Laparoscopic versus open resection for transverse and descending colon cancer: Short-term and long-term outcomes of a multicenter retrospective study of 1830 patients.
Previous randomized controlled trials demonstrated similar oncological outcomes between laparoscopic and open colectomies, except for cases involving transverse colon and splenic flexure colon cancer. The objective of this study was to confirm the oncological safety and advantages of the short-term results of laparoscopic surgery for transverse and descending colon cancer in comparison with open surgery. The study data were retrospectively collected from the databases of 45 hospitals. Patients with transverse or descending colon cancer who underwent laparoscopic or open R0 resection were registered. The primary end-points were the 3-year overall survival and relapse-free survival rates according to pathological stage. The secondary end-points were the short-term results, including blood loss, operative time, diet intake, hospital stay, and postoperative complications. Of the 1830 eligible patients, 872 underwent open colectomy and 958 underwent laparoscopic colectomy. The median follow-up period was 38.4 months. The conversion rate to open resection was 4.5%. The 3-year overall survival rate of the laparoscopic group was significantly higher than that of the open group for stage I patients (96.2% vs 99.2%; P = 0.04); it was also higher for stage II (94.0% vs 95.5%) and stage III (87.4% vs 90.2%) patients, but there were no significant differences. The 3-year relapse-free survival rate of the laparoscopic group was significantly higher than that of the open group for stage I patients; there were no differences between the open and laparoscopic groups among the stage II and III patients. In the multivariate analyses, laparoscopic resection was a significant factor in relapse-free survival. Laparoscopic patients had significantly lower blood loss and a significantly longer operative time than the open groups. Also, postoperative hospital stay was significantly shorter and postoperative morbidity was significantly lower in the laparoscopic group. Although this retrospective study has limitations, we can conclude that laparoscopic surgery for transverse and descending colon cancer is oncologically safe and yields better short-term results than open surgery.


A hydrophobic transmembrane segment at the carboxyl terminus of thy-1.
The mode of integration of the glycoprotein thy-1 within the cell membrane has been controversial due to an apparent lack of a transmembrane hydrophobic segment. Rat and mouse complementary DNA and genomic clones encoding the thy-1 molecule have been isolated and sequenced. These studies have enabled us to determine the intron-exon organization of the thy-1 gene. Furthermore, they have revealed the existence of a sequence which would encode an extra segment (31 amino acids) at the carboxyl terminus of the thy-1 molecule. These extra amino acids include a 20-amino acid hydrophobic segment which may be responsible for integration of thy-1 within the plasma membrane.


Burden of Sciatica on US Medicare Recipients.
This study evaluates the disease burden of sciatica on the US Medicare cohort. Sciatica is a common disability that has important physical, mental, and economic effects. The Medicare Health Outcomes Survey (HOS) is a demographic and outcomes survey used to monitor the performance of Medicare Advantage health plans in the United States. The HOS includes data on demographics, chronic medical conditions, and patient-reported outcomes. Medicare HOS data for cohorts from 2007 to 2013 were obtained. Patients were placed into two categories based on the survey results: with or without a history of sciatica. Baseline demographics, chronic medical conditions, and physical health symptoms were aggregated. In addition, average VR-12 physical component summary and mental component summary scores were calculated for each group at baseline and at 2-year follow-up. A Fisher exact test was used to assess significance for categorical variables, and a t-test was used for continuous variables. VR-12 changes as small as 1 to 2 units have been found to be clinically and socially relevant. The baseline cohort data of 1,000,952 patients yielded 250,869 patients (25%) who reported the diagnosis of sciatica, compared with 750,083 patients (75%) without sciatica. Patients with a history of sciatica tended to be younger, less educated, and notably with more medical comorbidities. Physical component summary outcomes were approximately 8 units lower in the sciatica group at baseline and 7 units lower at 2-year follow-up. Mental component summary outcomes were 6 units lower in the sciatica group at baseline and 5 units lower at 2-year follow-up. A large percentage of the US Medicare cohort suffers from symptomatic sciatica. Our study identified a 25% prevalence in the Medicare cohort. In addition, sciatica is associated with an increased incidence of comorbid medical conditions and poor health-related quality of life. Level III STUDY DESIGN:: Observational-Cohort Study.


Upregulation of axon guidance molecules in the adult central nervous system of Nogo-A knockout mice restricts neuronal growth and regeneration.
Adult central nervous system axons show restricted growth and regeneration properties after injury. One of the underlying mechanisms is the activation of the Nogo-A/Nogo receptor (NgR1) signaling pathway. Nogo-A knockout (KO) mice show enhanced regenerative growth in vivo, even though it is less pronounced than after acute antibody-mediated neutralization of Nogo-A. Residual inhibition may involve a compensatory component. By mRNA expression profiling and immunoblots we show increased expression of several members of the Ephrin/Eph and Semaphorin/Plexin families of axon guidance molecules, e.g. EphrinA3 and EphA4, in the intact spinal cord of adult Nogo-A KO vs. wild-type (WT) mice. EphrinA3 inhibits neurite outgrowth of EphA4-positive neurons in vitro. In addition, EphrinA3 KO myelin extracts are less growth-inhibitory than WT but more than Nogo-A KO myelin extracts. EphA4 KO cortical neurons show decreased growth inhibition on Nogo-A KO myelin as compared with WT neurons, supporting increased EphA4-mediated growth inhibition in Nogo-A KO mice. Consistently, in vivo, Nogo-A/EphA4 double KO mice show increased axonal sprouting and regeneration after spinal cord injury as compared with EphA4 KO mice. Our results reveal the upregulation of developmental axon guidance cues following constitutive Nogo-A deletion, e.g. the EphrinA3/EphA4 ligand/receptor pair, and support their role in restricting neurite outgrowth in the absence of Nogo-A.


Design and characterization of a synthetically accessible, photodegradable hydrogel for user-directed formation of neural networks.
Hydrogels with photocleavable units incorporated into the cross-links have provided researchers with the ability to control mechanical properties temporally and study the role of matrix signaling on stem cell function and fate. With a growing interest in dynamically tunable cell culture systems, methods to synthesize photolabile hydrogels from simple precursors would facilitate broader accessibility. Here, a step-growth photodegradable poly(ethylene glycol) (PEG) hydrogel system cross-linked through a strain promoted alkyne-azide cycloaddition (SPAAC) reaction and degraded through the cleavage of a nitrobenzyl ether moiety integrated into the cross-links is developed from commercially available precursors in three straightforward synthetic steps with high yields (>95%). The network evolution and degradation properties are characterized in response to one- and two-photon irradiation. The PEG hydrogel is employed to encapsulate embryonic stem cell-derived motor neurons (ESMNs), and in situ degradation is exploited to gain three-dimensional control over the extension of motor axons using two-photon infrared light. Finally, ESMNs and their in vivo synaptic partners, myotubes, are coencapsulated, and the formation of user-directed neural networks is demonstrated.


RUSHing to the Diagnosis: Aortic Abdominal Aneurysm Detected Using the Rapid Ultrasound for Shock and Hypotension (RUSH) Protocol in the Wards.
The rapid ultrasound for shock and hypotension (RUSH) protocol is a useful tool used in the emergency department (ED) when addressing the severity and etiology of shock. It was designed to be performed in under two minutes with evaluation of the pump (heart), tank (inferior vena cava, thoracic and abdominal compartments) and the pipes (large arteries and veins). However, its application or one similar should extend beyond the ED and into the hospital floor. Here we present an 80-year-old gentleman with a history of atrial fibrillation (A-Fib) on anticoagulation who arrived at the ED due to an episode of pre-syncope just prior to arrival. Initial EKG is concerning for A-Fib with rapid ventricular response (RVR) with a rate in the 130s. After fluid resuscitation patient improved and he was admitted to the telemetry floor for further cardiac workup and cardiology consultation. While waiting for a room in the ED, patient became hypotensive, diaphoretic and pale. After complaining of lower abdominal pain, the ED physician performed a RUSH which showed an abdominal aorta of 8 cm concerning for dissection. Diagnosis was confirmed with CT angiography of the abdomen and he was taken to the OR with successful repair of the abdominal aortic aneurysm (AAA). Patient made meaningful recovery and was discharged to in-patient rehab. The patient described in this vignette was delayed in the ED due to lack of beds on the floor. This allowed for quick ultrasound work-up by the ED physician which led to immediate recognition of the AAA and immediate response by the vascular surgery team. Should this patient have been on the hospital floor, it is unclear if such prompt steps would have occurred prior to patient's further hemodynamic demise.


Neopterin as discriminating and prognostic parameter in healthy homosexuals, ARC and AIDS patients.
Urinary neopterin, a sensitive marker for activation of cell mediated immunity, is compared to clinical and laboratory data in 5 patients with AIDS, in 15 patients with ARC and in 40 male subjects without AIDS-related clinical signs attending an AIDS outpatient clinic. The sensitivity of neopterin for AIDS-related diseases was higher than that of the CD4+/CD8+ subset ratio. The differences in neopterin levels between the controls, ARC and AIDS patients were found to be more significant than the T-cell subset data. The observation that the 3 AIDS patients with the highest neopterin levels have died whereas the two others with the lower levels are still alive also underlines the prognostic potential of urinary neopterin level determinations. Measurement of urinary neopterin is thus recommended as an additional criterion for monitoring ARC and AIDS patients.


Multifunctional dendronized peptide polymer platform for safe and effective siRNA delivery.
In this study, we designed and synthesized a biodegradable dendronized polypeptide (denpol) platform for delivery of small interfering RNA (siRNA). The novel denpol architecture combines the multivalency of dendrimers and conformational flexibility of linear polymers for optimal siRNA binding. Multifunctional amino acids were incorporated onto the dendrons and the structure was tuned both systematically and combinatorially to select optimal vectors. By screening a focused library, we identified several denpols that can effectively deliver siRNA to NIH 3T3 cells in vitro and exhibit minimal toxicity. For comparison, the best-performing denpol showed significantly improved transfection efficiency over Lipofectamine in serum-containing media. Fluorescence intracellular trafficking studies indicated that amphiphilicity is important for cell uptake and that the buffering capacity of histidine facilitates endosomal membrane rupture and therefore enhances the transfection efficiency. The combination of high delivery efficiency in serum and low cytotoxicity suggests the denpol system as a promising new carrier for siRNA delivery.


