, dimers and trimers.
We used a combination of Raman microscopy, AFM and TEM to quantify the influence of dimerization on the surface enhanced Raman spectroscopy (SERS) signal for gold and silver nanoparticles (NPs) modified with Raman reporters and situated on gold, silver, and aluminum films and a silicon wafer. The overall increases in the mean SERS enhancement factor (EF) upon dimerization (up by 43% on average) and trimerisation (up by 96% on average) of AuNPs and AgNPs on the studied metal films are within a factor of two, which is moderate when compared to most theoretical models. However, the maximum ratio of EFs for some dimers to the mean EF of monomers can be as high as 5.5 for AgNPs on a gold substrate. In contrast, for dimerization and trimerization of gold and silver NPs on silicon, the mean EF increases by 1-2 orders of magnitude relative to the mean EF of single NPs. Therefore, hot spots in the interparticle gap between gold nanoparticles rather than hot spots between Au nanoparticles and the substrate dominate SERS enhancement for dimers and trimers on a silicon substrate. However, Raman labeled noble metal nanoparticles on plasmonic metal films generate on average SERS enhancement of the same order of magnitude for both types of hot spot zones (e.g. NP/NP and NP/metal film).


Effect of a geriatric nurse education program on the knowledge, attitudes, and certification of hospital nurses.
Although nurses are the primary care providers for hospitalized and vulnerable older adults living in the community, they are generally not prepared in geriatric care. This study examined the effect of a 21-hour nursing education program on nurses' knowledge and attitudes regarding care of older adults and the geriatric nursing certification of the participants. The program was offered to 92 nurses in three cohorts over a 1-year period. Participants completed pre- and posttest measures of geriatric knowledge and attitudes. Significant differences were found between pretest and posttest knowledge and attitudes. Program evaluation and success in achieving certification in gerontological nursing also indicated a positive effect of the program.


Vaginal Mycoplasmataceae colonization and association with immune mediators in pregnancy.
To determine the prevalence of Mycoplasmataceae species in pregnant women and evaluate their association with immune system mediators. Women were prospectively enrolled between 16-22 weeks' gestation. Vaginal swabs were self-collected and analyzed with PCR for Mycoplasma hominis (MH) and Mycoplasma genitalium (MG) as well as Ureaplasma urealyticum (UU) and Ureaplasma parvum (UP) (collectively, Myc). Immune mediators were measured via Luminex multiplex assay. Women with vaginal Mycoplasmataceae were compared to women without Myc, and women with Mycoplasma species (MH or MG) were compared to women without MH or MG. Linear regression models were used to investigate the relationship of the presence of Mycoplasmataceae on log-transformed immune mediators while controlling for confounders using propensity scores. One-hundred-twenty women were enrolled and had complete lab data available. Colonization was 20.8, 2.5, 10.0, and 48.3% for MH, MG, UU, and UP, respectively. Women with any Mycoplasmataceae were more likely to be younger, of the Black race, and have public insurance. There were no significant differences in immune mediators between women with vaginal Mycoplasmataceae versus those without. After controlling for confounders, women with MH and/or MG had significantly elevated levels of IL-1β compared to women without MH or MG (estimate = 1.12; 95% CI = 0.33, 1.93). There were no other significant differences in immune mediators in women with MH and/or MG compared to those without. Colonization rates were highest for UP and lowest for MG. Higher IL-1β levels were seen in the presence of MH and/or MG, indicating that these less frequently encountered organisms may incite a stronger host response. There were no other significant differences in immune mediator levels.


Transformation of Streptococcus sanguis Challis by plasmid deoxyribonucleic acid from Streptococcus faecalis.
Plasmid deoxyribonucleic acid (DNA) from Streptococcus faecalis, strain DS5, was transferred to the Challis strain of Streptococcus sanguis by transformation. Two antibiotic resistance markers carried by the beta plasmid from strain DS5, erythromycin and lincomycin, were transferred to S. sanguis at a maximum frequency of 1.8 x 10-5/colony-forming unit. Approximately 70% of the covalently closed circular DNA isolated from transformant cultures by dye buoyant density gradients was shown to be hybridizable to beta plasmid DNA. Two major differences were observed between the beta plasmid from S. faecalis and the plasmid isolated from transformed S. sanguis: (i) the beta plasmid from strain DS5 sedimented in velocity gradients at 43S, whereas the covalently closed circular DNA from transformed Challis sedimented at 41S, suggesting a 1.5-Mdal deletion from the beta plasmid occurred; (ii) although the 43S beta plasmid remained in the supercoiled configuration for several weeks after isolation, the 41S plasmid was rapidly converted to a linear double-stranded molecule. Attempts to transform S. sanguis with the alpha plasmid from S. faecalis, strain DS5, were unsuccessful.


In vivo evidence for preferential role of dopamine D3 receptor in the presynaptic regulation of dopamine release but not synthesis.
Brain microdialysis was used to investigate the effects of the putative dopamine D3 receptor agonist (+/-)-7-hydroxy-N,N-di-n-propyl-2-aminotetralin (7-OH-DPAT) on dopamine release, metabolism and synthesis in the dorsal striatum and nucleus accumbens of awake rats. The drug administered i.p. dose dependently decreased the release, metabolism and synthesis of dopamine in both brain areas. The potency of 7-OH-DPAT to decrease dopamine release was found to be higher in the nucleus accumbens than in the dorsal striatum (ED50 for nucleus accumbens 0.0096 mg/kg, i.p.; for dorsal striatum 0.068 mg/kg, i.p.). Dopamine metabolism, assessed by measuring 3,4-dihydroxyphenylacetic acid extracellular levels, and dopamine synthesis, determined as 3,4-dihydroxyphenylalanine output following perfusion with the L-aromatic acid decarboxylase inhibitor 3-hydroxybenzylhydrazine (10(-5) M), were decreased at higher dose ranges of 7-OH-DPAT (ED50 for decrease of 3,4-dihydroxyphenylalanine output in nucleus accumbens 0.124 mg/kg, i.p.; in dorsal striatum 0.101 mg/kg, i.p.). The hypomotility of rats induced by 7-OH-DPAT in doses of 0.002-0.25 mg/kg, i.p., was shown to correlate with the decreased dopamine release in the nucleus accumbens. Pretreatment of animals with 7-OH-DPAT at the putative dopamine D3 receptor 'selective' dose of 0.05 mg/kg, i.p., was found to prevent the increase of dopamine release but not the increase in metabolism in the dorsal striatum of freely moving rats induced by (+)-AJ76, cis (+)-(1S,2R)-5-methoxy-1-methyl-1-2-(n-propylamino)tetralin HCI (7 mg/kg, i.p.) and haloperidol (0.1 mg/kg, i.p.). Local application of 7-OH-DPAT by addition into the perfusing medium also resulted in a preferential decrease of dopamine release in the nucleus accumbens as compared with the dorsal striatum (EC50 for nucleus accumbens 1.9 nM; for dorsal striatum 11.3 nM). The present results give further support to the hypothesis that the dopamine D3 autoreceptor is preferentially involved in the presynaptic regulation of dopamine release, while the D2 autoreceptor controls dopamine synthesis.


