Reasons for Tooth Extractions and Related Risk Factors in Adult Patients: A Cohort Study.
Background: The aim of this study was to evaluate oral status, the reasons for tooth extractions and related risk factors in adult patients attending a hospital dental practice. Methods: 120 consecutive patients ranging from 23 to 91 years in age (mean age of 63.3 ± 15.8) having a total of 554 teeth extracted were included. Surveys about general health status were conducted and potential risk factors such as smoking, diabetes and age were investigated. Results: a total of 1795 teeth were missing after extraction procedures and the mean number of remaining teeth after the extraction process was 16.8 ± 9.1 per patient. Caries (52.2%) was the most common reason for extraction along with periodontal disease (35.7%). Males were more prone to extractions, with 394 of the teeth extracted out of the total of 554 (71.1%). Male sex (β = 2.89; 95% CI 1.26, 4.53; p = 0.001) and smoking habit (β = 2.95; 95% CI 1.12, 4.79; p = 0.002) were related to a higher number of teeth extracted. Age (β = -0.24; 95% CI -0.31, -0.16; p < 0.001) and diabetes (β = -4.47; 95% CI -7.61, -1.33; p = 0.006) were related to a higher number of missing teeth at evaluation time. Moreover, periodontal disease was more common as a reason of extraction among diabetic patients than among non-diabetic ones (p = 0.04). Conclusions: caries and periodontal disease were the most common causes of extraction in a relatively old study population: further screening strategies might be required for the early interception of caries and periodontal disease.


Dexmedetomidine versus propofol/midazolam for long-term sedation during mechanical ventilation.
To compare dexmedetomidine (DEX) with standard care (SC, either propofol or midazolam) for long-term sedation in terms of maintaining target sedation and length of intensive care unit (ICU) stay. A pilot, phase III, double-blind multicenter study in randomized medical and surgical patients (n = 85) within the first 72 h of ICU stay with an expected ICU stay of >or=48 h and sedation need for >or=24 h after randomization. Patients were assigned to either DEX (<or=1.4 microg kg(-1) h(-1); n = 41) or SC (n = 44), with daily sedation stops. Non-inferiority of DEX versus SC was not confirmed. Target Richmond agitation-sedation score (RASS) was reached a median of 64% (DEX) and 63% (SC) of the sedation time (ns). The length of ICU stay was similar in DEX and SC. Patients with RASS target 0-3 (DEX 78%, SC 80%) were at target sedation 74% (DEX) and 64% (SC) of the time (ns), whereas those with RASS target -4 or less reached the target 42% (DEX) and 62% (SC) of the time (P = .006). Post hoc analyses suggested shorter duration of mechanical ventilation for DEX (P = 0.025). This pilot study suggests that in long-term sedation, DEX is comparable to SC in maintaining sedation targets of RASS 0 to -3 but not suitable for deep sedation (RASS -4 or less). DEX had no effect on length of ICU stay. Its effects on other relevant clinical outcomes, such as duration of mechanical ventilation, should be tested further.


Characterization of human cellular immune responses to Mycobacterium tuberculosis proteins encoded by genes predicted in RD15 genomic region that is absent in Mycobacterium bovis BCG.
RD15 is a genomic region of difference (RD) present in Mycobacterium tuberculosis H37Rv but absent in all strains of Mycobacterium bovis BCG. RD15 contains genes encoding proteins of mammalian cell entry (Mce3A-F), important for the invasion and survival of M. tuberculosis in host cells. In this study, we have evaluated cellular immune responses to RD15 proteins using peripheral blood mononuclear cells (PBMC) from pulmonary tuberculosis patients and M. bovis BCG-vaccinated healthy subjects. PBMC were tested for T-helper (Th) type 1 [antigen-induced proliferation and interferon (IFN)-gamma secretion] and anti-inflammatory [interleukin (IL)-10 secretion] responses to complex mycobacterial antigens and peptides corresponding to proteins of RD1 and RD15. In Th1 assays, complex mycobacterial antigens induced strong responses in both donor groups, and RD1 induced strong responses in tuberculosis patients and moderate responses in healthy subjects, whereas RD15 induced weak responses in tuberculosis patients and strong to moderate responses in healthy subjects. IL-10 secretion in both donor groups was strong to moderate in response to complex mycobacterial antigens, but weak in response to RD1 and RD15. Analysis of IFN-gamma : IL-10 ratios showed strong Th1 biases to complex mycobacterial antigens and RD1 in both donor groups, and to RD15 and RD1504 (Mce3A) in healthy subjects only. These results suggest that RD1504 is the best Th1-stimulating antigen present in RD15, and therefore may be a potential vaccine candidate against TB.


Co-factors, Microbes, and Immunogenetics in Celiac Disease to Guide Novel Approaches for Diagnosis and Treatment.
Celiac disease (CeD) is a frequent immune-mediated disease that affects not only the small intestine but also many extraintestinal sites. The role of gluten proteins as dietary triggers, HLA-DQ2 or -DQ8 as major necessary genetic predisposition, and tissue transglutaminase (TG2) as mechanistically involved autoantigen, are unique features of CeD. Recent research implicates many cofactors working in synergism with these key triggers, including the intestinal microbiota and their metabolites, nongluten dietary triggers, intestinal barrier defects, novel immune cell phenotypes, and mediators and cytokines. In addition, apart from HLA-DQ2 and -DQ8, multiple and complex predisposing genetic factors and interactions have been defined, most of which overlap with predispositions in other, usually autoimmune, diseases that are linked to CeD. The resultant better understanding of CeD pathogenesis, and its manifold manifestations has already paved the way for novel therapeutic approaches beyond the lifelong strict gluten-free diet, which poses a burden to patients and often does not lead to complete mucosal healing. Thus, supported by improved mouse models for CeD and in vitro organoid cultures, several targeted therapies are in phase 2-3 clinical studies, such as highly effective gluten-degrading oral enzymes, inhibition of TG2, cytokine therapies, induction of tolerance to gluten ingestion, along with adjunctive and preventive approaches using beneficial probiotics and micronutrients. These developments are supported by novel noninvasive markers of CeD severity and activity that may be used as companion diagnostics, allow easy-to perform and reliable monitoring of patients, and finally support personalized therapy for CeD.


Intravenous versus oral dexamethasone premedication in preventing Paclitaxel infusion hypersensitivity reactions in gynecological malignancies.
Dexamethasone premedication is required with paclitaxel to prevent infusion-related hypersensitivity reactions (HSRs). Both oral dexamethasone (PO-D; 20 mg 12 and 6 hours before paclitaxel) and intravenous dexamethasone (IV-D; 20 mg 30 minutes before paclitaxel) regimens are used. The optimal premedication regimen and management of patients after HSR are unclear. Data on HSRs in women receiving paclitaxel, 175 mg/m², every 3 weeks at Imperial College Healthcare Trust from May 2011 to February 2012 were obtained from the pharmacy database. During this period, dexamethasone premedication for paclitaxel was administered orally (PO-D; 20 mg 12 and 6 hours before paclitaxel) from May to August 2011, then changed to intravenous dexamethasone (IV-D; 20 mg 30 minutes before paclitaxel) for 3 months, and then reverted to PO-D from November 2011. There were 93 and 55 patients who received PO-D and IV-D before paclitaxel, respectively. Hypersensitivity reaction rates were pooled with those from published studies for analysis. Gynecologic oncology centers in the UK and Canada were surveyed regarding premedication and post-HSR management. A Markov Monte-Carlo simulation model compared costs and benefits of different strategies. Hypersensitivity reaction rates with PO-D and IV-D were 5.4% (5/93) versus 14.5% (8/55) (P = 0.07) in Imperial College Healthcare Trust patients, and 6.8% (20/290) versus 14.1% (30/212) (P = 0.009) on pooled analysis with data from 2 additional studies (502 patients), respectively. However, IV-D is the most common premedication regimen used in the UK and Canada (48.5% and 34.2% of centers). Post-HSR paclitaxel on a desensitization protocol is a cost-effective alternative to discontinuing paclitaxel altogether. Oral dexamethasone seems to be superior to IV-D in preventing HSRs. Post-HSR patients should be considered for desensitization.


Effects of pirenzepine and AF-DX 116 on ganglionic transmission in the cardiac sympathetic nerves of the dog: interaction of M1 and M2 receptors with nicotinic receptors.
Effects of the selective M1 receptor antagonist pirenzepine and the selective M2 receptor antagonist AF-DX 116 on ganglionic transmission were examined in anesthetized dogs, in order to elucidate a functional role of M1 and M2 receptors. Preganglionic or postganglionic stimulation of the cardiac sympathetic nerves (SNS, 0.5-16 Hz) produced frequency-dependent increases in heart rate. Pirenzepine (3-100 microgram/kg) caused dose-dependent and significant inhibition of positive chronotropic response to preganglionic SNS but not to postganglionic SNS. AF-DX 116 (10-100 micrograms/kg) had no effect on the preganglionic SNS-induced tachycardia. The simultaneous administration of pirenzepine (30 micrograms/kg) and hexamethonium (C6, 1 mg/kg), and the subsequent administration of 10 mg/kg of C6, inhibited more potently the tachycardic responses to preganglionic SNS than each dose of C6 did by itself. The enhancement by pirenzepine of the C6-induced inhibition was evident at high frequencies (8 and 16 Hz) of SNS. In contrast, the blocking effect of C6 (1 and 10 mg/kg) on ganglionic transmission was significantly attenuated by AF-DX 116 (30 micrograms/kg). The attenuation by AF-DX 116 was observed at a wide range of stimulation frequency (0.5-8 Hz). These results suggest that M1 receptors play a facilitatory role in ganglionic transmission but M2 receptors do not contribute to the transmission when nicotinic pathway is intact. However, the activation of M2 receptors would further suppress ganglionic transmission when nicotinic transmission is inhibited. Under these conditions, activation of M1 receptors would mediate the transmission elicited by high frequency of stimulation.


Hypnosis in surgery--why and when.
Hypnosis offers an excellent alternative to conventional anesthesia in certain selected or complicated cases. Ten different patients were selected from a group of 38 and were studied thoroughly. Good surgery and a smooth recovery were possible through hypnosis, alone or with conventional anesthesia.