The antimicrobial resistome in relation to antimicrobial use and biosecurity in pig farming, a metagenome-wide association study in nine European countries.
Previous studies in food-producing animals have shown associations between antimicrobial use (AMU) and resistance (AMR) in specifically isolated bacterial species. Multi-country data are scarce and only describe between-country differences. Here we investigate associations between the pig faecal mobile resistome and characteristics at the farm-level across Europe. A cross-sectional study was conducted among 176 conventional pig farms from nine European countries. Twenty-five faecal samples from fattening pigs were pooled per farm and acquired resistomes were determined using shotgun metagenomics and the Resfinder reference database, i.e. the full collection of horizontally acquired AMR genes (ARGs). Normalized fragments resistance genes per kilobase reference per million bacterial fragments (FPKM) were calculated. Specific farm-level data (AMU, biosecurity) were collected. Random-effects meta-analyses were performed by country, relating farm-level data to relative ARG abundances (FPKM). Total AMU during fattening was positively associated with total ARG (total FPKM). Positive associations were particularly observed between widely used macrolides and tetracyclines, and ARGs corresponding to the respective antimicrobial classes. Significant AMU-ARG associations were not found for β-lactams and only few colistin ARGs were found, despite high use of these antimicrobial classes in younger pigs. Increased internal biosecurity was directly related to higher abundances of ARGs mainly encoding macrolide resistance. These effects of biosecurity were independent of AMU in mutually adjusted models. Using resistome data in association studies is unprecedented and adds accuracy and new insights to previously observed AMU-AMR associations. Major components of the pig resistome are positively and independently associated with on-farm AMU and biosecurity conditions.


Antioxidant and angiostatic effect of Spirulina platensis suspension in complete Freund's adjuvant-induced arthritis in rats.
Currently, natural products have built a well-recognized role in the management of many degenerative diseases, mainly rheumatoid arthritis. Recent studies suggest that Spirulina, a unicellular blue-green alga, may have a variety of health benefits and curative properties and is also competent of acting as an anti-inflammatory, antioxidant and recently anti-angiogenic agent. In the present study, the antioxidant and the immunomodulatory effect of Spirulina platensis as well as its anti-angiogenic effect against complete Freund's adjuvant-induced arthritis (AIA) in rat model were tested. We found that the development of arthritis was concealed; moreover it successfully inhibited the development of macroscopic as well as microscopic and histopathological lesions in AIA rats when compared to control. Spirulina treated group showed a higher survival rate and moreover, it reduced the clinical score of RA in a dose dependent manner. Furthermore, Spirulina decreased serum levels of COX-2, TNF-α, IL-6, TBARS, VEGF and increased serum levels of GSH compared to the RA non-treated group. The present study concluded that Spirulina is able to restrain the changes produced through adjuvant-induced arthritis. The suppressing effect of Spirulina could be attributed, at least in part, to anti-inflammatory, antioxidant and anti-angiogenic properties.


Manipulating the 3D organization of the largest synthetic yeast chromosome.
Whether synthetic genomes can power life has attracted broad interest in the synthetic biology field. Here, we report de novo synthesis of the largest eukaryotic chromosome thus far, synIV, a 1,454,621-bp yeast chromosome resulting from extensive genome streamlining and modification. We developed megachunk assembly combined with a hierarchical integration strategy, which significantly increased the accuracy and flexibility of synthetic chromosome construction. Besides the drastic sequence changes, we further manipulated the 3D structure of synIV to explore spatial gene regulation. Surprisingly, we found few gene expression changes, suggesting that positioning inside the yeast nucleoplasm plays a minor role in gene regulation. Lastly, we tethered synIV to the inner nuclear membrane via its hundreds of loxPsym sites and observed transcriptional repression of the entire chromosome, demonstrating chromosome-wide transcription manipulation without changing the DNA sequences. Our manipulation of the spatial structure of synIV sheds light on higher-order architectural design of the synthetic genomes.


Novel in vitro diagnosis of equine allergies using a protein array and mathematical modelling approach: a proof of concept using insect bite hypersensitivity.
Insect bite hypersensitivity (IBH) is a seasonal recurrent skin allergy of horses caused by IgE-mediated reactions to allergens present in the saliva of biting insects of the genus Culicoides, and possibly also Simulium and Stomoxys species. In this work we show that protein microarrays containing complex extracts and pure proteins, including recombinant Culicoides allergens, can be used as a powerful technique for the diagnosis of IBH. Besides the obvious advantages such as general profiling and use of few microliters of samples, this microarray technique permits automation and allows the generation of mathematical models with the calculation of individual risk profiles that can support the clinical diagnosis of allergic diseases. After selection of variables on influence on the projection (VIP), the observed values of sensitivity and specificity were 1.0 and 0.967, respectively. This confirms the highly discriminatory power of this approach for IBH and made it possible to attain a robust predictive mathematical model for this disease. It also further demonstrates the specificity of the protein array method on identifying a particular IgE-mediated disease when the sensitising allergen group is known.


Prediction of drug-induced catalepsy based on dopamine D1, D2, and muscarinic acetylcholine receptor occupancies.
It is known that catalepsy serves as an experimental animal model of parkinsonism. In this study, the relationship between in vivo dopamine D1 and D2 receptor occupancies and catalepsy was investigated to predict the intensity of catalepsy induced by drugs that bind to D1 and D2 receptors nonselectively. 3H-SCH23390 and 3H-raclopride were used for the labeling of D1 and D2 receptors, respectively. The ternary complex model consisting of agonist or antagonist, receptor, and transducer was developed, and the dynamic parameters were determined. After coadministration of SCH23390 and nemonapride, catalepsy was stronger than sum of the values predicted by single administration of each drug, and it was intensified synergistically. This finding suggested the existence of interaction between D1 and D2 receptors, and the necessity for constructing the model including this interaction. To examine the validity of this model, catalepsy and in vivo dopamine receptor occupancy were measured after administration of drugs that induce or have a possibility to induce parkinsonism (haloperidol, flunarizine, manidipine, oxatomide, hydroxyzine, meclizine, and homochlorcycilzine). All of the tested drugs blocked both dopamine D1 and D2 receptors. Intensity of catalepsy was predicted with this dynamic model and was compared with the observed values. In contrast with haloperidol, flunarizine, manidipine, and oxatomide (which induced catalepsy), hydroxyzine, meclizine, and homochlorcyclizine failed to induce catalepsy. Intensities of catalepsy predicted with this dynamic model considering the interaction between D1 and D2 receptors overestimated the observed values, suggesting that these drugs have catalepsy-reducing properties as well. Because muscarinic acetylcholine (mACh) receptor antagonists inhibit the induction of catalepsy, the anticholinergic activities of the drugs were investigated. After SCH23390, nemonapride and scopolamine were administered simultaneously; catalepsy and in vivo mACh receptor occupancy were measured to evaluate quantitatively the anticholinergic activity. Relationship between mACh receptor occupancy and change in catalepsy was used as the measure of catalepsy-reducing effects of the drugs. Measurement of in vivo mACh receptor occupancy revealed a significant blockade of mACh receptor by all of the tested drugs except for haloperidol. The predicted values of catalepsy, when corrected for the mACh receptor-related reduction, approached the observed values. This finding indicates the possibility that mACh receptor antagonism of drugs may contribute to the reduction of catalepsy. In conclusion, the dynamic model considering D1, D2, and mACh receptor occupancies and synergism between D1 and D2 receptors may be useful for quantitative prediction of drug-induced catalepsy.


Cracking the shell on egg-hypersensitive patients and egg-containing vaccines.
Hens' eggs are a common food in the American diet. They are consumed as a primary food source and added as an ingredient to other foods. In individuals who are hypersensitive to eggs, egg-containing foods can cause mild to severe allergic reactions if ingested. These individuals may also have adverse reactions to vaccines produced on egg media. Vaccines that are created on egg media include those for measles, mumps, and rubella; rabies; yellow fever; and influenza. The authors discuss recent developments in the use of egg-containing vaccines in hypersensitive patients.


[Use of isoelectric focusing for assessing the composition of pollen allergens].
The preparations of allergens and allergoids obtained from ragweed, timothy and wormwood pollen, as well as the preparations of allergens from birch and orchard grass pollen differing in the method of their production, have been studied with the use of analytical isoelectric focusing in a thin gel layer. The composition of the preparations of allergoids differs from that of the allergenic preparations from the pollen of the same plant species by the decreased content of protein components detected in this investigation. The main proteins contained in the preparations of allergoids are distributed in the zone of pH 3.5-4.5. Differences in the composition of different batches of the same allergens, manifested by variations in some protein bands or by their absence, have been noted. Protein components with the isoelectric point in the alkaline zone have been detected only in the preparations of ragweed pollen allergens.


Blockade of the orexin-2 receptors within the ventral tegmental area facilitates the extinction and prevents the reinstatement of methamphetamine-seeking behavior.
Repeated use of methamphetamine (METH) causes severe effects on the central nervous system, associated with an increased relapse rate. The orexinergic system is highly implicated in the reward circuitry and may be a promising target for treating psychostimulant dependency. The present study aimed to investigate the involvement of the orexin system, mainly the orexin-2 receptors (OX2R) in the ventral tegmental area (VTA) in the extinction and reinstatement of METH-seeking behavior using a conditioned place preference (CPP) paradigm. To this end, animals received METH (1 mg/kg; sc) for a 5-day conditioning period. Then, in the first set of experiments, different groups of rats were given intra-VTA TCS OX2 29 (1, 3, 10, or 30 nmol/0.3 μl DMSO) as an OX2R antagonist over a 10-day extinction period. In another experiment, after the extinction period, a different set of animals received a single dose of TCS OX2 29 (1, 3, 10, or 30 nmol) before the priming dose of METH (0.25 mg/kg; sc) on the reinstatement day. The results revealed that TCS OX2 29 (10 and 30 nmol) remarkably facilitated the extinction of rewarding properties of METH (P < 0.001 for both doses). Furthermore, TCS OX2 29 (3, 10, or 30 nmol) significantly suppressed the METH-induced reinstatement (3 nmol; P < 0.05, 10 nmol; P < 0.01, and 30 nmol; P < 0.001). In conclusion, the current study revealed that the orexinergic system, specifically the VTA OX2R, is involved in METH-seeking behaviors and that manipulation of this system can be considered a potential therapeutics in treating METH dependency.


