Association between carriage of Streptococcus pneumoniae and Staphylococcus aureus in Children.
Widespread pneumococcal conjugate vaccination may bring about epidemiologic changes in upper respiratory tract flora of children. Of particular significance may be an interaction between Streptococcus pneumoniae and Staphylococcus aureus, in view of the recent emergence of community-acquired methicillin-resistant S aureus. To examine the prevalence and risk factors of carriage of S pneumoniae and S aureus in the prevaccination era in young children. Cross-sectional surveillance study of nasopharyngeal carriage of S pneumoniae and nasal carriage of S aureus by 790 children aged 40 months or younger seen at primary care clinics in central Israel during February 2002. Carriage rates of S pneumoniae (by serotype) and S aureus; risk factors associated with carriage of each pathogen. Among 790 children screened, 43% carried S pneumoniae and 10% carried S aureus. Staphylococcus aureus carriage among S pneumoniae carriers was 6.5% vs 12.9% in S pneumoniae noncarriers. Streptococcus pneumoniae carriage among S aureus carriers was 27.5% vs 44.8% in S aureus noncarriers. Only 2.8% carried both pathogens concomitantly vs 4.3% expected dual carriage (P =.03). Risk factors for S pneumoniae carriage (attending day care, having young siblings, and age older than 3 months) were negatively associated with S aureus carriage. Streptococcus pneumoniae carriage, specifically of vaccine-type strains, is negatively associated with S aureus carriage in children. The implications of these findings in the pneumococcal vaccine era require further investigation.


Altered contractility of circular and longitudinal muscle in TNBS-inflamed guinea pig ileum.
Intestinal motility disorders are often associated with gut inflammation. We evaluated, in vitro under isometric conditions, changes in contractility of longitudinal and circular muscle layers from guinea pig ileum after 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced ileitis. TNBS treatment did not modify length-active tension relationships for both muscle layers, whereas a significant increase in passive tension was observed in the circular muscle response to stretching. Moreover, in both control and inflamed strips at optimal stretch, concentration-response curves to KCl were similar for both layers. In contrast, contractile responses to receptor agonists were differentially altered in both layers in comparison with controls. Thus, in longitudinal strips from TNBS-treated ileum, there was a twofold increase in maximal response (Emax) induced by carbachol and histamine without modification of 50% effective concentration (EC50) values; responses to 5-hydroxytryptamine (5-HT) were not modified; both alpha 1- and alpha 2-adrenoceptor-mediated responses to epinephrine were abolished. In circular strips, inflammation did not affect the Emax induced by carbachol and histamine but led to increased EC50 values; Emax to 5-HT was reduced without change in EC50 values. Moreover, in the dose range used (0.1 nM to 0.1 mM), a maximal response to carbachol was not obtained in inflamed circular strips. The results indicate that in the guinea pig model of TNBS-induced ileitis, the in vitro contractile responses of circular and longitudinal smooth muscle to the stimulation of various receptors are differentially altered, whereas non-receptor-mediated contraction to KCl depolarization is not modified.


Mode-sorter design using continuous supersymmetric transformation.
We propose to use a continuous supersymmetric (SUSY) transformation of a dielectric permittivity profile in order to design a photonic mode sorter. The iso-spectrality of the SUSY transformation ensures that modes of the waveguide preserve their propagation constants while being spatially separated. This global matching of the propagation constants, in conjunction with the adiabatic modification of the refractive index landscape along the propagation direction, results in the negligible modal cross-talk and low scattering losses in the sorter. We show that a properly optimized SUSY mode sorter outperforms a standard asymmetric Y-splitter by reducing the cross-talk by at least two orders of magnitude. Moreover, the SUSY sorter is capable of sorting either transverse-electric or transverse-magnetic polarized modes and operates in a broad range of wavelengths. At the telecommunication wavelength, the 300-μm-long SUSY sorter provides the cross-talk of -35 dB and a broad operation bandwidth. The design proposed here paves the way toward efficient signal manipulation in integrated photonic devices.


How to promote knowledge transfer within R&D team? An evolutionary game based on prospect theory.
Knowledge transfer is the basis for R&D teams and enterprises to improve innovation performance, win market competition and seek sustainable development. In order to explore the path to promote knowledge transfer within the R&D team, this study considers the bounded rationality and risk preference of individuals, incorporates prospect theory into evolutionary game, constructs a perceived benefits matrix distinct from the traditional benefits matrix, and simulates the evolutionary game process. The results show that, R&D personnel's knowledge transfer decisions depend on the net income difference among strategies; only if perceived cost is less than the sum of perceived synergy benefit, perceived organization reward value, and perceived organization punishment value, can knowledge be fully shared and transferred within the R&D team. Moreover, R&D personnel's knowledge transfer decisions are interfered by the irrational psychological factors, including overconfidence, reflection, loss avoidance, and obsession with small probability events. The findings help R&D teams achieve breakthroughs in improving the efficiency of knowledge transfer, thereby enhancing the capacity of enterprises for collaborative innovation.


Vascular invasion and other invasive features in granular cell tumours of the skin: a multicentre study of 119 cases.
Incidental finding of vascular invasion has been described in some benign granular cell tumours. Malignancy in granular cell tumours is excessively rare and its assessment relies on necrosis and cytological criteria. To assess histopathological invasive features, particularly vascular invasion, in a large series of granular cell tumours of the skin. 119 granular cell tumours of the skin were collected in 114 patients between 2001 and 2011. Histopathological and epidemiological data were collected. Five step sections and one orcein staining were performed in all cases. Mean age of the patients was 43.7±18 years. Granular cell tumours were multiple in 7% of patients. They were classified as benign in 111 cases, and atypical in eight cases. No malignant granular cell tumour was present. Tumours had 1.48±1.3 cm mean diameter, showed peripheral invasive growth pattern in 71% of cases, had a mean depth of 8.8±4.7 mm, and reached the subcutis in 66% of cases. Infiltration of arrector pili muscle occurred in 23% (95% CI 16% to 32%), and perineural spread in 66% (95% CI 56% to 74%) of cases. Vascular invasion occurred in 23% (95% CI 16% to 32%) of cases, with subendothelial layers infiltration or vascular obliteration. No intraluminal embolus was found. No association was found between vascular invasion and clinical outcome. Histopathological features of local invasion are frequent in otherwise benign granular cell tumours. Vascular invasion consists of an infiltration of the subendothelial layers, without intraluminal cells, and may not be considered as a marker of adverse prognosis.


Mutation accumulation in transfer RNAs: molecular evidence for Muller's ratchet in mitochondrial genomes.
The accumulation of deleterious mutations is thought to be a major factor preventing the long-term persistence of obligately asexual lineages relative to their sexual ancestors. This phenomenon is also of potential relevance to sexual species that harbor asexually propagating organelle genomes. A comparative study of the transfer RNA genes in animal mitochondrial and nuclear genomes demonstrates that the former accumulate nucleotide substitutions much more rapidly than do the latter, and several lines of evidence are consistent with the idea that the excess substitutions are mildly deleterious. First, the average binding stability between complementary strands in the stems of mitochondrial tRNAs is less than half that in nuclear tRNAs. Second, most loop sizes in the mitochondrial tRNAs have experienced a net reduction in size over evolutionary time, and they are nearly 50 times more variable in the mitochondrial than in the nuclear genome. Third, although nearly 20% of the nucleotides in nuclear tRNA genes (particularly those involved in tertiary interactions) are invariant across all animal taxa and all tRNA species, there are no invariant sites in the mitochondrial tRNAs. These observations, as well as results from recent laboratory experiments, are consistent with the hypothesis that nonrecombining organelle genomes are subject to gradual loss of fitness due to the cumulative chance fixation of mildly deleterious mutations.


FIGalpha, a germ cell-specific transcription factor required for ovarian follicle formation.
Primordial follicles are formed perinatally in mammalian ovaries and at birth represent the lifetime complement of germ cells. With cyclic periodicity, cohorts enter into a growth phase that culminates in ovulation of mature eggs, but little is known about the regulatory cascades that govern these events. FIGalpha, a transcription factor implicated in postnatal oocyte-specific gene expression, is detected as early as embryonic day 13. Mouse lines lacking FIGalpha were established by targeted mutagenesis in embryonic stem cells. Although embryonic gonadogenesis appeared normal, primordial follicles were not formed at birth, and massive depletion of oocytes resulted in shrunken ovaries and female sterility. Fig(&agr;) (the gene for FIGalpha null males have normal fertility. The additional observation that null females do not express Zp1, Zp2 or Zp3 indicates that FIGalpha plays a key regulatory role in the expression of multiple oocyte-specific genes, including those that initiate folliculogenesis and those that encode the zona pellucida required for fertilization and early embryonic survival. The persistence of FIGalpha in adult females suggests that it may regulate additional pathways that are essential for normal ovarian development.


Principles and practice of disaster relief: lessons from Haiti.
Disaster relief is an interdisciplinary field dealing with the organizational processes that help prepare for and carry out all emergency functions necessary to prevent, prepare for, respond to, and recover from emergencies and disasters caused by all hazards, whether natural, technological, or human-made. Although it is an important function of local and national governing in the developed countries, it is often wanting in resource-poor, developing countries where, increasingly, catastrophic disasters tend to occur and have the greatest adverse consequences. The devastating January 12, 2010, Haiti earthquake is a case study of the impact of an extreme cataclysm in one of the poorest and most unprepared settings imaginable. As such, it offers useful lessons that are applicable elsewhere in the developing world. Emergency preparedness includes 4 phases: mitigation or prevention, preparedness, response, and recovery. Periods of normalcy are the best times to develop disaster preparedness plans. In resource-poor countries, where dealing with the expenses of daily living is already a burden, such planning is often neglected; and, when disasters strike, it is often with great delay that the assistance from international community can be deployed. In this increasingly interconnected world, the Haiti earthquake and the important international response to it make a strong case for a more proactive intervention of the international community in all phases of emergency management in developing countries, including in mitigation and preparedness, and not just in response and recovery. Predisaster planning can maximize the results of the international assistance and decrease the human and material tolls of inevitable disasters. There should be a minimum standard of preparedness that every country has to maintain and the international assistance to achieve that. International academic medical centers interested in global health could strengthen their programs by prospectively including in them contingency planning for international relief operations. Healthcare professionals of these institutions who travel to disaster zones should rigorously prepare themselves and make provisions for collecting and reporting data, which will enrich the knowledge of this growing activity.