Adult social outcomes of extremely low birth weight survivors of childhood sexual abuse.
Preterm birth and exposure to childhood sexual abuse (CSA) are early physiological and psychological adversities that have been linked to reduced social functioning across the lifespan. However, the joint effects of being born preterm and being exposed to CSA on adult social outcomes remains unclear. We sought to determine the impact of exposure to both preterm birth and CSA on adult social functioning in a group of 179 extremely low birth weight (ELBW; 2500 g) participants in the fourth decade of life. Social outcome data from a prospective, longitudinal, population-based Canadian birth cohort initiated between the years of 1977 and 1982 were examined. At age 29-36 years, ELBW survivors who experienced CSA reported poorer relationships with their partner, worse family functioning, greater loneliness, lower self-esteem and had higher rates of avoidant personality problems than those who had not experienced CSA. Birth weight status was also found to moderate associations between CSA and self-esteem (P=0.032), loneliness (P=0.021) and family functioning (P=0.060), such that the adverse effects of CSA were amplified in ELBW survivors. Exposure to CSA appears to augment the adult social risks associated with perinatal adversity. Individuals born preterm and exposed to CSA appear to be a group at particularly high risk for adverse social outcomes in adulthood.


Acquisition of Beta-Lactamase by Neisseria meningitidis through Possible Horizontal Gene Transfer.
We report the detection in France of a beta-lactamase-producing invasive meningococcal isolate. Whole-genome sequencing of the isolate revealed a ROB-1-type beta-lactamase gene that is frequently encountered in Haemophilus influenzae, suggesting horizontal transfer between isolates of these bacterial species. Beta-lactamases are exceptional in meningococci, with no reports for more than 2 decades. This report is worrying, as the expansion of such isolates may jeopardize the effective treatment against invasive meningococcal disease.


Frequency of Electrolyte Derangement after Transurethral Resection of Prostate: Need for Postoperative Electrolyte Monitoring.
Objective. To determine the electrolyte derangement following transurethral resection of prostate (TURP). Methods. All patients undergoing TURP from June 2012 to April 2013 were included. Preoperative electrolytes were performed within a week of procedures. Monopolar TURP using 1.5% glycine was performed. Serum Na(+) and K(+) were assessed within 1 hour postoperatively and subsequently if clinically indicated. Results. The study included 280 patients. Sixty-six patients (23.6%) had electrolyte derangement after TURP. Patients with deranged electrolytes were older (mean age of 73.41 ± 4.08 yrs. versus 68.93 yrs. ± 10.34) and had a longer mean resection time (42.5 ± 20.04 min versus 28.34 ± 14.64 min). Mean weight of tissue resected (41.49 ± 34.46 g versus 15.33 ± 9.74 g) and volume of irrigant used (23.55 ± 15.20 L versus 12.81 ± 7.57 L) were also significantly higher in patients with deranged electrolytes (all p = 0.00). On multivariate logistic regression analysis preoperative sodium level was found to be a significant predictor of postoperative electrolyte derangement (odds ratio 0.267, S.E. = 0.376, and p value = 0.00). Conclusion. Electrolyte derangement occurs in older patients, with larger amount of tissue and longer time of resection and higher volume of irrigant, and in those with lower serum preoperative sodium levels.


Energy-transfer from ultra-small Au nanoclusters to Er³⁺ ions: a short-range mechanism.
Sub-nanometric Au nanoclusters are known to act as very efficient sensitizers for the luminescent emission of Er(3+) ions in silica through a non-resonant broad-band energy-transfer mechanism. In the present work the energy-transfer process is investigated in detail by room temperature photoluminescence characterization of Er and Au co-implanted silica systems in which a different degree of coupling between Er(3+) ions and Au nanoclusters is obtained. The results allow us to definitely demonstrate the short-range nature of the interaction in agreement with non-radiative energy-transfer mechanisms. Moreover, an upper limit to the interaction length is also set by the Au-Au intercluster semi-distance which is smaller than 2.4 nm in the present case.


Over-expression of the triploid white poplar PtDrl01 gene in tobacco enhances resistance to tobacco mosaic virus.
A full-length cDNA, designated as the Populus tomentosa disease resistance-like 01 (PtDrl01) gene, was isolated from triploid white poplar [(Populus tomentosa × P. bolleana) × P. tomentosa]. The protein thought to be produced by the PtDrl01 gene contains a nuclear localisation sequence (NLS), a toll/interleukin-1 receptor (TIR) homologue region, a nucleotide binding site (NBS) and a leucine-rich repeat (LRR) domain. The protein also exhibits a considerable degree of homology to N-like resistance proteins. Real-time quantitative RT-PCR analysis revealed that expression of the PtDrl01 gene in triploid white poplar leaves could be induced by two defence signalling molecules: methyl jasmonate (MeJA) and salicylic acid (SA). Over-expression of the PtDrl01 gene in transgenic tobacco induced enhanced resistance to tobacco mosaic virus (TMV). Long-term resistance from the PtDrl01 gene to TMV infection was also observed in transgenic tobacco plants. Additionally, over-expression of the PtDrl01 gene resulted in transcriptional changes in genes expressing pathogenesis-related proteins in transgenic tobacco under non-stress conditions. These data strongly suggest that the PtDrl01 gene is involved in plant defence responses to pathogen infection.


Mechanical Metamaterials on the Way from Laboratory Scale to Industrial Applications: Challenges for Characterization and Scalability.
Mechanical metamaterials promise a paradigm shift in materials design, as the classical processing-microstructure-property relationship is no longer exhaustively describing the material properties. The present review article provides an application-centered view on the research field and aims to highlight challenges and pitfalls for the introduction of mechanical metamaterials into technical applications. The main difference compared to classical materials is the addition of the mesoscopic scale into the materials design space. Geometrically designed unit cells, small enough that the metamaterial acts like a mechanical continuum, enabling the integration of a variety of properties and functionalities. This presents new challenges for the design of functional components, their manufacturing and characterization. This article provides an overview of the design space for metamaterials, with focus on critical factors for scaling of manufacturing in order to fulfill industrial standards. The role of experimental and simulation tools for characterization and scaling of metamaterial concepts are summarized and herewith limitations highlighted. Finally, the authors discuss key aspects in order to enable metamaterials for industrial applications and how the design approach has to change to include reliability and resilience.


Pediatric Injuries at an Annual Motocross Competition: Rates and Severity.
Pediatric motocross injuries occur frequently and with significant associated morbidity; however, data are limited regarding injury occurring during competition. North Central Florida hosts an annual motocross competition with numerous pediatric competitors. We sought to evaluate the rate and severity of injuries sustained at this event over a multiyear period. Our primary hypothesis was that motocross injury rates and severity in competition exceed that of other competitive sports in this age group. A retrospective review was performed to identify pediatric patients (17 and younger) presenting to a single Level 1 Trauma Center as a result of injury sustained at an annual motocross competition from 2009 to 2012. Demographic, diagnostic, and treatment data were collected including injuries, procedural interventions, and length of hospital stay. Rate, type, and severity of injury were evaluated and compared with established norms of other competitive sports in a similar age group. Over a 4-year period, 51 pediatric motocross competitors sustained 75 injuries. Injured riders were 14.2±2.51 years (range, 8 to 17 y) old and 92% (47/51) male. Forty (78%) patients sustained at least 1 orthopaedic injury and 17 (33%) patients suffered polytrauma. The majority of injuries occurred in the upper extremity (36%). Twenty-four (47%) patients required procedural intervention, 15 of which were operative. Thirty-nine (76%) patients necessitated surgical specialty care. Average injury severity score was 6.5 (1 to 75) and 1 death occurred. The injury rate for pediatric motocross riders in this study was 19.9 injuries per 1000 competitors. High rates of injury occur among pediatric patients in competitive motocross, exceeding those seen in other competitive sports in a comparable age group. Significant morbidity and at least 1 fatality were demonstrated in the group studied. The majority of patients suffer at least 1 orthopaedic injury and nearly all patients require pediatric surgical specialist care. Polytrauma is common. Motocross participants and parents should be aware of these risks of competition and event organizers should arrange events near facilities with appropriate pediatric surgical specialty services. Level IV-case series.


Inhibition of local nitric oxide synthase increases homeostatic efficiency of tubuloglomerular feedback.
Nephron filtration rate (SNGFR) and proximal tubular reabsorption are coordinated by the processes of tubuloglomerular feedback (TGF) and glomerulotubular balance (GTB). We examined the role of nitric oxide (NO) in TGF and GTB, by delivering the NO synthase inhibitor, NG-monomethyl-L-arginine (L-NMMA) into late proximal nephrons by micropuncture in Inactin-anesthetized euvolemic rats. First, we determined the dependence of SNGFR on late proximal tubular flow rate (VLP) during orthograde microperfusion of Henle's loop downstream from an obstructing wax block (open-loop microperfusion). Second, we examined the homeostatic efficiency of the TGF-GTB system, (i.e., its ability to stabilize VLP) by perturbing VLP in free-flowing nephrons, while using a noninvasive optical technique to measure flow immediately upstream from the perturbation (closed-loop perturbation). Third, we tested whether L-NMMA could alter the afferent signal to TGF by changing the relationship between VLP and the flow rate (VED) or ionic content (CED) of early distal tubular fluid during open-loop microperfusion. During open-loop microperfusion, L-NMMA decreased SNGFR at VLP between 10 and 40 nl/min but did not alter the relationship between VLP and VED or CED. During closed-loop perturbation, L-NMMA did not affect ambient VLP but increased the homeostatic efficiency of the TGF-GTB system. The combined effects on SNGFR and ambient VLP suggest that L-NMMA reduces proximal reabsorption. However, this could account for only a small fraction of the increase in homeostatic efficiency, the greater share of which must be mediated within the juxtaglomerular apparatus. It appears that juxtaglomerular NO exerts an upward pressure on SNGFR and reduces the efficiency of the TGF-GTB system in stabilizing VLP at ambient flow rates under euvolemic conditions.


Effects of Constraint-Induced Movement Therapy and Application Time and Duration of Intervention for Lower Extremity in Stroke: A Systematic Review.
The primary aim of this review was to determine the effects of CIMT (constraint-induced movement therapy) on gait, balance, and motor functions of the lower extremity in stroke. The secondary aim was to determine the optimal dosage, application time, and duration of CIMT in the lower extremity in stroke. PubMed (1999-July 2021), Pedro (2000-December 2020), Google Scholar (1999-Febraury 2022), and Cochrane Library (2000-Febraury 2022) were searched in February 2022. The risk of bias was calculated through the criteria outlined in the (Cochrane-Handbook for Systematic-Reviews of Interventions). Eight RCTs were included in this review. CIMT was found to be effective in improving balance, gait, and motor functions of lower limbs; however, its superiority in comparison to the control group was not significant, no specific dosage was mentioned for lower limb CIMT as different studies used different durations and intensities of CIMT. Key Words: Cerebrovascular accident (CVA), Balance, Lower-extremity constraint-induced movement therapy (CIMT), Motor functions.