Intron features of key functional genes mediating nitrogen metabolism in marine phytoplankton.
Introns are widespread and variable in eukaryotic genomes. Although their histories and functions, or even whether all of them have any function, remain largely unknown, analysis of intron sequences and genomic contexts may shed light on the evolutionary history of genes and organisms. The number and frequency of introns vary widely in the small number of published genomes of protists and algae suggesting that the same is true of the vast diversity of protists and algae that remain uncultivated. The objective of this study were to investigate introns in sequences of functional genes of phytoplankton, both in published genomes and in sequences obtained from environmental clone libraries. We examined the introns of the genes involved in nitrogen uptake and assimilation pathways in the genome sequences of cultivated phytoplankton as well as in environmental clone libraries of nitrate reductases (NR), nitrite reductase (NiR), nitrate transporter (Nrt2) and ammonium transporter (AMT) genes constructed from pelagic phytoplankton communities in Monterey Bay (CA, USA) and Onslow Bay (NC, USA). Here we describe the most extensive set to date of intron sequences from uncultivated marine algae and report important differences for diatom vs. non-diatom sequences. The majority of the introns in NR, NiR, Nrt2 and AMT from cultured phytoplankton and environmental libraries showed canonical splice patterns. Introns found in diatom-like NR environmental libraries had lower GC content than the respective exons. The green algal-like NR and Nrt2 environmental sequences had introns and exons of much more similar GC content, and both higher than in diatoms. These patterns suggest a different evolutionary history and recent acquisition of diatom introns compared to other algae.


Differential diagnosis of acute pericarditis from normal variant early repolarization and left ventricular hypertrophy with early repolarization: an electrocardiographic study.
Differentiation of ST-segment elevation on electrocardiogram (ECG) from acute pericarditis (AP), normal variant early repolarization (ER) and early repolarization of left ventricular hypertrophy (ERLVH) can be problematic. Hence, the authors evaluated the accuracy of the ST/T ratio in ECG to more optimally differentiate between AP, ST-segment elevation, ER and ERLVH. Between September 2006 and July 2010, 80 patients were enrolled in this study consisting of 25 individuals with AP, 27 with ER and 28 with ERLVH. Each ECG was analyzed in a systematic manner including the measurement of PR interval, QRS duration, QT-segment duration, PR-segment deviation, ST-segment deviation and the height of T wave. The ratio of the height of ST segment to the height of T wave was measured in leads I, II, III, aVF and V2 through V6. The mean ages of the patients with AP, ER and ERLVH were 32 ± 16.5, 36 ± 15.4 and 53 ± 16 years, respectively. The ratio of the amplitude of ST segment to the amplitude of the T wave in leads I, V4, V5 and V6 proved to be a significant discriminator at a value of ≥0.25 (P < 0.05 for all). Leads I, V4, V5 and V6 can all be used to differentiate AP from ER and ERLVH. When ST elevation is present in lead I, the ST/T ratio has the best predictive value (0.82) to more accurately discriminate between AP, ER and ERLVH.


Envelope proteome changes driven by RamA overproduction in Klebsiella pneumoniae that enhance acquired β-lactam resistance.
In Klebsiella pneumoniae, overproduction of RamA results in reduced envelope permeability and reduced antimicrobial susceptibility but clinically relevant resistance is rarely observed. Here we have tested whether RamA overproduction can enhance acquired β-lactam resistance mechanisms in K. pneumoniae and have defined the envelope protein abundance changes upon RamA overproduction during growth in low and high osmolarity media. Envelope permeability was estimated using a fluorescent dye accumulation assay. β-Lactam susceptibility was measured using disc testing. Total envelope protein production was quantified using LC-MS/MS proteomics and transcript levels were quantified using real-time RT-PCR. RamA overproduction enhanced β-lactamase-mediated β-lactam resistance, in some cases dramatically, without altering β-lactamase production. It increased production of efflux pumps and decreased OmpK35 porin production, though micF overexpression showed that OmpK35 reduction has little impact on envelope permeability. A survey of K. pneumoniae bloodstream isolates revealed ramA hyperexpression in 3 of 4 carbapenemase producers, 1 of 21 CTX-M producers and 2 of 19 strains not carrying CTX-M or carbapenemases. Whilst RamA is not a key mediator of antibiotic resistance in K. pneumoniae on its own, it is potentially important for enhancing the spectrum of acquired β-lactamase-mediated β-lactam resistance. LC-MS/MS proteomics analysis has revealed that this enhancement is achieved predominantly through activation of efflux pump production.


Competition-based screening helps to secure the evolutionary stability of a defensive microbiome.
The cuticular microbiomes of Acromyrmex leaf-cutting ants pose a conundrum in microbiome biology because they are freely colonisable, and yet the prevalence of the vertically transmitted bacteria Pseudonocardia, which contributes to the control of Escovopsis fungus garden disease, is never compromised by the secondary acquisition of other bacterial strains. Game theory suggests that competition-based screening can allow the selective recruitment of antibiotic-producing bacteria from the environment, by providing abundant resources to foment interference competition between bacterial species and by using Pseudonocardia to bias the outcome of competition in favour of antibiotic producers. Here, we use RNA-stable isotope probing (RNA-SIP) to confirm that Acromyrmex ants can maintain a range of microbial symbionts on their cuticle by supplying public resources. We then used RNA sequencing, bioassays, and competition experiments to show that vertically transmitted Pseudonocardia strains produce antibacterials that differentially reduce the growth rates of other microbes, ultimately biassing the bacterial competition to allow the selective establishment of secondary antibiotic-producing strains while excluding non-antibiotic-producing strains that would parasitise the symbiosis. Our findings are consistent with the hypothesis that competition-based screening is a plausible mechanism for maintaining the integrity of the co-adapted mutualism between the leaf-cutting ant farming symbiosis and its defensive microbiome. Our results have broader implications for explaining the stability of other complex symbioses involving horizontal acquisition.


Strategies to manage permanent non-vital teeth with open apices: a clinical update.
If dental pulp injury occurs prior to complete root formation and apical closure, normal root development is halted. This condition produces several complications. Firstly, the apical diameter of the canal is often larger than the coronal diameter, so debridement is difficult. Secondly, the lack of an apical stop makes the obturation in all dimensions virtually impossible. And finally, the thin walls of the root canal are prone to fracture, so that surgical treatment is generally not a viable option. There are a number protocols to manage non-vital open-apex teeth such as apexification, apical barrier technique (one-visit apexification), orthograde root filling using MTA, triple antibiotic paste, and tissue engineering concept. The aim of this paper is to review these treatment protocols.


The TIR-domain containing effectors BtpA and BtpB from Brucella abortus impact NAD metabolism.
Brucella species are facultative intracellular Gram-negative bacteria relevant to animal and human health. Their ability to establish an intracellular niche and subvert host cell pathways to their advantage depends on the delivery of bacterial effector proteins through a type IV secretion system. Brucella Toll/Interleukin-1 Receptor (TIR)-domain-containing proteins BtpA (also known as TcpB) and BtpB are among such effectors. Although divergent in primary sequence, they interfere with Toll-like receptor (TLR) signaling to inhibit the innate immune responses. However, the molecular mechanisms implicated still remain unclear. To gain insight into the functions of BtpA and BtpB, we expressed them in the budding yeast Saccharomyces cerevisiae as a eukaryotic cell model. We found that both effectors were cytotoxic and that their respective TIR domains were necessary and sufficient for yeast growth inhibition. Growth arrest was concomitant with actin depolymerization, endocytic block and a general decrease in kinase activity in the cell, suggesting a failure in energetic metabolism. Indeed, levels of ATP and NAD+ were low in yeast cells expressing BtpA and BtpB TIR domains, consistent with the recently described enzymatic activity of some TIR domains as NAD+ hydrolases. In human epithelial cells, both BtpA and BtpB expression reduced intracellular total NAD levels. In infected cells, both BtpA and BtpB contributed to reduction of total NAD, indicating that their NAD+ hydrolase functions are active intracellularly during infection. Overall, combining the yeast model together with mammalian cells and infection studies our results show that BtpA and BtpB modulate energy metabolism in host cells through NAD+ hydrolysis, assigning a novel role for these TIR domain-containing effectors in Brucella pathogenesis.


Effects of thioacetamide treatment on nuclear envelope nucleoside triphosphatase activity and transport of RNA from rat liver nuclei.
An in vitro system was used for investigating transport of rapidly labeled RNA from liver nuclei isolated from control and thioacetamide-treated rats. An enhanced transport was observed in preparations from thioacetamide-treated animals. Analysis of transport at various temperatures indicated that it proceeded as an energy-requiring process, even in the absence of an exogenously added energy source, and was related to hydrolysis of high-energy nucleoside triphosphate esters. In control preparations, a nucleoside triphosphatase (NTPase) is present in nuclear envelopes, and this TNPase activity appears to be intimately related to in vitro RNA transport. Nuclear membranes were isolated from purified nuclear preparations taken from control and thioacetamide-treated rats and were examined for NPTase activity. The NTPase activity was significantly greater in the preparations from thioacetamide-treated animals. These observations indicate that an early change associated with carcinogen exposure is an enhanced NTPase activity. This may underlie the enhanced RNA transport observed in vitro.


Bowditch Lecture. Renal afferent inputs to ascending spinal pathways.
Studies of renal afferent fibers and their functions have continued since the work of Pines in 1959 (Fiziol. Zh. SSSR Im. I M Sechenova 45: 1339-1347, 1959). The kidney contains mechanoreceptors and chemoreceptors that appear to have two major functions. First, renal mechano- and chemoreceptors evoke a variety of renorenal reflexes, while more global cardiovascular reflexes are primarily evoked by renal mechanoreceptors. A second function of renal afferent fibers is to cause the pain of renal disease. Recent studies suggest that renal afferent fibers may also regulate secretion of vasopressin from the pituitary gland. Substantial evidence indicates that, although most renal afferent fibers enter the spinal cord, their functions depend to a large extent on supraspinal circuitry. Thus our research has focused on defining characteristics of spinal neurons that relay renal information to the brain. In the cat, neurons in the L2-T11 segments with excitatory responses to renal A delta and C fiber input project to the medial medullary reticular formation and to the caudal and rostral ventrolateral medulla. Renal afferent information reaches these cells by way of the least splanchnic nerve and by way of more than one dorsal root. In the monkey spinothalamic neurons in the L3-T10 segments respond to renal nerve stimulation. Excitatory responses predominate, but inhibitory responses occur in L2 and L3. These cells also respond to renal A delta and C fibers. Stimulation of renal mechanoreceptors by occlusion of the ureteropelvic junction or renal vein excites feline spinoreticular neurons. Graded increases in renal vein pressure produce graded increases in cell responses. Activation of renal chemoreceptors increases activity of spinal interneurons. Within the L2-T11 segments, cells responding to ureteral occlusion are located caudally, cells with responses to renal artery occlusion are located rostrally, and cells responding to renal vein occlusion are located in between. The differential locations of cells with these inputs suggests the existence of a coding mechanism for different renal receptor populations. Distention of the renal pelvis is a potent stimulator of primate spinothalamic neurons. These neurons encode renal pelvic pressures in the noxious range and appear to be important in mechanisms of renal pain.