Condition dependence of female choosiness in a field cricket.
Females generally choose mates that produce the loudest, brightest or most elaborate sexual displays, and these costly male displays are predicted to be condition dependent. However, mate choice itself is a costly behaviour also expected to be condition dependent. Male fall field crickets, Gryllus pennsylvanicus, produce a conspicuous long-distance calling song that attracts females and is condition dependent. In this study, we tested the condition dependence of female preferences (preference function and choosiness) for male calling effort in G. pennsylvanicus. We manipulated female condition by raising crickets from hatching on either a low- or high-quality diet. In a series of two-speaker phonotaxis trials, both low- and high-condition females preferred playbacks reflecting greater calling effort. However, relative to low-condition females, high-condition females took significantly longer to make a choice, were more likely to fail to choose within the time allotted for a phonotaxis trial and significantly increased their latency to choose over the course of multiple trials. We discuss these results with respect to the possibility that female G. pennsylvanicus may be foraging for direct benefits when they choose their mates.


Ergonomically designed kneeling chairs are they worth it? : Comparison of sagittal lumbar curvature in two different seating postures.
General agreement among researchers suggests that poor seating posture may predispose individuals to developing low back pain. A variety of methods such as ergonomically designed chairs have been developed to assist people to maintain good posture and preserve the 'natural' lumbar curve. The aim of this study was to compare lumbar curvature on an ergonomically designed kneeling chair (EKC) with that on a standard computer chair (SCC), with reference to the standing lumbar curvature. The study used a repeated measures, within-subjects design. A convenience sample of twenty participants was recruited aged 18-35 (9 male and 11 female). Lumbar curvature was measured using the 'Middlesbrough Integrated Assessment System' (MIDAS) postural assessment tool in three different postures; sitting on a SCC, sitting on an EKC set at +20 degrees inclination and standing as the reference measurement. Results were analysed by a repeated measures oneway ANOVA (1 factor) with 3 levels followed by the Bonferroni post hoc test. The results showed a statistically significant difference between standing lumbar curvature and lumbar curvature produced by both of the chairs (p<0.05). There was also a statistically significant difference between the two seated positions (p<0.05). This study suggests that ergonomically designed kneeling chairs set at +20 degrees inclination do maintain standing lumbar curvature to a greater extent than sitting on a standard computer chair with an overall mean difference of 7.633 degrees . Further research with a greater number of subjects and on different chair designs is warranted.


Four-year survival with nivolumab in patients with previously treated advanced non-small-cell lung cancer: a pooled analysis.
Phase 3 clinical data has shown higher proportions of patients with objective response, longer response duration, and longer overall survival with nivolumab versus docetaxel in patients with previously treated advanced non-small-cell lung cancer (NSCLC). We aimed to evaluate the long-term benefit of nivolumab and the effect of response and disease control on subsequent survival. We pooled data from four clinical studies of nivolumab in patients with previously treated NSCLC (CheckMate 017, 057, 063, and 003) to evaluate survival outcomes. Trials of nivolumab in the second-line or later setting with at least 4 years follow-up were included. Comparisons of nivolumab versus docetaxel included all randomised patients from the phase 3 CheckMate 017 and 057 studies. We did landmark analyses by response status at 6 months to determine post-landmark survival outcomes. We excluded patients who did not have a radiographic tumour assessment at 6 months. Safety analyses included all patients who received at least one dose of nivolumab. Across all four studies, 4-year overall survival with nivolumab was 14% (95% CI 11-17) for all patients (n=664), 19% (15-24) for those with at least 1% PD-L1 expression, and 11% (7-16) for those with less than 1% PD-L1 expression. In CheckMate 017 and 057, 4-year overall survival was 14% (95% CI 11-18) in patients treated with nivolumab, compared with 5% (3-7) in patients treated with docetaxel. Survival subsequent to response at 6 months on nivolumab or docetaxel was longer than after progressive disease at 6 months, with hazard ratios for overall survival of 0·18 (95% 0·12-0·27) for nivolumab and 0·43 (0·29-0·65) for docetaxel; for stable disease versus progressive disease, hazard ratios were 0·52 (0·37-0·71) for nivolumab and 0·80 (0·61-1·04) for docetaxel. Long-term data did not show any new safety signals. Patients with advanced NSCLC treated with nivolumab achieved a greater duration of response compared with patients treated with docetaxel, which was associated with a long-term survival advantage. Bristol-Myers Squibb.


Cardiac function is not significantly diminished in hearts isolated from young caveolin-1 knockout mice.
Matrix metalloproteinases (MMPs) are known to degrade components of the extracellular matrix. More recently, in myocardial oxidative stress injury including ischemia-reperfusion, MMP-2 is activated and degrades troponin I and α-actinin. MMP activity is regulated at several levels. We recently showed that MMP-2 is localized in the caveolae of cardiomyocytes and is negatively regulated by caveolin-1 (Cav-1). The caveolin scaffolding domain of Cav-1 inhibits MMP-2 proteolytic activity in vitro, and Cav-1(-/-) mouse hearts have increased MMP-2 activity compared with controls. Whether this increase in MMP-2 activity translates to impaired cardiac function is unknown. Hearts isolated from Cav-1(-/-) mice and their wild-type controls were perfused as isolated working hearts and physiologically challenged with increasing increments of left atrial preload (7-22.5 mmHg). The hearts were then pharmacologically challenged with increasing concentrations of isoproterenol (0.1-100 nM). Functionally, the Cav-1(-/-) hearts were similar to the controls in heart rate, peak systolic pressure, developed pressure, and rate pressure product. At higher preload pressures, the Cav-1(-/-) hearts outperformed the control hearts. Coronary flow was significantly higher in Cav-1(-/-) hearts under all conditions. The highest concentration of isoproternol increased the heart rate of Cav-1(-/-) hearts more than in controls. Western blot analysis revealed no significant changes in troponin I or α-actinin between Cav-1(-/-) hearts and their controls. There was a significant loss of MMP-2 from both knockout and control hearts during the perfusion. In summary, despite the loss of Cav-1, Cav-1(-/-) hearts show similar or better cardiac function compared with wild-type hearts following physiological challenge or β-adrenergic stimulation in vitro, and this appears unrelated to changes in MMP-2.


Tidal energy extraction: renewable, sustainable and predictable.
The tidal flow of sea water induced by planetary motion is a potential source of energy if suitable systems can be designed and operated in a cost-effective manner This paper examines the physical origins of the tides and how the local currents are influenced by the depth of the seabed and presence of land mass and associated coastal features. The available methods of extracting energy from tidal movement are classified into devices that store and release potential energy and those that capture kinetic energy directly. A survey is made of candidate designs and, for the most promising, the likely efficiency of energy conversion and methods of installing them are considered. Overall, the need to reduce CO2 emissions, a likely continued rise in fossil fuel cost will result in a significantly increased use of tidal energy. What is still required, especially for kinetic energy devices, is a much greater understanding of how they can be designed to withstand long-term immersion in the marine environment.