[Mechanical destruction of collagen].
Mechanical destruction of collagen was studied by means of electrophoresis in polyacrylamid gel and ESR-spectroscopy. It has been stated that the rates of the breakage of polypeptide chains measured by these two methods are in a good agreement. An analysis of ESR spectra of free radicals in collagen suggests that mechanical action induces the rupture of C alpha-N-bonds.


Functional analysis of the 5'-flanking region of FTA for expression of rat GDP-L-fucose:beta-D-galactoside 2-alpha-L-fucosyltransferase.
The tissue-specific and species-specific expression of the ABH antigens is well known among vertebrate species and it is regulated by the alpha(1,2)fucosyltransferase that forms the H antigen, a precursor of the A and B antigens. To investigate the mechanisms governing the tissue-specific and species-specific expression of this alpha(1,2)fucosyltransferase, we characterized the gene structure, including the promoter region, of FTA, a rat orthologous homolog of human FUT1 that encodes the H alpha(1, 2)fucosyltransferase and is responsible for the expression of the ABH antigens on human red blood cells. Northern blot and 5'-RACE analyses suggested that at least two forms of FTA mRNA (2.9 and 2.6 kb), which use alternative transcription start sites, are present in the cancer cell lines RCN-9 (rat colon cancer) and PC12 (rat pheochromocytoma), whereas only the 2.6 kb form was detected in normal colon, stomach and pancreas. Transcriptional activity of the 5'-flanking sequence, which contains three putative Sp1-binding sites, but lacks both TATA and CAAT boxes, was examined. Transient transfection experiments of promoter-reporter gene constructs showed high promoter activity in RCN-9, PC12 and human colon cancer (WiDr) cell lines, weak activity in human vascular endothelial (ECV304) cells and no activity in human erythroleukemia (HEL) cells. The results suggest that the 5'-flanking region of FTA contains a tissue-specific promoter. Deletional analysis of the 5'-flanking sequence revealed regions containing cell-type-specific positive acting element(s) and negative regulatory element(s), which are related to the promoter activity.


Suture of the right internal jugular vein catheter in a mitral valve replacement: a case report.
Central venous catheterization can be necessary for patients undergoing a cardiac operation. Accidental suturing of the catheter to the heart is a rare complication that is difficult to correct; excessive traction force on the central venous catheter can lead to heart breakage or even death. We describe the case of a 56-year-old Han Chinese woman who was scheduled to undergo mitral valve replacement. The central venous catheter placed into her right internal jugular vein was accidentally sutured to the left atrial suture line during the operation. The stuck catheter was successfully removed without having to perform a cardiopulmonary bypass. Attaching a catheter to the heart by cardiac sutures can occur when the tip of the catheter locates directly above the atrial-caval junction. Care should be taken when closing the cephalad end of a left atrial incision in a mitral valve replacement. Although rare, accidental suturing of the central venous catheter must be kept in mind, and an approach should be chosen to remove the catheter that best avoids additional insult to the heart function.


Effects of Provision of Concrete Information about Patient-controlled Analgesia in Hysterectomy Patients.
This study was to investigate the effects of the provision of concrete information about patient-controlled analgesia (PCA) in hysterectomy patients. Study design was a nonequivalent control group non-synchronized pre- and post-test design. Sixty subjects participated were assigned to experimental group (30 patients) or control group (30 patients) at one university hospital. Concrete information about PCA was composed of three sections: explanation with a leaflet, practice of using PCA, and question and answer session. The experimental group who received concrete information about PCA before surgery had statistically higher knowledge level about PCA, more positive attitude toward pain control analgesia, a lower pain score, and a higher satisfaction level of the use of PCA post-surgery compared to the control group who received general information before surgery. Provision of concrete information about PCA was an effective nursing intervention that reduced post-operative pain for patients and increased their satisfaction with using PCA. It is recommended that concrete information about PCA be provided by nurses to promote the use of PCA and consequently reduce patient's pain post-surgery.


Failure of falciparum malaria prophylaxis by mefloquine in travelers from West Africa.
Due to the spread of chloroquine-resistant strains of Plasmodium falciparum in French speaking parts of Africa, we have found it necessary to prescribe mefloquine for antimalaria prophylaxis to travelers to this area. Weekly doses of 125 or 250 mg have been recommended for short journeys. In spite of this regimen, 16 documented cases of falciparum malaria in travelers have been recorded in the Bordeaux hospital center since October, 1988. Fifteen of these patients were tourists returning from West African countries, and one was an Ivorian student who had been on vacation to his home country. Nine of these patients were evaluated and found to have high plasma mefloquine levels. This report strongly supports the existence of mefloquine-resistant falciparum malaria in West Africa, especially in Sierra Leone, Burkina Faso, and Cote d'Ivoire.


Clinical observation of soft palate-pharyngoplasty in the treatment of obstructive sleep apnea hypopnea syndrome in children.
Childhood obstructive sleep apnea hypopnea syndrome (OSAHS) is a common clinical disease that can cause serious complications if not treated in time. The preferred treatment for OSAHS in children is surgery. To observe the effects of soft palate-pharyngoplasty on postoperative outcome, pharyngeal formation, and possible complications. A total of 150 children with snoring, hernia, and mouth breathing were selected. A polysomnography test was performed to confirm the diagnosis of OSAHS. The children were randomly divided into experimental and control groups. The experimental group underwent adenoidectomy, tonsillectomy, and soft palate-pharyngoplasty. The control group underwent adenoidectomy and tonsillectomy. The t-test and chi-square test were used to compare conditions such as postoperative fever, postoperative hemorrhage, and pharyngeal reflux. Postoperative efficacy and complications were interrogated and observed in the form of outpatient follow-up and telephone follow-up at 6 mo and 1 year after surgery. The curative effects were divided into two groups: Cure (snoring, snoring symptoms disappeared) and non-cure. The effective rate of the experimental group was significantly higher than that of the control group, but the difference was not statistically significant (P > 0.05). The incidence of postoperative bleeding was lower in the experimental group. There was no postoperative pharyngeal reflux in either group. In the experimental group, the incidence of hyperthermia (body temperature exceeded 38.5 °C) was lower than that in the control group. The difference in postoperative swallowing pain scores between the experimental and control groups was significant. Soft palate-pharyngoplasty can more effectively enlarge the anteroposterior diameter and transverse diameter of the isthmus faucium. Compared with surgery alone, it can better treat OSAHS in children, improve the curative effect, reduce the risk of perioperative bleeding, close the surgical cavity, reduce the risk of postoperative infection, reduce the proportion of postoperative fever, and accelerate healing. Although this process takes more time, it is simple, safe, and effective.


Vascular effects of infused adenosine are not mediated by prostacyclin release in humans.
Adenosine may contribute to the regulation of tissue blood flow directly and via release of vasoactive substances. For example, in the isolated, perfused heart, the nucleoside has been reported to release prostacyclin, a potent vasodilator. In humans, minor variations in prostacyclin release into the circulation result in readily detectable changes in the urinary excretion of its metabolite, 2,3-dinor-6-ketoprostaglandin (PG) F1 alpha, as measured by negative ion-chemical ionization gas chromatography-mass spectrometry. To test the hypothesis that prostacyclin participates in or mediates the vascular effects of adenosine, we administered adenosine (5.1 mg/min) or vehicle to healthy volunteers in random order as a 2-h infusion into the femoral artery under double-blind conditions. The plasma levels of adenosine, inosine, and hypoxanthine increased significantly during infusion of active drug, but the urinary excretion of adenosine and uric acid were unchanged, implying efficient tissue uptake of the infused nucleoside. Adenosine, but not vehicle, significantly (P less than 0.01) increased leg blood flow (from 2.7 +/- 0.3 to 8.7 +/- 2.5 ml X 100 ml tissue-1 X min-1), heart rate (from 66 +/- 3 to 80 +/- 4 beats/min), and urinary epinephrine excretion (from 2.8 +/- 0.4 to 5.4 +/- 0.8 ng/mg creatinine). In contrast, the excretion of 2,3-dinor-6-keto-PGF1 alpha was unaltered by infusion of adenosine. We confirmed that biologically significant alterations in prostacyclin release in the lower limb vascular bed would be reflected by the urinary metabolite in experiments involving local infusion of prostacyclin at a rate below the threshold necessary to alter limb blood flow.(ABSTRACT TRUNCATED AT 250 WORDS)


Efficacy of Prophylactic Treatment for Oxycodone-Induced Nausea and Vomiting Among Patients with Cancer Pain (POINT): A Randomized, Placebo-Controlled, Double-Blind Trial.
Although opioid-induced nausea and vomiting (OINV) often result in analgesic undertreatment in patients with cancer, no randomized controlled trials have evaluated the efficacy of prophylactic antiemetics for preventing OINV. We conducted this randomized, placebo-controlled, double-blind trial to evaluate the efficacy and safety of prophylactic treatment with prochlorperazine for preventing OINV. Cancer patients who started to receive oral oxycodone were randomly assigned in a 1:1 ratio to receive either prochlorperazine 5 mg or placebo prophylactically, given three times daily for 5 days. The primary endpoint was the proportion of patients who had a complete response (CR) during the 120 hours of oxycodone treatment. CR was defined as no emetic episode and no use of rescue medication for nausea and vomiting during 5 days. Key secondary endpoints were the proportion of patients with emetic episodes, proportion of patients with moderate or severe nausea, quality of life, and proportion of treatment withdrawal. From November 2013 through February 2016, a total of 120 patients were assigned to receive prochlorperazine (n = 60) or placebo (n = 60). There was no significant difference in CR rates (69.5% vs. 63.3%; p = .47) or any secondary endpoint between the groups. Patients who received prochlorperazine were more likely to experience severe somnolence (p = .048). Routine use of prochlorperazine as a prophylactic antiemetic at the initiation of treatment with opioids is not recommended. Further research is needed to evaluate whether other antiemetics would be effective in preventing OINV in specific patient populations. Prophylactic prochlorperazine seems to be ineffective in preventing opioid-induced nausea and vomiting (OINV) and may cause adverse events such as somnolence. Routine use of prophylactic prochlorperazine at the initiation of treatment with opioids is not recommended. Further research is needed to evaluate whether other antiemetics would be effective in preventing OINV in specific patient populations.