TET2 and CSMD1 genes affect SBP response to hydrochlorothiazide in never-treated essential hypertensives.
Thiazide diuretics have been recommended as a first-line antihypertensive treatment, although the choice of 'the right drug in the individual essential hypertensive patient' remains still empirical. Essential hypertension is a complex, polygenic disease derived from the interaction of patient's genetic background with the environment. Pharmacogenomics could be a useful tool to pinpoint gene variants involved in antihypertensive drug response, thus optimizing therapeutic advantages and minimizing side effects. We looked for variants associated with blood pressure response to hydrochlorothiazide over an 8-week follow-up by means of a genome-wide association analysis in two Italian cohorts of never-treated essential hypertensive patients: 343 samples from Sardinia and 142 from Milan. TET2 and CSMD1 as plausible candidate genes to affect SBP response to hydrochlorothiazide were identified. The specificity of our findings for hydrochlorothiazide was confirmed in an independent cohort of essential hypertensive patients treated with losartan. Our best findings were also tested for replication in four independent hypertensive samples of European Ancestry, such as GENetics of drug RESponsiveness in essential hypertension, Genetic Epidemiology of Responses to Antihypertensives, NORdic DILtiazem intervention, Pharmacogenomics Evaluation of Antihypertensive Responses, and Campania Salute Network-StayOnDiur. We validated a polymorphism in CSMD1 and UGGT2. This exploratory study reports two plausible loci associated with SBP response to hydrochlorothiazide: TET2, an aldosterone-responsive mediator of αENaC gene transcription; and CSMD1, previously described as associated with hypertension in a case-control study.


Allergic manifestations in very low-birthweight infants: a 6-year follow-up.
The incidence of allergic manifestations was evaluated from birth until 6 y of age in 83 very low-birthweight infants (VLBWIs). In the same period 98 full-term babies were followed from birth to 24 mo of life. All the subjects were examined by paediatricians to establish the presence of atopic dermatitis (AD), gastrointestinal disturbances (GD) and asthma (AS). The incidence of total allergic manifestations (31.3%) in VLBWIs was significantly lower than that (52%) in 24-mo-old infants, born at full term. The incidence of allergies in VLBWIs did not differ at all at the subsequent checks, up to 6 y of age. AD (33.7%) was the most common symptom, statistically higher in full-term infants than in VLBWIs (7.2%). GD had a similar distribution (8.2% in full-term infants vs 7.2% in VLBWIs). AS (16.8%) was significantly higher in VLBWIs than in those born full term (10.2%). In the various VLBWI subgroups analysed, AD was more prevalent in babies weighing >1000 g and in babies >30 wk of age; the incidence of GD was higher in infants weighing <1000 g and in SGA infants, and AS was more prevalent in infants weighing <1000 g, in infants <30 wk of age and in babies appropriate for gestational age. A family history of allergy was related to a major incidence of allergies.


Toll-like receptor-7 agonist decoration enhances the adjuvanticity of chitosan-DNA nanoparticles.
In order to provide an adjuvant-equipped carrier system for plasmid deoxyribonucleic acid (pDNA) vaccines, we grafted for the first time a Toll-like receptor (TLR)-7 agonistic moiety [9-benzyl-8-hydroxyadenine (HA)] through a poly(ethylene glycol) (PEG) spacer onto a water-soluble chitosan derivative [final copolymer: 6-0-carboxymethyl-N,N,N-trimethylchitosan (CTC)-graft-PEG-HA (CTCPHA)]. Successful grafting was confirmed by spectroscopic (H NMR, mass, ultraviolet-visible, and Fourier transform infrared spectroscopy) and chromatographic (size-exclusion chromatography-multi-angle laser light scattering) methods. In this article, TLR-7 agonist-decorated CTCPHA nanoparticles (NPs) were formulated by complex coacervation with pDNA expressing the green fluorescence protein. Resulting NPs had a size of around 200 nm with a positive surface charge and high DNA encapsulation efficiency. In contrast to the use of DNA alone, NP protected DNA against enzymatic degradation and enabled transfection of alveolar A549 cells. Interestingly, TLR-7 agonist decoration increased significantly the interleukin-8-related immune stimulatory capacity of polymeric chitosan and chitosan-based NP in human THP-1 macrophages when compared with controls. In summary, we demonstrate here the proof-of-principle that covalent TLR-7 agonist functionalization of chitosan-DNA NPs enhances the carrier's adjuvanticity, representing a valuable concept for future polymer-based DNA vaccination.


Associations between malignancy and cardiovascular complications following emergency laparotomy - A retrospective cohort study.
Systemic inflammation in patients with malignant disease has been associated with increased risk of cardiovascular events. The pro-inflammatory perturbations following surgical trauma may further promote adverse perioperative cardiovascular events and increase the risk of patients with cancer undergoing major surgery. Our objective was to estimate the association between malignant disease and postoperative cardiovascular complications. Secondarily, we aimed to identify risk factors for postoperative cardiovascular complications. We conducted a retrospective cohort study of all patients ≥18 years undergoing emergency laparotomy between 2010 and 2016 at Department of Surgery, Zealand University Hospital, Denmark. Complications were graded according to Clavien-Dindo classification of surgical complications. Multivariate logistic regression analysis was performed to estimate association between malignant disease and cardiovascular complications within 30 days of emergency laparotomy and to identify other risk factors for postoperative cardiovascular complications after emergency laparotomy. We identified 1188 patients ≥18 years undergoing emergency laparotomy between 2010 and 2016, in which 254 (21%) had malignant disease. Within 30 days of emergency laparotomy, 89 (9.5%) of patients without malignancy died, as compared with 45 (18%) of patients with malignancy (p < 0.001). Cardiovascular death occurred in 17 (1.8%) and 5 (2.0%) patients in the non-malignant and malignant group, respectively. Severe cardiovascular complication graded CD 3-5 occurred in 93 (8%) of all patients within 30 days of emergency laparotomy. We found no association between malignancy and postoperative cardiovascular complications in patients undergoing emergency surgery (OR 0.8, 95% CI; 0.4, 1.5). Increasing age and ASA physical status classification system (ASA) score ≥ III were the only independent risk factors of cardiovascular complications graded CD 3-5. Malignancy was not associated with postoperative cardiovascular complications after emergency laparotomy. Risk factors for major cardiovascular complications after emergency abdominal surgery were age and ASA score ≥ III.


Reciprocal influence of the p53 and the hypoxic pathways.
When cells sense a decrease in oxygen availability (hypoxia), they develop adaptive responses in order to sustain this condition and survive. If hypoxia lasts too long or is too severe, the cells eventually die. Hypoxia is also known to modulate the p53 pathway, in a manner dependent or not of HIF-1 (hypoxia-inducible factor-1), the main transcription factor activated by hypoxia. The p53 protein is a transcription factor, which is rapidly stabilised by cellular stresses and which has a major role in the cell responses to these stresses. The aim of this review is to compile what has been reported until now about the interconnection between these two important pathways. Indeed, according to the cell line, the severity and the duration of hypoxia, oxygen deficiency influences very differently p53 protein level and activity. Conversely, p53 is also described to affect HIF-1α stability, one of the two subunits of HIF-1, and HIF-1 activity. The direct and indirect interactions between HIF-1α and p53 are described as well as the involvement in this complex network of their respective ubiquitin ligases von Hippel Lindau protein and murine double minute 2. Finally, the synergistic or antagonistic effects of p53 and HIF-1 on some important cellular pathways are discussed.


Glucose transporter-1 (GLUT-1): a potential marker of prognosis in rectal carcinoma?
The aim of the study is to evaluate the pattern and level of expression of glucose transporter-1 (GLUT-1) in rectal carcinoma in relation to outcome as a potential surrogate marker of tumour hypoxia. Formalin-fixed tumour sections from 43 patients with rectal carcinoma, who had undergone radical resection with curative intent, were immunohistochemically stained for GLUT-1. A mean of three sections per tumour (range 1-12) were examined. Each section was semiquantitatively scored; 0, no staining; 1, <10%; 2, 10-50%; 3, >50% and a score given for the whole section, the superficial (luminal) and deep (mural) part of the tumour. Staining was seen in 70% of tumours. Increased staining was noted adjacent to necrosis and ulceration. A diffuse and patchy pattern of staining, with and without colocalisation to necrosis was seen. Patients with high GLUT-1-expressing tumours (score 3 vs 0-2) had a significantly poorer overall survival (P=0.041), which was associated with poorer metastasis-free survival with no difference in local control. No significant correlation was seen with other prognostic factors. There was a strong correlation between the score for the superficial and deep parts of the tumour (r=0.81), but a significant relationship with outcome was only found in the deep part (P=0.003 vs P=0.46). In conclusion, increased GLUT-1 expression in rectal tumours was an adverse prognostic factor and is worth further evaluation as a predictive marker of response to therapy.


Activities of hexokinase, phosphofructokinase, 3-oxo acid coenzyme A-transferase and acetoacetyl-coenzyme A thiolase in nervous tissue from vertebrates and invertebrates.
1. The maximum activities of hexokinase and phosphofructokinase in nervous tissue from 18 different animals from different phyla range from 5.1 to 17.6 and from 24.0mumol/min per g fresh wt. respectively. In any one tissue the activities of these two enzymes are, in general, very similar. The rate of glucose utilization by the brain in vivo is much lower than the activities of hexokinase or phosphofructokinase. It is suggested that the high activities of these enzymes indicate a capacity for glycolysis which may be used by the brain during hypoxia or during conditions of extreme neuronal activity. 2. The activities of 3-oxo acid CoA-transferase and acetoacetyl-CoA thiolase in the nervous tissues range from 1.1 to 15.3 and from 0.7 to 4.5mumol/min per g fresh wt. respectively. Unfortunately the activities of these enzymes cannot be used to estimate maximal flux through the ketone-body-utilization pathway, since they may catalyse reactions that are close to equilibrium. Nonetheless, the presence of these enzymes in nervous tissue from a large variety of animals suggests that the importance of ketone bodies as a fuel for nervous tissue may be widespread in the animal kingdom.