Cannabidiol modulates expression of type I IFN response genes and HIV infection in macrophages.
Cannabis (Cannabis sativa) is a widely used drug in the United States and the frequency of cannabis use is particularly high among people living with HIV (PLWH). One key component of cannabis, the non-psychotropic (-)-cannabidiol (CBD) exerts a wide variety of biological actions, including anticonvulsive, analgesic, and anti-inflammatory effects. However, the exact mechanism of action through which CBD affects the immune cell signaling remains poorly understood. Here we report that CBD modulates type I interferon responses in human macrophages. Transcriptomics analysis shows that CBD treatment significantly attenuates cGAS-STING-mediated activation of type I Interferon response genes (ISGs) in monocytic THP-1 cells. We further showed that CBD treatment effectively attenuates 2'3-cGAMP stimulation of ISGs in both THP-1 cells and primary human macrophages. Interestingly, CBD significantly upregulates expression of autophagy receptor p62/SQSTM1. p62 is critical for autophagy-mediated degradation of stimulated STING. We observed that CBD treated THP-1 cells have elevated autophagy activity. Upon 2'3'-cGAMP stimulation, CBD treated cells have rapid downregulation of phosphorylated-STING, leading to attenuated expression of ISGs. The CBD attenuation of ISGs is reduced in autophagy deficient THP-1 cells, suggesting that the effects of CBD on ISGs is partially mediated by autophagy induction. Lastly, CBD decreases ISGs expression upon HIV infection in THP-1 cells and human primary macrophages, leading to increased HIV RNA expression 24 hours after infection. However, long term culture with CBD in infected primary macrophages reduced HIV viral spread, suggesting potential dichotomous roles of CBD in HIV replication. Our study highlights the immune modulatory effects of CBD and the needs for additional studies on its effect on viral infection and inflammation.


Astrophotonics: a new era for astronomical instruments.
Astrophotonics lies at the interface of astronomy and photonics. This burgeoning field has emerged over the past decade in response to the increasing demands of astronomical instrumentation. Early successes include: (i) planar waveguides to combine signals from widely spaced telescopes in stellar interferometry; (ii) frequency combs for ultra-high precision spectroscopy to detect planets around nearby stars; (iii) ultra-broadband fibre Bragg gratings to suppress unwanted background; (iv) photonic lanterns that allow single-mode behaviour within a multimode fibre; (v) planar waveguides to miniaturize astronomical spectrographs; (vi) large mode area fibres to generate artificial stars in the upper atmosphere for adaptive optics correction; (vii) liquid crystal polymers in optical vortex coronographs and adaptive optics systems. Astrophotonics, a field that has already created new photonic capabilities, is now extending its reach down to the Rayleigh scattering limit at ultraviolet wavelengths, and out to mid infrared wavelengths beyond 2500 nm.


Production and characterization of a new biodegradable fenugreek seed gum based active nanocomposite film reinforced with nanoclays.
In the present work, fenugreek seed gum (FSG)/clay nanocomposite films were prepared with nanoclays (Na+ montmorillonite [MMT], halloysite [HNT] and Nanomer® I.44 P [NM]) at different amounts (0, 2.5, 5.0 and 7.5g clay/100g FSG) by solution casting method and characterized. Increasing amount of nanoclay significantly (P<0.05) improved oxygen barrier and thermal properties of the biodegradable films. Agar diffusion tests revealed that FSG based nanocomposite films exhibited strong antimicrobial properties against foodborne pathogens namely Listeria monocytogenes, Escherichia coli O157:H7, Staphylococcus aureus and Bacillus cereus independently of clay type and concentration. In the case of mechanical properties, nanoclay incorporation up to 5% provided higher (P<0.05) tensile strength (TS) properties while elongation at break (EB) values of the films significantly (P<0.05) decreased in the presence of clay in the film matrix. SEM micrographs showed that especially lower levels (up to 5%) of nanoclay reinforcements provided a homogeneous and smooth film structure. In conclusion, FSG based nanocomposite films reinforced with nanoclays up to 5% showed a precious potential to be used in antimicrobial food packaging applications.


Agonist-induced desensitization/resensitization of human G protein-coupled receptor 17: a functional cross-talk between purinergic and cysteinyl-leukotriene ligands.
G protein-coupled receptor (GPR) 17 is a P2Y-like receptor that responds to both uracil nucleotides (as UDP-glucose) and cysteinyl-leukotrienes (cysLTs, as LTD(4)). By bioinformatic analysis, two distinct binding sites have been hypothesized to be present on GPR17, but little is known on their putative cross-regulation and on GPR17 desensitization/resensitization upon agonist exposure. In this study, we investigated in GPR17-expressing 1321N1 cells the cross-regulation between purinergic- and cysLT-mediated responses and analyzed GPR17 regulation after prolonged agonist exposure. Because GPR17 receptors couple to G(i) proteins and adenylyl cyclase inhibition, both guanosine 5'-O-(3-[(35)S]thio)triphosphate ([(35)S]GTPγS) binding and the cAMP assay have been used to investigate receptor functional activity. UDP-glucose was found to enhance LTD(4) potency in mediating activation of G proteins and vice versa, possibly through an allosteric mechanism. Both UDP-glucose and LTD(4) induced a time- and concentration-dependent GPR17 loss of response (homologous desensitization) with similar kinetics. GPR17 homologous desensitization was accompanied by internalization of receptors inside cells, which occurred in a time-dependent manner with similar kinetics for both agonists. Upon agonist removal, receptor resensitization occurred with the typical kinetics of G protein-coupled receptors. Finally, activation of GPR17 by UDP-glucose (but not vice versa) induced a partial heterologous desensitization of LTD(4)-mediated responses, suggesting that nucleotides have a hierarchy in producing desensitizing signals. These findings suggest a functional cross-talk between purinergic and cysLT ligands at GPR17. Because of the recently suggested key role of GPR17 in brain oligodendrogliogenesis and myelination, this cross-talk may have profound implications in fine-tuning cell responses to demyelinating and inflammatory conditions when these ligands accumulate at lesion sites.


Sumatriptan plus naproxen for the treatment of acute migraine attacks in adults.
This is an updated version of the original Cochrane review published in October 2013 on 'Sumatriptan plus naproxen for acute migraine attacks in adults'.Migraine is a common disabling condition and a burden for the individual, health services, and society. It affects two to three times more women than men, and is most common in the age range 30 to 50 years. Effective abortive treatments include the triptan and non-steroidal anti-inflammatory classes of drugs. These drugs have different mechanisms of action and combining them may provide better relief. Sumatriptan plus naproxen is now available in combination form for the acute treatment of migraine. To determine the efficacy and tolerability of sumatriptan plus naproxen, administered together as separate tablets or taken as a fixed-dose combination tablet, compared with placebo and other active interventions in the treatment of acute migraine attacks in adults. For this update we searched the Cochrane Central Register of Controlled Trials (CENTRAL) via The Cochrane Register of Studies Online (CRSO) to 28 October 2015, MEDLINE (via Ovid) from 1946 to 28 October 2015, and EMBASE (via Ovid) from 1974 to 28 October 2015, and two online databases (www.gsk-clinicalstudyregister.com and www.clinicaltrials.gov). We also searched the reference lists of included studies and relevant reviews. We included randomised, double-blind, placebo- or active-controlled studies, with at least 10 participants per treatment arm, using sumatriptan plus naproxen to treat a migraine headache episode. Two review authors independently assessed trial quality and extracted data. We used numbers of participants achieving each outcome to calculate risk ratio and numbers needed to treat for an additional beneficial outcome (NNT) or for an additional harmful outcome (NNH) compared with placebo or a different active treatment. For this update we identified one new study (43 participants), but it did not contribute any data for analysis. The review included 13 studies using sumatriptan 85 mg or 50 mg plus naproxen 500 mg to treat attacks of mild, moderate, or severe pain intensity. Twelve studies contributed data for analyses: 3663 participants received combination treatment, 3682 placebo, 964 sumatriptan, and 982 naproxen. We judged only one small study to be at high risk of bias for any of the criteria evaluated; it did not contribute to any analyses.Overall, the combination was better than placebo for the primary outcomes of pain-free and headache relief at two hours. The NNT for pain-free at two hours was 3.1 (95% confidence interval 2.9 to 3.5) when the baseline pain was mild (50% response with sumatriptan plus naproxen compared with 18% with placebo), and 4.9 (4.3 to 5.7) when baseline pain was moderate or severe (28% with sumatriptan plus naproxen compared with 8% with placebo) (high quality evidence). Using 50 mg of sumatriptan, rather than 85 mg, in the combination did not significantly change the result. Treating early, when pain was still mild, was significantly better than treating once pain was moderate or severe for pain-free responses at two hours and during the 24 hours post dose. Adverse events were mostly mild or moderate in severity and rarely led to withdrawal; they were more common with the combination than with placebo (moderate quality evidence).Where the data allowed direct comparison, combination treatment was superior to either monotherapy, but adverse events were less frequent with naproxen than with sumatriptan (moderate quality evidence). The conclusions of this review were not changed. Combination treatment was effective in the acute treatment of migraine headaches. The effect was greater than for the same dose of either sumatriptan or naproxen alone, but additional benefits over sumatriptan alone were not large. More participants achieved good relief when medication was taken early in the attack, when pain was still mild. Adverse events were more common with the combination and sumatriptan alone than with placebo or naproxen alone.


Characterization of two steroid hydroxylases from different Streptomyces spp. and their ligand-bound and -unbound crystal structures.
Bacterial cytochrome P450 (CYP) enzymes are involved in the hydroxylation of various endogenous substrates while using a heme molecule as a cofactor. CYPs have gained biotechnological interest as useful biocatalysts capable of altering chemical structures by adding a hydroxyl group in a regiospecific manner. Here, we identified, purified, and characterized two CYP154C4 proteins from Streptomyces sp. W2061 (StCYP154C4-1) and Streptomyces sp. ATCC 11861 (StCYP154C4-2). Activity assays showed that both StCYP154C4-1 and StCYP154C4-2 can produce 2'-hydroxylated testosterone, which differs from the activity of a previously described NfCYP154C5 from Nocardia farcinica in terms of its 16α-hydroxylation of testosterone. To better understand the molecular basis of the regioselectivity of these two CYP154C4 proteins, crystal structures of the ligand-unbound form of StCYP154C4-1 and the testosterone-bound form of StCYP154C4-2 were determined. Comparison with the previously determined NfCYP154C5 structure revealed differences in the substrate-binding residues, suggesting a likely explanation for the different patterns of testosterone hydroxylation, despite the high sequence similarities between the enzymes (54% identity). These findings provide valuable insights that will enable protein engineering for the development of artificial steroid-related CYPs exhibiting different regiospecificity.