Influences of sedimentary organic matter quality on the bioaccumulation of 4-nonylphenol by estuarine amphipods.
Nonylphenol (NP) is a moderately persistent, hydrophobic chemical with endocrine-disrupting and acute narcotic effects in aquatic biota. Concern exists about the ultimate fate of NP in aquatic ecosystems and the potential for bioaccumulation by benthic biota from the sediment with the potential for further transfer to higher trophic levels. Our goals were to determine if benthic amphipods bioaccumulate significant amounts of NP from sediment and to determine how additions of organic matter influence NP bioaccumulation by amphipods. Estuarine sediment was spiked with 14C-NP and enriched with two types of organic carbon (OC) sources of different nutritional qualities. Macrophytic algae (Ulva species) were used as a labile and nutritious OC source. Wood lignins were used as a refractory and low-nutrition OC source. Nonylphenol bioaccumulation was measured in Eohaustorius estuarius, Grandidierella japonica, and Corophium salmonis after 16 d of exposure. Nonylphenol accumulation was inversely proportional to OC quantity, but was unaffected by OC nutritional quality. Significant differences were found in the accumulation patterns between the three amphipod species. Mean biota-sediment accumulation factors ranged from 8.1 to 33.9 in E. estuarius, from 4.6 to 17.2 in G. japonica, and averaged 7.1 in male C. salmonis and 16.0 in female C. salmonis. These accumulation factors indicate that estuarine amphipods could constitute an important source of NP to higher trophic levels, such as juvenile fish.


The Acinetobacter baumannii Mla system and glycerophospholipid transport to the outer membrane.
The outer membrane (OM) of Gram-negative bacteria serves as a selective permeability barrier that allows entry of essential nutrients while excluding toxic compounds, including antibiotics. The OM is asymmetric and contains an outer leaflet of lipopolysaccharides (LPS) or lipooligosaccharides (LOS) and an inner leaflet of glycerophospholipids (GPL). We screened Acinetobacter baumannii transposon mutants and identified a number of mutants with OM defects, including an ABC transporter system homologous to the Mla system in E. coli. We further show that this opportunistic, antibiotic-resistant pathogen uses this multicomponent protein complex and ATP hydrolysis at the inner membrane to promote GPL export to the OM. The broad conservation of the Mla system in Gram-negative bacteria suggests the system may play a conserved role in OM biogenesis. The importance of the Mla system to Acinetobacter baumannii OM integrity and antibiotic sensitivity suggests that its components may serve as new antimicrobial therapeutic targets.


Molecular dynamics simulation, binding free energy calculation and unbinding pathway analysis on selectivity difference between FKBP51 and FKBP52: Insight into the molecular mechanism of isoform selectivity.
As co-chaperones of the 90-kDa heat shock protein(HSP90), FK506 binding protein 51 (FKBP51) and FK506 binding protein 52 (FKBP52) modulate the maturation of steroid hormone receptor through their specific FK1 domains (FKBP12-like domain 1). The inhibitors targeting FK1 domains are potential therapies for endocrine-related physiological disorders. However, the structural conservation of the FK1 domains between FKBP51 and FKBP52 make it difficult to obtain satisfactory selectivity in FK506-based drug design. Fortunately, a series of iFit ligands synthesized by Hausch et al exhibited excellent selectivity for FKBP51, providing new opportunity for design selective inhibitors. We performed molecular dynamics simulation, binding free energy calculation and unbinding pathway analysis to reveal selective mechanism for the inhibitor iFit4 binding with FKBP51 and FKBP52. The conformational stability evaluation of the "Phe67-in" and "Phe67-out" states implies that FKBP51 and FKBP52 have different preferences for "Phe67-in" and "Phe67-out" states, which we suggest as the determinant factor for the selectivity for FKBP51. The binding free energy calculations demonstrate that nonpolar interaction is favorable for the inhibitors binding, while the polar interaction and entropy contribution are adverse for the inhibitors binding. According to the results from binding free energy decomposition, the electrostatic difference of residue 85 causes the most significant thermodynamics effects on the binding of iFit4 to FKBP51 and FKBP52. Furthermore, the importance of substructure units on iFit4 were further evaluated by unbinding pathway analysis and residue-residue contact analysis between iFit4 and the proteins. The results will provide new clues for the design of selective inhibitors for FKBP51.


Temporary axial rotation stabilization for lumbar disc herniation surgery with the ARO® spinal system: a prospective analysis of safety and clinical efficacy.
Decompressive surgery has a failure rate of between 25% and 32% based on patient reported improvement in clinical symptoms. Significant back pain is associated with 53% of failures of decompressive surgery, while also being associated with abnormal axial rotation motion. We report on the clinical performance of subjects receiving a novel axial rotation stabilization implant (ARO Spinal System, ARO Medical), while undergoing a surgical decompression for a herniated lumbar disc, a condition associated with low back pain and abnormal movement. This Danish Medicines Agency and Ethics Committee approved clinical trial prospectively investigated the use of the ARO® Spinal System as part of lumbar discectomy surgery in 20 patients. All subjects had a single level posterolateral lumbar herniation with symptoms lasting more than 6 weeks. They underwent an open discectomy at the symptomatic level, and received the implant. No complications with the implant were observed. Four serious adverse events not related to the devices were reported, one subject had reoperation at 3 months. Leg pain median VAS score decreased from 70 to 2 at 1 year (P=0.01) back pain median VAS score from 48 to 6 (P=0.04). Satisfaction with surgery was 88%. Oswestry Disability Index scored likewise improvement going from 38 pre-operative to 5 at 1 year. Follow-up rate was excellent 100%. Discectomy with the ARO Spinal System proves equally safe as a standard discectomy at 1 year follow-up. The subjects had significant improvements in both leg and back pain. In addition, they did better than historical controls, though not statistically so in this patient sample.


Sacrococcygeal yolk sac carcinoma: imaging findings during treatment.
Sacrococcygeal teratomas and their malignant counterparts (germ cell tumors) are the most common solid tumors in neonates. Prompt diagnosis is essential because the frequency of malignant transformation increases from 10-20 % in neonates to 67 % in patients over 2 months of age. Cross-sectional imaging has largely replaced surgical exploration for staging these tumors and assessing their response to chemotherapy. Radiologists must be familiar with changes in the imaging findings of these tumors during and after treatment so that they can advise clinicians regarding the efficacy of therapy and the presence or absence of recurrent disease. From our study, magnetic resonance imaging appears to be a better modality for assessing sacral invasion and metastases and distinguishing fibrotic masses from recurrent tumor.


Formulations and processing of yogurt affect the microbial quality of carbonated yogurt.
Carbonation, flavor, culture type, pH, and storage time were varied to investigate the effects of these variables and their interactions on the growth of both typical and nontypical yogurt cultures and some contaminating bacteria. Two types of yogurt cultures (YC-470 and YC-180) were used as the source of typical yogurt bacteria, Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus. In addition, Lactobacillus acidophilus (LA-K) and Bifidobacterium longum ATCC 15707 were added as nontypical yogurt cultures to make sweetened low fat (1%) Swiss-style plain, strawberry, and lemon yogurts. Samples were incubated at 43 degrees C until pH values of 5.0 or 4.2 were reached. Strawberry yogurts at low (4.2) and high (5.0) pHs were divided into three portions, which were separately inoculated with contaminating bacteria, Bacillus licheniformis ATCC 14580, Escherichia coli ATCC 11775, and Listeria monocytogenes Scott A. After incorporation of carbon dioxide (1.10 to 1.27 volume of CO2 gas dissolved in water), the yogurt was stored at 4 degrees C for a 90-d period. Carbon dioxide did not affect the growth of typical or nontypical yogurt bacteria. Also, CO2 did not inhibit the growth of undesirable microorganisms. In general, low levels of CO2 did not affect the bacterial population in yogurt. The microflora of yogurt were influenced by culture type, pH, flavor type, and storage time or their interactions.


Rgma-Induced Neo1 Proteolysis Promotes Neural Tube Morphogenesis.
Neuroepithelial cell (NEC) elongation is one of several key cell behaviors that mediate the tissue-level morphogenetic movements that shape the neural tube (NT), the precursor of the brain and spinal cord. However, the upstream signals that promote NEC elongation have been difficult to tease apart from those regulating apico-basal polarity and hingepoint formation, due to their confounding interdependence. The Repulsive Guidance Molecule a (Rgma)/Neogenin 1 (Neo1) signaling pathway plays a conserved role in NT formation (neurulation) and is reported to regulate both NEC elongation and apico-basal polarity, through signal transduction events that have not been identified. We examine here the role of Rgma/Neo1 signaling in zebrafish (sex unknown), an organism that does not use hingepoints to shape its hindbrain, thereby enabling a direct assessment of the role of this pathway in NEC elongation. We confirm that Rgma/Neo1 signaling is required for microtubule-mediated NEC elongation, and demonstrate via cell transplantation that Neo1 functions cell autonomously to promote elongation. However, in contrast to previous findings, our data do not support a role for this pathway in establishing apical junctional complexes. Last, we provide evidence that Rgma promotes Neo1 glycosylation and intramembrane proteolysis, resulting in the production of a transient, nuclear intracellular fragment (NeoICD). Partial rescue of Neo1a and Rgma knockdown embryos by overexpressing neoICD suggests that this proteolytic cleavage is essential for neurulation. Based on these observations, we propose that RGMA-induced NEO1 proteolysis orchestrates NT morphogenesis by promoting NEC elongation independently of the establishment of apical junctional complexes.SIGNIFICANCE STATEMENT The neural tube, the CNS precursor, is shaped during neurulation. Neural tube defects occur frequently, yet underlying genetic risk factors are poorly understood. Neuroepithelial cell (NEC) elongation is essential for proper completion of neurulation. Thus, connecting NEC elongation with the molecular pathways that control this process is expected to reveal novel neural tube defect risk factors and increase our understanding of NT development. Effectors of cell elongation include microtubules and microtubule-associated proteins; however, upstream regulators remain controversial due to the confounding interdependence of cell elongation and establishment of apico-basal polarity. Here, we reveal that Rgma-Neo1 signaling controls NEC elongation independently of the establishment of apical junctional complexes and identify Rgma-induced Neo1 proteolytic cleavage as a key upstream signaling event.