Chromosome-level de novo assembly of Coprinopsis cinerea A43mut B43mut pab1-1 #326 and genetic variant identification of mutants using Nanopore MinION sequencing.
The homokaryotic Coprinopsis cinerea strain A43mut B43mut pab1-1 #326 is a widely used experimental model for developmental studies in mushroom-forming fungi. It can grow on defined artificial media and complete the whole lifecycle within two weeks. The mutations in mating type factors A and B result in the special feature of clamp formation and fruiting without mating. This feature allows investigations and manipulations with a homokaryotic genetic background. Current genome assembly of strain #326 was based on short-read sequencing data and was highly fragmented, leading to the bias in gene annotation and downstream analyses. Here, we report a chromosome-level genome assembly of strain #326. Oxford Nanopore Technology (ONT) MinION sequencing was used to get long reads. Illumina short reads was used to polish the sequences. A combined assembly yield 13 chromosomes and a mitochondrial genome as individual scaffolds. The assembly has 15,250 annotated genes with a high synteny with the C. cinerea strain Okayama-7 #130. This assembly has great improvement on contiguity and annotations. It is a suitable reference for further genomic studies, especially for the genetic, genomic and transcriptomic analyses in ONT long reads. Single nucleotide variants and structural variants in six mutagenized and cisplatin-screened mutants could be identified and validated. A 66 bp deletion in Ras GTPase-activating protein (RasGAP) was found in all mutants. To make a better use of ONT sequencing platform, we modified a high-molecular-weight genomic DNA isolation protocol based on magnetic beads for filamentous fungi. This study showed the use of MinION to construct a fungal reference genome and to perform downstream studies in an individual laboratory. An experimental workflow was proposed, from DNA isolation and whole genome sequencing, to genome assembly and variant calling. Our results provided solutions and parameters for fungal genomic analysis on MinION sequencing platform.


Global, regional and national burden of rheumatoid arthritis 1990-2017: a systematic analysis of the Global Burden of Disease study 2017.
To provide the level and trends of prevalence, incidence and disability adjusted life years (DALYs) for rheumatoid arthritis (RA) in 195 countries from 1990 to 2017 by age, sex, Socio-demographic Index (SDI; a composite of sociodemographic factors) and Healthcare Access and Quality (an indicator of health system performance) Index. Data from the Global Burden of Diseases, Injuries, and Risk Factors study (GBD) 2017 were used. GBD 2017 modelled the burden of RA for 195 countries from 1990 to 2017, through a systematic analysis of mortality and morbidity data to estimate prevalence, incidence and DALYs. All estimates were presented as counts and age-standardised rates per 100 000 population, with uncertainty intervals (UIs). Globally, the age-standardised point prevalence and annual incidence rates of RA were 246.6 (95% UI 222.4 to 270.8) and 14.9 (95% UI 13.3 to 16.4) in 2017, which increased by 7.4% (95% UI 5.3 to 9.4) and 8.2% (95% UI 5.9 to 10.5) from 1990, respectively. However, the age-standardised rate of RA DALYs per 100 000 population was 43.3 (95% UI 33.0 to 54.5) in 2017, which was a 3.6% (95% UI -9.7 to 0.3) decrease from the 1990 rate. The age-standardised prevalence and DALY rates increased with age and were higher in females; the rates peaked at 70-74 and 75-79 age groups for females and males, respectively. A non-linear association was found between age-standardised DALY rate and SDI. The global age-standardised DALY rate decreased from 1990 to 2012 but then increased and reached higher than expected levels in the following 5 years to 2017. The UK had the highest age-standardised prevalence rate (471.8 (95% UI 428.9 to 514.9)) and age-standardised incidence rate (27.5 (95% UI 24.7 to 30.0)) in 2017. Canada, Paraguay and Guatemala showed the largest increases in age-standardised prevalence rates (54.7% (95% UI 49.2 to 59.7), 41.8% (95% UI 35.0 to 48.6) and 37.0% (95% UI 30.9 to 43.9), respectively) and age-standardised incidence rates (48.2% (95% UI 41.5 to 55.1), 43.6% (95% UI 36.6 to 50.7) and 36.8% (95% UI 30.4 to 44.3), respectively) between 1990 and 2017. RA is a major global public health challenge. The age-standardised prevalence and incidence rates are increasing, especially in countries such as Canada, Paraguay and Guatemala. Early identification and treatment of RA is vital especially among females, in order to reduce the ongoing burden of this condition. The quality of health data needs to be improved for better monitoring of disease burden.


Probing macroscopic realism via Ramsey correlation measurements.
We describe a new and experimentally feasible protocol for performing fundamental tests of quantum mechanics with massive objects. In our approach, a single two-level system is used to probe the motion of a nanomechanical resonator via multiple Ramsey interference measurements. This scheme enables the measurement of modular variables of macroscopic continuous-variable systems; we show that correlations thereof violate a Leggett-Garg inequality and can be applied for tests of quantum contextuality. Our method can be implemented with a variety of different solid-state or photonic qubit-resonator systems, and it provides a clear experimental signature to distinguish the predictions of quantum mechanics from those of other alternative theories at a macroscopic scale.


Smoking cessation is associated with decreased mortality and improved amputation-free survival among patients with symptomatic peripheral artery disease.
Although smoking cessation is recommended for all patients with peripheral artery disease, there are little data regarding the prevalence of smoking among patients at the time of angiography or the effect of smoking cessation on clinical outcomes. Consecutive patients with claudication or critical limb ischemia who underwent peripheral angiography from 2006 to 2013 were included in an observational cohort analysis. Smoking status was assessed at the time of angiography and during follow-up clinic visits. Kaplan-Meier analysis was used to assess the relationship between smoking cessation, mortality, and amputation-free survival. Among 739 patients (423 men and 316 women; mean age, 60 ± 12 years), 204 (28%) remained active smokers at the time of lower extremity angiography. At the time of angiography, the mean number of cigarettes smoked per day was 16 ± 10, and the mean pack-years was 40 ± 25. During the course of the subsequent year, 61 patients (30%) successfully quit smoking and maintained continued abstinence. Baseline medication use between groups did not differ significantly. The mean ankle-brachial index was also similar for quitters vs nonquitters (0.53 ± 24 vs 0.49 ± 0.22; P = .3). During follow-up to 5 years, patients who quit smoking had significantly lower all-cause mortality (14% vs 31%; hazard ratio, 0.40; 95% confidence interval, 0.18-0.90) and improved amputation-free survival (81% vs 60%; hazard ratio, 0.43, 95% confidence interval, 0.22-0.86) compared with patients who continued smoking, with most of the difference driven by reduced mortality among patients who quit smoking. The findings remained significant on multivariable analysis. Approximately one-third of active smokers with peripheral artery disease successfully quit smoking ≤ 1 year after lower extremity angiography. Patients who quit smoking have lower mortality and improved amputation-free survival compared with patients who continue smoking.