Development of Kinase Inactive PD173955 Analogues for Reducing Production of Aβ Peptides.
Compound 3a, DV2-103, is a kinase inactive analogue of a potent Abl1/Src kinase inhibitor, PD173955, 2. Both compounds, 2 and 3a, are known to reduce production of beta amyloid (Aβ) peptide in cells and animal models. We have now prepared and evaluated a series of PD-173955 analogues, several of which reduced Aβ production potently. This occurs in cells expressing human full-length amyloid precursor protein (APP) and not in cells expressing APP β-C terminal fragment (APP-C99), suggesting that the kinase inactive analogues strongly affect β-secretase (BACE1) cleavage of APP, similarly to Gleevec. A combination of the kinase inactive analogues of PD173955 with a BACE1 inhibitor (BACEi), namely, BACE IV, strongly reduced Aβ levels in cells, as noted previously with Gleevec and analogues. Several potent compounds also penetrated and accumulated in mouse brain in high nanomolar to low micromolar concentration.


Clinical genetic testing of periodic fever syndromes.
Periodic fever syndromes (PFSs) are a wide group of autoinflammatory diseases. Due to some clinical overlap between different PFSs, differential diagnosis can be a difficult challenge. Nowadays, there are no universally agreed recommendations for most PFSs, and near half of patients may remain without a genetic diagnosis even after performing multiple-gene analyses. Molecular analysis of periodic fevers' causative genes can improve patient quality of life by providing early and accurate diagnosis and allowing the administration of appropriate treatment. In this paper we focus our discussion on effective usefulness of genetic diagnosis of PFSs. The aim of this paper is to establish how much can the diagnostic system improve, in order to increase the success of PFS diagnosis. The mayor expectation in the near future will be addressed to the so-called next generation sequencing approach. Although the application of bioinformatics to high-throughput genetic analysis could allow the identification of complex genotypes, the complexity of this definition will hardly result in a clear contribution for the physician. In our opinion, however, to obtain the best from this new development a rule should always be kept well in mind: use genetics only to answer specific clinical questions.


Factors affecting recall of different types of personal genetic information about Alzheimer's disease risk: the REVEAL study.
Data were obtained through a multisite clinical trial in which different types of genetic risk-related information were disclosed to individuals (n = 246) seeking a risk assessment for Alzheimer's disease. Six weeks after disclosure, 83% of participants correctly recalled the number of risk-increasing APOE alleles they possessed, and 74% correctly recalled their APOE genotype. While 84% of participants recalled their lifetime risk estimate to within 5 percentage points, only 51% correctly recalled their lifetime risk estimate exactly. Correct recall of the number of APOE risk-increasing alleles was independently associated with higher education (p < 0.001), greater numeracy (p < 0.05) and stronger family history of Alzheimer's disease (p < 0.05). Before adjustments for confounding, correct recall of APOE genotype was also associated with higher education, greater numeracy and stronger family history of Alzheimer's disease, as well as with higher comfort with numbers and European American ethnicity (all p < 0.05). Correct recall of the lifetime risk estimate was independently associated only with younger age (p < 0.05). Recall of genotype-specific information is high, but recall of exact risk estimates is lower. Incorrect recall of numeric risk may lead to distortions in understanding risk. Further research is needed to determine how best to communicate different types of genetic risk information to patients, particularly to those with lower educational levels and lower numeracy. Health-care professionals should be aware that each type of genetic risk information may be differentially interpreted and retained by patients and that some patient subgroups may have more problems with recall than others.


Cirrhosis mortality and per capita consumption of distilled spirits, United States, 1949-1994: trend analysis.
To describe, evaluate, and suggest interpretations for an observed aggregate-level relation between trends in mortality from cirrhosis and per capita consumption of distilled spirits in the United States. Trend analysis using data on US cirrhosis mortality and per capita alcohol consumption. There is a consistent long-term trend relation between mortality from cirrhosis and per capita consumption of distilled spirits in the United States from 1949 to 1994. Two instances of comparatively sharp drops in the consumption of spirits in the 1940s generated mixed results in predicting changes in cirrhosis mortality. An aggregate-level relation between trends in long-term cirrhosis mortality and the consumption of spirits falls considerably short of establishing a direct causal link between the two for individuals. Moreover, two sharp drops in the consumption of spirits generated only mixed results with respect to the short-term trend in cirrhosis. Nevertheless, the observed relation between the consumption of spirits and cirrhosis mortality merits further investigation.


Use of antibiotic and analgesic drugs during lactation.
During lactation, multiple situations can arise that require maternal pharmacological treatment. Because of the many health advantages of human milk to infants, breast feeding should be interrupted only when the needed drug might be harmful to the nursing child and exposure via the breast milk will be sufficient to pose a risk. Since the majority of drugs have not been shown to cause adverse effects when used during lactation, and even temporary interruption of breast feeding can be difficult for the nursing dyad, decisions regarding maternal medication use during breast feeding should be based on accurate and up-to-date information. This article reviews available data on the most commonly used antibiotics and analgesics. The use of most antibiotics is considered compatible with breast feeding. Penicillins, aminopenicillins, clavulanic acid, cephalosporins, macrolides and metronidazole at dosages at the low end of the recommended dosage range are considered appropriate for use for lactating women. Fluoroquinolones should not be administered as first-line treatment, but if they are indicated, breast feeding should not be interrupted because the risk of adverse effects is low and the risks are justified. Paracetamol (acetaminophen), low-dose aspirin (acetylsalicylic acid) [up to 100 mg/day] and short-term treatment with NSAIDs, codeine, morphine and propoxyphene are considered compatible with breast feeding. Safer alternatives should be considered instead of dipyrone, aspirin at a dosage >100 mg/day and pethidine (meperidine). In the light of the many safe alternatives for pain control, breast-feeding mothers should not be allowed to experience pain or be made to feel that they must choose between analgesia and breast feeding.


Two-step immunological approaches for imaging and therapy.
Radiolabeled monoclonal antibodies and their F(ab')2 and Fab fragments have been successful for imaging and therapy. However, their prolonged circulation and nonspecific accumulation in metabolic organs have resulted in high background radioactive levels and delayed imaging times. Current approaches are two- and three-step procedures consisting of first, the injection of a targeting moiety, which has specific binding affinity for both a lesion and a small molecular weight radiolabeled agent, and, second, a subsequently injected radioactive diagnostic or therapeutic agent. The rapid blood clearance of the free radiolabeled agent greatly reduces background levels and radiation dose and increases target-to-background ratios. This review will focus on two different approaches: 1) bispecific monoclonal antibodies and 2) monoclonal antibodies in conjunction with the streptavidin (avidin)/biotin system.


[Anterior recurrent luxation of the shoulder. Postoperative recurrences].
Seventy-nine recurrences have been seen after surgical procedures for recurrent dislocation of the shoulder. There were 58 "true" recurrences, 17 shoulders with anterior instability and 4 with instability in all directions. The "true" recurrences were more frequent in young people with a mean age of 20 years at the time of the first surgical procedure. Two-thirds of them occurred on the dominant side. Most of the cases had been treated by distal displacement or lengthening of the coracoid process (38 Trillat procedures, 8 Latarjet procedures and 8 Oudart procedures). Only two had been treated by a Bankart procedure. The main cause of recurrence was a failure of repair of the antero-inferior part of the joint. Twenty three cases were operated on again, the most usual procedures being the Trillat procedure and rarely the Latarjet or Eden Hybinette procedures. Six cases recurred once again. Cases with anterior instability were observed after the Trillat procedure. They were due to capsular laxity and did not have to be operated on again. Instability in multiple directions was observed in young women who continued to dislocate their shoulder inferiorly or posteriorly, despite two or three surgical procedures.


A machine learning method for fast and accurate characterization of depth-of-interaction gamma cameras.
Measuring the depth-of-interaction (DOI) of gamma photons enables increasing the resolution of emission imaging systems. Several design variants of DOI-sensitive detectors have been recently introduced to improve the performance of scanners for positron emission tomography (PET). However, the accurate characterization of the response of DOI detectors, necessary to accurately measure the DOI, remains an unsolved problem. Numerical simulations are, at the state of the art, imprecise, while measuring directly the characteristics of DOI detectors experimentally is hindered by the impossibility to impose the depth-of-interaction in an experimental set-up. In this article we introduce a machine learning approach for extracting accurate forward models of gamma imaging devices from simple pencil-beam measurements, using a nonlinear dimensionality reduction technique in combination with a finite mixture model. The method is purely data-driven, not requiring simulations, and is applicable to a wide range of detector types. The proposed method was evaluated both in a simulation study and with data acquired using a monolithic gamma camera designed for PET (the cMiCE detector), demonstrating the accurate recovery of the DOI characteristics. The combination of the proposed calibration technique with maximum- a posteriori estimation of the coordinates of interaction provided a depth resolution of ≈1.14 mm for the simulated PET detector and ≈1.74 mm for the cMiCE detector. The software and experimental data are made available at http://occiput.mgh.harvard.edu/depthembedding/.


Characterization of Sensory Differences in Mixing and Premium Rums Through the Use of Descriptive Sensory Analysis.
This study identified and quantitated perceived sensory differences between 7 premium rums and 2 mixing rums using a hybrid of the Quantitative Descriptive Analysis and Spectrum methods. In addition, the results of this study validated the previously developed rum flavor wheel created from web-based materials. Results showed that the use of the rum flavor wheel aided in sensory term generation, as 17 additional terms were generated after the wheel was provided to panelists. Thirty-eight sensory terms encompassing aroma, aroma-by-mouth, mouthfeel, taste and aftertaste modalities, were generated and evaluated by the panel. Of the finalized terms, only 5 did not exist previously on the rum flavor wheel. Twenty attributes were found to be significantly different among rums. The majority of rums showed similar aroma profiles with the exception of 2 rums, which were characterized by higher perceived intensities of brown sugar, caramel, vanilla, and chocolate aroma, caramel, maple, and vanilla aroma-by-mouth and caramel aftertaste. These results demonstrate the previously developed rum flavor wheel can be used to adequately describe the flavor profile of rum. Additionally, results of this study document the sensory differences among premium rums and may be used to correlate with analytical data to better understand how changes in chemical composition of the product affect sensory perception.