NIR Rapid Assessments of Blumea balsamifera (Ai-na-xiang) in China.
Blumea balsamifera (Ai-na-xiang) is used as an important plant source of natural borneol, which is widely used in the pharmaceutical industry. The aim of this study was to establish the methods based on near infrared (NIR) spectroscopy for determining the geographical origin of B. balsamifera, as well as developing a method for the quantitative rapid analysis of the active pharmaceutical ingredients (APIs). A total of 109 samples were collected from China in 2013 and arbitrarily divided into calibration and prediction sets using the Kennard-Stone algorithm. The l-borneol and total flavone contents of the samples were measured by gas chromatography and ultraviolet-visible spectroscopy, respectively. The NIR spectra were acquired using an integrating sphere and a partial least squares (PLS) model was built using the optimum wavelength regions, which were selected using a synergy interval partial least-squares (SiPLS) algorithm. The root mean square errors of prediction of the l-borneol and total flavone models were 0.0779 and 2.2694 mg/g, with R² of 0.9069 and 0.8013, respectively. A discriminant model to determine the geographical origin of B. balsamifera (Guizhou and Hainan) was also established using a partial least squares discriminant analysis method with an optimum pretreatment method. The prediction accuracy rate of the model was 100%. NIR spectroscopy can be used as a reliable and environmentally friendly method to determine the API and the origin of different B. balsamifera samples.


[Biological treatment options for Graves' ophthalmopathy].
The current medical treatment options for Graves' ophthalmopathy (GO) are unsatisfactory. Recent treatment of GO patients with the B-lymphocyte depleting monoclonal antibody rituximab or with the anti-tumor necrosis factor-alpha agents etanercept and infliximab has shown promising results. We discuss the use of these and other biological agents targeting B lymphocytes, T-lymphocyte interaction with antigen-presenting cells, or cytokines in the future treatment of GO.


Optical Assets of In situ Electro-assembled Platinum Black Nanolayers.
Optoelectronic technology has been increasingly driven towards miniaturization. In this regard, maintaining the optical properties of the bulk materials while reducing their size is a critical need. How thin must the film be to preserve the bulk material´s optical absorbance and reflectance characteristics? This is the central question for our study of the in situ electro-assembly broad band optical absorber films of platinum in non-aqueous solution of PtCl4. By reducing the in situ constructed film to sub-visible-wavelength thicknesses, the measured reflectance in the region from the ultraviolet to the infrared remained close to that exhibited by the micrometre-width films. These platinum black films broadly absorb electromagnetic waves at a sub-incident-wavelength thickness owing to their plasmonically increased absorbance cross-section. Simulation of various incident energy electron trajectories gives insights into the electron depth through the porous platinum black of ρ = 1.6 g/cm3 and previews the optical behaviour close to the atomic thickness.


Non-SMC condensin I complex subunit H enhances proliferation, migration, and invasion of hepatocellular carcinoma.
Non-SMC condensing I complex subunit H (NCAPH) is a member of the Barr protein family and part of the condensin I complex. The upregulation of NCAPH is associated with poor prognosis in patients with colon cancer. However, the relationship between NCAPH and hepatocellular carcinoma (HCC) remains unclear. This study aimed to explore NCAPH expression in HCC tissues and to investigate NCAPH functions in HCC cells. In this study, we found that high expression of NCAPH in HCC indicated worse prognosis via bioinformatics analysis. Consistently, quantitative real-time polymerase chain reaction assays in 20 pairs of HCC specimens and the immunohistochemical analysis of 100 HCC tissues showed the upregulation of NCAPH. We established stable NCAPH-overexpressing and NCAPH knockdown cell lines. Cell Counting Kit-8 assays and colony formation assay were performed to analyze cell proliferation. Migration and invasion were analyzed by Transwell assays. Subcutaneous xenograft models were used to explore the role of NCAPH in tumor formation in vivo. Our results showed that NCAPH promoted tumor proliferation, migration, and invasion in vitro and in vivo. In conclusion, our findings indicate that NCAPH could serve as a novel prognostic biomarker and a potential therapeutic target for patients with HCC.


Innate immune activation by checkpoint inhibition in human patient-derived lung cancer tissues.
Although Pembrolizumab-based immunotherapy has significantly improved lung cancer patient survival, many patients show variable efficacy and resistance development. A better understanding of the drug's action is needed to improve patient outcomes. Functional heterogeneity of the tumor microenvironment (TME) is crucial to modulating drug resistance; understanding of individual patients' TME that impacts drug response is hampered by lack of appropriate models. Lung organotypic tissue slice cultures (OTC) with patients' native TME procured from primary and brain-metastasized (BM) non-small cell lung cancer (NSCLC) patients were treated with Pembrolizumab and/or beta-glucan (WGP, an innate immune activator). Metabolic tracing with 13C6-Glc/13C5,15N2-Gln, multiplex immunofluorescence, and digital spatial profiling (DSP) were employed to interrogate metabolic and functional responses to Pembrolizumab and/or WGP. Primary and BM PD-1+ lung cancer OTC responded to Pembrolizumab and Pembrolizumab + WGP treatments, respectively. Pembrolizumab activated innate immune metabolism and functions in primary OTC, which were accompanied by tissue damage. DSP analysis indicated an overall decrease in immunosuppressive macrophages and T cells but revealed microheterogeneity in immune responses and tissue damage. Two TMEs with altered cancer cell properties showed resistance. Pembrolizumab or WGP alone had negligible effects on BM-lung cancer OTC but Pembrolizumab + WGP blocked central metabolism with increased pro-inflammatory effector release and tissue damage. In-depth metabolic analysis and multiplex TME imaging of lung cancer OTC demonstrated overall innate immune activation by Pembrolizumab but heterogeneous responses in the native TME of a patient with primary NSCLC. Metabolic and functional analysis also revealed synergistic action of Pembrolizumab and WGP in OTC of metastatic NSCLC.


Sustained reductions of invasive infectious disease following general infant Haemophilus influenzae type b and pneumococcal vaccination in a Swedish Arctic region.
Vaccine-preventable pathogens causing severe childhood infections include Haemophilus influenzae type b (Hib), Streptococcus pneumoniae and Neisseria meningitidis. In this study conducted in a Swedish Arctic region, we evaluated the effects of general infant Hib and pneumococcal vaccination on invasive infectious diseases among children and assessed the need of meningococcal vaccination. We identified cases of bacterial meningitis and sepsis from diagnosis and laboratory registers in the Västerbotten Region, Sweden, during 1986-2015. We then reviewed medical records to confirm the diagnosis and extract data for assessing incidence changes, using an exploratory data analysis and a time-series analysis. Invasive Haemophilus disease declined by 89.1% (p < 0.01), Haemophilus meningitis by 95.3% (p < 0.01) and all-cause bacterial meningitis by 82.3% (p < 0.01) in children aged 0 to four years following general infant Hib vaccination. Following pneumococcal vaccination, invasive pneumococcal disease declined by 84.7% (p < 0.01), pneumococcal meningitis by 67.5% (p = 0.16) and all-cause bacterial meningitis by 48.0% (p = 0.23). Incidence of invasive meningococcal disease remained low during the study period. Remarkable sustained long-term declines of invasive infectious diseases in younger children occurred following infant Hib and pneumococcal vaccinations in this Swedish Arctic region. Despite not offering general infant meningococcal vaccination, incidence of invasive meningococcal disease remained low.


Signet ring cell carcinoma of the stomach.
Between 1965 and 1985, 51 of 1500 patients (3.4%) with gastric cancer who had gastric resection had signet ring cell gastric cancer. Patients with this form of cancer tended to be younger and female; the tumors were smaller and involved the stomach body, serosal invasion was less prominent, and lymph node metastases were less likely to be present. Early mucosal and submucosal cancer was present in 54.9% of the patients with the signet ring cell and in 24.6% with other types of gastric cancer. In 15.7% of patients with signet ring cell cancer, a noncurative resection was performed. The 5-year survival rate was 74.5% for patients with signet ring cell cancer and 52.4% for those with other types of gastric cancer (P less than 0.01). In patients with signet ring cell gastric cancer, the lesion is less extensive; thus, these patients probably can expect a longer survival time.


Unraveling the Cave: A Seventy-Year Journey into the Caveolar Network, Cellular Signaling, and Human Disease.
In the mid-1950s, a groundbreaking discovery revealed the fascinating presence of caveolae, referred to as flask-shaped invaginations of the plasma membrane, sparking renewed excitement in the field of cell biology. Caveolae are small, flask-shaped invaginations in the cell membrane that play crucial roles in diverse cellular processes, including endocytosis, lipid homeostasis, and signal transduction. The structural stability and functionality of these specialized membrane microdomains are attributed to the coordinated activity of scaffolding proteins, including caveolins and cavins. While caveolae and caveolins have been long appreciated for their integral roles in cellular physiology, the accumulating scientific evidence throughout the years reaffirms their association with a broad spectrum of human disorders. This review article aims to offer a thorough account of the historical advancements in caveolae research, spanning from their initial discovery to the recognition of caveolin family proteins and their intricate contributions to cellular functions. Furthermore, it will examine the consequences of a dysfunctional caveolar network in the development of human diseases.


Primers for Castilleja and their utility across Orobanchaceae: II. Single-copy nuclear loci.
We developed primers targeting nuclear loci in Castilleja with the goal of reconstructing the evolutionary history of this challenging clade. These primers were tested across other major clades in Orobanchaceae to assess their broader utility. We assembled low-coverage genomes for three taxa in Castilleja and developed primer combinations for the single-copy conserved ortholog set (COSII) and the pentatricopeptide repeat (PPR) gene family. These primer combinations were designed to take advantage of the Fluidigm microfluidic PCR platform and are well suited for high-throughput sequencing applications. Eighty-seven primers were designed for Castilleja, and 27 were found to have broader utility in Orobanchaceae. These results demonstrate the utility of these primers, not only across Castilleja, but for other lineages within Orobanchaceae as well. This expanded molecular toolkit will be an asset to future phylogenetic studies in Castilleja and throughout Orobanchaceae.


Expression profiling of genes regulated by Fra-1/AP-1 transcription factor during bleomycin-induced pulmonary fibrosis.
The Fra-1/AP-1 transcription factor regulates the expression of genes controlling various processes including migration, invasion, and survival as well as extracellular remodeling. We recently demonstrated that loss of Fra-1 leads to exacerbated bleomycin-induced pulmonary fibrosis, accompanied by enhanced expression of various inflammatory and fibrotic genes. To better understand the molecular mechanisms by which Fra-1 confers protection during bleomycin-induced lung injury, genome-wide mRNA expression profiling was performed. We found that Fra-1 regulates gene expression programs that include: 1) several cytokines and chemokines involved in inflammation, 2) several genes involved in the extracellular remodeling and cell adhesion, and 3) several genes involved in programmed cell death. Loss of Fra-1 leads to the enhanced expression of genes regulating inflammation and immune responses and decreased the expression of genes involved in apoptosis, suggesting that this transcription factor distinctly modulates early pro-fibrotic cellular responses.