Experimental evaluation of commercially available semi-permeable membranes for use with parallel-plate dialysers in flow injection systems.
Mass transfer in on-line analytical parallel-plate dialysers was studied in the continuous and flow injection (FI) modes in order to characterise the nature of the dialysis process and its dependence on experimental variables. A number of different semi-permeable membrane surfaces were evaluated in the laminar-flow and plug-flow configurations. The fraction of analyte transferred from the donor to the acceptor stream depends on parameters such as type of membrane used, membrane surface, membrane line-length, membrane porosity, concentration of analyte in the donor stream, the use of concurrent and countercurrent flow between the donor and acceptor streams, and flow-rates of the donor and acceptor streams. The dialysis of calcium and chloride ions, in the absence of protein, was studied with different membrane types and dialysers with different dimensions in continuous flow and FI, and the influence of the above mentioned parameters on the dynamic dialysis process of these ions is described.


"No one left abandoned": Cuba's national health system since the 1959 revolution.
In spite of the economic hardships during the 1990s, Cuba has achieved health indicators that are among the best in the world. This article describes the development of the Cuban health system over more than four decades and analyzes its dynamics. Four stages can be identified. The system's foundations were laid during the first post-revolutionary decade (1959--1970) and consolidated during the succeeding decade (1970--1979). In the third stage, from 1980 onward, the system reached its full expansion with the development of family medicine. Following the crisis of the 1990s, a fourth stage began with reforms and adjustments to the new situation after the collapse of the Soviet Union. Today, health care continues to be of high quality and free for all Cubans. It remains exclusively in the hands of the public sector, and privatization is not an option. This is exactly the opposite of what is happening in other parts of the world where public services are underfunded and people are made to believe that privatization is the only way to ensure high-quality care.


Maintenance of Alveolar Ridge Dimensions Utilizing an Extracted Tooth Dentin Particulate Autograft and PlateletRich Fibrin: A Retrospective Radiographic ConeBeam Computed Tomography Study.
This study utilized radiographic comparative analysis in order to evaluate dimensional ridge changes four months after tooth extraction and immediate grafting with mineralized dentin particulate autograft and chopped plateletrich fibrin. Fiftyeight extraction sockets with up to 2 mm of missing buccal bone in the coronal aspect compared to the lingual bone were included. Graft material was covered with either a plateletrich fibrin membrane or collagen sponge with no effort to achieve primary closure. The dimensional changes of the ridge were assessed on conebeam computed tomography (CBCT) images acquired prior to extraction and four months later. The reduction in the buccal bone plate thickness 1 mm, 3 mm, and 5 mm below the buccal crest was -0.87 ± 0.84 mm, -0.60 ± 0.70 mm, and -0.41 ± 0.55 mm, respectively. The mean ridge width changes 1 mm, 3 mm, and 5 mm below the crest were -1.38 ± 1.24 mm, -0.82 ± 1.13 mm, and -0.43 ± 0.89 mm, respectively. The average midbuccal bone height gain was +1.1%, while the midlingual height gain was 5.6%. A mineralized dentin autograft with plateletrich fibrin is effective in preserving postextraction alveolar ridge dimensions.


PKC{alpha} is essential for the proper activation and termination of rod bipolar cell response.
Protein kinase (PKC)-α is abundant in retinal bipolar cells. This study was performed to explore its role in visual processing. PKCα-knockout (Prkca(-/-)) mice and control animals were examined by using electroretinography (ERG), light microscopy, and immunocytochemistry. The Prkca(-/-) mice showed no signs of retinal degeneration up to 12 months of age, but ERG measurements indicated a decelerated increase in the ascending limb of the scotopic (rod-sensitive) b-wave as well as a delayed return to baseline. These results suggest that PKCα is an important modulator that affects bipolar cell signal transduction and termination. Confocal microscopy of retinal sections showed that PKCα co-localized with calbindin, which indicates a PKCα localization in close proximity to the horizontal cell terminals. In addition, the implicit time of the ERG c-wave originating from the retinal pigment epithelium (RPE) and the recovery of photoreceptors from bleaching conditions were substantially faster in the knockout mice than in the wild-type control animals. These results suggest that PKCα is a modulator of rod-bipolar cell function by accelerating glutamate-driven signal transduction and termination. This modulation is of importance in the switch between scotopic and photopic vision. Furthermore, PKCα seems to play a role in RPE function.


Tumor necrosis factor receptor I from patients with tumor necrosis factor receptor-associated periodic syndrome interacts with wild-type tumor necrosis factor receptor I and induces ligand-independent NF-kappaB activation.
To investigate the molecular consequences of expressing mutated forms of tumor necrosis factor receptor I (TNFRI) as found in patients with TNFR-associated periodic syndrome (TRAPS). We cloned and expressed full-length wild-type (WT) and T50K and P46L variants of TNFRI using a new tightly regulated doxycycline-dependent expression system. This system enabled the study of molecular interactions between these receptors at both physiologic and pathophysiologic levels of expression. We used chemical crosslinking on the cell surface to show that WT and mutant forms of TNFRI, derived from TRAPS patients, interact in the absence of TNF ligand. Doxycycline-controlled up-regulation of one TNFRI allele, either WT or mutant, caused down-regulation of the other allele, indicating dynamic control of cell surface assembly. We also demonstrated that increased expression of mutant TNFRI (T50K) was associated with a parallel increase in NF-kappaB p65 (RelA) subunit activation, which did not occur with increased expression of WT TNFRI. The T50K TRAPS-related variant is capable of sustaining inappropriate NF-kappaB activation, resulting in persistent auto-inflammation in target organs such as skin, synovial membrane, and the central nervous system. We conclude that some of the inflammatory processes seen in TRAPS do not involve direct interaction of TNF with its receptors, but that other proinflammatory mechanisms capable of up-regulating TNFRI expression may cause cellular activation through the NF-kappaB signaling pathway.


IFN-β Plays Both Pro- and Anti-inflammatory Roles in the Rat Cardiac Fibroblast Through Differential STAT Protein Activation.
Cardiac fibroblasts (CFs) contribute to theinflammatory response to tissue damage, secreting both pro- and anti-inflammatory cytokines and chemokines. Interferon beta (IFN-β) induces the phosphorylation of signal transducer and activator of transcription (STAT) proteins through the activation of its own receptor, modulating the secretion of cytokines and chemokines which regulate inflammation. However, the role of IFN-β and STAT proteins in modulating the inflammatory response of CF remains unknown. CF were isolated from adult male rats and subsequently stimulated with IFN-β to evaluate the participation of STAT proteins in secreting chemokines, cytokines, cell adhesion proteins expression and in their capacity to recruit neutrophils. In addition, in CF in which the TRL4 receptor was pre-activated, the effect of INF-β on the aforementioned responses was also evaluated. Cardiac fibroblasts stimulation with IFN-β showed an increase in STAT1, STAT2, and STAT3 phosphorylation. IFN-β stimulation through STAT1 activation increased proinflammatory chemokines MCP-1 and IP-10 secretion, whereas IFN-β induced activation of STAT3 increased cytokine secretion of anti-inflammatory IL-10. Moreover, in TLR4-activated CF, IFN-β through STAT2 and/or STAT3, produced an anti-inflammatory effect, reducing pro-IL-1β, TNF-α, IL-6, MCP-1, and IP-10 secretion; and decreasing neutrophil recruitment by decreasing ICAM-1 and VCAM-1 expression. Altogether, our results indicate that IFN-β exerts both pro-inflammatory and anti-inflammatory effects in non-stimulated CF, through differential activation of STAT proteins. When CF were previously treated with an inflammatory agent such as TLR-4 activation, IFN-β effects were predominantly anti-inflammatory.