Temporally stable rotational coherent states for molecular simulations. I. Spherical and linear rotor cases.
We reformulate a previous rotational coherent state (CS) to obtain temporally stable (TS) CSs for the spherical rotor (SR) and linear rotor (LR): TSSR and TSLR CSs, respectively. Being TS, the new CSs remain within their own classes during dynamics by evolving exclusively through their CS parameters. The new TS CSs are now appropriate to reconstruct quantum rotational properties from classical-mechanics simulations of chemical reactions. Following literature precedents, we enforce temporal stability by incorporating action-angle-related phase factors into the parameters of the original CS. In addition, to elucidate CS quantum reconstruction procedures, we derive one more rotational CS from a quantum electron nuclear dynamics description of a diatomic rotor (DR). The DR CS and the TSLR CS are not identical but display similar structures and properties. We rigorously demonstrate and examine the key properties of the three CSs: continuity, resolution of unity, temporal stability, action identity, minimum uncertainty relationships, and quasi-classical behavior. Finally, we present computer simulations of the CSs dynamics and an application of them to predict CO rotational excitation probabilities in the Li+ + CO reaction. CS results agree satisfactorily with experimental ones and encourage future applications in chemical dynamics, statistical mechanics, spectroscopy, nuclear physics, quantum coherence, and quantum computing.


Agglutination of African primate and rodent erythrocytes by adenoviruses, reoviruses, and enteroviruses.
African nonhuman primate and rodent erythrocytes were tested for agglutination by adenoviruses, reoviruses, and enteroviruses. Squirrel erythrocytes were agglutinated by reovirus serotypes and adenovirus types 3, 11, 16, and 21. Adenoviruses also agglutinated brazza monkey erythrocytes to the same titers as those obtained with either rhesus or grey monkey cells. Prototype reovirus types 1 and 2 agglutinated grey monkey erythrocytes to much lower titers than either squirrel or human group O red cells. Among the enteroviruses tested, only echovirus types 7 and 12 agglutinated grey, red-tail, brazza, and rhesus monkey erythrocytes. The specificity of agglutination of squirrel, grey, and brazza monkey erythrocytes by reoviruses, echoviruses, and adenoviruses, respectively, was confirmed by hemagglutination-inhibition tests. The titers obtained were similar to those obtained with erythrocytes usually used in these tests. Erythrocytes of bush babies, potto unstriped grass mice, swamp rat, rusty-nosed rat, bush rat, harsh-furred mice, soft-furred rat, and giant rat were not agglutinated by adenoviruses, reoviruses, or enteroviruses.


Transcriptomic analysis identifies organ-specific metastasis genes and pathways across different primary sites.
Metastasis is the most devastating stage of cancer progression and often shows a preference for specific organs. To reveal the mechanisms underlying organ-specific metastasis, we systematically analyzed gene expression profiles for three common metastasis sites across all available primary origins. A rank-based method was used to detect differentially expressed genes between metastatic tumor tissues and corresponding control tissues. For each metastasis site, the common differentially expressed genes across all primary origins were identified as organ-specific metastasis genes. Pathways enriched by these genes reveal an interplay between the molecular characteristics of the cancer cells and those of the target organ. Specifically, the neuroactive ligand-receptor interaction pathway and HIF-1 signaling pathway were found to have prominent roles in adapting to the target organ environment in brain and liver metastases, respectively. Finally, the identified organ-specific metastasis genes and pathways were validated using a primary breast tumor dataset. Survival and cluster analysis showed that organ-specific metastasis genes and pathways tended to be expressed uniquely by a subgroup of patients having metastasis to the target organ, and were associated with the clinical outcome. Elucidating the genes and pathways underlying organ-specific metastasis may help to identify drug targets and develop treatment strategies to benefit patients.


A Cam Morphology Develops in the Early Phase of the Final Growth Spurt in Adolescent Ice Hockey Players: Results of a Prospective MRI-based Study.
Cam morphologies seem to develop with an increased prevalence in adolescent boys performing high-impact sports. The crucial question is at what age the cam morphology actually develops and whether there is an association with an aberration of the shape of the growth plate at the cam morphology site. (1) What is the frequency of cam morphologies in adolescent ice hockey players, and when do they appear? (2) Is there an association between an extension of the physeal growth plate and the development of a cam morphology? (3) How often do these players demonstrate clinical findings like pain and lack of internal rotation? A prospective, longitudinal MRI study was done to monitor the proximal femoral development and to define the appearance of cam morphologies in adolescent ice hockey players during the final growth spurt. Young ice hockey players from the local boys' league up to the age of 13 years (mean age 12 ± 0.5 years) were invited to participate. From 35 players performing on the highest national level, 25 boys and their parents consented to participate. None of these 25 players had to be excluded for known disease or previous surgery or hip trauma. At baseline examination as well as 1.5 and 3 years later, we performed a prospective noncontrast MRI scan and a clinical examination. The three-dimensional morphology of the proximal femur was assessed by one of the authors using radial images of the hip in a clockwise manner. The two validated parameters were: (1) the alpha angle for head asphericity (abnormal > 60°) and (2) the epiphyseal extension for detecting an abnormality in the shape of the capital physis and a potential correlation at the site of the cam morphology. The clinical examination was performed by one of the authors evaluating (1) internal rotation in 90° of hip and knee flexion and (2) hip pain during the anterior impingement test. Cam morphologies were most apparent at the 1.5-year follow-up interval (10 of 25; baseline versus 1.5-year follow-up: p = 0.007) and a few more occurred between 1.5 and 3 years (12 of 23; 1.5-year versus 3-year follow-up: p = 0.14). At 3-year follow-up, there was a positive correlation between increased epiphyseal extension and a high alpha angle at the anterosuperior quadrant (1 o'clock to 3 o'clock) (Spearman correlation coefficient = 0.341; p < 0.003). The prevalence of pain on the impingement test and/or restricted internal rotation less than 20° increased most between 1.5-year (1 of 25) and the 3-year follow-up (6 of 22; 1.5-year versus 3-year follow-up: p = 0.02). Our data suggest that a cam morphology develops early during the final growth spurt of the femoral head in adolescent ice hockey players predominantly between 13 to 16 years of age. A correlation between an increased extension of the growth plate and an increased alpha angle at the site of the cam morphology suggests a potential underlying growth disturbance. This should be further followed by high-resolution or biochemical MRI methods. Considering the high number of cam morphologies that correlated with abnormal clinical findings, we propose that adolescents performing high-impact sports should be screened for signs of cam impingement, such as by asking about hip pain and/or examining the patient for limited internal hip rotation. Level I, prognostic study.


The Effect of RANKL/OPG Balance on Reducing Implant Complications.
Despite the phenomenal success of implants particularly in the realms of dentistry and orthopaedics, there are still challenges to overcome. The failure of implants resulting from infection, prosthetic loosening, and non-union continue to be the most notorious examples. The cascade of fracture healing and bone repair, especially with the presence of an implant, is complex because it involves a multifaceted immune response alongside the intricate process of bone formation and remodelling. Bone loss is a serious clinical problem that is frequently accompanied by chronic inflammation, illustrating that there is a convoluted relationship between inflammation and bone erosion. The effects of pro-inflammatory factors play a significant role in initiating and maintaining osteoclastogenesis that results in bone resorption by osteoclasts. This is because there is a disruption of the relative ratio between Receptor Activator of Nuclear Factor κB-Ligand (RANKL) and osteoprotegerin (OPG), which is central to modulating bone repair and remodelling. This review aims to provide a background to the bone remodelling process, the bone repair cascade post-implantation, and the associated complications. Furthermore, current clinical solutions that can influence bone formation via either internal or extrinsic mechanisms will be described. These efficacious treatments for osteolysis via targeting the RANKL/OPG ratio may be crucial to reducing the incidence of related implant failures in the future.


Stroke risk among adult patients with third, fourth or sixth cranial nerve palsy: a Nationwide Cohort Study.
This study sought to determine whether isolated third, fourth and sixth cranial nerve palsies (NPs) are associated with increased short- and long-term risk of a subsequent stroke. This was a nationwide retrospective propensity score-matched cohort study. A cohort of patients with NP (n = 466) and a randomly selected, propensity-matched control cohort (n = 2281) were extracted from the Korean national insurance claim database. Subjects were tracked for 5 years total, subdivided into periods of 0-1 years, 1-3 years and 3-5 years. We assessed the risk of stroke using hazard ratios (HRs) and confidence intervals (CIs) after adjustments using Cox regression at different time intervals. The median follow-up was 3.1 years. Stroke developed in 18.9% of the NP cohort and 7.5% of the control cohort. Stroke risk after NP was highest in the first year [14.7 per 100 person-year at 0-1 years (HR = 6.6), 3.1 per 100 person-year at 1-3 years (HR = 1.6) and 4.3 per 100 person-year at 3-5 years (HR = 2.8)]. Each type of NP was also associated with stroke risk: within 0-1 years, stroke risk was increased in third (HR = 7.6), fourth (HR = 6.0) and sixth (HR = 5. 84) NPs. In the 3- to 5-year period, risk was increased in sixth (HR = 4.7) and fourth (HR = 3.3) NPs, but not third (HR = 0.6) NPs. Patients in the NP cohort were more likely to have a stroke than those in the matched control cohort; the increased risk was both time- and cranial nerve-dependent.


BRAFV600E cooperates with PI3K signaling, independent of AKT, to regulate melanoma cell proliferation.
Mutationally activated BRAF(V600E) cooperates with PTEN silencing in the conversion of normal melanocytes to metastatic melanoma cells, but the mechanism underlying this cooperation is poorly understood. Here, the consequences of pharmacologic blockade of BRAF(V600E) or phosphoinositide 3-kinase (PI3K) signaling were explored using pathway-targeted inhibitors and a panel of human BRAF-mutated melanoma-derived cell lines. Blockade of BRAF(V600E) → MEK1/2 → ERK1/2 or class I PI3K inhibited melanoma proliferation, whereas inhibition of AKT had only modest effects, even in cells with mutated or amplified AKT. Although single-agent inhibition of either BRAF(V600E) or PI3K signaling elicited antiproliferative effects, combinatorial inhibition was more potent. Analysis of signaling downstream of BRAF(V600E) or PI3K revealed that these pathways cooperated to regulate protein synthesis through AKT-independent, mTOR complex 1 (mTORC1)-dependent effects on p70(S6K), ribosomal protein S6, and 4E-BP1 phosphorylation. Moreover, inhibition of mTORC1/2 inhibited cell proliferation as profoundly as single-agent inhibition of either BRAF(V600E) or PI3K signaling. These data reveal a mechanism by which BRAF(V600E) and PI3K signaling cooperate to regulate melanoma proliferation through AKT-independent effects on protein translation. Furthermore, this study provides a potential foundation for pathway-targeted combination therapy designed to enhance the therapeutic benefit to patients with melanoma that contain combined alterations in BRAF and PI3K signaling. PI3K, but not AKT, represent potential targets for melanoma therapy.