Effect of dietary conjugated linoleic acid on marbling and intramuscular adipocytes in pork.
Dietary CLA has been reported to decrease backfat and increase marbling in pigs. Our objective was to determine whether the increase in marbling involved changes in intramuscular adipocyte number or size or both. Twenty barrows (53 kg) were penned in pairs and pens were randomly assigned to receive diets containing either 1% soybean oil (SBO) or CLA (60% CLA isomers) for 6 wk. Body weight and feed intake were determined weekly. At slaughter, loin samples were obtained and flash frozen for RNA extraction and real-time reverse-transcription PCR analysis of gene expression. After a 24-h chill, loin eye area and backfat depth were measured and subjective marbling and color scores were assigned. Loin, backfat, and belly fat samples were obtained for fatty acid analysis by gas chromatography. Loin samples were also frozen in ice-cold isopentane for histological analysis of intramuscular adipocytes. Dietary CLA did not affect BW or feed intake at any point (P > 0.10), nor did treatment groups differ in HCW (P = 0.417) or loin color (P = 0.500). The CLA-fed pigs did have less (P = 0.018) backfat and smaller (P = 0.047) loin eye area than SBO-fed pigs and had a trend for an increase (P = 0.069) in marbling score. Relative gene expression for markers of preadipocytes (preadipocyte factor 1; Pref-1), differentiating adipocytes (PPARγ), and mature adipocytes [fatty acid binding protein 4 (FABP4) and perilipin (PLIN)] were determined and normalized to the expression of acidic ribosomal phosphoprotein. No significant differences were detected, but the expression of PPARγ (P = 0.265), PLIN (P = 0.265), and FABP4 (P = 0.148) was numerically greater in CLA-fed pigs than in SBO-fed pigs. Loin samples were stained with Oil Red O to identify intramuscular adipocytes. The average cell area was increased (P = 0.030) in CLA-fed pigs. The cis-9,trans-11 and trans-10,cis-12 CLA isomers were incorporated (P = 0.006) into backfat and belly fat, but only trans-10,cis-12 CLA was increased in the loin (P = 0.004) of CLA-fed pigs. The proportion of SFA was increased (P = 0.006) by CLA in all tissues. These results indicate that the increase in marbling in pigs fed CLA may be related to increased intramuscular adipocyte size, and the combination of increased marbling and degree of saturation could improve the eating quality of CLA-fed pork.


Analysis of 15 different genome types of adenovirus type 7 isolated on five continents.
A total of 15 different genome types of adenovirus type 7 (Ad7), i.e., Ad7p, Ad7p1, Ad7a, Ad7a1 to Ad7a5, Ad7b, Ad7c, Ad7d, Ad7d1, Ad7e, Ad7f, and Ad7g, were identified among 40 selected strains isolated in Europe, Asia, North America, South America, and Australia by using restriction endonucleases BamHI, BclI, BglI, BglII, BstEII, EcoRI, HindIII, HpaI, SalI, SmaI, XbaI, and XhoI. Eight of them, Ad7p1, Ad7a1 to Ad7a5, Ad7d1, and Ad7g, are newly discovered. All 15 genome types could be distinguished by the four restriction endonucleases BamHI, BclI, BglI, and XbaI. At least four restriction sites differed between Ad7d and Ad7g. Pairwise analyses of comigrating DNA restriction fragments of all 15 Ad7 genome types were performed and presented in a schematic fashion. According to the degree of comigration of DNA restriction fragments, the 15 genome types could be divided into three clusters. Ad7b was the dominant genome type in different parts of the world and may have evolved in China into Ad7d and further to Ad7d1.


Negative subtraction hybridization: an efficient method to isolate large numbers of condition-specific cDNAs.
The construction of cDNA libraries is a useful tool to understand gene expression in organisms under different conditions, but random sequencing of unbiased cDNA collections is laborious and can give rise to redundant EST collections. We aimed to isolate cDNAs of messages induced by switching Aspergillus nidulans from growth on glucose to growth on selected polysaccharides. Approximately 4,700 contigs from 12,320 ESTs were already available from a cDNA library representing transcripts isolated from glucose-grown A. nidulans during asexual development. Our goals were to expand the cDNA collection without repeated sequencing of previously identified ESTs and to find as many transcripts as possible that are specifically induced in complex polysaccharide metabolism. We have devised a Negative Subtraction Hybridization (NSH) method and tested it in A. nidulans. NSH entails screening a plasmid library made from cDNAs prepared from cells grown under a selected physiological condition with labeled cDNA probes prepared from another physiological condition. Plasmids with inserts that failed to hybridize to cDNA probes through two rounds of screening (i.e. negatives) indicate that they are transcripts present at low concentration in the labeled probe pool. Thus, these transcripts will be predominantly condition-specific, along with some rare transcripts. In a screen for transcripts induced by switching the carbon source from glucose to 12 selected polysaccharides, 3,532 negatives were isolated from approximately 100,000 surveyed colonies using this method. Negative clones were end-sequenced and assembled into 2,039 contigs, of which 1,722 were not present in the previously characterized glucose-grown cDNA library. Single-channel microarray hybridization experiments confirmed that the majority of the negatives represented genes that were differentially induced by a switch from growth in glucose to one or more of the polysaccharides. The Negative Subtraction Hybridization method described here has several practical benefits. This method can be used to screen any existing cDNA library, including full-length and pooled libraries, and does not rely on PCR or sequence information. In addition, NSH is a cost-effective method for the isolation of novel, full-length cDNAs for differentially expressed transcripts or enrichment of rare transcripts.


Induction of tumor necrosis factor production from monocytes stimulated with mannuronic acid polymers and involvement of lipopolysaccharide-binding protein, CD14, and bactericidal/permeability-increasing factor.
Well-defined polysaccharides, such as beta1-4-linked D-mannuronic acid (poly[M]) derived from Pseudomonas aeruginosa, induce monocytes to produce tumor necrosis factor (TNF) through a pathway involving membrane CD14. In this study we have investigated the effects of soluble CD14 (sCD14), lipopolysaccharide-binding protein (LBP), and bactericidal/permeability-increasing factor (BPI) on poly(M) binding to monocytes and induction of TNF production. We show that LBP increased the TNF production from monocytes stimulated with poly(M). Addition of sCD14 alone had only minor effects, but when it was added together with LBP, a rise in TNF production was seen. BPI was found to inhibit TNF production from monocytes stimulated with poly(M) in the presence of LBP, LBP-sCD14, or 10% human serum. Binding studies showed that poly(M) bound to LBP- and BPI-coated immunowells, while no significant binding of poly(M) to sCD14-coated wells in the absence of serum was observed. Binding of poly(M) to monocytes was also examined by flow cytometry, and it was shown that the addition of LBP or 10% human serum clearly increased the binding of poly(M) to monocytes. BPI inhibited the binding of poly(M) to monocytes in the presence of LBP, LBP-sCD14, or 10% human serum. Our data demonstrate a role for LBP, LBP-sCD14, and BPI in modulating TNF responses of defined polysaccharides.


IAA stimulates pollen tube growth and mediates the modification of its wall composition and structure in Torenia fournieri.
The effects of several hormones on pollen tube growth were compared in Torenia fournieri and it was found that IAA was the most effective, stimulating pollen tube growth and causing the shank part of pollen tubes to be slender and straighter. The role of IAA was investigated by studying the changes in ultrastructure and PM H(+)-ATPase distribution in the pollen tubes and the modification of the tube wall. Using the fluorescent marker FM4-64, together with transmission electron microscopy, it was shown that secretory vesicles and mitochondria increased in IAA-treated tubes. Immunolocalization and fluorescence labelling, together with Fourier-transform infrared analysis, detected that IAA enhanced the level of PM H(+)-ATPase and the synthesis of pectins, and reduced the cellulose density in pollen tubes. Importantly, to observe the orientation of cellulose microfibrils in pollen tubes in situ, atomic force microscopy was used to examine the 'intact' tube wall. Atomic force microscopy images showed that cellulose microfibrils were parallel to each other in the subapical region of IAA-treated tubes, but disorganized in control tubes. All results provided new insights into the functions of cellulose microfibrils in pollen tube growth and direction, and revealed that the IAA-induced changes of pollen tubes were attributed to the increase in secretory vesicles, mitochondria, and PM H(+)-ATPase, and the modification of pectin and cellulose microfibrils in the tube wall.


Novel Approach for Obtaining Variable Domain of New Antigen Receptor with Different Physicochemical Properties from Japanese Topeshark (Hemitriakis japanica).
Diverse candidate antibodies are needed to successfully identify therapeutic and diagnostic applications. The variable domain of IgNAR (VNAR), a shark single-domain antibody, has attracted attention owing to its favorable physicochemical properties. The phage display method used to screen for optimal VNARs loses sequence diversity because of the bias caused by the differential ease of protein expression in Escherichia coli. Here, we investigated a VNAR selection method that combined panning with various selection pressures and next-generation sequencing (NGS) analyses to obtain additional candidates. Drawing inspiration from the physiological conditions of sharks and the physicochemical properties of VNARs, we examined the effects of NaCl and urea concentrations, low temperature, and preheating at the binding step of panning. VNAR phage libraries generated from Japanese topeshark (Hemitriakis japanica) were enriched under these conditions. We then performed NGS analysis and attempted to select clones that were specifically enriched under each panning condition. The identified VNARs exhibited higher reactivity than those obtained by panning without selection pressure. Additionally, they possess physicochemical properties that reflect their respective selection pressures. These results can greatly enhance our understanding of VNAR properties and offer guidance for the screening of high-quality VNAR clones that are present at low frequencies.