Genetic differentiation and recombination among geographic populations of the fungal pathogen Colletotrichum truncatum from chili peppers in China.
Colletotrichum truncatum is an extremely important fungal pathogen. It can cause diseases both in humans and in over 460 plant species. However, little is known about its genetic diversity within and among populations. One of the major plant hosts of C. truncatum is pepper, and China is one of the main pepper-producing countries in the world. Here, we propose the hypotheses that geography has a major influence on the relationships among populations of C. truncatum in China and that infections in different populations need to be managed differently. To test these hypotheses, we obtained and analyzed 266 C. truncatum isolates from 13 regions representing the main pepper-growing areas throughout China. The analysis based on nine microsatellite markers identified high intrapopulation genetic diversity, evidence of sexual recombination, and geographic differentiation. The genetic differentiation was positively correlated with geographic distance, with the southern and northern China populations grouped in two distinct clusters. Interestingly, isolates collected from the pepper-breeding center harbored the most private alleles. The results suggest that the geographic populations of C. truncatum on peppers in China are genetically differentiated and should be managed accordingly. Our study also provides a solid foundation from which to further explore the global genetic epidemiology of C. truncatum in both plants and humans.


Morphology of cell injury: An approach to the EDIT programme by the use of tobacco pollen tubes. Evaluation-guided Development of New In Vitro Test Batteries.
Tobacco pollen tubes were used as a standard in vitro system to investigate cell growth aberrations caused by some of the Multicentre Evaluation of In Vitro Cytotoxicity (MEIC) programme chemicals and other toxic compounds. Changes in cytoskeletal pattern were observed in the tube cells by using tubulin immunofluorescence and rhodamin-phalloidin fluorescence for the localisation of microtubules and actin filaments, respectively. Four different types of cell malformation were found: screw-like growth, isodiametric tip swelling, hook formation, and pollen grain enlargement. We suggest that these malformations resulted from an interference by the chemicals with the cytosolic calcium gradient which controls tip growth and the orientation of the pollen tube. The results may contribute to a general understanding of toxicity-based cell malformations.


A trial to increase mammography utilization among Los Angeles Hispanic women.
The objective of this program was to increase mammography screening rates among Hispanic women through a series of targeted community-wide interventions. A diverse array of outreach efforts was offered by the program to increase awareness and use of screening mammography. Before the program, 12 percent of the Hispanic women surveyed in the intervention community had been screened, compared with 27 percent after the program. There was no change in screening among Hispanic women in the control community (23 percent before and 24 percent after the program). The program demonstrated that the awareness and behavior of "hard-to-reach" underscreened Hispanic women can be changed through intensive targeted outreach and that a church-based cancer control program can play an effective role in the process. This finding has national health policy implications.


Internet and cell phone usage patterns among young adults with intellectual disabilities.
The risks and opportunities associated with the use of technologies are of growing research interest. Patterns of technology usage illuminate these opportunities and risks. However, no studies have assessed the usage patterns (frequency, duration, and intensity) and related factors in young people with intellectual disabilities. Questionnaires on Internet and cell phone usage patterns, the Internet Over-Use Scale and the Cell-Phone Over-Use Scale, as well as the Beck Depression Inventory were filled out in one-on-one interviews of 216 youth with intellectual disabilities. Young people with disabilities make more social and recreational rather than educational use of these tools, and show higher rates of excessive use of both technologies than a comparison group of 410 young people without disabilities. Also, their overuse is associated with other unhealthy behaviors. The framework of support needs of people with disabilities should be considered to promote healthy Internet and cell phone use.


Twisting the arm: structural constraints in bicyclic expanded-ring N-heterocyclic carbenes.
A series of diaryl, mono-aryl/alkyl and dialkyl mono- and bicyclic expanded-ring N-heterocyclic carbenes (ER-NHCs) have been prepared and their complexation to Au(i) investigated through the structural analysis of fifteen Au(NHC)X and/or [Au(NHC)2]X complexes. The substituted diaryl 7-NHCs are the most sterically encumbered with large buried volume (%VB) values of 40-50% with the less flexible six-membered analogues having %VB values at least 5% smaller. Although the bicyclic systems containing fused 6- and 7-membered rings (6,7-NHCs) are constrained with relatively acute NCN bond angles, they have the largest %VB values of the dialkyl derivatives reported here, a feature related to the fixed conformation of the heterocyclic rings and the compressional effect of a pre-set methyl substituent.


In vivo noninvasive optical imaging of receptor-mediated RGD internalization using self-quenched Cy5-labeled RAFT-c(-RGDfK-)(4).
We reported that regioselectively addressable functionalized template (RAFT)-c(-RGDfK-)(4 )presenting four cyclic (Arg-Gly-Asp) (cRGD) peptides targets integrin alpha(V)beta(3) with an improved specificity compared with monomeric cRGD. In this study, we improved this vector by creating a "stealth" molecule in which a fluorescence quencher (Q) is linked to Cy5 via a disulfide bond (-SS-). RAFT-c(-RGDfK-)(4)-Cy5-SS-Q fluorescence is quenched unless activated by reduction during internalization. RAFT-c(-RGDfK-)(4)-Cy5-SS-Q fluorescence was negligible when compared with the control but totally recovered after cleavage of the disulfide bridge. Confocal microscopy showed that only the intracellular Cy5 signal could be detected using RAFT-c(-RGDfK-)(4)-Cy5-SS-Q, confirming that uncleaved extracellular molecules are not visible. Whole-body imaging of mice bearing subcutaneous tumors injected intravenously with RAFT-c(-RGDfK-)(4)-Cy5-SS-Q showed a very significant enhancement of the fluorescent contrast in tumors compared with the unquenched molecule. Histology of the tumor confirmed the intracellular accumulation of Cy5. These results demonstrate that the presence of a labile disulfide bridge between the targeting vector and a drug mimetic is an efficient way to deliver a dye, or a drug, intracellularly. In addition, this quenched RAFT-c(-RGDfK-)(4)-Cy5-SS-Q probe is a very powerful vector for imaging tumor masses and investigating in vivo RGD-mediated internalization.