Food & meal decision making in lockdown: How and who has Covid-19 affected?
The Covid-19 lockdown resulted in all but essential shops closing in many countries, with inevitable and immediate impact on food availability and choice. Reasons for specific food choices influence diet and mealtimes and can affect psychological and physical well-being. The current study aimed first to investigate whether individuals and families in the UK have changed their food choice motivations over lockdown and second to identify sub-groups in particular need of support in the event of future lockdowns. Two hundred and forty adult participants from the UK completed an online survey, consisting of a series of demographic questions, the Food Choice Questionnaire, the Family Mealtime Goals Questionnaire and some short open-ended questions. They were asked to consider their goals and motivations around food and mealtimes at two timepoints: before lockdown and Summer 2020. Results indicated that the sample placed more importance on health, weight control and mood when choosing their food after lockdown than they had before, and less importance on familiarity. A number of sub-groups were identified who may be particularly vulnerable to food-related challenges in future lockdowns including younger adults, parents and carers of children, those self-isolating and individuals who do not live within close proximity to food shops. These results are preliminary and larger sub-group sample sizes will be necessary to draw firm conclusions. Future research should consider the nature and impact of these challenges in more detail across a more varied population.


Bone and joint diseases around the world. Sweden: a brief update on burden and priority.
Musculoskeletal conditions are increasingly common with advancing age. The life expectancy increased early and rapidly in Sweden, and today 17% of the population is over age 65. Musculoskeletal problems are therefore common but in general are receiving low attention, despite enormous costs for society. The indirect costs for musculoskeletal conditions vastly exceed direct costs (80% vs 20%). This is obviated by the fact that in 2001, of all persons receiving disability pension or taking longterm sick leave, 60% had a diagnosis related to the musculoskeletal system. Further, in a population of 9 million, 70,000 fragility fractures occur each year, 18,000 of the hip and 25,000 of the forearm, corresponding to a hip fracture incidence of 20.14/10,000, among the highest in the world. Government policy is implemented through several agencies, national and local. State of the art reports are currently available in 12 areas of musculoskeletal condition, including osteoarthritis, rheumatoid arthritis, osteoporosis, and hip fracture, while no national evidence-based guidelines have been developed for these conditions. The Swedish Council on Technology Assessment in Health Care provides the scientific foundation of present methods for treatment through evidence-based evaluations. National registers are continuously evaluating orthopedic implant procedures, of which the Swedish Hip Register has provided valuable information on total hip replacements since 1979. For the future, there is a need for setting of priorities with regard to musculoskeletal conditions, including development of guidelines of mechanism of implementation.


Earning stripes: myosin binding protein-C interactions with actin.
Myosin binding protein-C (MyBP-C) was first discovered as an impurity during the purification of myosin from skeletal muscle. However, soon after its discovery, MyBP-C was also shown to bind actin. While the unique functional implications for a protein that could cross-link thick and thin filaments together were immediately recognized, most early research nonetheless focused on interactions of MyBP-C with the thick filament. This was in part because interactions of MyBP-C with the thick filament could adequately explain most (but not all) effects of MyBP-C on actomyosin interactions and in part because the specificity of actin binding was uncertain. However, numerous recent studies have now established that MyBP-C can indeed bind to actin through multiple binding sites, some of which are highly specific. Many of these interactions involve critical regulatory domains of MyBP-C that are also reported to interact with myosin. Here we review current evidence supporting MyBP-C interactions with actin and discuss these findings in terms of their ability to account for the functional effects of MyBP-C. We conclude that the influence of MyBP-C on muscle contraction can be explained equally well by interactions with actin as by interactions with myosin. However, because data showing that MyBP-C binds to either myosin or actin has come almost exclusively from in vitro biochemical studies, the challenge for future studies is to define which binding partner(s) MyBP-C interacts with in vivo.


The inter-rater reliability of the modified finger goniometer for measuring forearm rotation.
Prospective cohort study. To compare the inter-rater reliability of using a modified finger goniometer (MFG) for the measurement of isolated forearm rotation for patients with distal radius fractures to the currently accepted technique for isolated forearm measurement. The currently accepted method of forearm measurement requires the assessor to visually estimate vertical for the stationary arm and placement of the moveable arm while placing a straight edge along a curved surface. Inter-rater reliability may be limited as assessors may estimate the placement of the goniometer arms differently depending on their experience, posture, and even their positioning relative to the patient. Rather than continue to place a straight edge on a round surface, we evaluate a new technique using an MFG for measuring isolated forearm rotation. Patients with clinically healed distal radius fractures were enrolled in the study. Measurement of active forearm pronation and supination was recorded using 2 separate measurement techniques. These measurements were taken by 2 separate hand therapists with more than 10 years of clinical experience in a tertiary care setting at the beginning and end of hand therapy sessions for 3 consecutive weekly visits. Intraclass correlation coefficients (ICCs), standard error of measurement, and minimal detectable change were calculated for each technique. The point estimates for the MFG method demonstrated a slightly higher ICC than the standard method for pronation (0.86 vs 0.82). For supination, both measurement techniques displayed equally high pooled ICCs (0.95). The standard error of measurements for the MFG were 2.1 for pronation and 1.2 for supination compared with 2.9 (pronation) and 1.2 (supination) for the standard technique. These translate into 90% minimal detectable changes of 5° and 3° for the MFG pronation/supination compared with 7° (pronation) and 3° (supination) for the standard technique, respectively. Although the point estimates for the ICCs of the MFG method are equal or higher than the standard method, the confidence intervals for the ICCs overlap, indicating that the MFG is at least equivalent to the standard method in terms of inter-rater reliability. 2b.


Male sterility at extreme temperatures: a significant but neglected phenomenon for understanding Drosophila climatic adaptations.
The thermal range for viability is quite variable among Drosophila species and it has long been known that these variations are correlated with geographic distribution: temperate species are on average more cold tolerant but more heat sensitive than tropical species. At both ends of their viability range, sterile males have been observed in all species investigated so far. This symmetrical phenomenon restricts the temperature limits within which permanent cultures can be kept in the laboratory. Thermal heat sterility thresholds are very variable across species from 23 degrees C in heat sensitive species up to 31 degrees C in heat tolerant species. In Drosophila melanogaster, genetic variations are observed among geographic populations. Tropical populations are more tolerant to heat induced sterility and recover more rapidly than temperate ones. A genetic analysis revealed that about 50% of the difference observed between natural populations was due to the Y chromosome. Natural populations have not reached a selection limit, however: thermal tolerance was still increased by keeping strains at a high temperature, close to the sterility threshold. On the low temperature side, a symmetrical reverse phenomenon seems to exist: temperate populations are more tolerant to cold than tropical ones. Compared to Mammals, drosophilids exhibit two major differences: first, male sterility occurs not only at high temperature, but also at a low temperature; second, sterility thresholds are not evolutionarily constrained, but highly variable. Altogether, significant and sometimes major genetic variations have been observed between species, between geographic races of the same species, and even between strains kept in the laboratory under different thermal regimes. In each case, it is easily argued that the observed variations correspond to adaptations to climatic conditions, and that male sterility is a significant component of fitness and a target of natural selection.


Structural Proteins in Severe Acute Respiratory Syndrome Coronavirus-2.
What began with a sign of pneumonia-related respiratory disorders in China has now become a pandemic named by WHO as Covid-19 known to be caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). The SARS-CoV-2 are newly emerged β coronaviruses belonging to the Coronaviridae family. SARS-CoV-2 has a positive viral RNA genome expressing open reading frames that code for structural and non-structural proteins. The structural proteins include spike (S), nucleocapsid (N), membrane (M), and envelope (E) proteins. The S1 subunit of S protein facilitates ACE2 mediated virus attachment while S2 subunit promotes membrane fusion. The presence of glutamine, asparagine, leucine, phenylalanine and serine amino acids in SARS-CoV-2 enhances ACE2 binding. The N protein is composed of a serine-rich linker region sandwiched between N Terminal Domain (NTD) and C Terminal Domain (CTD). These terminals play a role in viral entry and its processing post entry. The NTD forms orthorhombic crystals and binds to the viral genome. The linker region contains phosphorylation sites that regulate its functioning. The CTD promotes nucleocapsid formation. The E protein contains a NTD, hydrophobic domain and CTD which form viroporins needed for viral assembly. The M protein possesses hydrophilic C terminal and amphipathic N terminal. Its long-form promotes spike incorporations and the interaction with E facilitates virion production. As each protein is essential in viral functioning, this review describes the insights of SARS-CoV-2 structural proteins that would help in developing therapeutic strategies by targeting each protein to curb the rapidly growing pandemic.


Concurrent and longitudinal effects of ethnic identity and experiences of discrimination on psychosocial adjustment of Navajo adolescents.
In this study, we examined concurrent and longitudinal relations among Navajo adolescents' ethnic identity, experiences of discrimination, and psychosocial outcomes (i.e., self-esteem, substance use, and social functioning). At Time 1, 137 Navajo adolescents (67 male, 70 female), primarily in Grades 9 and 10, completed a written survey assessing ethnic identity, discrimination experiences, and a range of internalizing and externalizing behaviors. Two years later, 92 participants completed the same survey again. Ethnic and cultural identification was assessed via the Multiethnic Ethnic Identity Measure (MEIM; Phinney, 1992) and the Orthogonal Cultural Identification Scale (OCIS; Oetting & Beauvais, 1990). At Time 1, MEIM Affirmation and Belonging, MEIM Exploration, and OCIS White American identification all demonstrated strong, positive associations with adaptive functioning for male and female adolescents, whereas discrimination experiences were linked to lower self-esteem and social functioning for male adolescents. By Time 2, fewer significant concurrent associations between ethnic identity and psychosocial functioning scores remained, and discrimination experiences emerged as the most consistent correlate of poorer psychosocial functioning for male adolescents. Controlling for Time 1 psychosocial functioning, relatively few direct effects of ethnic and cultural identification variables predicted psychosocial functioning longitudinally, but discrimination experiences demonstrated strong and consistent longitudinal links with boys' substance use. Finally, interaction effects assessing the moderating influence of ethnic and cultural identification on negative links between discrimination and psychosocial functioning suggested that embeddedness in and connection to Navajo culture and, in some cases, connection to White American culture, served as a buffer to the negative effects of discrimination experiences.