Agonism of Gpr40 Protects the Capacities of Epidermal Stem Cells (ESCs) Against Ultraviolet-B (UV-B).
Skin damage due to overexposure to ultraviolet B (UV-B) radiation can lead to the development of cancers and reduce the skin's functionality as a vital protective barrier. Epidermal stem cells (ESCs) are pluripotent cells responsible for skin regeneration and healing. Upon exposure to UV-B radiation, ESCs produce excess amounts of reactive oxygen species (ROS) and inflammatory cytokines. However, the functional protection of ESCs is not fully explored. G-protein coupled G protein-coupled receptor 40 (Gpr40) is a free fatty acid receptor that is emerging as a potential treatment target for various diseases. Gpr40 has been found to be expressed in various cell types. ESCs were exposed to UV-B at the intensities of 25, 50, and 100 mJ/cm2 for 24 h using TL 20 W/12 RS UV lamps. ESCs were treated with UV-B at 50 mJ/cm2 in the presence or absence of 25 or 50 µM of the Gpr40 agonist GW9508 for 24 h. The gene expression of the Wnt1 pathway and proinflammatory cytokines were evaluated. To antagonize Gpr40 expression, ESCs were treated with 10 µM GW1100. Our findings demonstrate that Gpr40 agonism can reduce the production of ROS as well as the expression of interleukins 1β and 8, two key proinflammatory cytokines. We demonstrate that agonism of Gpr40 can rescue the reduction in integrin β1 and Krt19 induced by UV-B exposure, thereby improving the capacities of ESCs to resist UV-B damage. Moreover, we show that the effects of Gpr40 agonism observed in our experiments are mediated through the Wnt/β-catenin canonical signaling pathway, as evidenced by the expression of Wnt1 and cyclin D1. Our findings present evidence of the role of Gpr40 agonism in mediating the protective capacities of ESCs against insult from UV-B radiation.


The use of census data for determining race and education as SES indicators: a validation study.
Little research has examined the validity of using census data to determine an individual's socio-economic status (SES), as measured by race and educational level. This study assessed the accuracy of using aggregate level data from United States Census Block Groups in determining race and education SES indicators in a cohort of women from North Carolina. The study analyzed patient data from the Carolina Mammography Registry and 1990 United States Census in 21 North Carolina counties. Women (n = 39,546) were geocoded to their census block group and their block group characteristics (surrogate measures) were validated with their self-reported values on race and education. An analysis was performed to explore whether using these surrogate measures would affect measured associations with the self-reported values. Whites were accurately identified (84.8%) more consistently than Blacks (14.1%) regardless of their urban/rural status. Women without a high school diploma or equivalent were accurately identified (56.2%) more often than those with higher education levels (45.9%). Analyses using the surrogate measures were significantly different than the true values according to chi-square statistics. Use of census data to derive SES indicators tends to be more accurate for the majority than the minority population. Researchers must be sensitive to the ecologic fallacy when using aggregate level data such as the census to determine individual level characteristics.


[Heart involvement in multiple myeloma].
The results of the myocardial functional capacity study in 54 patients with multiple myeloma were compared with the data of clinical and anatomopathologic investigations. No specific clinico-functional signs of cardiac affection in multiple myeloma, having differential diagnostic importance, were revealed. Specific affection of the heart was detected in 30% of cases during anatomopathologic investigations.


Regulation of the program of DNA replication by CDK: new findings and perspectives.
Progression through the cell cycle is driven by the activities of the cyclin-dependent kinase (CDK) family of enzymes, which establish an ordered passage through the cell cycle phases. CDK activity is crucial for the cellular transitions from G1 to S and G2 to M, which are highly controlled to promote the faithful duplication of the genetic material and the transmission of the genome into daughter cells, respectively. While oscillations in CDK activity are essential for cell division, how its specific dynamics may shape cellular processes remains an open question. Recently, we have investigated the potential role of CDK in establishing the profile of replication initiation along the chromosomes, also referred to as the replication program. Our results demonstrated that the timing and level of CDK activity at G1/S provide two critical and independent inputs that modulate the pattern of origin usage. In this review, we will present the conclusions of our study and discuss the implications of our findings for cellular function and physiology.


Spectral analysis of arterial pressure lability in rats with sinoaortic deafferentation.
Traditionally, the standard deviation (SD) of the mean arterial pressure (AP) has been used as an index for the AP lability produced by interruption of baroreceptor afferents. Although a useful measure of variance about the mean, the SD does not provide any information about the temporal characteristics of this variability. We employed two different spectral analytic techniques to characterize AP waveforms in rats with sinoaortic deafferentation (SAD) and in sham-operated (Sham) rats to determine if the AP waveform in SAD animals was qualitatively and/or quantitatively different from that of Sham animals. The SAD and Sham animals exhibited qualitatively different spectral profiles, suggesting that lability of AP in SAD animals is not simply an exaggeration of normal fluctuations. In addition, a low-frequency (0.3-0.5 Hz) spectral peak was found in Sham but not SAD animals, suggesting that it is associated with the baroreflex. Finally, we observed in both normal rats and rats without intact baroreceptors that the spectral components of AP are not static but rather vary continuously across time.


Surface-Enhanced Raman Spectroscopy: A New Modality for Cancer Imaging.
Although surface-enhanced Raman scattering (SERS) spectroscopy has traditionally been used as an in vitro analytic tool, in the past few years the first reports of the feasibility of in vivo imaging of cancer with biocompatible SERS probes have emerged. SERS imaging has great potential in the field of medical imaging because it offers several major advantages over other molecular imaging methods. Medical imaging using SERS nanoprobes can yield higher sensitivity and higher signal specificity than other imaging modalities, while also offering multiplexing capabilities that allow for unique applications. This article reviews the principles of SERS and highlights recent advances for in vivo cancer imaging. To present the abilities of this method as accurately as possible, the discussion is limited to studies in which the imaging data were confirmed by histological correlation.


Activated carbon adsorption of trichloroethylene (TCE) vapor stripped from TCE-contaminated water.
Ground water contaminated with trichloroethylene (TCE) used in electronic, electric, dry cleaning and the like industries is often treated by air-stripping. In this treatment process, TCE in its vapor form is stripped from ground water by air stream and sometimes emitted into the atmosphere without any additional treatments. Activated carbon adsorption is one of the practical and useful processes for recovering the TCE vapor from the exhaust air stream. However, adsorption of the TCE vapor from the stripping air stream onto activated carbons is not so simple as that from dry air, since in the exhaust air stream the TCE vapor coexists with water vapor with relatively high concentrations. The understanding of the adsorption characteristics of the TCE vapor to be adsorbed on activated carbon in the water vapor-coexisting system is essential for successfully designing and operating the TCE recovery process. In this work, the adsorption equilibrium relations of the TCE vapor adsorption on activated carbons were elucidated as a function of various relative humidity. Activated carbon fibers (ACFs) were used as model activated carbon. The adsorption equilibrium relations were studied by the column adsorption method. The adsorption isotherms of TCE vapor adsorbed on sample ACFs were successfully correlated by the Dubinin-Radushkevich equation for both cases with and without coexistent water vapor. No effects of coexistent water vapor were found on the limiting adsorption volume. However, the adsorption characteristic energy was significantly reduced by the coexistence of water vapor and its reduction was successfully correlated with the equilibrium amount of water vapor adsorbed under the dynamic condition.


Activation of the nucleotide oligomerization domain signaling pathway by the non-bacterially derived xanthone drug 5'6-dimethylxanthenone-4-acetic acid (Vadimezan).
The cytosolic nucleotide-binding oligomerization domain 1 (NOD1)/CARD4 and NOD2/CARD15 proteins are members of NOD-like receptors recognizing specific motifs within peptidoglycans of both Gram-negative and Gram-positive bacteria. NOD1 and NOD2 signal via the downstream adaptor serine/threonine kinase RIP2/CARDIAK/RICK to initiate NF-kappaB activation and the release of inflammatory cytokines/chemokines. In this report, we show that 5,6-dimethylxanthenone-4-acetic acid (DMXAA), a cell-permeable, small molecule that has anti-tumor activity, can also activate NOD1 and NOD2. This was demonstrated: 1) by using human embryonic kidney epithelial (HEK) 293 cells transfected with a NF-kappaB reporter plasmid in combination with NOD1 or NOD2 expression plasmids; 2) by inhibiting DMXAA-induced chemokine (CXCL10) mRNA and protein production in the AB12 mesothelioma cell line using a pharmacological inhibitor of RICK kinase, SB20358; and 3) by using small interfering RNA to knock down NOD2 and lentiviral short hairpin RNA to knock down RICK. These findings expand the potential ligands for the NOD-like receptors, suggesting that other xanthone compounds may act similarly and could be developed as anti-tumor agents. This information also expands our knowledge on the mechanisms of action of the anti-tumor agent DMXAA (currently in clinical trials) and may be important for its biological activity.


Differences in heterocycle basicity distinguish homocysteine from cysteine using aldehyde-bearing fluorophores.
We report the detection of homocysteine over cysteine based upon characteristic differences between 5- and 6-membered heterocyclic amines formed upon reaction with aldehyde-bearing compounds. Homocysteine-derived thiazinane-4-carboxylic acids are more basic than cysteine-derived thiazolidines-4-carboxylic acids. Fluorescence enhancement in response to homocysteine is achieved by tuning pH and excitation wavelength.


Effect of massage therapy with lavender oil on severity of restless legs syndrome and quality of life in hemodialysis patients.
This study assessed the effects of massage therapy using 5% lavender oil on the severity of restless legs syndrome (RLS) and the quality of life (QoL) of patients on hemodialysis (HD). This is a randomized placebo-controlled study with a pretest-posttest design. This study was conducted from January 30, 2019, to May 6, 2019, at HD centers in Turkey, and it includes 58 participants-31 study patients and 27 controls. Data were collected using patient identification form, RLS severity rating scale, Kidney Disease Quality of Life Scale (KDQOLTM -36) and patient follow-up charts. As per the massage therapy protocol, the patients in the study and control groups received massage therapy with lavender oil and baby oil, respectively. RLS severity significantly decreased in all follow-up weeks in the study group and in the first, second, and third follow-up weeks in the control group. There were significant differences between the groups in terms of KDQOLTM -36 subscales and total scores at the initial and final follow-ups. In HD patients, massage with lavender oil lessened the severity of RLS and improved the QoL. Accordingly, this therapy can be recommended to HD patients. Massage therapy during HD sessions is easy, inexpensive, and patient-friendly with no side effects. It is known to reduce symptoms and enable the patients to easily perform daily activities of living. Massage therapy with lavender oil is effective and can be easily applied to patients with RLS by nurses. This study was registered under the Clinical Trials protocol registration system (NCT04630470) upon completion.