Promise of molecular epidemiology--epidemiologic reasoning, biological rationale and risk assessment.
Molecular epidemiology has emerged as a natural outgrowth of attempts to apply information derived from the explosion in molecular biology to disease in human populations. The incorporation of biomarkers into classical epidemiologic designs holds the promise of unraveling mechanisms, elucidating gene-environment interactions, and dissecting heterogeneity. The primary interest of molecular epidemiology is in the identification of factors in the physical and social environment which affect the risk for disease and which are amenable to preventive intervention. The explosion in molecular technology has not, however, resulted in radical, widespread improvements in epidemiologic results and therefore has led to a sense of frustration in the public health community. As experience accumulates, there is new appreciation that attention to study design, infrastructure, and biomarker validation can improve the results.


Development and validation of a brief screening instrument for psychosocial risk associated with genetic testing: a pan-Canadian cohort study.
To develop a brief, reliable and valid instrument to screen psychosocial risk among those who are undergoing genetic testing for Adult-Onset Hereditary Disease (AOHD). A prospective two-phase cohort study. 5 genetic testing centres for AOHD, such as cancer, Huntington's disease or haemochromatosis, in ambulatory clinics of tertiary hospitals across Canada. 141 individuals undergoing genetic testing were approached and consented to the instrument development phase of the study (Phase I). The Genetic Psychosocial Risk Instrument (GPRI) developed in Phase I was tested in Phase II for item refinement and validation. A separate cohort of 722 individuals consented to the study, 712 completed the baseline package and 463 completed all follow-up assessments. Most participants were female, at the mid-life stage. Individuals in advanced stages of the illness or with cognitive impairment or a language barrier were excluded. Phase I: GPRI items were generated from (1) a review of the literature, (2) input from genetic counsellors and (3) phase I participants. Phase II: further item refinement and validation were conducted with a second cohort of participants who completed the GPRI at baseline and were followed for psychological distress 1-month postgenetic testing results. GPRI, Hamilton Depression Rating Scale (HAM-D), Hamilton Anxiety Rating Scale (HAM-A), Brief Symptom Inventory (BSI) and Impact of Event Scale (IES). The final 20-item GPRI had a high reliability-Cronbach's α at 0.81. The construct validity was supported by high correlations between GPRI and BSI and IES. The predictive value was demonstrated by a receiver operating characteristic curve of 0.78 plotting GPRI against follow-up assessments using HAM-D and HAM-A. With a cut-off score of 50, GPRI identified 84% of participants who displayed distress postgenetic testing results, supporting its potential usefulness in a clinical setting.


Assessing the intrinsic precision of 3D/3D rigid image registration results for patient setup in the absence of a ground truth.
To assess the precision and robustness of patient setup corrections computed from 3D/3D rigid registration methods using image intensity, when no ground truth validation is possible. Fifteen pairs of male pelvic CTs were rigidly registered using four different in-house registration methods. Registration results were compared for different resolutions and image content by varying the image down-sampling ratio and by thresholding out soft tissue to isolate bony landmarks. Intrinsic registration precision was investigated by comparing the different methods and by reversing the source and the target roles of the two images being registered. The translational reversibility errors for successful registrations ranged from 0.0 to 1.69 mm. Rotations were less than 1 degrees. Mutual information failed in most registrations that used only bony landmarks. The magnitude of the reversibility error was strongly correlated with the success/ failure of each algorithm to find the global minimum. Rigid image registrations have an intrinsic uncertainty and robustness that depends on the imaging modality, the registration algorithm, the image resolution, and the image content. In the absence of an absolute ground truth, the variation in the shifts calculated by several different methods provides a useful estimate of that uncertainty. The difference observed by reversing the source and target images can be used as an indication of robust convergence.


Transcription of metallothionein isoform promoters is differentially regulated in cadmium-sensitive and -resistant CHO cells.
Transcription regulation of metallothionein (MT) isoform promoters was investigated in Chinese hamster ovary (CHO) K1 and MT gene amplified, cadmium-resistant (CdR) cells. The transfected promoter of Chinese hamster MTI and MTII genes can be activated in both cell lines by stimulation with Cd or Zn ions, although no MT mRNA can be detected in CHO K1 cells after challenge with metal ions. Neither MT promoter used in this study can be activated by induction with dexamethasone, regardless of whether a sequence homologous to glucocorticoid responsive element is present. During induction by metal ions, differential promoter activities of the MT genes occurs in both CHO K1 and CdR cells where MTII promoter has a stronger activity than that of MTI. As indicated by a time course study in both cell lines, the relative induction ratios of both MTI and MTII promoters are similar at each time interval. This result is consistent with a differential transcriptional factor-promoter interaction for the two MT promoters. By using the CHO K1 and CdR cells as a model system, the occurrence of autoregulation for yeast CUP1 (MT) gene was examined in mammalian cells. Both MT promoters consistently show a lower basal activity but a higher induction ratio in CHO K1 than CdR cells; a result different from that of yeast CUP1 gene. When MTF-1 mRNA was examined, no difference in relative quantity was observed in CHO K1 and in CdR cells treated with metal ions or with metal ions absent.


Screening Children for Caries: An Ethical Dilemma in Nigeria.
Dental caries is the main oral health challenge for children in Nigeria. Concern about its negative impact makes screening for caries in children an attractive public health strategy. The ability to detect the preclinical phase of caries, the availability of screening tools with high accuracy, and the possibility of treatment before onset of clinical symptoms with significant cost and health benefits, makes it appropriate for screening. However in Nigeria, the poor availability of highly specific and sensitive screening tools, poor access to oral health care and concerns with pre-screening consent, raise the question of the appropriateness of conducting screening programmes for children. We argue that a number of structural challenges associated with poor uptake of oral health care services need to be addressed before screening for caries can be considered ethically appropriate. These include facilitating access of children to quality oral health care and a systematic national approach to oral health implementation. Failure to address challenges associated with dental service utilization by children in Nigeria increases the risk of screening programmes promoting inequitable access to oral health care services.


A cluster randomised controlled trial protocol of an adapted intervention for alcohol use disorders in people living with HIV and AIDS: impact on alcohol use, general functional ability, quality of life and adherence to HAART.
Interventions for alcohol use disorders (AUDs) in HIV infected individuals have been primarily targeted at HIV risk reduction and improved antiretroviral treatment adherence. However, reduction in alcohol use is an important goal. Alcohol use affects other key factors that may influence treatment course and outcome. In this study the authors aim to administer an adapted intervention for AUDs to reduce alcohol use in people living with HIV/AIDS (PLWHA). This study is a cluster randomised controlled trial at 16 HIV care clinics. A motivational interviewing and cognitive behavioural therapy based intervention for AUDs, developed through adaptation and piloted in Zimbabwe, will be administered to PLWHA with AUDs recruited at HIV clinics. The intervention will be administered over 16 sessions at 8 HIV clinics. This intervention will be compared with an equal attention control in the form of the World Health Organization Mental Health Gap Action Programme (WHO mhGAP) guide, adapted for the Zimbabwean context. General function, quality of life, and adherence to highly active antiretroviral treatment (HAART) will be secondary outcomes. Booster sessions will be administered to both groups at 3 and 6 months respectively. The primary outcome measure will be the Alcohol Use Disorder Identification Test (AUDIT) score. The World Health Organisation Disability Assessment Schedule 2.0 (WHODAS 2.0), World Health Organisation Quality of Life (WHOQoL) HIV, viral load, and CD4 counts will be secondary outcome measures. Outcome assessments will be administered at baseline, 3, 6, and 12 months. Moderating factors such as perceived social support, how people cope with difficult situations and post-traumatic exposure and experience will be assessed at baseline. Trained research assistants will recruit participants. The outcome assessors who will be trained in administering the outcome and moderating tools will be blinded to the treatment arms allocated to the participants. However, the principal investigator, participants and intervention staff will be unblinded. Data will be analysed using STATA Version 14. Primary and secondary outcomes will be measured at four time points that is; at baseline, 3, 6, and 12 months respectively. All participants will be included in the analysis of primary and secondary outcome measures. The mean AUDIT scores will be compared between groups using student t-tests. Multilevel logistic regression analysis will be performed for binominal variables and multilevel linear regression for continuous variables. Descriptive statistics will be computed for baseline and follow-up assessments. The study will be the first to address problematic alcohol use in PLWHA in Zimbabwe. It seeks to use local resources in delivering a modified, brief, evidence-based, and culturally contextualised intervention. The study results will determine the effectiveness of adapting psychological interventions for AUDs in HIV infected adults using a task-sharing framework. Pan African Clinical Trial Registry, PACTR201509001211149 . Registered 22 July 2015.


Very low excess noise Al0.75Ga0.25As0.56Sb0.44 avalanche photodiode.
AlxGa1-xAsySb1-y grown lattice-matched to InP has attracted significant research interest as a material for low noise, high sensitivity avalanche photodiodes (APDs) due to its very dissimilar electron and hole ionization coefficients, especially at low electric fields. All work reported to date has been on Al concentrations of x = 0.85 or higher. This work demonstrates that much lower excess noise (F = 2.4) at a very high multiplication of 90 can be obtained in thick Al0.75Ga0.25As0.56Sb0.44 grown on InP substrates. This is the lowest excess noise that has been reported in any III-V APD operating at room temperature. The impact ionization coefficients for both electrons and holes are determined over a wide electric field range (up to 650 kV/cm) from avalanche multiplication measurements undertaken on complementary p-i-n and n-i-p diode structures. While these ionization coefficients can fit the experimental multiplication over three orders of magnitude, the measured excess noise is significantly lower than that expected from the β/α ratio and the conventional local McIntyre noise theory. These results are of importance not just for the design of APDs but other high field devices, such as transistors using this material.