Functional analyses reveal an important role for tyrosine residues in the staphylococcal multidrug efflux protein QacA.
The staphylococcal QacA multidrug efflux protein confers resistance to an exceptional number of structurally unrelated antimicrobial compounds. Aromatic amino acid residues have been shown to be highly important for the transport function of several multidrug transporters and are intimately involved in multidrug binding. This study investigated the structural and functional importance of the seven tyrosine residues in QacA by examining the phenotypic effect of incorporating conservative (aromatic) and non-conservative (non-aromatic) substitutions for these residues. Determination of the resistance profiles and analysis of drug transport assays revealed that non-conservative substitutions for most tyrosine residues influenced the QacA drug recognition spectrum. However, an aromatic residue at three tyrosine positions, 63, 410 and 429, was of importance for QacA-mediated transport and resistance to the majority of substrates tested. A tyrosine or phenylalanine residue at amino acid positions corresponding to 63 of QacA in related drug efflux proteins is found to be highly conserved. Therefore, an aromatic side chain at this position is likely to partake in a function common to these drug transporters, such as proton translocation or essential intramolecular contacts, whereas aromatic residues at the non-conserved 410 and 429 positions are expected to mediate a QacA-specific function, possibly forming or stabilising part of the QacA drug binding region.


Necrosis in breast cancer patients with skin metastases receiving bevacizumab-based therapy.
Bevacizumab has entered daily practice in advanced breast cancer patients, in whom skin metastases occurrence is a common event. Wound healing impairment has been described with bevacizumab, and this study looks at possible deleterious side effects of bevacizumab in patients with skin metastases. We retrospectively reviewed a series of 12 patients with advanced breast cancer presenting extensive skin metastases, and who received bevacizumab based therapy. Nine patients who initially presented with erosive skin lesions developed extensive and durable skin necrosis, as well as delayed healing of surgical flaps, despite early discontinuation of bevacizumab therapy and intensive skin care in a specialised unit. Skin necrosis was usually associated with extensive tumoural involvement of the skin. Bevacizumab may be harmful in selected breast cancer patients with metastatic cutaneous involvement.


Magnetic resonance imaging of the transverse atlantal ligament for the evaluation of atlantoaxial instability.
Twenty normal human subjects and 14 patients with upper cervical spine pathology were studied with axial high-field magnetic resonance (MR) imaging to examine the transverse atlantal ligament. Gradient-echo MR imaging pulse sequences provided reliable visualization of the transverse ligament, which exhibited low signal intensity and extended behind the dens between the medial portions of the lateral masses of C-1. The MR imaging characteristics of the transverse ligament were verified in clinical studies and in postmortem specimens. The clinical MR examinations defined 27 normal ligaments, three ligament disruptions, and four stretched rheumatoid ligaments. Atlantoaxial instability associated with transverse ligament rupture or ligamentous laxity required internal fixation. In contrast, fractures of C-1 or C-2 or atlantoaxial rotatory dislocations associated with an intact transverse ligament healed without instability or nonunion. The transverse ligament is the primary stabilizing component of C-1. The treatment of atlantoaxial instability has previously been based on criteria drawn from computerized tomography or plain radiographic studies, which only indirectly assess the probability of rupture of the transverse ligament. It is concluded that MR imaging accurately depicts the anatomical integrity of the transverse ligament. After transverse ligament failure, the remaining ligaments of the craniovertebral junction are inadequate to maintain stability. The presence of ligament disruption should be considered as a criterion for early fusion.


Inactivation of an invertebrate acetylcholinesterase by sulfhydryl reagents: the roles of two cysteines in the catalytic gorge of the enzyme.
We have used site-directed mutagenesis and molecular modeling to investigate the inactivation of an invertebrate acetylcholinesterase (AChE), ChE2 from amphioxus, by the sulfhydryl reagents 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and N-ethylmaleimide (NEM), creating various mutants, including C310A and C466A, and the double mutants C310A/C466A and C310A/F312I, to assess the relative roles of the two cysteines and a proposal that the increased rate of inactivation in the F312I mutant is due to increased access to Cys310. Our results suggest that both cysteines may be involved in inactivation by sulfhydryl reagents, but that the cysteine in the vicinity of the acyl pocket is more accessible. We speculate that the inactivation of aphid AChEs by sulfhydryl reagents is due to the presence of a cysteine homologous to Cys310. We also investigated the effects of various reversible cholinergic ligands, which bind to different subsites of the active site of the enzyme, on the rate of inactivation by DTNB of wild type ChE2 and ChE2 F312I. For the most part the inhibitors protect the enzymes from inactivation by DTNB. However, a notable exception is the peripheral site ligand propidium, which accelerates inactivation in the wild type ChE2, but retards inactivation in the F312I mutant. We propose that these opposing effects are the result of an altered allosteric signal transduction mechanism in the F312I mutant compared to the wild type ChE2.


Allergic reactions in the community: a questionnaire survey of members of the anaphylaxis campaign.
Allergic reactions to food are well recognized in both children and adults, but because of their relative infrequency their typical features may not be readily recognized by patients and their medical care givers who are not allergists. We sought to investigate the circumstances and clinical characteristics of food allergies in adults and children in the community. Self-completed questionnaire responses over a 6-month period from 109 members of the Anaphylaxis Campaign, the major British patient resource group for people who have suffered severe allergic reactions. One hundred and nine respondents reported 126 reactions during the study period. Seventy-five were children (under 16 years, median age 6 years at the time of reaction). Predictably more boys than girls were reported to have had reactions but more women reported reactions than men (P<0.05). Although the groups were equally aware of their food allergies the children had undergone diagnostic tests more often (P<0.001). Foods were implicated in 112 (89%) of reports. Restaurants were implicated less often (14%) than in other series, probably reflecting British eating habits. Children with asthma reported more severe reactions than those without asthma (P=0.008), although frequency or severity of recent asthma symptoms was not associated with severity of allergic reaction reported. When available, self-injectable adrenaline was used in 35% of severe reactions and 13% of non-severe reactions (P=0.01). A quarter of adults who received one dose of adrenaline also received a second dose. The allergens implicated in this report reflect previous data from similar patient groups in North America. Asthmatic children suffer more severe reactions than non-asthmatic children. It appears that British adults need better access to expert care of their allergies. Even when it is prescribed and available self-injectable adrenaline appears under-used in severe reactions. The incidence of severe but non-fatal allergic reactions in the UK may have been underestimated in the past.


Antitumor Activity of Ziziphus Jujube Fruit Extracts in KG-1 and NALM-6 Acute Leukemia Cell Lines.
Background : Ziziphus jujube Mill. belongs to the Rhamnaceae family. It has been reported to have a variety of biological activities such as antitumor, antioxidant, and anti-inflammatory effects. This study investigates the antiproliferative effect of Ziziphus jujube on KG-1 and NALM-6 acute leukemia cell lines. Materials and Methods : In this experimental study, the aqueous, ethyl acetate, and hydroalcoholic extracts of the Ziziphus jujube were prepared. Total phenolic and flavonoid components were detected because the presence of these compounds is associated with antioxidant and anticancer effects. Different concentrations of extracts were prepared, and KG-1 and NALM-6 cell lines were treated with them at 12, 24, 36, and 48 hours. Cell viability and IC50 values of the extracts were calculated using MTT assays. BD Cycle TEST PLUS DNA Kit was used for cell cycle progression analysis. Bcl2, Bax, and caspase-3 mRNA expressions were also assessed. Results : Cell viability decreased in a concentration-dependent manner. The best efficacy belonged to the ethyl acetate extract. Investigation of cell cycle progression demonstrated that the number of G0/G1 cells enhanced and the number of G2/M cells decreased when the ethyl acetate extract was applied in its IC50 concentration. A considerable increase in Caspase-3 and Bax and a decrease in Bcl2 gene expression were detected in molecular examination. Conclusion : According to our research, Ziziphus jujube ethyl acetate extract has antitumor properties on KG-1 and NALM-6 cell lines, possibly through induction of apoptosis and cell cycle regulation.


[Restriction fragment length polymorphism assays of clinical isolated human cytomegalovirus strain genomic DNA].
Ten clinical isolated strains of Human cytomegalovirus (HCMV) were obtained from 73 urine specimens of different people. Isolations from urine were carried out in human embryolung fibroblasts. Viral isolates were passaged four times. HCMV DNAs of laboratory strain AD169 and 10 clinical isolated strains were extracted with Hirt method, digested with each of the restriction enzymes EcoRI, Hind II. Comparison of restriction fragment length polymorphism (RFLP) of AD169 and isolated strains were made by hybridizing digested DNA with 32P labeled with HCMV Hind II cloned subgenomic fragments as the probe (pCM1035, pCM1015). pCM1035 is located in the joining region between the long(L) and short (S) unique sequences of the virus (L-S junction) pCM1015 is located in the terminal sequences of the virus. The results showed the genomic high degree of homology existed among all strains and the variable restriction site was in the L-S junction and terminal portion. The RFLP patterns of the clinical isolates which did not have relation in epidemiology were different, but the patterns of clinical isolates related in epidemiology were quite similar. Polymorphism frequently occurred in this case of EcoRI digested fragment hybridized with the probe of pCM1035. Southern hybridization of HCMV isolations is useful to researches into the molecular epidemiology and pathogenesis of HCMV infection.


Self-healing gels based on constitutional dynamic chemistry and their potential applications.
As representative soft materials with widespread applications, gels with various functions have been developed. However, traditional gels are vulnerable to stress-induced formation of cracks. The propagation of these cracks may affect the integrity of network structures of gels, resulting in the loss of functionality and limiting the service life of the gels. To address this challenge, self-healing gels that can restore their functionalities and structures after damage have been developed as "smart" soft materials. In this paper, we present an overview of the current strategies for synthesizing self-healing gels based on the concept of constitutional dynamic chemistry, which involves molecular structures capable of establishing dynamic networks based upon physical interactions or chemical reactions. The characterization methods of self-healing gels and the key factors that affect self-healing properties are analyzed. We also illustrate the emerging applications of self-healing gels, with emphasis on their usage in industry (coatings, sealants) and biomedicine (tissue adhesives, agents for drug or cell delivery). We conclude with a perspective on challenges facing the field, along with prospects for future development.