COYOTES (CANIS LATRANS) IN ARIZONA, USA, EXHIBIT IMMUNE AND GENETIC EVIDENCE OF RICKETTSIAL INFECTIONS.
Rocky Mountain spotted fever (RMSF), caused by the bacterium Rickettsia rickettsii, was recognized as endemic in Arizona, US after a 2002 outbreak and has since been a public health concern. The brown dog tick (Rhipicephalus sanguineus sensu lato) is the principal vector of this pathogen in Arizona. Domesticated dogs (Canis lupus familiaris) are the tick's main host, so free-roaming dogs in peridomestic areas have been named the primary risk factor for human cases of RMSF. However, the sudden emergence and long-distance dispersal of the pathogen have not been adequately explained, and one possible mechanism could include wildlife. Coyotes (Canis latrans) are wide ranging in Arizona and closely related to dogs, so it is possible that brown dog ticks parasitize coyotes and infect them. Although R. rickettsii is the most severe spotted fever group (SFG) rickettsial pathogen in humans, others occur in Arizona, and antibodies raised against them are cross-reactive, so we more-broadly hypothesized that coyotes in Arizona are exposed to SFG rickettsiae. We collected coyote tissues in spring 2016 and 2017. We tested sera for antibodies to R. rickettsii and found 9% (8/94) of samples were antibody-positive with titers of ≥256. Subsequent quantitative PCR analyses of skin showed evidence for Rickettsia spp. in 2.9% (4/138) of samples. These data suggest that coyotes have a role in the maintenance of SFG rickettsiae in Arizona. Further investigation is warranted to reveal which specific pathogen-vector complexes act on coyotes in the region and whether they represent a risk to human health.


[The role of the nucleus accumbens in psychiatric disorders].
The nucleus accumbens is the most inferior part of the striatum and is mainly connected to the limbic system. It is neurochemically and immunohistochemically divided into a shell laterally and a core medially. As a functionally central structure between amygdala, basal ganglia, mesolimbic dopaminergic regions, mediodorsal thalamus and prefrontal cortex, the nucleus accumbens appears to play a modulative role in the flow of the information from the amygdaloid complex to these regions. Dopamine is a major neurotransmitter of the nucleus accumbens and this nucleus has a modulative function to the amygdala-basal ganglia-prefrontal cortex circuit. Together with the prefrontal cortex and amygdala, nucleus accumbens consists a part of the cerebral circuit which regulates functions associated with effort. It is anatomically located in a unique way to serve emotional and behavioral components of feelings. It is considered as a neural interface between motivation and action, having a key-role in food intake, sexual behavior, reward-motivated behavior, stress-related behavior and substance-dependence. It is involved in several cognitive, emotional and psychomotor functions, altered in some psychopathology. Moreover it is involved in some of the commonest and most severe psychiatric disorders, such as depression, schizophrenia, obsessive-compulsive disorder and other anxiety disorders, as well as in addiction, including drugs abuse, alcoholism and smoking. Nucleus accumbens has also a role in other psychiatric disorders such as bipolar disorder, attention deficit/ hyperactivity disorder and post-traumatic stress disorder. Because of its rich dopaminergic projections, this nucleus has been subject of many studies in animals as well as in humans, connecting its malfunction with the disturbed reward process observed in depression. Neuromodulation interventions targeting the nucleus accumbens are nowadays applied in strictly selected patients suffering from treatment-resistant depression, obsessive-compulsive disorder, Tourette syndrome and addiction to drugs or alcohol. Specifically, bilateral and unilateral (right) deep brain stimulation of the nucleus accumbens has been applied in obsessive-compulsive patients resulting into significant improvement of their symptoms and their quality of life. Nucleus accumbens deep brain stimulation has been also associated with antidepressant and anxiolytic effect, as well as quality of life improvement in patients suffering from severe resistant depression. Finally, this minimally invasive stereotactic procedure has been proved beneficial for all phenotypic components of the Tourette syndrome, with remarkable reduction of the syndrome's motor manifestations, including tics.


Study on the Effect of Early Comprehensive Intervention of Skin Contact Combined with Breastfeeding on Improving Blood Glucose in Early Birth of Newborns with Gestational Diabetes Mellitus.
To explore the value of early comprehensive intervention of skin contact combined with breastfeeding on improving early blood glucose in newborns with gestational diabetes mellitus (GDM). A total of 300 newborns from pregnant women with gestational diabetes who were hospitalized in Wuxi People's Hospital from January 2021 to December 2021 were randomly assigned into the observation group (n = 150) and the control group (n = 150). The former group received early comprehensive intervention of skin contact combined with breastfeeding, and the latter group received postnatal naked contact, physical examination after late navel severing, and routine nursing intervention such as early contact and early sucking in 30 min. The peripheral blood microglucose value at 1 and 2 hours after birth, neonatal hospitalization rate, ear temperature of 30 min, 60 min, 90 min, and 120 min after birth, neonatal crying, incidence of postpartum hemorrhage, uterine contraction/wound pain index, lactation before delivery, immediately after delivery, early sucking 15 min, and 2 hours postpartum were observed. Compared to the control group, the values of trace blood glucose at 1 hour and 2 hours after birth in the observation group were higher, and the difference between groups was statistically significant (P < 0.05), the neonatal hospitalization rate in the observation group was lower, and the difference between groups was statistically significant (P < 0.05); the ear temperature of 30 min, 60 min, 90 min and 120 min after birth in the observation group was higher, and the difference between groups was statistically significant (P < 0.05). The crying frequency of newborns in the observation group was lower, and the difference between groups was statistically significant (P < 0.05). The incidence of postpartum hemorrhage in the observation group was lower, and the difference between groups was statistically significant (P < 0.05). The rate of uterine contraction/wound pain index grade 1 in the observation group was higher, and the difference between groups was statistically significant (P < 0.05). The rates of uterine contraction/wound pain index grade 2 and grade 3 in the observation group were lower, and the difference between groups was statistically significant (P < 0.05). The rate of lactation at 2 hours postpartum in the observation group was higher, and the difference between groups was statistically significant (P < 0.05). Early comprehensive intervention of skin contact combined with breastfeeding can significantly increase the early blood glucose of newborns with GDM, effectively promote the occurrence of early hypoglycemia of GDM newborns, avoid a series of serious complications caused by excessive fluctuation of blood sugar, promote the stability of vital signs of newborns, reduce the hospitalization rate of newborns, improve the success rate of breastfeeding, reduce uterine contraction/wound pain, and reduce the incidence of postpartum hemorrhage. My clinical registration number is chictr220059454.


Using phase II data for the analysis of phase III studies: An application in rare diseases.
Clinical research and drug development in orphan diseases are challenging, since large-scale randomized studies are difficult to conduct. Formally synthesizing the evidence is therefore of great value, yet this is rarely done in the drug-approval process. Phase III designs that make better use of phase II data can facilitate drug development in orphan diseases. A Bayesian meta-analytic approach is used to inform the phase III study with phase II data. It is particularly attractive, since uncertainty of between-trial heterogeneity can be dealt with probabilistically, which is critical if the number of studies is small. Furthermore, it allows quantifying and discounting the phase II data through the predictive distribution relevant for phase III. A phase III design is proposed which uses the phase II data and considers approval based on a phase III interim analysis. The design is illustrated with a non-inferiority case study from a Food and Drug Administration approval in herpetic keratitis (an orphan disease). Design operating characteristics are compared to those of a traditional design, which ignores the phase II data. An analysis of the phase II data reveals good but insufficient evidence for non-inferiority, highlighting the need for a phase III study. For the phase III study supported by phase II data, the interim analysis is based on half of the patients. For this design, the meta-analytic interim results are conclusive and would justify approval. In contrast, based on the phase III data only, interim results are inconclusive and require further evidence. To accelerate drug development for orphan diseases, innovative study designs and appropriate methodology are needed. Taking advantage of randomized phase II data when analyzing phase III studies looks promising because the evidence from phase II supports informed decision-making. The implementation of the Bayesian design is straightforward with public software such as R.


Specificity of protein phosphatases in the dephosphorylation of protein kinase C.
Protein kinase C can autophosphorylate in vitro and has also been shown to be phosphorylated in vivo. In order to investigate the factors that may determine the phosphorylation state of protein kinase C in vivo, we determined the ability of the ATP + Mg2+-dependent phosphatase and the polycation-stimulated (PCS) phosphatases to dephosphorylate protein kinase C in vitro. These studies show that all the oligomeric forms of the PCS phosphatases (PCSH1, PCSH2, PCSM and PCSL phosphatases) are effective in the dephosphorylation of protein kinase C, showing 34-82% of the activity displayed with phosphorylase a as substrate. In contrast both the catalytic subunit of the PCS phosphatase and that of the ATP+Mg2+-dependent phosphatase showed only weak activity with protein kinase C as substrate. All these phosphatases, however, were activated by protamine (Ka 14-16 micrograms/ml) through what appears to be a substrate-directed effect. The relative role of these phosphatases in the control of protein kinase C is discussed.


Chemical abuse in nurses. We must no longer enable, ignore, deny, or excuse this growing public and professional epidemic.
Chemical abuse is a symptom of a deep and complex physical, psychosocial, and spiritual problem which, in the nurse, can have particularly disastrous results. Clearly there is no "quick fix," nor one solution. Education is a crucial element in the prevention, early identification, and timely treatment of chemical abuse. When awareness is increased, nurses will be better prepared to recognize, prevent, and confront the growing problem of chemical abuse in the profession and in society at large.