Prognostic value of molecular markers, sub-stage and European Organisation for the Research and Treatment of Cancer risk scores in primary T1 bladder cancer.
What's known on the subject? and What does the study add? The stakes are high when making treatment decisions in T1 bladder cancer (BC). Conservative management may lead to progression and possibly death from BC. Conversely, radical cystectomy could be over-treatment of non-progressive disease. The problem for clinicians is that reliable prognostic indices are lacking. We performed a head-to-head comparison of two substaging systems, European Organisation for the Research and Treatment of Cancer (EORTC) risk scores and four molecular markers in T1 carcinomas of the bladder treated conservatively with BCG. T1 sub-stage according to a new system (micro-invasive [T1m] and extensive-invasive [T1e]) was the most important clinical variable for predicting progression to carcinoma invading bladder muscle. The performance of the EORTC risk scores was disappointing for this T1 sub-group. Molecular markers were not significant in multivariable analysis for predicting progression. Future studies may lead to the incorporation of sub-stage (T1m/T1e) in the TNM classification system for urinary BC to guide clinical decision-making in T1 BC. To evaluate the prognostic significance of four molecular markers, sub-stage and European Organisation for the Research and Treatment of Cancer (EORTC) risk scores in primary T1 bladder cancer (BC) treated with adjuvant bacille Calmette-Guérin. The slides of 129 carcinomas of the bladder from two university hospitals were reviewed and the T1 diagnosis was confirmed. T1 sub-staging was done in two separate rounds, using a new system that identifies micro-invasive (T1m) and extensive-invasive (T1e) T1BC, and then according to invasion of the muscularis mucosae (T1a/T1b/T1c). The EORTC risk scores for recurrence and progression were calculated. Uni- and multivariable analyses for recurrence and progression were performed using clinicopathological variables, T1 sub-stage, EORTC risk scores and molecular markers (fibroblast growth factor receptor 3 gene mutation and Ki-67, P53, P27 expression). The median follow-up was 6.5 years. Forty-two patients remained recurrence-free (33%). Progression to T2 or metastasis was observed in 38 (30%) patients. In multivariable analysis for recurrence, multiplicity was significant. In multivariable analysis for progression, female gender, sub-stage (T1m/T1e) and carcinoma in situ (CIS) were significant. Molecular markers were significant in univariable and in multivariable analyses for recurrence. EORTC risk scores were not significant. CIS, female gender and sub-stage (T1m/T1e) were the most important variables for progression. The additional value of molecular markers was modest. Sub-stage (T1m/T1e) could potentially be incorporated in future tumour-node-metastasis classifications.


Multiple explosion pathways of the deuterated benzene trication in 9-fs intense laser fields.
The fragmentation of deuterated benzene (C6D6) in ultrashort intense laser fields (9 fs, 1 x 10(15) W/cm2) is studied by the ion-coincidence momentum imaging technique. Five two-body and eight three-body Coulomb explosion pathways from the trication (C6D6(3+)), associated with the deprotonation and ring-opening reactions, are identified. It is found from the fragment momentum correlation that all the observed three-body explosion processes proceed sequentially via the two-body Coulomb explosion forming molecular dications, C(m)D(n)(2+), with (m,n) = (6,5), (5,5), (5,4), (4,4), (4,3), and (3,3), which further dissociate into pairs of monocations. The branching ratio of the fragmentation pathways estimated from the number of the observed coincidence events indicates that the fragmentation is nonstatistical.


Tyrosine phosphorylation is an early and specific event involved in primary keratinocyte differentiation.
Very little is known about early molecular events triggering epithelial cell differentiation. We have examined the possible role of tyrosine phosphorylation in this process, as observed in cultures of primary mouse keratinocytes after exposure to calcium or 12-O-tetradecanoylphorbol-13-acetate (TPA). Immunoblotting with phosphotyrosine-specific antibodies as well as direct phosphoamino acid analysis revealed that induction of tyrosine phosphorylation occurs as a very early and specific event in keratinocyte differentiation. Very little or no induction of tyrosine phosphorylation was observed in a keratinocyte cell line resistant to the differentiating effects of calcium. Treatment of cells with tyrosine kinase inhibitors prevented induction of tyrosine phosphorylation by calcium and TPA and interfered with the differentiative effects of these agents. These results suggest that specific activation of tyrosine kinase(s) may play an important regulatory role in keratinocyte differentiation.


Ethnobotanical, Phytochemistry, and Pharmacological Activity of Onosma (Boraginaceae): An Updated Review.
The genus Onosma belongs to the Boraginaceae family and contains over 230 species. The present review sheds light on the ethnopharmacology, phytoconstituents, bioactivity, and toxicology of the Onosma species from previous investigations. Furthermore, the paper also highlights the unresolved issues for the future investigations. The review included previous studies of the genus Onosma available from Google Scholar and Baidu Scholar, Science Direct, SciFinder, Wiley Online Library, and Web of Science. Until now, more than 200 chemical compounds have been detected from the genus Onosma, including naphthoquinone (33), flavonoids (30), hydrocarbon (23), phenolic (22), ester (17), alkaloids (20), aromatics (12), carboxylic acid (11), fatty acids (9), terpenoids (10), while the most important ones are rosmarinic, ferulic, protocatechuic, chlorogenic, caffeic, p-coumaric acids, and apigenin. The Onosma species are reported as traditional medicine for wound healing, heart disease, and kidney disorders, while the pharmacological investigations revealed that the extracts and the phytochemicals of Onosma species have different therapeutic properties including antioxidant, enzyme inhibitory, antitumor, hepatoprotective, antiviral, anti-inflammatory, and antimicrobial actions. The summarized knowledge in this review provides valuable ideas for the current and future drug discovery and a motivation for further investigation on the genus Onosma.


The E. coli S30 lysate proteome: A prototype for cell-free protein production.
Protein production using processed cell lysates is a core technology in synthetic biology and these systems are excellent to produce difficult toxins or membrane proteins. However, the composition of the central lysate of cell-free systems is still a "black box". Escherichia coli lysates are most productive for cell-free expression, yielding several mgs of protein per ml of reaction. Their preparation implies proteome fractionation, resulting in strongly biased and yet unknown lysate compositions. Many metabolic pathways are expected to be truncated or completely removed. The lack of knowledge of basic cell-free lysate proteomes is a major bottleneck for directed lysate engineering approaches as well as for assay design using non-purified reaction mixtures. This study is starting to close this gap by providing a blueprint of the S30 lysate proteome derived from the commonly used E. coli strain A19. S30 lysates are frequently used for cell-free protein production and represent the basis of most commercial E. coli cell-free expression systems. A fraction of 821 proteins was identified as the core proteome in S30 lysates, representing approximately a quarter of the known E. coli proteome. Its classification into functional groups relevant for transcription/translation, folding, stability and metabolic processes will build the framework for tailored cell-free reactions. As an example, we show that SOS response induction during cultivation results in tuned S30 lysate with better folding capacity, and improved solubility and activity of synthesized proteins. The presented data and protocols can serve as a platform for the generation of customized cell-free systems and product analysis.


Oviposition of fruit flies (Diptera: Tephritidae) and its relation with the pericarp of citrus fruits.
Fruit flies (Diptera: Tephritidae) represent a threat to fruit growing worldwide, mainly the citrus culture, however, biological studies show that fruit flies are not perfectly adapted to this host. This study investigated oviposition of Anastrepha fraterculus (Wiedemann, 1830) and Ceratitis capitata (Wiedemann, 1824) and its relation with the pericarp of citrus fruits. We evaluated the relationship between depth of oviposition of A. fraterculus and C. capitata and epicarp thickness of orange [Citrus sinensis (L.) Osbeck)] 'Navelina' and tangerine [C. reticulata (L.)] 'Clemenules' and the influence of fruit mesocarp of tangerine 'Clemenules' on oviposition of these species. The study was conducted under controlled conditions of temperature (25 ± 2 °C), relative humidity (70 ± 10% RH) and photophase (12 h). A. fraterculus and C. capitata laid their eggs in the flavedo region of orange 'Navelina' and between the albedo and flavedo of tangerine 'Clemenules'. When fruits with mesocarp exposed were offered, there was no oviposition by both fruit fly species. The results show that epicarp thickness of citrus fruits did not influence oviposition of A. fraterculus and C. capitata as oviposition did not occur only in the presence of the mesocarp, suggesting that other factors are involved in oviposition of these species.


Illusory faces are more likely to be perceived as male than female.
Despite our fluency in reading human faces, sometimes we mistakenly perceive illusory faces in objects, a phenomenon known as face pareidolia. Although illusory faces share some neural mechanisms with real faces, it is unknown to what degree pareidolia engages higher-level social perception beyond the detection of a face. In a series of large-scale behavioral experiments (ntotal = 3,815 adults), we found that illusory faces in inanimate objects are readily perceived to have a specific emotional expression, age, and gender. Most strikingly, we observed a strong bias to perceive illusory faces as male rather than female. This male bias could not be explained by preexisting semantic or visual gender associations with the objects, or by visual features in the images. Rather, this robust bias in the perception of gender for illusory faces reveals a cognitive bias arising from a broadly tuned face evaluation system in which minimally viable face percepts are more likely to be perceived as male.


Immunolocalization of PTHrP in the parotid glands of three rodents species: Clethrionomys glareoulus, Microtus arvalis and white Swiss mice.
The current study was inspired by the fact that since 2004 no report had appeared on the occurrence of this peptide in healthy parotid glands of humans and animals. The objective of the current study was to investigate the immunolocalization of PTHrP in the parotid gland of three male rodents: 6 common voles (Microtus arvalis, Pallas, 1779), 6 bank voles (Clethrionomys glareoulus, Schreber, 1780) and 6 white Swiss mice, as well as to find out any species differences in the distribution of this peptide in various types of cells of the parotid gland. Immunocytochemical reactions were performed using the ABC technique with specific rabbit antibodies against human PTHrP (34-53) (CALBIOCHEM), diluted 1:70 and 1:50. We observed positive PTHrP expression in the epithelial cells of the striated duct in all the three animal species. The expression was strong in white mouse and very strong in common vole and bank vole. In all the rodent species studied, the reaction for PTHrP was granular in nature and irregularly distributed in the cytoplasm, being definitely stronger at the base and weaker at the apex of the cells. The PTHrP expression was negative in the epithelium of the intercalated duct, interlobular duct, main excretory duct, as well as in the myoepithelial cells surrounding the excretory ducts or serous acini.


Central band reconstruction for the treatment of Essex-Lopresti injury : A novel technique using the brachioradialis tendon.
Longitudinal stability of the forearm is mainly provided by three structures: the radiocapitellar contact, which acts as the primary stabilizer, the central band of the interosseous ligamentous complex (IOLC) and the intact triangular fibrocartilage complex (TFCC). In an Essex-Lopresti lesion the forearm becomes fully destabilized, since all of these three components are injured. Fixation or replacement of the radial head with a metallic prosthesis along with repair of the TFCC and stabilization of the distal radioulnar joint (DRUJ) are well-established treatment goals. However the reconstruction of the central band of the IOLC remains to some extent controversial. The authors believe that the reconstruction of the central band, particularly in active patients, is crucial in order to restore normal load distribution through the forearm, thus ensuring both transverse and longitudinal stability. In this article, we present a case with an Essex-Lopresti lesion, which was effectively treated acutely with restoration of all three components of the injury (radial head prosthesis, DRUJ repair and reconstruction of the central band of the IOLC). A novel technique by rerouting the brachioradialis tendon is described in detail.