Multicenter Evaluation of the QIAstat-Dx Respiratory Panel for Detection of Viruses and Bacteria in Nasopharyngeal Swab Specimens.
The QIAstat-Dx Respiratory Panel (QIAstat-Dx RP) is a multiplex in vitro diagnostic test for the qualitative detection of 20 pathogens directly from nasopharyngeal swab (NPS) specimens. The assay is performed using a simple sample-to-answer platform with results available in approximately 69 min. The pathogens identified are adenovirus, coronavirus 229E, coronavirus HKU1, coronavirus NL63, coronavirus OC43, human metapneumovirus A and B, influenza A, influenza A H1, influenza A H3, influenza A H1N1/2009, influenza B, parainfluenza virus 1, parainfluenza virus 2, parainfluenza virus 3, parainfluenza virus 4, rhinovirus/enterovirus, respiratory syncytial virus A and B, Bordetella pertussis, Chlamydophila pneumoniae, and Mycoplasma pneumoniae This multicenter evaluation provides data obtained from 1,994 prospectively collected and 310 retrospectively collected (archived) NPS specimens with performance compared to that of the BioFire FilmArray Respiratory Panel, version 1.7. The overall percent agreement between QIAstat-Dx RP and the comparator testing was 99.5%. In the prospective cohort, the QIAstat-Dx RP demonstrated a positive percent agreement of 94.0% or greater for the detection of all but four analytes: coronaviruses 229E, NL63, and OC43 and rhinovirus/enterovirus. The test also demonstrated a negative percent agreement of ≥97.9% for all analytes. The QIAstat-Dx RP is a robust and accurate assay for rapid, comprehensive testing for respiratory pathogens.


The eyes have it: the neuroethology, function and evolution of social gaze.
Gaze is an important component of social interaction. The function, evolution and neurobiology of gaze processing are therefore of interest to a number of researchers. This review discusses the evolutionary role of social gaze in vertebrates (focusing on primates), and a hypothesis that this role has changed substantially for primates compared to other animals. This change may have been driven by morphological changes to the face and eyes of primates, limitations in the facial anatomy of other vertebrates, changes in the ecology of the environment in which primates live, and a necessity to communicate information about the environment, emotional and mental states. The eyes represent different levels of signal value depending on the status, disposition and emotional state of the sender and receiver of such signals. There are regions in the monkey and human brain which contain neurons that respond selectively to faces, bodies and eye gaze. The ability to follow another individual's gaze direction is affected in individuals with autism and other psychopathological disorders, and after particular localized brain lesions. The hypothesis that gaze following is "hard-wired" in the brain, and may be localized within a circuit linking the superior temporal sulcus, amygdala and orbitofrontal cortex is discussed.


Fronto-ethmoidal osteoma. Open treatment.
Osteoma is a benign tumour, composed of mature compact or cancellous bone, which can arise in any facial bone. Among the paranasal sinuses, the frontal and ethmoid sinuses are most frequently involved. Osteoma grows very slowly and small lesions are often not symptomatic. On the other hand, sometimes patients present with symptoms such as sinusitis, headache or ophtalmologic manifestations. These findings are secondary to obstruction of the involved sinus cavity with secondary mucocele formation or occasionally an expansile lesion with distortion of the facial contour. From 2005 to 2010, twenty-one (21) patients affected with non-syndromic fronto-ethmoidalosteomas were evaluated in our Department. Collected data include patients' age at the time of disorders, gender, presenting signs and symptoms, primary diagnosis, type and characteristics of the treatment performed, radiological findings and post-treatment results. All patients were investigated by CT scans in axial and coronal planes. The treatment and outcomes of this group were reviewed. All patients of the study underwent surgery and had a follow-up of at least 5 years. Surgical excision of the tumour was undertaken. Postoperative CT scans in axial and coronal planes showed complete removal of the tumour in all cases. No complications or recurrences were observed. Small, asymptomatic osteomas probably do not need to be treated but should be observed periodically. Surgical management remains the mainstay of treatment for these tumours. It requires total excision via an adequate approach, depending upon the site of presentation. A purely endoscopic endonasal approach has the risk of incomplete excision. In our experience open technique provides a wide exposure and better control. Even frontal osteomas can be safely removed by careful open surgery Ethmoidal sinus, Frontal sinus, Open treatment, Osteoma, Paranasal sinuses.


Helicobacter pylori antibody titres in serum, plasma and successively thawed specimens: implications for epidemiological and clinical studies.
Agreement between Helicobacter pylori IgG antibodies measured using the Pylori-set EIA-G kit in serum, plasma and successively thawed specimens was studied and the implications for epidemiological and clinical studies assessed. Plasma titres may differ from serum titres by -6% to +8% and therefore may be substituted for serum. The change in titre around the cut off value was -0.31 (se = 5.7, p = 0.96) per thaw. The estimated maximum drop after three thawings, 34.5, would result in only a small decrease in sensitivity (1.3%). For qualitative epidemiological studies, this additional misclassification rate is relatively small. However, positive titres did reduce over successive thawings, with the estimated maximum drop being 11.4% per thaw. Therefore, thawing does need to be considered as a contributing factor when interpreting titre drops in eradication trials. Baseline and follow up specimens from clinical studies should be thawed once only and tested concurrently.


Modulating Direct Growth of Copper Cobaltite Nanostructure on Copper Mesh as a Hierarchical Catalyst of Oxone Activation for Efficient Elimination of Azo Toxicant.
As cobalt (Co) has been the most useful element for activating Oxone to generate SO4•-, this study aims to develop a hierarchical catalyst with nanoscale functionality and macroscale convenience by decorating nanoscale Co-based oxides on macroscale supports. Specifically, a facile protocol is proposed by utilizing Cu mesh itself as a Cu source for fabricating CuCo2O4 on Cu mesh. By changing the dosages of the Co precursor and carbamide, various nanostructures of CuCo2O4 grown on a Cu mesh can be afforded, including nanoscale needles, flowers, and sheets. Even though the Cu mesh itself can be also transformed to a Cu-Oxide mesh, the growth of CuCo2O4 on the Cu mesh significantly improves its physical, chemical, and electrochemical properties, making these CuCo2O4@Cu meshes much more superior catalysts for activating Oxone to degrade the Azo toxicant, Acid Red 27. More interestingly, the flower-like CuCo2O4@Cu mesh exhibits a higher specific surface area and more superior electrochemical performance, enabling the flower-like CuCo2O4@Cu mesh to show the highest catalytic activity for Oxone activation to degrade Acid Red 27. The flower-like CuCo2O4@Cu mesh also exhibits a much lower Ea of Acid Red 27 degradation than the reported catalysts. These results demonstrate that CuCo2O4@Cu meshes are advantageous heterogeneous catalysts for Oxone activation, and especially, the flower-like CuCo2O4@Cu mesh appears as the most effective CuCo2O4@Cu mesh to eliminate the toxic Acid Red 27.


Births: Final Data for 2018.
Objectives-This report presents 2018 data on U.S. births according to a wide variety of characteristics. Trends in fertility patterns and maternal and infant characteristics are described and interpreted. Methods-Descriptive tabulations of data reported on the birth certificates of the 3.79 million births that occurred in 2018 are presented. Data are presented for maternal age, live-birth order, race and Hispanic origin, marital status, tobacco use, prenatal care, source of payment for the delivery, method of delivery, gestational age, birthweight, and plurality. Selected data by mother's state of residence and birth rates by age also are shown. Trend data for 2010 through 2018 are presented for selected items. Trend data by race and Hispanic origin are shown for 2016-2018. Results-3,791,712 births were registered in the United States in 2018, down 2% from 2017. Compared with rates in 2017, the general fertility rate declined to 59.1 births per 1,000 women aged 15-44. The birth rate for females aged 15-19 fell 7% in 2018. Birth rates declined for women aged 20-34 and increased for women aged 35-44. The total fertility rate declined to 1,729.5 births per 1,000 women in 2018. Birth rates for both married and unmarried women declined from 2017 to 2018. The percentage of women who began prenatal care in the first trimester of pregnancy rose to 77.5% in 2018; the percentage of all women who smoked during pregnancy declined to 6.5%. The cesarean delivery rate decreased to 31.9% in 2018 following an increase in 2017. Medicaid was the source of payment for 42.3% of all 2018 births, down 2% from 2017. The preterm birth rate rose for the fourth straight year to 10.02% in 2018; the rate of low birthweight was unchanged at 8.28%. Twin and triplet and higher-order multiple birth rates declined in 2018 (Figure 1).


Sonography of the lateral antebrachial cutaneous nerve with magnetic resonance imaging and anatomic correlation.
Abnormalities of the lateral antebrachial cutaneous nerve (LABCN) are associated with antecubital elbow conditions, such as distal biceps brachii tendon tears and traumatic cephalic vein phlebotomy. These can lead to lateral forearm, elbow, and wrist symptoms that can mimic other disease processes. The purpose of this study was to characterize the sonographic appearance of the LABCN using cadaveric dissection and retrospective analysis of sonographic examinations of symptomatic patients with magnetic resonance imaging correlation. For the first part of this study, a cadaveric elbow specimen was examined, and sonography was performed after dissection to identify the LABCN. Subsequently, 26 elbows in 13 patients with LABCN abnormalities were identified with sonography and retrospectively evaluated to characterize the appearance of the LABCN in both symptomatic and asymptomatic elbows. The symptomatic LABCNs showed fusiform enlargement, increased echogenicity, and loss of the normal fascicular echo texture. The mean cross-sectional area of the symptomatic nerves was 12.0 mm(2) (range, 6.1-17.2 mm(2)), with a maximum thickness of 3.5 mm (range, 2.3-5.9 mm), compared to 3.3 mm(2) (range, 1.9-5.2 mm(2)), with a maximum thickness of 1.3 mm (range, 0.9-2.2 mm), in the contralateral normal elbows. The close proximity of the LABCN to the distal biceps tendon and the cephalic vein makes it vulnerable to compression and injury in the setting of distal biceps tendon tears and traumatic phlebotomy, which may cause nerve enlargement and increased echogenicity. Awareness of the location and appearance of the LABCN on sonography is important for determining potential causes of lateral elbow and forearm pain.