Quantitative relaxometry and diffusion MRI for lateralization in MTS and non-MTS temporal lobe epilepsy.
We developed novel methodology for investigating the use of quantitative relaxometry (T1 and T2) and diffusion tensor imaging (DTI) for lateralization in temporal lobe epilepsy. Patients with mesial temporal sclerosis confirmed by pathology (N=8) and non-MTS unilateral temporal lobe epilepsy (N=6) were compared against healthy controls (N=19) using voxel-based analysis restricted to the anterior temporal lobes, and laterality indices for each MRI metric (T1, T2, fractional anisotropy (FA), mean diffusivity, axial and radial diffusivities) were computed based on the proportion of significant voxels on each side. The diffusivity metrics were the most lateralizing MRI metrics in MTS and non-MTS subsets, with significant differences also seen with FA, T1 and T2. Patient-specific multi-modal laterality indices were also computed and were shown to clearly separate the left-onset and right-onset patients. Marked differences between left-onset and right-onset patients were also observed, with left-onset patients exhibiting stronger laterality indices. Finally, neocortical abnormalities were found to be more common in the non-MTS patients. These preliminary results on a small sample size support the further investigation of quantitative MRI and multi-modal image analysis in clinical determination of seizure onset. The presence of more neocortical abnormalities in the non-MTS group suggests a role in seizure onset or propagation and motivates the investigation of more sensitive histopathological analysis to detect and delineate potentially subtle neocortical pathology.


Levothyroxine or minimally invasive therapies for benign thyroid nodules.
Thyroid nodules (TN) are common in the adult population. Some physicians use suppressive levothyroxine (LT4) therapy to achieve a reduction in the number and volume of TN. In addition, minimally invasive treatments, such as percutaneous ethanol injection (PEI) sclerotherapy, laser photocoagulation (LP), and microwave (MW), radiofrequency (RF) and high-intensity focused ultrasound (HIFU) ablation, have been proposed, especially for pressure symptoms and cosmetic complaints, as an alternative to surgery. However, the risk to benefit ratio of all treatments for benign TN is currently unknown. To assess the effects of LT4 or minimally invasive therapies (PEI, LP, and RF/HIFU/MW ablation) on benign TN. We identified studies from computerised searches of The Cochrane Library, MEDLINE, EMBASE and LILACS (all performed up to April 2014). We also searched trial registers, examined reference lists of included randomised controlled trials (RCTs) and systematic reviews, and contacted study authors. We included studies if they were RCTs of LT4, PEI, LP, RF, HIFU or MW therapy in participants with an established diagnosis of benign TN. We excluded trials investigating the prevention of recurrence of thyroid disease after surgery, irradiation or treatment with radioiodine. Two review authors independently extracted data, assessed studies for risk of bias and evaluated overall study quality utilising the GRADE instrument. We assessed the statistical heterogeneity of included studies by visually inspecting forest plots and quantifying the diversity using the I² statistic. We synthesised data using random-effects model meta-analysis or descriptive analysis, as appropriate. Thirty-one studies randomised 2952 outpatients to investigate the effects of different therapies on benign TN. Studies on LT4, PEI, LP and RF ablation therapy randomised 2083, 607, 192 and 70 participants, respectively. We found no RCTs of HIFU or MW ablation therapy in benign TN. The duration of treatment varied according to the applied therapies: up to five years for LT4 and one to three PEI ablations, one to three LP sessions and one or two RF sessions. Median follow-up was 12 months for LT4 and six months for minimally invasive therapies. Evidence was of low-to-moderate quality, and risk of performance and detection bias for subjective outcomes was high in most trials.No study evaluated all-cause mortality or health-related quality of life. Only one LT4 study provided some data on the development of thyroid cancer, reporting no abnormal cytological findings. One LP study provided limited information on costs of treatment.LT4 compared with no treatment or placebo was associated with a nodule volume reduction of 50% or more in 16% compared with 10% of participants after 6 to 24 months of follow-up (risk ratio (RR) 1.57 (95% confidence interval (CI) 1.04 to 2.38); P = 0.03; 958 participants; 10 studies; moderate-quality evidence). Pressure symptoms or cosmetic complaints were not investigated in LT4 studies. LT4 therapy was generally well tolerated: three studies provided quantitative data on signs and symptoms of hyperthyroidism, which were observed in 25% of LT4-treated versus 7% of placebo-treated participants at 12 to 18 months of follow-up (269 participants; 3 trials; low-quality evidence).PEI compared with cyst aspiration only was associated with a nodule volume reduction of 50% or more in 83% compared with 44% of participants after 1 to 24 months of follow-up (RR 1.83 (95% CI 1.32 to 2.54); P = 0.0003; 105 participants; 3 studies; low-quality evidence). Improvements in neck compression symptoms after 6 to 12 months of follow-up were seen in 78% of participants receiving PEI versus 38% of those in comparator groups. No reliable summary effect estimate could be established, RR ranged from 1.0 to 3.06 in favour of PEI (370 participants; 3 trials; low-quality evidence). In all trials, participants experienced periprocedural cervical tenderness and light-to-moderate pain usually lasting from minutes to several hours. As a result of the PEI procedure, 26% of participants reported slight-to-moderate pain compared with 12% of those receiving cyst aspiration only (RR 1.78 (95% CI 0.62 to 5.12); P = 0.28; 104 participants; 3 studies; low-quality evidence).One study comparing LP with LT4 showed a nodule volume reduction of 50% or more in favour of LP after 12 months of follow-up in 33% of LP participants versus 0% of LT4 participants, respectively (62 participants; 1 trial; low-quality evidence). A total of 82% of LP-treated versus 0% of untreated participants showed improvements in pressure symptoms after 6 to 12 months of follow-up (RR 26.65 (95% CI 5.47 to 129.72); P < 0.0001; 92 participants; 3 trials; low-quality evidence). Around 20% of LP-treated participants reported light-to-moderate cervical pain lasting 48 hours or more (97 participants; 3 trials; low-quality evidence).One trial with 40 participants, comparing RF with no treatment, resulted in a mean nodule volume reduction of 76% in the RF group compared with 0% of those in the no-treatment group at six months of follow-up (low-quality evidence). These RF-treated participants had fewer pressure symptoms and cosmetic complaints after 12 months of follow-up compared with untreated participants (a 2.8 decrease versus a 1.1 increase on a six-point scale, respectively, with higher values indicating more severe symptoms; low-quality evidence). All participants complained of pain and discomfort during RF, which disappeared when the energy was reduced or turned off (low-quality evidence). No study evaluated all-cause mortality, health-related quality of life or provided systematic data on the development of thyroid cancer. Longest follow-up was five years and median follow-up was 12 months. Nodule volume reductions were achieved by PEI, LP and RF, and to a lesser extent, by LT4. However, the clinical relevance of this outcome measure is doubtful. PEI, LP and RF led to improvements in pressure symptoms and cosmetic complaints. Adverse events such as light-to-moderate periprocedural pain were seen after PEI, LP and RF. Future studies should focus on patient-important outcome measures, especially health-related quality of life, and compare minimally invasive procedures with surgery. RCTs with follow-up periods of several years and good-quality observational studies are needed to provide evidence on the development of thyroid cancer, all-cause mortality and long-term adverse events.


Impact of Patient Education on Compliance with Positive Airway Pressure Treatment in Obstructive Sleep Apnea.
BACKGROUND We addressed the impact of patient education followed by frequent visits on compliance with positive airway pressure (PAP) treatment in patients with obstructive sleep apnea (OSA) in a Turkish sleep clinic cohort. MATERIAL AND METHODS This single-center, randomized, controlled study was conducted in Istanbul, Turkey between June 2014 and April 2015. Among 115 eligible OSA patients (mean age 51.0±9.3 years; 75.5% men), 63 were randomized to standard support (SS) group (general information about OSA and PAP treatment at baseline), and 52 to educational support (ES) group (additional polysomnography chart viewing from both diagnostic and titration nights). All patients were scheduled to five PAP control visits between two weeks and six months after the PAP prescription. Primary outcome was the PAP compliance (4 hours/night for 70% of all the nights) at the last visit. RESULTS Average PAP usage was 4.2±2.5 hours/night in the SS group, and 5.2±2.1 hours/night in the ES group (p=0.027). PAP compliance was achieved among 68.3% in the SS group, and 86.5% in the ES group (p=0.021). In a multivariate analysis, ES strategy followed by frequent visits predicted PAP compliance (odds ratio [OR] 3.6, 95% confidence interval [CI] 1.2-10.6; p=0.020). Other predictors were obesity (OR 3.4, 95% CI 1.2-9.7; p=0.019) and severe OSA (apnea-hypopnea index ≥30/hour) at baseline (OR 4.7, 95% CI 1.2-17.6; p=0.023). Primary school education level was inversely related with PAP compliance (OR 0.3, 95% CI 0.1-0.9; p=0.036). CONCLUSIONS Patient education with polysomnography chart view followed by frequent visits increased long-term compliance with PAP treatment.


Computationally Designed ACE2 Decoy Receptor Binds SARS-CoV-2 Spike (S) Protein with Tight Nanomolar Affinity.
Even with the availability of vaccines, therapeutic options for COVID-19 still remain highly desirable, especially in hospitalized patients with moderate or severe disease. Soluble ACE2 (sACE2) is a promising therapeutic candidate that neutralizes SARS CoV-2 infection by acting as a decoy. Using computational mutagenesis, we designed a number of sACE2 derivatives carrying three to four mutations. The top-predicted sACE2 decoy based on the in silico mutagenesis scan was subjected to molecular dynamics and free-energy calculations for further validation. After illuminating the mechanism of increased binding for our designed sACE2 derivative, the design was verified experimentally by flow cytometry and BLI-binding experiments. The computationally designed sACE2 decoy (ACE2-FFWF) bound the receptor-binding domain of SARS-CoV-2 tightly with low nanomolar affinity and ninefold affinity enhancement over the wild type. Furthermore, cell surface expression was slightly greater than wild-type ACE2, suggesting that the design is well-folded and stable. Having an arsenal of high-affinity sACE2 derivatives will help to buffer against the emergence of SARS CoV-2 variants. Here, we show that computational methods have become sufficiently accurate for the design of therapeutics for current and future viral pandemics.


Unprecedented rearrangement of diketopyrrolopyrroles leads to structurally unique chromophores.
Diketopyrrolopyrroles possessing thienyl, furyl and benzofuryl substituents undergo unprecedented skeletal rearrangement in the presence of trimethylsilyl bromide resulting in the formation of thieno[2,3-f]isoindole-5,8-diones and furo[2,3-f]isoindole-5,8-diones. These relatively small dyes possess favorable photophysical properties with the emission maxima within the range of 573-624 nm, large fluorescence quantum yields, moderate sensitivity of emission to solvent polarity and a HOMO-LUMO gap of ca. 1.8 eV.