Experimental study on the effect of scanning path on skin tissue properties of femtosecond laser welding.
To study the influence pattern of femtosecond laser scanning path on the welding effect of skin tissue, this experiment analyzed the influence of scanning path on the surface morphology, degree of thermal damage, tensile strength, and microstructure of skin samples after skin attachment by designing nine scanning paths to weld skin tissue. The results showed that the skin samples connected by interrupted parallel mattress eversion sewing method with d = 0.2 mm showed no obvious color changes in morphology, the skin samples were connected on both front and back sides, the tensile strength was the highest, reaching 12.80 N/cm2 , the thermal damage parameter was low at 1.08 × 10-2 , the microstructure had obvious directionality, and the texture was clear and uniformly distributed.


Beta-lactam-specific resistant mutants of Staphylococcus aureus.
In an approach to understanding the origin of methicillin resistance in clinical isolates of staphylococci, a series of Staphylococcus aureus mutants resistant to various beta-lactam antibiotics were isolated in the laboratory by antibiotic selection. Mutants with low- and intermediate-level resistance showed considerable specificity for the particular antibiotic used in the selection process (methicillin, cefotaxime, cephalexin, and amdinocillin), and resistance in such mutants also showed alterations in the antibiotic binding capacities of penicillin-binding proteins (PBPs). In each case the isolation of mutants resistant to high concentrations of antibiotics required sequential passage in gradually increasing concentrations of the drug. The acquisition of increasing levels of methicillin resistance was paralleled by a gradual decrease in the binding capacities of PBPs 2, 3, and, possibly, 1. In a highly methicillin-resistant mutant (MIC, 150 micrograms/ml), PBPs 2 and 3 were no longer detectable by the penicillin binding assay. Instead, a new PBP of poor binding capacity and anomalous molecular size (about 78 kilodaltons [kDa]) appeared in these cells. This corresponds to the molecular size of PBP 2a, the unique PBP that appears to be the biochemical correlate of resistance in clinical isolates of methicillin-resistant S. aureus. Also, similar to the case of resistant clinical isolates, high-level beta-lactam resistance was highly pH dependent in the laboratory mutants. We compared the patterns of radioactive peptides generated by partial proteolysis from the penicillin-labeled PBP 2 of antibiotic-susceptible staphylococci and from the 78-kDa PBP 2a of a resistant clinical strain. Although the patterns were clearly different, seven of the eight characteristic peptides generated from PBP 2 of the susceptible strain were also detectable among the peptides released from PBP 2a. The results suggest that the 78-kDa PBP 2a of the resistant clinical strain evolved from PBP 2 of antibiotic-susceptible staphylococci and that in PBP 2a of the clinical isolate mutational changes have resulted in extensive alterations near the beta-lactam binding site.


ClpXP protease targets long-lived DNA translocation states of a helicase-like motor to cause restriction alleviation.
We investigated how Escherichia coli ClpXP targets the helicase-nuclease (HsdR) subunit of the bacterial Type I restriction-modification enzyme EcoKI during restriction alleviation (RA). RA is a temporary reduction in endonuclease activity that occurs when Type I enzymes bind unmodified recognition sites on the host genome. These conditions arise upon acquisition of a new system by a naïve host, upon generation of new sites by genome rearrangement/mutation or during homologous recombination between hemimethylated DNA. Using recombinant DNA and proteins in vitro, we demonstrate that ClpXP targets EcoKI HsdR during dsDNA translocation on circular DNA but not on linear DNA. Protein roadblocks did not activate HsdR proteolysis. We suggest that DNA translocation lifetime, which is elevated on circular DNA relative to linear DNA, is important to RA. To identify the ClpX degradation tag (degron) in HsdR, we used bioinformatics and biochemical assays to design N- and C-terminal mutations that were analysed in vitro and in vivo. None of the mutants produced a phenotype consistent with loss of the degron, suggesting an as-yet-unidentified recognition pathway. We note that an EcoKI nuclease mutant still produces cell death in a clpx- strain, consistent with DNA damage induced by unregulated motor activity.


Evaluating an Intervention Program Using WeChat for Patients With Chronic Obstructive Pulmonary Disease: Randomized Controlled Trial.
The application of telemedicine in home pulmonary rehabilitation interventions for the management of patients with chronic obstructive pulmonary disease (COPD) has achieved promising results. This study aimed to develop a WeChat official account (Pulmonary Internet Explorer Rehabilitation [PeR]) based on social media. It further evaluated the effect of PeR on the quality of life, symptoms, and exercise self-efficacy of patients with COPD. The functional modules of PeR were developed by a multidisciplinary team according to the electronic health-enhanced chronic care model (eCCM) components. A total of 106 patients were randomly selected (53 in the PeR group and 53 in the outpatient face-to-face group [FtF]). Pulmonary rehabilitation intervention was conducted for 3 months, and the outcome was observed for 3 months. The primary outcome was patient quality of life measured with the COPD assessment test (CAT). The secondary outcomes were evaluated using the modified Medical Research Council scale (mMRC), exercise self-regulatory efficacy scale (Ex-SRES), and St George's Respiratory Questionnaire (SGRQ). The intention-to-treat analysis was used in the study. A total of 94 participants completed the 6-month pulmonary rehabilitation program. No statistically significant differences were observed in CAT (F1,3=7.78, P=.001), Ex-SRES (F1,3=21.91, P<.001), and mMRC scores (F1,3=29.64, P<.001) between the two groups with the variation in time tendency. The Ex-SRES score had a significant effect on the CAT score (P=.03). The partial regression coefficient of Ex-SRES to CAT was 0.81, and Exp (B) was 2.24. The telemedicine technology was effective using the eCCM combined with a behavioral intervention strategy centering on self-efficacy. Pulmonary rehabilitation at home through PeR and FtF could improve the sense of self-efficacy and quality of life and alleviate symptoms in patients with COPD. Chinese Clinical Trial Registry ChiCTR1900022770; https://tinyurl.com/tmmvpq3.


Inhibition of experimental autoimmune encephalomyelitis induction in SJL/J mice by using a peptide with high affinity for IAs molecules.
Blocking of the Ag presenting function of MHC by peptides capable of high affinity binding to this molecule has been proposed as a potential immunotherapeutic intervention in MHC-associated diseases. Recent studies have used this strategy to prevent the induction of experimental allergic encephalomyelitis (EAE) in mice. However, because of the close structural relationship between the inhibitor and encephalitogenic peptides, the results of these previous studies have been difficult to interpret with regard to whether MHC blockade was the mechanism by which the inhibitory peptides functioned. In our study, we have determined the capacity of unrelated peptides capable of binding with high affinity to IAs in inhibiting the induction of EAE in SJL/J mice after immunization with the autoantigenic peptide PLP 139-151. Prevention of the disease was accomplished by two methods: 1) when inhibitor was administered together with the encephalitogenic peptide at the time of immunization, as in previous studies, and 2) when inhibitor was administered at a separate site from the autoantigen 1 day before the immunization with that Ag. Inhibition was due to binding of the inhibitor to IAs, as evidenced by the fact that a control peptide incapable of binding to this MHC had no effect on the course of the disease. The finding that inhibitor could also be efficacious when administered at a separate site has implications for potential use of such a strategy to reverse ongoing autoimmune diseases. The inhibitor had to be present during the time of Ag stimulation, and had no long term inhibitory effects, in that a secondary immune response to the encephalitogenic peptide was not inhibited in animals given the inhibitory peptide before the induction of a primary response. This is compatible with the conclusion that MHC blockade was, in fact, the mechanism of the inhibition, rather than as a result of any long term suppressive effects on immunoreactive T cells. Finally, not only did administration of the inhibitory peptide lead to a prevention of the induction of EAE, but it could also be shown to decrease the T cell proliferative response in vitro to the autoantigen.


Ultrasonic Aspirator for Tissue Contact Detection: An Online Classification on Time-Series.
The goal of neurosurgical tumor surgery is to remove the tumor completely without damaging healthy brain structures and thereby impairing the patient's neurological functions. This requires careful planning and execution of the operation by experienced neurosurgeons using the latest intraoperative technologies to achieve safe and rapid tumor reduction without harming the patient. To achieve this goal, a standard ultrasonic aspirator designed for tissue removal is equipped with additional intraoperative tissue detection using machine learning methods.Since decision-making in a clinical context must be fast, online contact detection is critical. Data are generated on three types of artificial tissue models in a CNC machine-controlled environment with four different ultrasonic aspirator settings. Contact classification on artificial tissue models is evaluated on four classification algorithms: change point detection (CPD), random forest (RF), recurrent neural network (RNN) and temporal convolutional network (TCN). Data preprocessing steps are applied, and their impacts are investigated. All methods are evaluated on five-fold cross-validation and provide generally good results with a performance of up to 0.977±0.007 in mean F1-score. Preprocessing the data has a positive effect on the classification processes for all methods and consistently improves the metrics. Thus, this work indicates in a first step that contact classification is feasible in an online context for an ultrasonic aspirator. Further research is necessary on different tissue types, as well as hand-held use to more closely resemble the intraoperative clinical conditions.


Emerging concepts in the pathogenesis of antineutrophil cytoplasmic antibody-associated vasculitis.
Antineutrophil cytoplasmic antibodies (ANCAs) remain central to our current understanding of the pathogenesis of ANCA-associated vasculitis (AAV), and this review considers recent developments in the context of four key questions: are there targets for ANCA beyond myeloperoxidase (MPO) and proteinase 3 (PR3); are all ANCA pathogenic; how are ANCAs generated; and how do ANCA cause disease? B-cell epitope mapping raises the possibility that only a subset of ANCA may be pathogenic. Anti-lysosomal-associated membrane protein 2 autoantibodies have recently emerged as a novel form of ANCA and can be found in anti-MPO and anti-PR3 negative disease. These also provide recent evidence for molecular mimicry in the pathogenesis of AAV, but a definitive proof in human AAV remains elusive. Neutrophil extracellular traps may represent an important mechanism by which MPO and PR3 are taken up by dendritic cells for presentation to the adaptive immune system, and the role of the alternative pathway of complement in AAV has recently been emphasized, with therapeutic implications. Our current understanding of the pathogenesis of AAV not only reinforces the central role of neutrophils but also provides a sound rationale for B-cell and complement-directed therapies.