Perioperative outcomes and complications related to teaching residents and fellows in scoliosis surgery.
Single-surgeon retrospective case series of 303 consecutive operative patients with idiopathic scoliosis (IS). The purpose of this study is to evaluate the perioperative outcomes in patients undergoing surgery for IS as a function of the experience level of the surgical assistant. The experience level of the surgical assistant, who is often a resident or fellow, has never before been evaluated as an independent factor in predicting perioperative outcomes and morbidity in scoliosis surgery. We hypothesize that there is no difference in perioperative outcomes with varying experience level of the surgical assistant. We evaluated the clinical, radiographic, and operative records from 303 consecutive operative patients from consecutive patients with IS. Group I was comprised of residents or spine fellows as assistants (teaching service, n = 175), and Group II consisted of junior or senior attendings as assistants (private practice service, n = 128). Multivariable linear regression was used to evaluate the relationship between experience level of the assistant and curve correction, operative time, estimated blood loss (EBL), complications, transfusions, and length of stay. In the posterior spinal fusion group (PSF, n = 164), there were no statistically significant differences in operative times between Groups I and II. Group I operative time was significantly increased, however, in patients undergoing anterior spinal surgery (ASF, P = 0.01), video-assisted thoracoscopic surgery (P = 0.0004), and combined anterior/posterior surgeries (ASF/PSF, P = 0.0063). There were no differences in EBL in ASF, video-assisted thoracoscopic surgery, or PSF surgeries, however, Group I had significantly higher EBL in the ASF/PSF group (P = 0.0016). No group differences were detected with respect to curve correction, transfusion rates, length of stay, or early complication rates. The experience level of surgical assistant had little bearing on perioperative morbidity or radiographic outcomes in scoliosis surgery. Marginally increased operative times and EBL, without an increase in transfusions or complications, is an acceptably safe tradeoff for educating orthopedic residents and fellows.


Evaluation of lung epithelial permeability.
During the last few years a number of factors affecting the measurement of the rate of absorption of 99mTc-DTPA across the alveolar-capillary membrane have been identified. These have helped to provide insights into the significance of lung epithelial permeability (LEP) measurements and their potential limitations. Pulmonary absorption of 99mTc-DTPA has been studied in a variety of clinical conditions and the results are discussed.


Quality analysis of selective microparticle deposition on electrically programmable surfaces.
Image processing and pattern analysis can evaluate the deposition quality of triboelectrically charged microparticles on charged surfaces. The image processing method presented in this paper aims at controlling the quality of peptide arrays generated by particle based solid phase Merrifield combinatorial peptide synthesis. Incorrectly deposited particles are detected before the amino acids therein are coupled to the growing peptide. The calibration of the image acquisition is performed in a supervised training step in which all parameters of the quality analyzing algorithm are learnt given one representative image. Then, the correct deposition pattern is determined by a linear support vector machine. Knowing the pattern, contaminated areas can be detected by comparing the pattern with the actual deposition. Taking into account the resolution of the image acquisition system and its magnification factor, the number and size of contaminating particles can be calculated out of the number of connected foreground pixels.


Proliferating cell nuclear antigen (PCNA) is required for cell cycle-regulated silent chromatin on replicated and nonreplicated genes.
In Saccharomyces cerevisiae, silent chromatin is formed at HMR upon the passage through S phase, yet neither the initiation of DNA replication at silencers nor the passage of a replication fork through HMR is required for silencing. Paradoxically, mutations in the DNA replication processivity factor, POL30, disrupt silencing despite this lack of requirement for DNA replication in the establishment of silencing. We tested whether pol30 mutants could establish silencing at either replicated or non-replicated HMR loci during S phase and found that pol30 mutants were defective in establishing silencing at HMR regardless of its replication status. Although previous studies tie the silencing defect of pol30 mutants to the chromatin assembly factors Asf1p and CAF-1, we found pol30 mutants did not exhibit a gross defect in packaging HMR into chromatin. Rather, the pol30 mutants exhibited defects in histone modifications linked to ASF1 and CAF-1-dependent pathways, including SAS-I- and Rtt109p-dependent acetylation events at H4-K16 and H3-K9 (plus H3-K56; Miller, A., Yang, B., Foster, T., and Kirchmaier, A. L. (2008) Genetics 179, 793-809). Additional experiments using FLIM-FRET revealed that Pol30p interacted with SAS-I and Rtt109p in the nuclei of living cells. However, these interactions were disrupted in pol30 mutants with defects linked to ASF1- and CAF-1-dependent pathways. Together, these results imply that Pol30p affects epigenetic processes by influencing the composition of chromosomal histone modifications.


DMBT1 functions as pattern-recognition molecule for poly-sulfated and poly-phosphorylated ligands.
Deleted in malignant brain tumors 1 (DMBT1) is a secreted glycoprotein displaying a broad bacterial-binding spectrum. Recent functional and genetic studies linked DMBT1 to the suppression of LPS-induced TLR4-mediated NF-kappaB activation and to the pathogenesis of Crohn's disease. Here, we aimed at unraveling the molecular basis of its function in mucosal protection and of its broad pathogen-binding specificity. We report that DMBT1 directly interacts with dextran sulfate sodium (DSS) and carrageenan, a structurally similar sulfated polysaccharide, which is used as a texturizer and thickener in human dietary products. However, binding of DMBT1 does not reduce the cytotoxic effects of these agents to intestinal epithelial cells in vitro. DSS and carrageenan compete for DMBT1-mediated bacterial aggregation via interaction with its bacterial-recognition motif. Competition and ELISA studies identify poly-sulfated and poly-phosphorylated structures as ligands for this recognition motif, such as heparansulfate, LPS, and lipoteichoic acid. Dose-response studies in Dmbt1(-/-) and Dmbt1(+/+) mice utilizing the DSS-induced colitis model demonstrate a differential response only to low but not to high DSS doses. We propose that DMBT1 functions as pattern-recognition molecule for poly-sulfated and poly-phosphorylated ligands providing a molecular basis for its broad bacterial-binding specificity and its inhibitory effects on LPS-induced TLR4-mediated NF-kappaB activation.


Effect of ozone exposure and infection on bronchoalveolar lavage: sex differences in response patterns.
Female mice exhibit a better survival rate than males after infection, but if infection follows an ozone-induced oxidative stress, male survival exceeds that of females. Our goal was to study bronchoalveolar lavage factors that contribute to these sex differences in outcome. We studied parameters at 4, 24, and 48 h after ozone exposure and infection, including markers of inflammation, oxidative stress, and tissue damage, and surfactant phospholipids and surfactant protein A (SP-A). A multianalyte immunoassay at the 4h time point measured 59 different cytokines, chemokines, and other proteins. We found that: (1) Although some parameters studied revealed sex differences, no sex differences were observed in LDH, total protein, MIP-2, and SP-A. Males showed more intragroup significant differences in SP-A between filtered air- and ozone-exposed mice compared to females. (2) Oxidized dimeric SP-A was higher in FA-exposed female mice. (3) Surfactant phospholipids were typically higher in males. (4) The multianalyte data revealed differences in the exuberance of responses under different conditions - males in response to infection and females in response to oxidative stress. These more exuberant, and presumably less well-controlled responses associate with the poorer survival. We postulate that the collective effects of these sex differences in response patterns of lung immune cells may contribute to the clinical outcomes previously observed.


Kinetic analyses for species differences in P-glycoprotein-mediated drug transport.
P-glycoprotein (P-gp) plays an important role in the pharmacokinetics of drugs. There is little information on the species differences in P-gp-mediated drug transport activity. The purpose of the present study was to clarify the differences in the kinetic parameters and the existence of species differences in the P-gp-mediated drug transport activity using seven multidrug resistence1 (MDR1) transfected cell lines, in which the cDNA was from human, monkey, canine, rat (MDR1a and MDR1b), and mouse (mdr1a and mdr1b). The transcellular transport of diltiazem, cyclosporin A, and dexamethasone across monolayers of MDR1 transfected cells. The apparent K(m) values of diltiazem exhibited approximately 16.5-fold differences among the seven cell lines. Concerning the diltiazem transport, the V(max)/K(m) value of human P-gp corrected by the P-gp expression level was similar to that of monkey P-gp, but was 5.6-fold higher than that of canine P-gp. On the other hand, the corrected V(max)/K(m) value of human P-gp for cyclosporin A transport was 3.8-fold higher than that of monkey P-gp. The present study would be valuable to evaluate the P-gp function of various animals in the same experimental condition. It was clarified that the species differences in P-gp-mediated drug transport activity evaluated by the corrected V(max)/K(m) value differed according to the substrate.


Reactivity of rhodium and iridium peroxido complexes towards hydrogen in the presence of B(C6F5)3 or [H(OEt2)2][B{3,5-(CF3)2C6H3}4].
The peroxido complexes trans-[M(4-C5F4N)(O2)(CNtBu)(PR3)2] (1: M = Rh, R = Et; 2a: M = Ir, R = iPr) can be used in the metal-mediated hydrogenation of O2. The reaction of trans-[Rh(4-C5F4N)(O2)(CNtBu)(PEt3)2] (1) with B(C6F5)3 and H2 gave trans-[Rh(4-C5F4N)(CNtBu)(PEt3)2] (3), OPEt3 and (H2O)·B(C6F5)3, whereas treatment of [H(OEt2)2][B{3,5-(CF3)2C6H3}4] with 1 in the presence of H2 yielded trans-[Rh(4-C5F4N)(CNtBu)(PEt3)2] (3) and H2O2. The reactivity of 2a towards B(C6F5)3 and BClCy2 was also studied and an intermediate was detected which is assigned to be trans-[Ir(4-C5F4N)(Cl)(OOBCy2)(CNtBu)(PiPr3)2] (4a).


Nanomedicine, a valuable tool for skeletal muscle disorders: Challenges, promises, and limitations.
Muscular dystrophies are a group of rare genetic disorders characterized by progressive muscle weakness, which, in the most severe forms, leads to the patient's death due to cardiorespiratory problems. There is still no cure available for these diseases and significant effort is being placed into developing new strategies to either correct the genetic defect or to compensate muscle loss by stimulating skeletal muscle regeneration. However, the vast anatomical extension of the target tissue poses great challenges to these goals, highlighting the need for complementary strategies. Nanomedicine is an actively evolving field that merges nanotechnologies with biomedical and pharmaceutical sciences. It holds great potential in regenerative medicine, both in supporting tissue engineering and regeneration, and in optimizing drug and oligonucleotide delivery and gene therapy strategies. In this review, we will summarize the state-of-the-art in the field of nanomedicine applied to skeletal muscle regeneration. We will discuss the recent work toward the development of nanopatterned scaffolds for tissue engineering, the efforts in the synthesis of organic and inorganic nanoparticles for gene therapy and drug delivery applications, as well as their use as immune modulators. Although nanomedicine holds great promise for muscle and other degenerative diseases, many challenges still need to be systematically addressed to assure a smooth transition from the bench to the bedside. This article is categorized under: Implantable Materials and Surgical Technologies > Nanotechnology in Tissue Repair and Replacement.