Discriminative touch and emotional touch.
Somatic sensation comprises four main modalities, each relaying tactile, thermal, painful, or pruritic (itch) information to the central nervous system. These input channels can be further classified as subserving a sensory function of spatial and temporal localization, discrimination, and provision of essential information for controlling and guiding exploratory tactile behaviours, and an affective function that is widely recognized as providing the afferent neural input driving the subjective experience of pain, but not so widely recognized as also providing the subjective experience of affiliative or emotional somatic pleasure of touch. The discriminative properties of tactile sensation are mediated by a class of fast-conducting myelinated peripheral nerve fibres--A-beta fibres--whereas the rewarding, emotional properties of touch are hypothesized to be mediated by a class of unmyelinated peripheral nerve fibres--CT afferents (C tactile)--that have biophysical, electrophysiological, neurobiological, and anatomical properties that drive the temporally delayed emotional somatic system. CT afferents have not been found in the glabrous skin of the hand in spite of numerous electrophysiological explorations of this area. Hence, it seems reasonable to conclude that they are lacking in the glabrous skin. A full understanding of the behavioural and affective consequences of the differential innervation of CT afferents awaits a fuller understanding of their function.


Quantitation of flavor-related alkenylbenzenes in tobacco smoke particulate by selected ion monitoring gas chromatography-mass spectrometry.
Little is known about the possible health effects associated with inhaling alkenylbenzenes through cigarette smoking, even though these flavor-related compounds have known toxic effects in animals. We developed a rapid and sensitive solid-phase extraction (SPE) method to quantify seven alkenylbenzenes and piperonal in mainstream cigarette smoke particulate. The smoke particulate fraction of a single cigarette was collected on Cambridge filter pads, solvent extracted, concentrated, purified with SPE, and analyzed by selected ion monitoring gas chromatography-mass spectrometry. We positively identified and quantified five alkenylbenzenes compounds (eugenol, isoeugenol, methyleugenol myristicin, and elemicin) and piperonal in the smoke particulate from eight U.S. brands with mean levels (measured in triplicate) ranging from 6.6 to 4210 ng per cigarette. Additionally, complete blocking of nearly invisible ventilation holes in the cigarette filter increased 2- to 7-fold the percent transfer of alkenylbenzenes from tobacco to the particulate fraction of mainstream smoke.


Mechanisms of myocardial infarction without obstructive coronary artery disease.
Angiography in patients with myocardial infarction (MI) most commonly reveals one or more significantly narrowed coronary arteries, but a substantial minority of patients with spontaneous MI have no obstructive coronary artery disease (CAD) at angiography. This review summarizes evidence for the most commonly hypothesized mechanisms, including plaque disruption, plaque erosion, vasospasm, embolism, and spontaneous coronary dissection. In addition, tako-tsubo syndrome and myocarditis are discussed. The best treatment of MI without obstructive CAD is likely to differ substantially based on the underlying cause. Additional mechanistic research is needed to facilitate the design of research studies aimed at documenting the best treatments for these patients, numbering as many as 225,000 per year in the US.


Dry-Jet Wet Spinning of Thermally Stable Lignin-Textile Grade Polyacrylonitrile Fibers Regenerated from Chloride-Based Ionic Liquids Compounds.
In this paper, we report on the use of amorphous lignin, a waste by-product of the paper industry, for the production of high performance carbon fibers (CF) as precursor with improved thermal stability and thermo-mechanical properties. The precursor was prepared by blending of lignin with polyacrylonitrile (PAN), which was previously dissolved in an ionic liquid. The fibers thus produced offered very high thermal stability as compared with the fiber consisting of pure PAN. The molecular compatibility, miscibility, and thermal stability of the system were studied by means of shear rheological measurements. The achieved mechanical properties were found to be related to the temperature-dependent relaxation time (consistence parameter) of the spinning dope and the diffusion kinetics of the ionic liquids from the fibers into the coagulation bath. Furthermore, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and dynamic mechanical tests (DMA) were utilized to understand in-depth the thermal and the stabilization kinetics of the developed fibers and the impact of lignin on the stabilization process of the fibers. Low molecular weight lignin increased the thermally induced physical shrinkage, suggesting disturbing effects on the semi-crystalline domains of the PAN matrix, and suppressed the chemically induced shrinkage of the fibers. The knowledge gained throughout the present paper allows summarizing a novel avenue to develop lignin-based CF designed with adjusted thermal stability.


Ca2+ uptake by corpus-luteum plasma membranes. Evidence for the presence of both a Ca2+-pumping ATPase and a Ca2+-dependent nucleoside triphosphatase.
Plasma-membrane vesicles from rat corpus luteum showed an ATP-dependent uptake of Ca2+. Ca2+ was accumulated with a K1/2 (concn. giving half-maximal activity) of 0.2 microM and was released by the bivalent-cation ionophore A23187. A Ca2+-dependent phosphorylated intermediate (Mr 100,000) was detected which showed a low decomposition rate, consistent with it being the phosphorylated intermediate of the transport ATPase responsible for Ca2+ uptake. The Ca2+ uptake and the phosphorylated intermediate (E approximately P) displayed several properties that were different from those of the high-affinity Ca2+-ATPase previously observed in these membranes. Both Ca2+ uptake and E approximately P discriminated against ribonucleoside triphosphates other than ATP, whereas the ATPase split all the ribonucleoside triphosphates equally. Both Ca2+ uptake and E approximately P were sensitive to three different Hg-containing inhibitors, whereas the ATPase was inhibited much less. Ca2+ uptake required added Mg2+ (Km = 2.2 mM), whereas the ATPase required no added Mg2+. The maximum rate of Ca2+ uptake was about 400-fold less than that of ATP splitting; under different conditions, the decomposition rate of E approximately P was 1,000 times too slow to account for the ATPase activity observed. All of these features suggested that Ca2+ uptake was due to an enzyme of low activity, whose ATPase activity was not detected in the presence of the higher-specific-activity Ca2+-dependent ATPase.


Southeast asian ovalocytosis: the need for a carefull observation of red cell indices and blood smear.
Southeast asian ovalocytosis (SAO) is characterized by macro-ovalocytes and ovalo-stomatocytes on blood smear. SAO is common in Malaisia and Papua-New-Guinea where upwards to 40 per cent of the population is affected in some coastal region. Inherited in an autosomal dominant way, illness results from deletion of codons 400-408 in SLC4A1 gene which encodes for band 3 erythrocyte membrane protein. This deletion is responsible for an unusual erythrocyte stiffness and oval shape of the cells on blood smear. Heterozygous carriers are usually asymptomatic whereas homozygous are not viable without an intensive antenatal care. Here, we describe 4 patients diagnosed incidentally by cytogram appearance of the Advia® 2120i (Siemens) representing hemoglobin concentration according to red blood mean cellular volume (GR/VCH).


Effect of polyelectrolyte complex formation on the antibacterial activity of copolymer of alkylated 4-vinylpyridine.
Polymers bearing quaternized 4-vinylpyridine (QVP) groups are known for their antibacterial activities and these polymers can form polyelectrolyte complexes (PEC) with polyanions through electrostatic interactions. PEC formation can be used to adjust the antibacterial activity of polymers of QVP, deliver active molecules, or design antibacterial supramolecular structures. However, the antibacterial activity of PECs of QVP polymers has not been investigated. In this study, a copolymer of QVP was mixed with polyacrylic acid in various molar ratios of components to form PECs. Hydrodynamic diameters and zeta potentials of formed PECs were determined by dynamic and electrophoretic light scattering spectroscopy techniques. The zeta potentials of PECs changed between -24 and +16 mV with variation in the ratio of components. Antibacterial assays against E. coli revealed a relation of PEC formation with antibacterial activity since MIC values changed between 125-1000 μg/mL according to the ratio of components.


Cutting edge: Rapamycin augments pathogen-specific but not graft-reactive CD8+ T cell responses.
Recent evidence demonstrating that exposure to rapamycin during viral infection increased the quantity and quality of Ag-specific T cells poses an intriguing paradox, because rapamycin is used in transplantation to dampen, rather than enhance, donor-reactive T cell responses. In this report, we compared the effects of rapamycin on the Ag-specific T cell response to a bacterial infection versus a transplant. Using a transgenic system in which the Ag and the responding T cell population were identical in both cases, we observed that treatment with rapamycin augmented the Ag-specific T cell response to a pathogen, whereas it failed to do so when the Ag was presented in the context of a transplant. These results suggest that the environment in which an Ag is presented alters the influence of rapamycin on Ag-specific T cell expansion and highlights a fundamental difference between Ag presented by an infectious agent as compared with an allograft.


A longitudinal study of reproductive performance and management of 82 dairy herds in the Waikato region with differing policies on the routine use of induction of parturition.
To describe the reproductive performance and management practices over three lactations in dairy herds that had different policies on the routine use of induction of parturition; to describe any change in outcomes in herds that ceased inductions during the study period; and to investigate other herd-level factors associated with reproductive performance. From 2002 to 2004 inclusive, a longitudinal study was undertaken in 82 herds in the Waikato region of New Zealand. The main outcome variables were the percentage of cows pregnant after 8 weeks of mating (8-week in-calf rate) and the percentage of cows not pregnant at the end of mating (final empty rate). Data were obtained from pregnancy diagnosis records from a veterinary practice and from an electronic database, and a questionnaire. Herds were classified as either not using inductions in the three lactations (Nil; n=14), ceased inductions in the second or third lactation (Transitional; n=12), or continuing to induce (Continuing; n=56). Nil herds had a higher 8-week in-calf rate (p=0.01) than Transitional or Continuing herds (83%, 78% and 79%, respectively). The final empty rate in Nil and Transitional herds was similar (10.2% and 9.9%), tended to be greater in Nil than in Continuing herds (10.2% vs 9.0%; p=0.06) and was greater in Transitional than in Continuing herds (9.9% vs 9.0%; p=0.04), respectively. Multivariable modelling found that 8-week in-calf rates were additionally associated with year of lactation, interval from the planned start of calving (PSC) to the median calving date, duration of artificial insemination, and 21-day submission rate; and that final empty rates were associated with year of lactation, predominant breed, percentage of the herd calving within 40 days of the planned start of mating (PSM), the total duration of mating, and the 21-day submission rate, but not the herd induction policy. A change to ceasing routine use of induction tended to be associated (p=0.07) with a 2.5% increase in the final empty rate in the lactation of change. The reproductive performance of herds was associated with induction policy, in addition to other factors. Herds that ceased induction had a greater 8-week in-calf rate but a greater final empty rate than those that continued. The higher empty rate in Nil herds was partly due to their shorter duration of mating. Herdowners and their advisors planning to cease routine use of induction should plan for an increase in the final empty rate in both the lactation of change and in the longer term, and undertake measures to mitigate against this effect.


A Genome-Wide CRISPR/Cas9-Based Screen Identifies Heparan Sulfate Proteoglycans as Ligands of Killer-Cell Immunoglobulin-Like Receptors.
While human leukocyte antigen (HLA) and HLA-like proteins comprise an overwhelming majority of known ligands for NK-cell receptors, the interactions of NK-cell receptors with non-conventional ligands, particularly carbohydrate antigens, is less well described. We previously found through a bead-based HLA screen that KIR3DS1, a formerly orphan member of the killer-cell immunoglobulin-like receptor (KIR) family, binds to HLA-F. In this study, we assessed the ligand binding profile of KIR3DS1 to cell lines using Fc fusion constructs, and discovered that KIR3DS1-Fc exhibited binding to several human cell lines including ones devoid of HLA. To identify these non-HLA ligands, we developed a magnetic enrichment-based genome-wide CRISPR/Cas9 knock-out screen approach, and identified enzymes involved in the biosynthesis of heparan sulfate as crucial for the binding of KIR3DS1-Fc to K562 cells. This interaction between KIR3DS1 and heparan sulfate was confirmed via surface plasmon resonance, and removal of heparan sulfate proteoglycans from cell surfaces abolished KIR3DS1-Fc binding. Testing of additional KIR-Fc constructs demonstrated that KIR family members containing a D0 domain (KIR3DS1, KIR3DL1, KIR3DL2, KIR2DL4, and KIR2DL5) bound to heparan sulfate, while those without a D0 domain (KIR2DL1, KIR2DL2, KIR2DL3, and KIR2DS4) did not. Overall, this study demonstrates the use of a genome-wide CRISPR/Cas9 knock-out strategy to unbiasedly identify unconventional ligands of NK-cell receptors. Furthermore, we uncover a previously underrecognized binding of various activating and inhibitory KIRs to heparan sulfate proteoglycans that may play a role in NK-cell receptor signaling and target-cell recognition.


Recruiting mechanism of the AAA peroxins, Pex1p and Pex6p, to Pex26p on the peroxisomal membrane.
A peroxisomal C-tail-anchored type-II membrane protein, Pex26p, recruits AAA ATPase Pex1p-Pex6p complexes to peroxisomes. We herein attempted to gain mechanistic insight into Pex26p function. Pex26pΔ33-40 truncated in amino-acid residues at 33-40 abolishes the recruiting of Pex1p-Pex6p complex to peroxisomes and fails to complement the impaired phenotype of pex26 CHO cell mutant ZP167, thereby suggesting that peroxisomal localization of Pex1p and Pex6p is indispensable for the transport of matrix proteins. In in vitro transport assay using semipermeabilized CHO cells, Pex1p is targeted to peroxisomes in a manner dependent on ATP hydrolysis, while Pex6p targeting requires ATP but not its hydrolysis. This finding is confirmed by the assay using Walker-motif mutants. Transport of Pex1p and Pex6p is temperature-dependent. In vitro binding assays with glutathione-S-transferase-fused Pex26p, Pex1p and Pex6p bind to Pex26p in a manner dependent on ATP binding but not ATP hydrolysis. These results suggest that ATP hydrolysis is required for stable localization of Pex1p to peroxisomes, but not for binding to Pex26p. Moreover, Pex1p and Pex6p are altered to a more compact conformation upon binding to ATP, as verified by limited proteolysis. Taken together, Pex1p and Pex6p are most likely regulated in their peroxisomal localization onto Pex26p via conformational changes by the ATPase cycle.


Early surgery of multiple versus single aneurysms after subarachnoid hemorrhage: an increased risk for cerebral vasospasm?
As many as 33% of patients suffering from subarachnoid hemorrhage (SAH) present with multiple intracranial aneurysms (MIAs). It is believed that aneurysm surgery has the potential to increase the risk of cerebral vasospasm due to surgical manipulations of the parent vessels and brain tissue. Consequently, 1-stage surgery of MIAs, which usually takes longer and requires more manipulation, could even further increase the risk of vasospasm. The aim of this study is to define the correlation between vasospasm and the operative treatment of single intracranial aneurysms versus MIAs in a 1-stage operation. The authors analyzed a database including 1016 patients with SAH, identified retrospectively between 1989 and 1996 and prospectively collected between 1997 and 2004. Exclusion criteria were endovascular treatment, surgery after SAH Day 3, and, in patients with MIAs, undergoing more than 1 operation. Cerebral vasospasm was diagnosed by transcranial Doppler (TCD) ultrasonography and was defined as a maximum mean blood flow velocity > 120 cm/second. The diagnosis of symptomatic vasospasm was made if a new neurological deficit occurred that could not be explained by concomitant complications. A total of 643 patients who experienced 810 aneurysms were included. Four hundred twenty-four patients were female (65.9%) and 219 were male (34.1%) with an average age of 53.1 years. One hundred twenty-one patients (18.8%) were diagnosed with MIAs. Maximum mean flow velocities measured by TCD were 131 cm/second in patients with MIAs and 129.5 cm/second in patients with single intracranial aneurysms. The incidence of TCD vasospasm (p = 0.561) as well as of symptomatic vasospasm (p = 0.241) was not significantly different in the 2 groups. Clipping of more than 1 aneurysm in a 1-stage operation within 72 hours after SAH can be performed without increasing the risk of cerebral (TCD) vasospasm and symptomatic vasospasm.


Clinical and serological variation between patients infected with different Hepatitis B virus genotypes.
Hepatitis B virus (HBV) has eight genotypes which have distinct geographical distributions. Studies comparing differences in the clinical outcomes of infections caused by strains with genotype-related variations in the HBV genome have largely compared genotypes B and C and genotypes A and D but not all four genotypes. The present study included 196 HBV-infected patients attending an infectious diseases outpatient clinic in Sweden. The age and geographic origin, liver function, HBeAg and anti-HBe status, and the presence or absence of HBV DNA were analyzed for each patient. HBV DNA was detected in 144 patients, and the HBV genotype and the core promoter and precore sequences were determined for the isolates from 101 of these patients. Among the patients who might be considered most likely to be nonviremic, namely, anti-HBe-positive HBV carriers with normal alanine aminotransferase (ALT) levels, 65% had detectable HBV DNA and were thus viremic. Among the viremic patients, HBeAg-positive patients were more likely to have elevated ALT levels than anti-HBe-positive patients. HBV genotypes A to F were represented in the study, and their distributions coincided accurately with the origin of the patient. A significantly higher number of genotype D-infected patients were anti-HBe positive and had elevated ALT levels (42% of genotype D-infected patients but 0% of patients infected with genotypes B and C). Genotype D strains with mutations in the core promoter and precore regions were significantly correlated with elevated ALT levels in the patients. The differences were not age related. Therefore, in this large-scale cross-sectional study, genotype D appears to be associated with more active disease.


[Estimating prevalence of inadequate nutrient intake].
In assessing food intake patterns in groups of individuals, it is usually important to know how many of these individuals present nutrient intakes above or below a determined criterion. This information is relevant for planning healthcare actions, both in monitoring and in assessing such interventions, and also for the purposes of regulating commercial activities. This paper discusses a new methodology for estimating the prevalence of inadequate nutrient intake, with the Dietary Reference Intakes as the reference values. The utilization of diet surveys that allow the most accurate quantitative estimate possible is dealt with. These should use repeated measurements of food intake, on non-consecutive days. The food intake should be adjusted for between-person and within-person variance, so as to obtain a corrected distribution for the nutrient intake values. After constructing this distribution curve based on the corrected estimated parameters (average and standard deviation), the EAR method (Estimated Average Requirement) can be used as a cutoff point for verifying the proportion of individuals that has inadequate nutrient intake. This corresponds to the area of the curve below this reference value. It is concluded that this new approach minimizes the error in the calculation of the prevalence of inadequate nutrient intake, since it takes into consideration the random characteristics of the diet.


Salvage Endoscopic Argon Plasma Coagulation After Chemoradiotherapy for Inoperable Esophageal Cancer.
The purpose of this study is to evaluate the effectiveness of salvage endoscopic argon plasma coagulation (APC) therapy for local failure occurring after definitive chemoradiotherapy (CRT) in patients with inoperable esophageal cancer. A total of 47 patients with esophageal squamous cell carcinoma received CRT from January 2009 through November 2011 in our hospital. Thirty-five of these patients were excluded because of long-term disease-free status or because they received salvage surgery, systemic chemotherapy, or best supportive care. The other 12 patients had local relapse (n=8) or residual lesions (n=4) after definitive CRT and received salvage APC therapy. Local failures after definitive CRT were divided into 2 groups: the superficial growth group (n=6) and the submucosal growth group (n=6). The primary endpoints were relapse-free survival and overall survival after 60 months of follow-up. Six patients in the superficial growth group and 1 patient in submucosal growth group had a complete response after salvage APC. The rates of both relapse-free survival and overall survival were higher in the superficial growth group than in the submucosal growth group. No complications occurred after APC. Salvage APC is a potentially curative treatment for patients who have local failure associated with superficial tumors after CRT for esophageal squamous cell carcinoma.


Chromatic blur perception in the presence of luminance contrast.
Hel-Or showed that blurring the chromatic but not the luminance layer of an image of a natural scene failed to elicit any impression of blur. Subsequent studies have suggested that this effect is due either to chromatic blur being masked by spatially contiguous luminance edges in the scene (Journal of Vision 13 (2013) 14), or to a relatively compressed transducer function for chromatic blur (Journal of Vision 15 (2015) 6). To test between the two explanations we conducted experiments using as stimuli both images of natural scenes as well as simple edges. First, we found that in color-and-luminance images of natural scenes more chromatic blur was needed to perceptually match a given level of blur in an isoluminant, i.e. colour-only scene. However, when the luminance layer in the scene was rotated relative to the chromatic layer, thus removing the colour-luminance edge correlations, the matched blur levels were near equal. Both results are consistent with Sharman et al.'s explanation. Second, when observers matched the blurs of luminance-only with isoluminant scenes, the matched blurs were equal, against Kingdom et al.'s prediction. Third, we measured the perceived blur in a square-wave as a function of (i) contrast (ii) number of luminance edges and (iii) the relative spatial phase between the colour and luminance edges. We found that the perceived chromatic blur was dependent on both relative phase and the number of luminance edges, or dependent on the luminance contrast if only a single edge is present. We conclude that this Hel-Or effect is largely due to masking of chromatic blur by spatially contiguous luminance edges.